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EP4326892A1 - Method for selecting ecobiological topical compositions and ingredients capable of restoring or reinforcing the skin microbiota - Google Patents

Method for selecting ecobiological topical compositions and ingredients capable of restoring or reinforcing the skin microbiota

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Publication number
EP4326892A1
EP4326892A1 EP22723166.9A EP22723166A EP4326892A1 EP 4326892 A1 EP4326892 A1 EP 4326892A1 EP 22723166 A EP22723166 A EP 22723166A EP 4326892 A1 EP4326892 A1 EP 4326892A1
Authority
EP
European Patent Office
Prior art keywords
microbiota
composition
ascorbyl
biomass
skin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP22723166.9A
Other languages
German (de)
French (fr)
Inventor
Jean-Noël THOREL
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Naos Institute of Life Science SAS
Original Assignee
Naos Institute of Life Science SAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Naos Institute of Life Science SAS filed Critical Naos Institute of Life Science SAS
Publication of EP4326892A1 publication Critical patent/EP4326892A1/en
Pending legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06Quantitative determination
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/045Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/047Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates having two or more hydroxy groups, e.g. sorbitol
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/185Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/205Amine addition salts of organic acids; Inner quaternary ammonium salts, e.g. betaine, carnitine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/41641,3-Diazoles
    • A61K31/4172Imidazole-alkanecarboxylic acids, e.g. histidine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7016Disaccharides, e.g. lactose, lactulose
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/19Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/46Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing sulfur
    • A61K8/466Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing sulfur containing sulfonic acid derivatives; Salts
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A61K8/4946Imidazoles or their condensed derivatives, e.g. benzimidazoles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60Sugars; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/645Proteins of vegetable origin; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73Polysaccharides
    • A61K8/735Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/10Washing or bathing preparations
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
    • CCHEMISTRY; METALLURGY
    • C40COMBINATORIAL TECHNOLOGY
    • C40BCOMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00Methods of screening libraries
    • C40B30/06Methods of screening libraries by measuring effects on living organisms, tissues or cells
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52Stabilizers
    • A61K2800/522Antioxidants; Radical scavengers

Definitions

  • the present invention relates to the study of the skin microbiota and its use for the restoration and/or maintenance of healthy skin microbiota. STATE OF THE ART
  • the skin the largest human organ, is colonized by billions of microorganisms such as bacteria, yeasts, fungi or viruses collectively called microbiota or microflora.
  • the human skin flora can be subdivided into two groups: -
  • the resident flora is made up of commensal germs, ie living at the expense of their host, without causing them any harm.
  • the resident human flora is dominated by Proteobacteria, including about ten types of Gram-positive aerobic bacteria such as Staphylococcus epidermidis and Staphylococcus hominis (Otto, 2010).
  • the resident flora is also composed of Gram-positive anaerobic bacteria belonging to the Actinobacteria division (genera Cutibacterium, Corynebacterium, Dermabacter and Micrococcus) (Kligman et al., 1976). Among the microorganisms of the other kingdoms, Malassezia is the fungal species most frequently found on the skin. Finally, the mites Demodex folliculorum and Demodex brevis, or "eyelash mites", are also considered members of the resident human flora.
  • Transient flora is composed of mostly harmless fungi, viruses and bacteria, called saprophytes, that is to say, which feed on decomposing organic matter from the environment.
  • This flora can also consist of opportunistic pathogenic bacteria that can cause disease in the host.
  • the transient flora does not establish itself permanently on the surface of the skin, varying during the day, according to the activities carried out and the surrounding conditions. However, it can persist for hours or even days. Its density is low on dry areas and particularly high on hairy areas, folds and areas subject to perspiration.
  • the most common transient species are Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and several species of the genus Bacillus.
  • MRSA methicillin-resistant Staphylococcus aureus
  • the management of MRSA requires the use of new antibiotics, which can however further destabilize the entire human microbiota by slowing down the recovery of its diversity.
  • indices making it possible to evaluate the diversity of the microbiota are developed, among which one can quote the indices of Shannon or Simpson, the most used being the index of Shannon which makes it possible at the same time to evaluate the number of species of this environment (specific richness) and the distribution of individuals within these species (specific equitability; Lemos et al., 2011).
  • topical products can have a significant impact on the composition and balances of cutaneous microbial communities (Kong, 2011; Kong et al., 2012a).
  • Antiseptic products i.e. fungicides, bactericides and/or virucides
  • ethanol in particular, impoverish not only the quantity, but also the diversity of the skin microbiota and this potentially more durably (San Miguel et al., 2018).
  • biocides to the surface of the skin was relatively rare and limited to the hospital setting until 2020.
  • the global pandemic of COVID-19 the use of this category of products has become commonplace and widespread. to levels never before achieved.
  • the present invention is based on the demonstration of the interest, under non-pathological conditions, of promoting the increase in the biomass of the cutaneous microbiota (multiplication of the microbiota), without reducing its overall diversity (conservation or increase in the diversity of said microbiota).
  • the healthy skin microbiota and its balance are then reinforced and/or reconditioned to better resist potential opportunistic dysregulation and colonization.
  • the Applicant has thus found a way that is simple, effective, affordable and usable on an industrial scale, in particular in cosmetics or dermatology, for selecting ingredient(s) and/or composition(s) suitable to restore and/or strengthen the healthy skin microbiota, by seeking to increase the biomass of the skin microbiota, in particular without reducing its overall diversity.
  • two methods are combined: a method for measuring the biomass on a skin area and a method for measuring the microbial diversity on this same area.
  • the “microbiota” concerns all of the microbial flora, including microbes, bacteria, yeasts, fungi and viruses.
  • the microbiota relates to all the genes or genomes present in the microbiota.
  • biodiversity or “diversity” or “diversity of the microbiota” or “diversity of the microbiome” or “bacterial diversity”, one refers to all bacterial populations or species present in the skin microbiota.
  • This biodiversity can be characterized in particular by the number of different bacterial genetic sequences within said microbiome (also denoted OTU, for “Operational Taxonomic Unit”) and/or their relative abundance.
  • OTU Orderal Taxonomic Unit
  • Several indices making it possible to evaluate the diversity of the microbiota have been developed and are well known to those skilled in the art, among which mention may be made of the Shannon or Simpson indices.
  • biomass of the skin microbiota” or “biomass” or “bacterial load” is meant the number or quantity of bacteria on a skin area.
  • This biomass can be characterized in particular by the amount of bacterial genetic material, in particular 16S rDNA, from said microbiota.
  • the biomass and diversity levels of a microbiota can be measured by any appropriate technique well known to those skilled in the art, such as quantitative amplification and 16S rDNA sequencing.
  • “healthy” is understood to mean non-pathological.
  • by “healthy skin area”, is meant a skin area not exhibiting dermatosis.
  • by “healthy skin microbiota” or “healthy skin microbiota”, is meant a stable and diversified skin microbiota on a given skin area, such as on the same healthy skin area not subjected to stress conditions or particular aggressions (such as, but not limited to, treatments or care or non-ecobiological environments, in particular dermatological, cosmetics, hygiene, pollution, etc.).
  • the microbiota of healthy skin protects in particular against invasive pathogens, but also against drying out of the skin.
  • stress or “aggression” is meant any event, of any nature whatsoever, single or repeated, in particular of external origin, which may be the cause of destabilization or modifying a system.
  • stress can in particular lead to a reduction in the biomass and/or the diversity of the cutaneous microbiota.
  • it may be linked to non-ecobiological treatments or care or environments, in particular dermatological, cosmetic, hygiene, pollution, etc., and, more particularly, to the action of bactericides or virucides or fungicides or soaps or hygiene or care products that are aggressive towards the microbiota.
  • the present invention relates to a method for selecting or screening ingredients and compositions, preferably ecobiological, cosmetic or dermopharmaceutical or dermatological, advantageously applicable topically, according to their effect on the cutaneous microbiota, in particular their ability to induce the increasing the biomass of the cutaneous microbiota, in particular without reducing its biodiversity.
  • the present invention also relates to a method for preparing a composition, preferably topical, capable of inducing an increase in the biomass of the cutaneous microbiota, as well as its use for the preparation of care compositions, preferably topical, advantageously ecobiological and ecobiological care methods, preferably topically, based on the use of the aforementioned compositions or ingredients.
  • the present invention also relates to a composition for its use for maintaining and/or restoring a healthy skin microbiota via an increase in the biomass of the skin microbiota, in particular without reducing its biodiversity.
  • the invention aims for the first time to analyze and quantify the biomass of the cutaneous microbiota, as well as, where appropriate, its biodiversity and to select the substances and compositions which make it possible to increase the biomass of the cutaneous microbiota, in particular without diminishing its biodiversity.
  • the present invention therefore relates to a method for selecting or screening ingredients, substances or topical compositions making it possible to maintain and/or restore, preferably rapidly, a healthy microbiota.
  • the present invention relates in particular to a screening method whose objective is to select ingredients, preferably topically acceptable, individually and/or compositions comprising a mixture of several aforementioned ingredients, optionally combined with excipients, preferably topically acceptable, capable of increase the biomass of the cutaneous microbiota, in particular without reducing its biodiversity.
  • the present invention also relates to a composition, preferably topical, selected by the screening method.
  • the present invention also relates to one or more ingredients, optionally in the form of a composition, preferably topical, to be used for the manufacture of a composition, preferably topical, for the care of the skin, mucous membranes or appendages capable of increase the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, to maintain and/or restore the healthy nature of the skin, mucous membranes or appendages.
  • the present invention finally relates to a care method, preferably by topical route, using one or more ingredients, advantageously topically acceptable, capable of increasing the biomass of the cutaneous microbiota, by particular without reducing its biodiversity, to maintain and/or restore the healthy character of the skin, mucous membranes or appendages.
  • the invention relates to a method, preferably in vitro, for selecting an ingredient or composition, preferably ecobiological, advantageously applicable topically, in particular cosmetic or dermatological, useful in maintaining or/and the restoration of a healthy skin microbiota, comprising the steps consisting in: a) bringing an ingredient or composition to be tested into contact with bacteria of a reference microbiota, b) analyzing the quantity of biomass and, advantageously the biodiversity of the microbiota after bringing into contact, and c) selecting as an ingredient or composition useful in maintaining and/or restoring a healthy skin microbiota, an ingredient or a composition capable of increasing the biomass of the skin microbiota, advantageously without reduce its biodiversity, compared to the quantification of the biomass and, where applicable, the biodiversity of the reference microbiota.
  • the method according to the invention is personalized, based on the conditions, specificities and reactivity of the skin microbiota of a given individual.
  • step a is carried out in a single or repeated manner and/or step b is carried out from 1 hour to 31 days, preferably 4 days, after step a.
  • the invention relates to a method, preferably in vitro, for screening or evaluating or selecting the activity of at least one ingredient or composition, preferably applicable topically, in particular an ingredient or cosmetic composition and/or dermatological agent, optionally formulated in the form of a topical composition, characterized in that the method comprises the following steps, preferably in vitro: i) before the application of the ingredient or of the composition, optionally formulated in the form of a topical composition, on at least one skin area to be studied presenting a so-called reference microbiota, from a first sample taken from said skin area, the analyzes of the amount of biomass of the skin microbiota and, where appropriate, of its biodiversity, thus obtaining a first so-called reference measurement, for quantifying the biomass and, where appropriate, a first so-called reference measurement, of the biodiversity of the bacteria of said reference microbiota; and ii) after at least one application of the ingredient or of the composition, optionally formulated in the form of a topical composition, on the said
  • the invention relates to a method, preferably in vitro, of screening or selecting at least one ingredient or composition, optionally formulated in the form of a topical composition, to induce an increase in the biomass of a cutaneous microbiota, advantageously without reduce its biodiversity including the following steps: i) quantifying the biomass of the skin microbiota and possibly measuring the biodiversity of a first sample from a skin area, thus making it possible to obtain a first quantification measurement of the biomass and, where appropriate, a first measurement of the biodiversity of a so-called reference skin microbiota; and ii) quantifying the biomass of the skin microbiota and optionally measuring the biodiversity of a second sample from said skin area to which the at least one ingredient or composition, optionally formulated in the form of a topical composition, has been applied, thus making it possible to obtain a second biomass quantification measurement and, if necessary, a second biodiversity measurement; and iii) comparing the second biomass quantification measurement with the first biomass quantification measurement, and where appropriate the second biodiversity measurement
  • the predetermined action time of the ingredient can vary between 1 hour and several days, in particular 31 days, advantageously 4 days.
  • a “reference skin microbiota” or “so-called reference skin microbiota” or “reference microbiota” or “so-called reference microbiota” is for example a microbiota characteristic of a skin zone to be studied.
  • a reference microbiota can also be a medium comprising different bacteria in various proportions which may or may not correspond to a microbiota existing in nature.
  • the reference microbiota can be obtained in particular from a sample of a skin zone of an individual or by preparing a medium comprising the different bacteria in the proportions corresponding to said reference microbiota.
  • the levels of biomass and diversity of a reference microbiota characteristic of a skin area is an average of the levels of biomass and diversity and/or of microbiota diversity profile measured on samples from said area of skin in several individuals.
  • said reference microbiota is that of a healthy skin area and/or is depleted, advantageously following stress or cutaneous aggression, more advantageously following the action of bactericides or virucides or soaps or hygiene or care products that are aggressive towards the microbiota.
  • a method for screening or selecting or evaluating the activity of at least one ingredient which is preferentially applicable topically, in particular a cosmetic and/or dermatological ingredient, optionally formulated in the form of a topical composition characterized in that the method comprises: a) before any operation, the selection of a skin area to be studied from one or more human beings, b) the taking of at least a first sample of biological or genetic material of the microbiota present on this selected area, of a or several human beings, constituting a so-called reference microbiota, followed by the analysis of the quantity of biomass and the biodiversity of the skin microbiota, thus obtaining first so-called reference measurements respectively for quantification of the biomass of the skin microbiota and of biodiversity of the cutaneous microbiota, c) the application of at least one ingredient or a composition to this area, in a single or repeated manner, d) after a period of 1 hour to 31 days, advantageously 4 days, after the application single or repeated applications
  • said ingredient or said composition to be tested in the selection or screening method according to the invention is chosen from the group comprising: a) at least one carbon source which is capable of being used by bacteria such as substrate for growth and multiplication, and preferably chosen from: - partially or totally hydrolyzed proteins, from cereals (wheat, corn, barley, rye, oats), legumes (soya, fava beans, broad beans, peas), milk ( caseins, albumins), yeast, as well as peptides and amino acids alone or as a mixture; - short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) at least one carbon source which is capable of
  • the invention also relates to a method for preparing a composition, advantageously topical, in particular cosmetic or dermatological, preferably ecobiological, characterized in that it comprises (i) the implementation of a method of selection or screening according to the invention (ii) the selection of at least one composition, advantageously topical, in particular a cosmetic and/or dermatological composition, comprising at least one ingredient selected by the selection or screening method according to the invention and capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity and (iii) the conditioning of the said composition.
  • the invention relates to a composition, advantageously topical, preferentially ecobiological, in particular cosmetic and/or dermatological, comprising at least one ingredient or a composition, selected by the selection or screening method according to the invention, for its use for its use in maintaining and/or restoring a healthy skin microbiota.
  • the composition for its use according to the invention comprises at least one carbon source, an antioxidant and an inorganic or organic salt, more advantageously said carbon source, said antioxidant and/or said salt being selected by selection or screening method according to the invention.
  • the at least one carbon source is capable of being used by bacteria as a substrate for growth and multiplication, and, preferentially, is chosen from: - partially or totally hydrolyzed proteins, from cereals (wheat, corn, barley, rye, oats), legumes (soya, fava beans, beans, peas), milk (caseins, albumins), yeast, as well as peptides and amino acids alone or as a mixture; - short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) The at least one antioxidant exhibits activity on reactive oxygen species (ROS), advantageously free radicals, and, in
  • the composition for its use according to the invention comprises sodium citrate and carnosine.
  • the composition for its use according to the invention is an aqueous composition, comprising at least 60% by weight of the total composition of water, in particular at least 60%, better still at least 70%, or at least 80% or at least 90% or at least 95%.
  • the aqueous phase represents at least 60% by weight of the total composition, better still at least 70%, or at least 80% or at least 90% or at least 95%.
  • the aqueous phase is characterized by: - a resistivity, at ambient temperature, of between 12.5 and 12500 Ohms.cm, in particular between 80 and 8000 Ohms.cm; and/or - an osmotic pressure between 70 and 2500 mOsmol/l and in particular between 100 and 800 mOsmol/l as well as a pH between 5.0 and 9.0 and in particular between 6.0 and 8, 0.
  • the composition for its use according to the invention has a pH comprised between 5.0 and 9.0 and in particular comprised between 6.0 and 8.0.
  • the composition for its use according to the invention is applied to a healthy skin area, before or following a skin attack, advantageously the action of bactericides or virucides or soaps or hygiene products or aggressive treatments for the microbiota.
  • said cutaneous aggression according to the invention can be single or repeated, preferentially repeated.
  • the composition for its use according to the invention is for its use for combating cutaneous aggression on healthy skin, by stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, for increase the amount of this biomass of said microbiota in order to avoid the arrival and/or installation of new microbial strains linked to the environment.
  • the composition according to the invention is for its use in combating cutaneous aggression on healthy skin by stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, to restore the quantity of this biomass of said microbiota, greatly reduced during a skin attack and for example after the action of bactericides or virucides or soaps or hygiene or care products that are aggressive towards the microbiota.
  • the invention relates to a method of treatment, advantageously topical, in particular non-therapeutic or dermatological cosmetic, preferably ecobiological, for maintaining and/or restoring a healthy skin microbiota, characterized by the use of a composition such as previously described, allowing the biomass of the skin microbiota to be increased, in particular without reducing its biodiversity.
  • the invention relates to the use of at least one ingredient, or at least one substance, or at least one at least one composition, selected by the selection or screening method according to the invention for the preparation of a composition, preferably topical, in particular ecobiological, for maintaining and/or restoring a healthy skin microbiota.
  • the invention relates to a method of topical care, in particular ecobiological, advantageously cosmetic or dermatological, for treating the skin requiring maintenance or restoration of its healthy microbiota, by increasing the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, characterized in that at least one area of the skin requiring this care is selected, and one applies to at least this area of the skin an ingredient, advantageously formulated in a composition, or a composition selected through the selection or screening method described previously or in the following description.
  • the invention also relates to a topical care method, preferably ecobiological, characterized in that it comprises (i) the implementation of a selection or screening method defined previously or in the description following, and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing the increase in the biomass of the cutaneous microbiota described previously, in particular without reducing its biodiversity ( ii) the formulation of the ingredients or compositions thus selected in a formula for cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota without reducing its diversity (iii) the packaging of said formula (iv) the topical application of said formulation to an area of skin or a surface of the selected appendages.
  • a topical care method preferably ecobiological, characterized in that it comprises (i) the implementation of a selection or screening method defined previously or in the description following, and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatolog
  • the invention relates to a treatment method, advantageously topical, preferably cosmetic or dermatological, in particular ecobiological, for combating cutaneous aggression on healthy skin, characterized in that at least one zone is selected of the skin requiring this care and the administration is carried out, preferably by topical application, of at least one ingredient or composition, advantageously applicable topically, in particular a cosmetic and/or dermatological ingredient or composition, in particular formulated in the form of topical composition, selected by the screening or selection method described previously or in the following description, stimulating the biomass of the microbiota present in the skin, or present on a keratin material, to increase the quantity of this biomass of said microbiota in order to avoid the arrival of new microbial strains linked to the environment.
  • a treatment method advantageously topical, preferably cosmetic or dermatological, in particular ecobiological, for combating cutaneous aggression on healthy skin, characterized in that at least one zone is selected of the skin requiring this care and the administration is carried out, preferably by topical application, of at least one ingredient or composition, advantageous
  • the invention relates to a treatment method, advantageously topical, preferably cosmetic or dermatological, in particular ecobiological, for combating cutaneous aggression on healthy skin, characterized in that at least one area of the skin requiring this care and the application is carried out, preferably topically, of at least one ingredient which can be applied topically, in particular a cosmetic and/or dermatological ingredient, preferably formulated in the form of a topical composition, selected by the method of selection or screening described previously or in the following description, stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, to restore the quantity of this biomass of said microbiota against its reduction in quantity by a cutaneous aggression and for example after the action of bactericides or virucides or soaps or aggressive hygiene or care products vis-à-vis the mic robot.
  • keratinous materials is meant according to the invention the skin, the semi-mucous membranes (in particular the lips), the mucous membranes and the appendages including the skin appendages.
  • method for quantifying the biomass of the microbiota we mean according to the invention, the use of a quantitative or semi-quantitative method which is based for example: - on quantitative PCR (or qPCR) techniques, or - on flow cytometry, or - on agar counting methods, or - on any related method.
  • method for measuring the biodiversity of the microbiota is meant according to the invention, the use: - of a method which is based on the analysis of part of the hypervariable gene of the 16S ribosome of bacteria after their targeted amplification , or - a method using the metagenomic shotgun technique (on microbiome DNA), or - a so-called metatranscriptomic shotgun sequencing method (on microbiome RNA), or - any related method, followed by a method for calculating the microbial diversity from the data of the previous method used, such as the calculation: - of the Shannon index, - of the Simpson index, - of the Inverse-Simpson index, - the number of species or OTU, - the Evenness or the Pielou index, - the Hurlbert index, - the Hill index.
  • the sample is taken by carrying out 10 return trips in Z by executing a slight pressure on the swab. At this stage, it is possible to freeze the swabs, which can be stored at -80°C. From the swabs, the bacterial pellets can be recovered by centrifugation.
  • the extraction of genomic DNA (gDNA) can be carried out according to methods known to those skilled in the art, for example with the Nucleospin Microbial DNA kit (Macherey Nagel, France).
  • the samples can be taken from several areas of the human body, such as, for example, the cheek, forehead, forearm, in order to study the response of the microbiota of the different biogeographical areas to the ingredients or compositions applied topically.
  • the sample is taken from the face, which makes it possible to test the ingredients or compositions on a realistic area exposed to the environment, or on the back, which makes it possible to test a set of different conditions. on a relatively homogeneous microbiota (Grice et al., 2009).
  • the area concerned can be marked by a mask, advantageously delimiting a sampling surface of between 10 and 20 cm 2 , advantageously 15 cm 2 .
  • this biomass is evaluated starting from the biological or genetic material recovered from the keratin materials, but it can also be quantified directly according to the methods known to those skilled in the art, for example by immunolabeling or flow cytometry on labeled bacteria.
  • standardization within the meaning of the invention, is meant the standardization of the quantity of biomass sampled.
  • amplification within the meaning of the invention is meant any in vitro procedure which produces multiple copies of a target nucleic acid sequence, or of its complementary sequence, or of fragments thereof (i.e. i.e. an amplified sequence containing less than the complete target nucleic acid).
  • this amplification is carried out by polymerase chain reaction or polymerase chain reaction (PCR), but other methods known to those skilled in the art, such as isothermal amplification, can also be used.
  • quantitative amplification is meant within the meaning of the invention any in vitro procedure which produces multiple copies of a target nucleic acid sequence and which at the same time makes it possible to determine the concentration of target sequence initially present in the reaction mixture. .
  • quantitative amplification reactions are described. We can distinguish quantitative amplifications based on the use of an external standard, competitive amplifications, using an internal standard, and finally kinetic amplifications, which consist in measuring in real time, the increase in the quantity of the target sequence .
  • the quantitative amplification is a kinetic amplification carried out by PCR and corresponds to the technique known to those skilled in the art under the name of quantitative PCR (qPCR) or even real-time PCR.
  • target sequence refers to the particular nucleotide sequence of the target nucleic acid which is to be amplified and/or detected.
  • target sequence includes complexing sequences to which oligonucleotides (eg, primer oligonucleotides and/or promoter oligonucleotides) complex during amplification processes (eg, PCR).
  • target sequence When the target nucleic acid is originally single-stranded, the term “target sequence” will also designate the sequence complementary to the “target sequence” as present in the target nucleic acid. When the target nucleic acid is originally double-stranded, the term “target sequence” refers to both sense (+) and antisense (-) strands.
  • region as used in the present invention, means a portion of a nucleic acid, said part being smaller than the entire nucleic acid.
  • region V1, V2, V3, V4 hypervariable regions of the bacterial 16S ribosomal DNA (rDNA) defined for example in Gray et al., 1984, which constitutes a portion of the gDNA and which allows the identification of the species to which they belong during sequencing.
  • rDNA bacterial 16S ribosomal DNA
  • amplicon or amplification product is meant a nucleic acid molecule generated in an amplification reaction, whether quantitative or not, of nucleic acid and which is derived from a target nucleic acid.
  • An amplicon or amplification product contains a target nucleic acid sequence which may be of the same or opposite sense to the target nucleic acid.
  • sequencing within the meaning of the invention is meant the decoding of at least one strand of a target sequence, and the reading of the sequence of nucleotides composing it in their order.
  • Several methods for sequencing and assembling the sequences read are known to those skilled in the art.
  • the sequencing is carried out using the Illumina MiseqTM platform.
  • the documents WO03048387A2, WO2005024010A1 WO2006064199A1 and WO2007123744A1 or also WO2012096703A1 and the references cited in these documents describe sample preparation and sequencing methods suitable for the present invention.
  • the sequencing can be carried out directly on the amplicons, or alternatively on banks of amplicons.
  • the sequencing is carried out using a library of amplicons, advantageously the libraries of amplicons obtained according to the MetabioteTM solution described in Bryan et al., 2014.
  • a “primer” is an oligomer which is 'hybridizes to a nucleic acid and has a 3' end which is extended by polymerization.
  • a primer can optionally be modified, for example by including a 5' region that is not complementary to the target sequence. Such modification may include functional additions, such as tags, promoters or other non-specific target sequences, used or useful to manipulate or amplify the primer or target oligonucleotide. Primers can be "degenerate”.
  • degenerate primer denotes primers allowing incorporation of different nucleotides at determined positions.
  • linear 16S primers within the meaning of the invention, is meant oligonucleotides allowing the amplification of highly conserved regions of the 16S rDNA.
  • these are the primers corresponding to SEQ ID.18F (5'-AGAGTTTGATCCTGGCTCAG-3') and SEQ ID.21391R (5'-GACGGGCGGTGWGTRCA-3').
  • SEQ ID.3533F 5'-GTGCCAGCAGCCGCGGTAA-3'
  • SEQ ID.4902R 5'-GTCAATTCITTTGAGTTTYARYC-3'
  • 16S specific primers within the meaning of the invention is meant oligonucleotides allowing the amplification of hypervariable regions of 16S rDNA.
  • these are the V1-V4 regions of 16S rDNA, preferably the V1-V3 and/or V3-V4 regions, advantageously the V3-V4 regions.
  • the 16S specific primers make it possible to obtain amplicons of size between 100 and 500 base pairs, advantageously between 200 and 400 base pairs.
  • these are the primers for the V1-V3 regions corresponding to SEQ. ID 527F (5'-AGAGTTTGATCCTGGCTCAG-3') and SEQ.
  • a DNA library can be done according to the methods known to those skilled in the art, for example by using kits such as the NEBNextTM UltraTM DNA Library Prep Kit. According to a particular embodiment, it is considered within the scope of the invention that the selected ingredient or composition applied topically has a significant effect on the increase in the biomass of the cutaneous microbiota, if the biomass increases statistically significant between two samples, in particular between the first sample and the second sample mentioned above, in particular after 1 hour to 31 days of treatment.
  • the evaluation of the activity of the ingredient or composition applied topically on the biomass of the sampled microbiota can be carried out by comparing the total bacterial genetic material present on the sampled area before and after the treatment with the ingredient or composition through quantitative amplification with universal 16S primers, which makes it possible to estimate the genetic material present at the time of sampling before and after treatment.
  • the at least one amplification of the standardized amounts of genetic material is a quantitative amplification carried out using the universal 16S primers. For this, bacterial lysis followed by a genomic DNA extraction step with the PureLink Mini kit (Life Technologies, Grand Island, NY) is carried out.
  • 16S rRNA gene-targeted qPCR is performed using specific primers (Bact-8F: AGAGTTTGATCCT GGCTCAG, Bacte338R: CTGCTGCCTCCCGTAGGAGT) with SYBR Green detection (Applied Biosystems, Foster City, CA) on equipment Mx300P (Agilent Technology).
  • Standard curves are prepared by amplifying known amounts of DNA from E. coli. By comparing the number of cycles to the standard curve it is possible to calculate the number of copies of the RNA16S gene per microliter of sample.
  • the evaluation of the biomass of the cutaneous microbiota is carried out by quantification in flow cytometry.
  • the evaluation of the biomass of the cutaneous microbiota is carried out by quantification by counting on agar.
  • the bacterial suspensions are deposited on nutrient agar in a Petri dish, and are incubated for 48 hours at 35°C. A count of the bacterial colonies makes it possible to obtain the bacterial concentration by multiplying the number of CFU by the dilution factor, and is expressed in CFU/ml.
  • the evaluation of the biomass of the cutaneous microbiota is carried out by quantification by direct counting under a microscope. For this, the bacterial suspensions are inoculated in a nutrient broth such as Mueller-Hinton or Brain Heart Infusion. After incubation, a direct enumeration under the microscope can be carried out on a Thoma cell.
  • the evaluation of the activity of the ingredient or composition applied topically on the diversity of the sampled microbiota can be performed by comparing the bacterial species present in the different experimental conditions through amplification with specific 16S primers and sequencing of the amplicons obtained.
  • the index is the so-called Shannon index, which is calculated by following the method described in Lemos et al., 2011.
  • the Shannon index integrates two distinct parameters: the number of operational taxonomic units ( Operational Taxonomic Units or OTUs) and the distribution of these OTUs in each sample. This last parameter can be calculated by taking into account the abundance of the 15 most represented bacterial genera, or alternatively, representing the 90% of the amplicons obtained in the amplification with the specific 16S primers.
  • the bioinformatics processing of the target sequences can be carried out according to the methods known to those skilled in the art, for example by using a PERL script.
  • the analysis by amplification with the specific 16S primers then, optionally, sequencing is carried out only for the samples of genetic material from a sample making it possible to obtain, during the quantitative amplification with the 16S universal primers in a reaction volume of 4.38 ⁇ l, at least 400 copies of 16S rDNA amplicon/ ⁇ L after 30 cycles of amplification and/or at least 500 copies of amplicon of 16S/ ⁇ L rDNA after 35 cycles of amplification.
  • the analysis of the diversity of the cutaneous microbiota is carried out by metagenomics in the following way: after extraction of the gDNA, the microbial diversity is determined for each sample by targeted amplification of part of the gene ribosomal.
  • a fragment of the 16S rRNA gene comprising the hypervariable regions V3 to V4 (or V1-V3) is amplified with primers 338F (ACTCCTACGGGAGGCAGCAG) and 806R (GGACTACHVGGGTWTCTAAT) (Munyaka et al., 2015).
  • the amplification is carried out and the products resulting from PCR are purified and normalized using Agencourt AMPure XP magnetic beads (Beckman Coulter). Once purified, amplicons are pooled for Illumina sequencing, performed in 2x300 bp on an Illumina MiSeq platform using MiSeq Reagent Kit v3 (600 cycles).
  • the sequences are grouped into operational taxonomic units (OTUs) with an identity threshold of 97%.
  • Taxonomic assignment is performed using the Ribosomal Database Project (RDP) classification on the Greengenes V13_8 database. The analysis is carried out with the R software.
  • the Shannon-Weaver index, the number of OTUs observed and the Bray-Curtis dissimilarity index are calculated with the Phyloseq and vegan packages.
  • Statistical significance is determined using the Wilcoxon test. A p-value ⁇ 0.05 is considered statistically significant.
  • the analysis of the diversity of the cutaneous microbiota is carried out by Shotgun: after extraction of the gDNA, the microbial diversity is determined for each sample by Whole metagenome sequencing/WMS or Shotgun metagenome sequencing, i.e. i.e. complete sequencing of the entire bacterial genome.
  • the libraries are prepared using the NexteraXT preparation kit (Illumina). Sequencing is performed at the Penn Next Generation Sequencing Core on Illumina MiSeq chemistry to obtain final paired reads of 150 bp. Sequencing data is obtained in fastq format. Sequences shorter than 80 nucleotides are removed from the analysis.
  • DeconSeq is used to filter fastq files and one of the paired reads (SE1) is entered into MetaPhlAn version 1.7.7 for taxonomic classification.
  • SE1 One of the matched ends (SE1) of the MCC sample is used to perform a BLAST against a personalized database of genomes among the 20 expected bacterial species (blastn, max_target_seqs 1, e ⁇ 10 ⁇ 10; alignment length > 50) to determine the composition of bacterial communities.
  • the alpha diversity index is then calculated using the R software and the Phyloseq and Vegan packages using the Shannon index.
  • the analysis of the diversity of the cutaneous microbiota is carried out by Metatranscriptomic shotgun sequencing (MTS): for this the total RNA is extracted from the bacterial sample.
  • MTS Metatranscriptomic shotgun sequencing
  • the polyadenylated mRNAs are separated from the non-polyadenylated ones (rRNA, mitochondrial RNA, and tRNA) by affinity on a poly-dT column (Dynabeads Oligo (dT) kit, Dynal).
  • the enriched mRNA is mechanically fragmented to a size range of +/- 200 bp with an ultrasonicator. mRNA fragmentation is assessed using the Agilent RNA 6000 Pico Kit on a 2100 Bioanalyzer instrument (Agilent Technologies, Inc.).
  • Metatranscriptome libraries are prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs Inc).
  • the quality and quantity of all final libraries are analyzed with an Agilent DNA 1000 kit on the 2100 Bioanalyzer and Qubit instrument and are quantified and validated by a qPCR assay using the PerfeCTa NGS library quantification kit for Illumina (Quanta Biosciences, Inc.) using the CFX Connect real-time PCR detection system (Rad Laboratories, Inc.). Sequencing of one of the MT libraries is performed on an Illumina HiSeq 2000 using TruSeq SBS v3 reagent for 100 pair read length (BGI Americas) (labeled HS100) and on Illumina MiSeq using the v3-600 cycle kit for 301 base pairs (labeled MS301).
  • Another set of twelve libraries is sequenced on Illumina MiSeq using 151 “paired-end chemistry” (marked MS151).
  • the manufacturer's recommended protocol is used to perform the sequencing reaction on the HiSeq and MiSeq platforms.
  • BLASTN searches default settings, except word size set to 7 are performed against the GenBank nr nucleotide database (http://www.ncbi.nlm.nih.gov /).
  • the 16S sequences are then assigned to major phyla to perform phylogenetic analyzes (BioNJ and PhyM) using the Best Blast Hit (BBH) algorithm and a set of reference sequences for each of the different phyla.
  • the method according to the invention comprises at least two samples of biological or genetic material from the microbiota: - a sample of biological or genetic material from the microbiota present in the skin, or present on a keratin material of said being human, said sample being taken from an untreated area before the treatment, - a sample of biological or genetic material from the microbiota present in the skin, or present on a keratin material of said human being, said sample being taken from an area treated with the ingredient or composition, preferably applicable topically, after the treatment.
  • the method according to the invention comprises at least four samples of biological or genetic material of the microbiota present in the skin, or present on a keratin material, from one or more human beings: - A sample carried out in an area to be treated with the ingredient or composition, preferably applied topically, before the treatment; - A sample taken from an untreated area before treatment; - A sample taken from an area treated with the ingredient or composition, preferably applicable topically, after the treatment; - A sample taken in an untreated area after the treatment period.
  • the “untreated zone” according to the invention is a skin zone equivalent or close, functionally and/or in microbial composition, to said “zone to be treated”.
  • the invention relates to a method for selecting or screening or evaluating the activity of at least one ingredient or composition, preferably applicable topically, in particular a cosmetic ingredient and /or dermatological, optionally formulated in the form of a topical composition, characterized in that the method comprises the following steps: - a sample of biological or genetic material of the microbiota present in the skin, of one or more human beings, said sample being performed in an area to be treated with the topically applicable ingredient or composition prior to treatment; - a sample of biological or genetic material of the microbiota present in the skin, or present on a keratinous material, of one or more human beings, said sample being taken in an untreated area before the treatment; - a sample of biological or genetic material of the microbiota present on the skin, or present on a keratin material, of one or more human beings, said sample being taken from an area treated with the ingredient or composition applicable topically after the treatment ; - a sample of biological or genetic material of the micro
  • the duration of the treatment with the ingredient or the composition is variable.
  • the application of the product can be done daily or several times a day or over the test period.
  • several samples can be taken from the treated zone, at, for example, 5 minutes, 1 hour, 24 hours, 2 days, 4 days, 7 days and 31 days from the start of the treatment.
  • the sample is taken from the treated zone after 2 to 7 days of daily treatment.
  • the application of the product is done due to a volume of between 0.1 and 100 ⁇ L/cm 2 , advantageously between 1 and 10 ⁇ l/cm 2 , standardizing the application as much as possible, for example by using a robotic arm.
  • at least one sample is taken from several human beings.
  • the particular group of subjects at the origin of the samples can share characteristics (ethnicity, age, phototype, chronic dermatological conditions, type of skin, diet, lifestyle, rate of hydration of the skin, geolocation and other similar parameters) which allow the development of treatment methods, preferably cosmetic and/or dermatological, specifically targeting said groups of particular subjects, also called “populations".
  • the population at the origin of the samples is characterized by an impoverished skin microbiota at the time of the samples.
  • depleted skin microbiota within the meaning of the invention is meant any decline in the biomass of the skin microbiota.
  • the depletion can be linked to a skin aggression according to the invention, more particularly to the action of bactericides or virucides or fungicides or soaps or aggressive hygiene or care products vis-à-vis of the microbiota.
  • an antiseptic agent that is to say fungicide, bactericide and/or virucide
  • a composition comprising a C1-C8 alcohol chosen from ethanol, isopropanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, benzyl alcohol and mixtures thereof; and/or an antiseptic agent other than an alcohol, such as for example benzalkonium chloride, chlorhexidine gluconate, sodium hypochlorite, a soap produced by saponification of a fatty acid; and/or also a composition comprising an acid
  • the sampling of biological or genetic material of the microbiota present in the skin, or present on a keratinous material is carried out on a single human being.
  • This method makes it possible to develop care methods, advantageously cosmetic and/or dermatological, preferably ecobiological, personalized, based on the conditions, specificities and responsiveness to care, of the skin microbiota of each individual.
  • the invention also relates to a personalized method for selecting or screening or evaluating, preferably in vitro, the activity of at least one ingredient or composition, preferably applicable topically, in particular a cosmetic and/or dermatological ingredient, optionally formulated in the form of a composition, advantageously topical, characterized in that the method comprises the following steps: - the selection of a skin area or skin appendages to be studied, - at least one sample of biological or genetic material of the microbiota present in this area, of a single human being, - the application of at least one ingredient or a composition to this area, in a single or repeated manner, - after a period of 1 hour at 7 days, at least one sample of biological or genetic material from the microbiota present in this area, from a single human being, - at least one method for quantifying the biomass present in the samples, - at least one method making it possible to measure the microbial diversity present in the samples, - the evaluation of the activity of the ingredient or composition applied topically on the biomass
  • the personalized method according to the invention can be accompanied by an analysis of the dermatological profile of the single human being at the origin of the at least one sample on the basis of his ethnicity, age, phototype, chronic dermatological conditions, skin type, diet, lifestyle, skin hydration level, geolocation and other similar parameters.
  • This information can be collected by direct analysis or through a form.
  • the methods for weighting cutaneous, capillary, or nail characteristics and the computer program described in document FR 2983328 can be combined with the results of the personalized screening method mentioned above to select an ecobiological dermatological treatment capable of inducing the increase in the biomass of the cutaneous microbiota of an individual, in particular without reducing its diversity, and in accordance with the overall dermatological needs of said individual.
  • the invention also relates to a method for preparing a composition, preferably topical, characterized in that it comprises (i) the implementation of a screening or selection method defined previously and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing an increase in the biomass of the cutaneous microbiota in particular without reducing its biodiversity (ii) the formulation of the ingredients or compositions thus selected in a formula with cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity (iii) the packaging of the said formula.
  • the invention also relates to a topical care method, preferably cosmetic or dermatological, advantageously ecobiological, characterized in that it comprises (i) the implementation of a screening method defined previously or in the following description and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing the increase in the biomass of the cutaneous microbiota described above, in particular without reducing its biodiversity (ii) the formulation of the ingredients or compositions thus selected in a composition for cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity (iii) the packaging of said formula (iv) the topical application of said composition to an area of skin or a surface of the selected appendages.
  • a topical care method preferably cosmetic or dermatological, advantageously ecobiological, characterized in that it comprises (i) the implementation of a screening method defined previously or in the following description and the selection of at least one ingredient or a composition, in particular
  • the advantageously ecobiological topical care method can target specific populations, or be personalized and adapted to the microbiota specific to each individual.
  • the population to be treated is characterized by an impoverished skin microbiota.
  • it is a personalized method capable of treating an individual characterized by an impoverished cutaneous microbiota.
  • the invention relates to an advantageously cosmetic or dermatological topical care method, preferably ecobiological, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without impact or reduction on its biodiversity, characterized in that a composition selected through the selection or screening method mentioned above or described below is applied to the skin.
  • the invention relates to a care method, preferably cosmetic or dermatological, for combating a skin attack (for example against a bactericidal and/or virucidal and/or fungicidal action), and/or maintaining and /or restore a healthy microbiota, characterized in that the method comprises the topical application of at least one topical composition selected in the screening or selection method described previously or in the following description, said composition being capable of inducing the increase of the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, on the skin or on a keratin material.
  • the composition selected through the screening or selection method mentioned above or in the following description and capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity is associated with the action of maintaining and/or restoring a healthy skin microbiota.
  • the composition to be used in particular in the implementation of the preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its diversity, according to the The invention is an aqueous composition, comprising in particular at least 60%, better still at least 70%, or at least 80%, or at least 90%, or even at least 95% of water.
  • the invention relates to a preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota in particular without reducing its diversity, according to the invention, with an aqueous composition, advantageously comprising at least 60% of water, preferably 70%, 80%, 90%, or even at least 95% water.
  • the aqueous composition to be used in particular in the care method mentioned above or in the following description comprises water which is characterized by a resistivity of between 12.5 and 12,500 Ohms.cm, in particular between 80 and 8000 Ohms.cm, at room temperature
  • the composition to be used in particular in the care method mentioned above or in the following description has an osmotic pressure of between 70 and 2500 mOsmol/l, and in particular between 100 and 800 mOsmol/l.
  • the composition to be used in particular in the implementation of the preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity comprises at least one antioxidant and at least one inorganic or organic salt, each or at least one of these components or ingredients having, advantageously, been selected beforehand by the screening method described above or in the following description.
  • antioxidant is meant a compound which exhibits an antiradical activity, in particular on reactive oxygen (ROS), carbonyl (RCS) or nitrogen (RNS) species.
  • the at least one antioxidant is chosen from the group comprising octadecyl di-t-butyl-4-hydroxyhydrocinnamate; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t- butylnitrone; ergothioneine; melatonin; acetyl cyste
  • the at least one antioxidant is chosen from the group comprising tocopherol, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose.
  • the at least one antioxidant according to the invention is carnosine.
  • the invention relates to a composition
  • a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine.
  • a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine.
  • the invention relates to a composition
  • a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, chloride of sodium, sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine for its use in maintaining and/or restoring a healthy skin microbiota.
  • the invention relates to a composition comprising: - sodium citrate, - carnosine.
  • the invention relates to a composition
  • a composition comprising: - sodium citrate, - carnosine for its use in maintaining and/or restoring a healthy skin microbiota.
  • the composition according to the invention, or for its use according to the invention also comprises at least one carbon source.
  • Carbon source means a compound likely to be used by bacteria as a substrate for growth and multiplication, such as: - partially or totally hydrolyzed proteins from cereals (wheat, corn, barley, rye, oats), legumes (soya, broad bean, broad bean, pea), milk (caseins, albumins), yeast, as well as peptides and amino acids and their derivatives alone or in mixtures; - oligosaccharides or polysaccharides, advantageously short-chain, such as fructooligosaccharides, hyaluronic acid, pullulan, alginate, cellulose, advantageously microcrystalline cellulose, carrageenan, chitosan, dextran and dextran sulphate, galactoarabinan, amylose, amylopectin, maltodextrins, pectins, and their respective hydrolysates, as well as sugars, oses, osides and their derivatives such as glucose, galacto
  • Fructo oligosaccharides or FOS can be derived from plant materials such as beets, artichokes or corn.
  • the raw material corresponding to the INCI designation fructooligosaccharides can be used in the context of the present invention, for example the product marketed by the company Shaanxi Bolin Biotechnology Co under the name of FRUCTOOLIGOSACCHARIDES.
  • the FOS represent from 0.001% to 20% by total weight of the composition, advantageously from 0.01% to 10%, even more advantageously from 0.1% to 5%.
  • the invention relates to a composition
  • a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously cellulose microcrystalline.
  • a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine -
  • the invention relates to a composition
  • a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine; - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose for its use in maintaining and/or restoring a healthy skin microbiota.
  • a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphat
  • the invention relates to a composition
  • a composition comprising: - sodium citrate, - carnosine. - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose
  • the invention relates to a composition comprising: - sodium citrate, - carnosine - au least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose for its use in maintaining and/or restoring a healthy skin microbiota.
  • a pure or highly purified form of carnosine is used in the composition according to the invention.
  • the carnosine is obtained by chemical synthesis.
  • the cosmetic material L-Carnosine corresponding to the INCI name carnosine and marketed by the company MERCK KgaA.
  • the carnosine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%.
  • a pure or highly purified form of hypotaurine can be used in the composition according to the invention.
  • the hypotaurine is obtained by chemical synthesis.
  • the hypotaurine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%.
  • the tocopherol used is a mixture of purified natural tocopherols, in particular ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, and ⁇ -tocopherol; this mixture can be used in particular in an oil chosen from vegetable, mineral, silicone, and preferably vegetable oils.
  • a mixture of natural tocopherols that can be used according to the invention, mention may be made of the mixture sold by the company Nisshin OilliO Group, Ltd under the name Tocopherol 80 and corresponding to the INCI name “Tocopherol”.
  • the tocopherol represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%.
  • Betaine is an osmolyte and an antioxidant found in particular in the plant kingdom. Betaine can be of vegetable origin or obtained by biotechnology.
  • a highly purified form of betaine, with a purity of at least 95%, is used in the compositions according to the invention.
  • a source of betaine that can be used according to the invention, mention may be made of the raw material sold by the company Evonik Nutrition & Care GmbH under the name of Tego Natural Betaine (Evonik Nutrition & Care GmbH) and corresponding to the INCI name “Betaine”.
  • the betaine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%.
  • the xylitol can be powdered xylitol, for example of vegetable origin, in particular obtained from wood by hydrogenation of xyloses, having in particular a purity greater than 95%.
  • the company Orient Star LCC markets the product OriStar XLT corresponding to the INCI name “Xylitol” which can be implemented within the framework of the invention.
  • the xylitol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%.
  • the mannitol can for example be powdered mannitol, of plant origin (corn, potato, wheat), preferably having a purity greater than 98%.
  • the company American Biochemicals, Inc. markets the product D-mannitol corresponding to the INCI name mannitol which can be used in the context of the invention.
  • the mannitol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%.
  • Inositol is a cyclic compound derived from cyclohexane and carrying six secondary alcohol functions.
  • inositol of plant origin is used, synthesized naturally from glucose-6-phosphate (G6P), resulting from photosynthesis or from the hydrolysis of the carbon reserves of the plant.
  • G6P glucose-6-phosphate
  • inositol extracted from rice by techniques known to those skilled in the art is used. It is also possible to use inositol prepared by chemical or enzymatic synthesis.
  • the company Dupont Industrial Biosciences markets the product Genencare OSMS MI corresponding to the INCI name inositol which can be used in the context of the invention.
  • the inositol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%.
  • All cosmetically or pharmaceutically acceptable inorganic or organic salts are suitable for use in the composition according to the invention or for its use according to the invention.
  • it is at least one inorganic or organic salt chosen from the group consisting of a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulfate, potassium or sodium phosphate, copper or zinc gluconate, and mixtures thereof, more particularly sodium citrate.
  • the at least one inorganic or organic salt is chosen from the group comprising calcium chloride; potassium chloride; potassium phosphate; magnesium sulfate; sodium chloride; sodium hydrogen carbonate; sodium hydrogen phosphate; sodium citrate; magnesium gluconate.
  • mineral salts are available from several suppliers of cosmetic raw materials such as, for example, COOPER INDUSTRIE or BRENNTAG FRANCE.
  • the composition according to the invention comprises at least one of the following mineral salts, in percentage by weight: - From 0.000001% to 0.1% by total weight of the composition of calcium chloride , in particular from 0.0001% to 0.1%; - From 0.0001% to 0.1% by total weight of the potassium chloride composition, in particular from 0.001% to 0.1%; - From 0.00001% to 0.1% by total weight of the phosphate composition of potassium, in particular from 0.0001% to 0.1%; - From 0.0001% to 0.1% by total weight of the magnesium sulphate composition, in particular from 0.001% to 0.1%; - From 0.001 to 4% by total weight of the composition of sodium chloride, in particular from 0.1 to 4%; - From 0.0001 to 0.1% by total weight of the sodium hydrogencarbonate composition; - From 0.0001% to 0.1% by total weight of the sodium hydrogen phosphate composition; - From 0.0001% to 1% by total weight of the citric acid composition, in particular from 0.001% to 1%; - From 0.000001% to 0.
  • the total weight of the mineral and/or organic salts is less than or equal to 1% by weight, in particular is less than or equal to 0.1% by weight of the total weight of the composition.
  • the inorganic and organic salts are used at a concentration which makes it possible to be within the range of osmotic pressure sought.
  • the sodium citrate represents from 0.001% to 4% by total weight of the composition and the carnosine represents from 0.0001% to 5% by total weight of the composition, advantageously from 0.001% to 2%.
  • Several cosmetic sources of hyaluronic acid are known.
  • the hyaluronic acid has a molecular weight comprised between comprised between 20 and 50 kDa.
  • the raw materials PRIMALHYALTM 50 or HyaCare 50 marketed respectively by the companies SOLIANCE and EVONIK and corresponding to hyaluronic acid with a molecular weight (Mw) of between 20 and 50 kDa can be used in the context of the present invention.
  • the hyaluronic acid represents from 0.001 to 4% by total weight of the composition, advantageously from 0.001% to 2%.
  • microcrystalline cellulose suitable for topical and cosmetic application can be used in the compositions according to the invention.
  • the microcrystalline cellulose represents from 0.001 to 5% by total weight of the composition, advantageously from 0.001% to 3%.
  • the composition according to the invention or for its use according to the invention or to be used in particular in the care method mentioned above or in the following description is intended for populations characterized by an impoverished cutaneous microbiota.
  • FIG.5] and [Fig.6] show the effect of the pure ingredients petrolatum [Fig.5] and silicone [Fig.6] after 4 days of contact versus untreated area (ZNT), on the bacterial diversity of the microbiota cutaneous, measured by the Shannon index, in the volunteers taken into account for the analysis.
  • ZNT untreated area
  • FIG.7] and [Fig.8] show the effect of formula 1 (fle 1; Fig.7) and formula 2 (fle 2; Fig.8) after 4 days of contact versus untreated area ( ZNT), on the bacterial diversity of the skin microbiota, measured by the Shannon index, in the volunteers taken into account for the analysis.
  • Statistical significance is determined by a paired Wilcoxon test. (ns, not significant: p>0.05; significant*: p ⁇ 0.05; significant**: p ⁇ 0.01; significant***: p ⁇ 0.001). Examples 1 of the Evaluation of the Effect of Two Cosmetic Compositions and Two Pure Ingredients on the Skin Microbiota of Healthy Volunteers
  • the objective of the study is to evaluate the effect at 4 days of 2 cosmetic ingredients
  • the methodological approach is based on a process consisting of the following steps: a) Extraction of the gDNA ( genomic DNA) from swabs b) Analysis of the quantity of biomass using a qPCR approach c) Analysis of microbial diversity based on:
  • step a) of extracting the genomic DNA from the swabs consists of an optimized bacteria recovery step (consisting of wiping the swab and recovering the bacterial pellets by centrifugation) which is carried out before the extraction step.
  • the genomic DNA extraction was then carried out with the Nucleospin Microbial DNA kit (Macherey Nagel, France).
  • the extracted gDNAs are not “assayable”, a quantification of the biomass and a quality control was therefore carried out by 16S qPCR.
  • step b) of analysis of the biomass and quality control by qPCR16S consists of carrying out a standard range under standardized experimental conditions.
  • the negative control called “Tneg” is the negative control for qPCR carried out with sterile water.
  • the quantitative amplifications are carried out with 16S universal primers [SEQ ID. 18F (5 AGAGTTT G ATCCT GGCT CAG-3') and SEQ ID. 2,139 IR (5'-GACGGGCGGTGWGTRCA-3')] (Kong et al. 2012b).
  • SEQ ID. 18F 5 AGAGTTT G ATCCT GGCT CAG-3'
  • SEQ ID. 2,139 IR 5'-GACGGGCGGTGWGTRCA-3'
  • step c) of analysis of the diversity of the cutaneous microbiome a maximum quantity of material is used for the creation of specific Illumina MiSeq libraries .
  • the amplification step is carried out with the primers for the V1-V3 regions corresponding to SEQ. ID 5 27F (5'-AGAGTTTTGATCCT GGCT CAG-3') and SEQ. ID 6534R (5-ATTACCGCGGCTGCTGTGG-3) according to San Miguel et al. 2018, with the following modifications.
  • Two additional controls are integrated during the preparation of the Metabiote® libraries.
  • the second control is a negative control called "BF", this one is also integrated into the preparation process and makes it possible to evaluate the background noise inherent to the reagents used after the extraction step.
  • the quantitative control of the final libraries is carried out by a fluorometric method.
  • the ABC control community whose composition is known (17 bacteria including 2 archaebacteria) is introduced into the sequencing; it serves as a positive control during our MetaBiote® process, but also as a positive control when performing the taxonomic affiliation of the sequences.
  • the negative library creation control (BF.V1V3) is used to identify what profile can be obtained if no initial biomass is introduced and the MetaBiote® preparation process is pushed. This control makes it possible to distinguish the samples to be excluded from the comparisons.
  • the alpha diversity indices of the samples as well as the measures of rarefaction are defined, thus making it possible to observe the taxonomic diversity (in number of OTUs) of the samples.
  • formula 1 (fle 1) induces an average increase in biomass of 134% after 4 days of application (figure 3) in the volunteers taken into account for the analysis, while formula 2 (fle 2 ) induces an average increase of 150% (Figure 4).
  • the difference between the untreated zone (ZNT) and the treated zone (ZT) is significant in both cases (test of
  • Amplification with specific 16S primers followed by sequencing of the amplicons obtained shows that petroleum jelly and silicone have, in practice, no effect on the diversity of the skin microbiota (figures 5 and 6; p>0.05 in both case therefore difference not significant; see tables 4 and 5). This observation is consistent with the results of the biomass analysis and indicates that these ingredients are substantially inert with respect to the skin microbiota.
  • Example 1 The techniques described in Example 1 are reproduced identically.
  • the ingredients listed in Table 11 are placed at a percentage indicated in grams per 100 grams (qsp 100 g of water), the compositions are applied neat.
  • the products are applied to the back of series of 10 volunteers. Applications are daily for 4 days and samples of biological material from the skin are taken before the first application and after the 4th day.
  • the biomass analysis is carried out by qPCR (16S universal primers) and the analysis of the microbial diversity by amplification with specific 16S primers, then analysis of the results by measuring the Shannon index.
  • the summary results are grouped in the following table 11:
  • Example 3 Study on a cutaneous area characterized by a reduced cutaneous microbiota biomass after the action of an ethanolic solution
  • the study is carried out on the forehead of 5 volunteers.
  • a biomass sample is taken at T0 on the forehead of the volunteers.
  • disinfection of the entire forehead with a compress soaked in 5 mL of 70° alcohol (modified alcohol from Cooper, batch 013199) is carried out by rubbing for 30 seconds and leaving for an additional 3 minutes.
  • 2 samples are taken: T5 min on the left (G) and T5 min on the right (D).
  • a compress soaked in 5 mL of the above composition is placed on the right half of the forehead for 3 min.
  • a 2 sec mist of the above composition is applied to the right half of the forehead.
  • 2 samples are taken: T4h on the alcohol side and T4h on the composition to be tested side.
  • the samples are taken using swabs (Sigma Trans Swab liquid Amies ref MW176S from Elitech) at times T0, T5min, T2h and T4h, by performing 10 round trips in Z over an area of 3.75 cm 2 and exerting light pressure on the swab in a standardized manner. The swab is then kept on ice. Effect on biomass:
  • This suspension is then diluted 1/2, 1/10 and 1/50 in nutrient broth.
  • a seeding of 1 mL of each dilution is then carried out on TSA agar in a Petri dish and incubated at 30-35° C. for 48 hours. After 48 hours of incubation the colonies (CFU) are counted in full on the dishes.
  • the bacterial biomass is obtained by multiplying the number of CFU counted by its dilution factor and is expressed in CFU/mL.
  • Disinfection with 70° alcohol induced a sharp decrease in the amount of biomass of cultivable bacteria by 99.8%, 5 min after application of the alcohol.
  • Application of the product to be tested after 4 hours of contact on the depleted microbiota increases this biomass by 160% compared to the 70° alcohol control alone.
  • Example 4 Study on a skin area characterized by a reduced skin microbiota biomass after the action of a hygiene product based on Marseille soap
  • the study is carried out on the forehead of 5 volunteers.
  • a biomass sample is taken at T0 on the forehead of the volunteers.
  • washing the entire forehead with a compress soaked in 5 mL of a 2% by weight Marseille soap solution (Marseille soap, Persavon) is carried out by rubbing for 30 seconds.
  • Two rinses are then carried out with compresses soaked in water. After 5 min of drying, a sample is taken: T5min.
  • a compress soaked in 5 mL of the above composition is placed on the right half of the forehead for 3 min. After 2 hours, a 2 sec mist of the above composition is applied to the right half of the forehead.
  • 2 samples are taken: T4h on the Marseille soap side and T4h on the composition to be tested side.
  • the samples are taken using swabs (Sigma Trans Swab liquid Amies ref MW176S from Elitech) at times T0, T5min, T2h and T4h, by performing 10 round trips in Z over an area of 3.75 cm 2 and exerting light pressure on the swab in a standardized manner. The swab is then kept on ice.
  • swabs Sigma Trans Swab liquid Amies ref MW176S from Elitech
  • This suspension is then diluted 1/2, 1/10 and 1/50 in nutrient broth.
  • a seeding of 1 mL of each dilution is then carried out on TSA agar in a Petri dish and incubated at 30-35° C. for 48 hours. After 48 hours of incubation, the colonies (CFU) are counted in their entirety on the dishes.
  • the bacterial biomass is obtained by multiplying the number of CFU counted by its dilution factor and is expressed in CFU/ml.
  • Kong HH, Segre JA (2012a) Skin microbiome: looking back to move forward Review J Invest Dermatol 132(3 Pt 2): 933-939.
  • 2012b Temporal shifts in the skin microbiome associated with disease flares and treatment in children with atopy dermatitis Genome Res. 22(5): 850-9.

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Abstract

The invention relates to a method of selection which combines quantifying the biomass of the skin microbiota, and analysing the biodiversity thereof, for selecting the ecobiological substances or compositions which make it possible to increase the biomass of the skin microbiota, in particular without reducing the biodiversity thereof, for the ability of said substances or compositions to maintain and/or re-establish a normal skin microbiota. The invention makes it possible in particular to efficiently reinforce the skin against attacks, in particular repeated attacks, on the microbiota of said skin, in particular by topical virucidal or bactericidal, hygiene or care products.

Description

[Description] METHODE DE SELECTION D’INGREDIENTS ET DE COMPOSITIONS TOPIQUES ECOBIOLOGIQUES APTES A RESTAURER OU RENFORCER LE MICROBIOTE CUTANE Domaine technique de l’invention La présente invention concerne l'étude du microbiote cutané et son utilisation pour le rétablissement et/ou le maintien d'un microbiote cutané sain. Etat de la technique La peau, plus grand organe humain, est colonisée par des milliards de microorganismes tels que bactéries, levures, champignons ou virus collectivement appelés microbiote ou microflore. Ces microorganismes habitent l'épiderme, et se retrouvent principalement dans les couches supérieures du stratum corneum ainsi que dans les conduits des glandes sudoripares et des follicules pilo-sébacés (Kong et al., 2012a). Historiquement, l'étude du microbiote humain s'est focalisée d'abord sur la microflore intestinale, dont le rôle crucial dans le maintien de l'équilibre physiologique humain est désormais reconnu de façon indiscutable. Le microbiote cutané, beaucoup moins abondant, a dû attendre le développement et la mise au point de méthodes métagénomiques permettant sa caractérisation détaillée. Ces techniques, maintenant disponibles, permettent l'amplification et le séquençage du matériel génétique des communautés microbiennes de la peau. A ce jour, la composition de la flore cutanée n'est que partiellement connue. Seule la composition bactérienne de la peau est étudiée en détail. Le nombre de bactéries présentes sur la peau, que ce soit à sa surface, au niveau des glandes ou sur les phanères , peut atteindre près d'1 milliard par cm2 (Kong et al., 2012a). La flore cutanée humaine peut être subdivisée en deux groupes : - La flore résidente est composée de germes commensaux, c'est-à-dire vivant aux dépens de leur hôte, sans leur causer de dommage. La flore résidente humaine est dominée par les Proteobacteria, notamment une dizaine de types de bactéries aérobies à Gram positif telles que Staphylococcus epidermidis et Staphylococcus hominis (Otto, 2010). La flore résidente est aussi composée de bactéries anaérobies à Gram positif appartenant à la division des Actinobactéries (genres Cutibacterium, Corynebacterium, Dermabacter et Micrococcus) (Kligman et al., 1976). Parmi, les microorganismes des autres règnes, Malassezia est l'espèce fongique la plus fréquemment retrouvée sur la peau. Enfin, les acariens Demodex folliculorum et Demodex brevis, ou « acariens des cils », sont aussi considérés comme des membres de la flore résidente humaine. - La flore transitoire est composée de champignons, virus et bactéries pour la plupart inoffensifs, dites saprophytes, c'est-à-dire qui se nourrissent de matières organiques en décomposition provenant de l'environnement. Cette flore peut aussi être constituée de bactéries pathogènes opportunistes pouvant entraîner des maladies chez l'hôte. La flore transitoire ne s'établit pas de façon permanente à la surface de la peau, variant dans la journée, selon les activités réalisées et les conditions environnantes. Elle peut néanmoins persister des heures voire des jours. Sa densité est faible sur les zones sèches et particulièrement élevée sur les zones poilues, les plis et des zones sujettes à transpiration. Les espèces transitoires les plus communes sont Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa et plusieurs espèces du genre Bacillus. Plusieurs études ont montré que le microbiote cutané présente des différences considérables entre sexes, groupes ethniques et même individus et que la répartition des résidents microbiens dans les différentes zones du corps est également très variable (Grice et al., 2009). Malgré ces différences interpersonnelles et biogéographiques marquées, il est solidement établi que l'ensemble de la microflore résidente joue un rôle important dans la santé de la peau, notamment au niveau de ses défenses immunitaires et de sa résistance à la colonisation par des microorganismes pathogènes, impliquant par ailleurs des déséquilibres dans le microbiote et l'apparition ou l'aggravation de conditions dermatologiques comme la dermatite atopique ou le psoriasis (Nakatsuji et al., 2017 ; Gao et al., 2008 ; Kong et al., 2012b). En effet, tout comme dans les écosystèmes à large échelle, une perte de diversité du microbiote s'accompagne d'une dérégulation parfois irréversible de ses équilibres, dans lesquels un nombre limité d'espèces opportunistes prolifèrent de façon incontrôlée aux dépens des autres. Dans le cas du microbiote cutané, il est par exemple possible de citer en particulier les espèces responsables d'infections graves, souvent d'origine nosocomiale, Staphylococcus aureus et Pseudomonas aeruginosa. Il est délicat de traiter les infections cutanées causées par ces souches qui ont parfois acquis au cours de leur évolution, une ou plusieurs résistances aux antibiotiques classiques, en particulier les beta-lactames. Par exemple, le staphylocoque doré résistant à la méticilline (SARM) représente à ce jour un risque concret pour la santé publique, ayant été en 2020 responsable de plus d'un tiers de toutes les infections communautaires et hospitalières associées à une mortalité et une morbidité élevée. La prise en charge du SARM requiert l'utilisation de nouveaux antibiotiques, qui peuvent cependant déstabiliser encore davantage l'ensemble du microbiote humain en ralentissant le rétablissement de sa diversité. Plusieurs indices permettant d'évaluer la diversité du microbiote sont développés, parmi lesquels on peut citer les indices de Shannon ou de Simpson, le plus employé étant l'indice de Shannon qui permet à la fois d'évaluer le nombre d'espèces de ce milieu (richesse spécifique) et la répartition des individus au sein de ces espèces (équitabilité spécifique ; Lemos et al., 2011). Il est connu que l'utilisation de produits topiques peut avoir un impact important sur la composition et les équilibres des communautés microbiennes cutanées (Kong, 2011 ; Kong et al., 2012a). Les produits antiseptiques (c'est-à-dire fongicides, bactéricides et/ou virucides) en particulier, parmi lesquels figure notamment l'éthanol, appauvrissent non seulement la quantité, mais aussi la diversité du microbiote cutané et ce potentiellement plus durablement (San Miguel et al., 2018). L'application de biocides à la surface de la peau était relativement rare et limitée au contexte hospitalier jusqu'en 2020. Cependant, depuis la pandémie planétaire de COVID-19, l'utilisation de cette catégorie de produits est banalisée et s'est répandue jusqu'à des niveaux jamais atteints auparavant. Pour les produits à base d'alcool, l'inhibition indiscriminée de la croissance de l'ensemble du microbiote s'additionne à son effet de destruction du film hydrolipidique, ce qui entraine une altération de son effet barrière, avec une pénétration accrue d'allergènes et d'irritants. La réduction de la qualité du microbiote cutané, notamment causée par les produits biocides et par les traitements antibiotiques, requiert le développement d'une nouvelle classe de soins topiques écobiologiques permettant de maintenir et/ou rétablir rapidement un microbiote cutané sain et diversifié. Il subsiste donc un besoin évident de mise au point de méthodes permettant la sélection d'ingrédients, voire de compositions topiques, aptes à maintenir et/ou rétablir, préférentiellement rapidement, un microbiote sain. Description de l'invention La présente invention repose sur la mise en évidence de l'intérêt, dans des conditions non pathologiques, de favoriser l'augmentation de la biomasse du microbiote cutané (multiplication du microbiote), sans réduction de sa diversité globale (conservation ou augmentation de la diversité dudit microbiote). Le microbiote cutané sain et son équilibre sont alors renforcés et/ou remis en condition de mieux résister aux dérégulations et colonisations opportunistes potentielles. De façon surprenante, la demanderesse a ainsi trouvé une façon simple, efficace, abordable et utilisable à l'échelle industrielle, notamment en cosmétique ou dermatologie, de sélection d'ingrédient(s) et/ou de composition(s) apte(s) à restaurer et/ou renforcer le microbiote cutané sain, en recherchant l'augmentation de la biomasse du microbiote cutané, en particulier sans réduction de sa diversité globale. Pour cela, dans un mode de réalisation de l'invention, deux méthodes sont combinées : une méthode de mesure de la biomasse sur une zone cutanée et une méthode de mesure de la diversité microbienne sur cette même zone. Selon l'invention, le « microbiote » concerne l'ensemble de la flore microbienne, dont les microbes, bactéries, levures, champignons et virus. Selon l'invention, par « microbiote cutané » ou de(s) « matière(s) kératinique(s) », on entend le microbiote de la peau, des semi-muqueuses (en particulier les lèvres), des muqueuses et des annexes dont les phanères. Selon l'invention, le « microbiome » concerne l'ensemble des gènes ou génomes présents dans le microbiote. Selon l'invention, par « biodiversité » ou « diversité » ou « diversité du microbiote » ou « diversité du microbiome » ou « diversité bactérienne », on entend l'ensemble des populations ou espèces bactériennes présentes dans le microbiote cutané. Cette biodiversité peut être caractérisée notamment par le nombre de séquences génétiques bactériennes différentes au sein dudit microbiome (noté également OTU, pour « Operational Taxonomic Unit ») et/ou leur abondance relative. Plusieurs indices permettant d'évaluer la diversité du microbiote sont développés et bien connus de l'homme du métier, parmi lesquels on peut citer les indices de Shannon ou de Simpson. Selon l'invention, par « biomasse du microbiote cutané » ou « biomasse » ou « charge bactérienne », on entend le nombre ou la quantité de bactéries sur une zone de peau. Cette biomasse peut être caractérisée notamment par la quantité de matériel génétique bactérien, en particulier d'ADNr 16S, issu dudit microbiote. Les niveaux de biomasse et de diversité d'un microbiote peuvent être mesurés par toutes techniques appropriées bien connues de l'homme du métier, telles que l'amplification quantitative et le séquençage de l'ADNr 16S. Selon l'invention, par « sain », on entend non pathologique. Selon l'invention, par « zone cutanée saine », on entend une zone cutanée ne présentant pas de dermatose. Selon l'invention, par « microbiote cutané sain » ou « microbiote de la peau saine », on entend un microbiote cutané stable et diversifié sur une zone cutanée donnée, tel que sur une même zone cutanée saine non soumises à des conditions de stress ou d'agression particulières (telles que, mais non limité à, des traitements ou soins ou environnements non écobiologiques, notamment dermatologiques, cosmétiques, d'hygiène, de pollution …). Le microbiote de la peau saine protège notamment des pathogènes invasifs, mais également d'un dessèchement de la peau. Selon l'invention, par « stress » ou « agression », on entend tout événement, de quelque nature que ce soit, unique ou répété, notamment d'origine externe, pouvant être à l'origine d'une déstabilisation ou d'une modification d'un système. Au niveau cutané, une telle agression peut notamment entrainer une diminution de la biomasse et/ou de la diversité du microbiote cutanée. En particulier, elle peut être liée à des traitements ou soins ou environnements non écobiologiques, notamment dermatologiques, cosmétiques, d'hygiène, de pollution etc, et, plus particulièrement, à l'action de bactéricides ou de virucides ou de fongicides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Par « écobiologique », on entend respectueux de la personne, de ses interactions avec le monde et de la planète. La présente invention concerne une méthode de sélection ou criblage d'ingrédients et de compositions, de préférence écobiologiques, cosmétiques ou dermopharmaceutiques ou dermatologiques, avantageusement applicables par voie topique, en fonction de leur effet sur le microbiote cutané, notamment de leur aptitude à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans réduire sa biodiversité. La présente invention concerne également une méthode de préparation d'une composition, de préférence topique, apte à induire l'augmentation de la biomasse du microbiote cutané, ainsi que son utilisation pour la préparation de compositions de soins, de préférence topiques, avantageusement écobiologiques et de méthodes de soin écobiologique, de préférence par voie topique, basées sur l'utilisation des compositions ou ingrédients susmentionnés. La présente invention vise encore une composition pour son utilisation pour maintenir et/ou restaurer un microbiote cutané sain via une augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité. Autrement dit, l'invention vise pour la première fois à analyser et quantifier la biomasse du microbiote cutané, ainsi que, le cas échéant, sa biodiversité et à sélectionner les substances et compositions qui permettent d'augmenter la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité. La présente invention concerne donc une méthode de sélection ou criblage d'ingrédients, de substances ou de compositions topiques permettant de maintenir et/ou rétablir, de préférence rapidement, un microbiote sain. La présente invention concerne notamment une méthode de criblage dont l'objectif est de sélectionner des ingrédients, préférentiellement topiquement acceptables, individuellement et/ou des compositions comprenant un mélange de plusieurs ingrédients précités, éventuellement associés à des excipients, préférentiellement topiquement acceptables, capables d'augmenter la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité. La présente invention concerne par ailleurs une composition, de préférence topique, sélectionnée par la méthode de criblage. La présente invention concerne également un ou des ingrédients, éventuellement sous forme d'une composition, de préférence topique, à utiliser pour la fabrication d'une composition, de préférence topique, pour le soin de la peau, des muqueuses ou des phanères capables d'augmenter la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité, pour maintenir et/ou restaurer le caractère sain de la peau, des muqueuses ou des phanères. La présente invention concerne enfin une méthode de soin, de préférence par voie topique, utilisant un ou des ingrédients, avantageusement topiquement acceptables, capables d'augmenter la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité, pour maintenir et/ou restaurer le caractère sain de la peau, des muqueuses ou des phanères. Ainsi, selon un premier aspect, l'invention concerne une méthode, préférentiellement in vitro, de sélection d'un ingrédient ou composition, de préférence écobiologique, avantageusement applicable par voie topique, en particulier cosmétique ou dermatologique, utile dans le maintien ou/et la restauration d'un microbiote cutané sain, comprenant les étapes consistant à : a) mettre un ingrédient ou composition à tester en contact avec des bactéries d'un microbiote de référence, b) analyser la quantité de biomasse et, avantageusement la biodiversité du microbiote après la mise en contact, et c) sélectionner en tant qu'ingrédient ou composition utile dans le maintien et/ou la restauration d'un microbiote cutané sain, un ingrédient ou une composition capable d'augmenter la biomasse du microbiote cutané, avantageusement sans en diminuer la biodiversité, par rapport à la quantification de la biomasse et, le cas échéant, la biodiversité du microbiote de référence. Selon un mode de réalisation particulier, la méthode selon l'invention est personnalisée, basée sur les conditions, les spécificités et la réactivité du microbiote cutané d'un individu donné. Selon un mode de réalisation particulier, l'étape a est réalisée de façon unique ou répétée et / ou l'étape b est réalisée de 1 h à 31 jours, préférentiellement 4 jours, après l'étape a. Selon une variante de réalisation, l'invention concerne une méthode, préférentiellement in vitro, de criblage ou d'évaluation ou de sélection de l'activité d'au moins un ingrédient ou composition, préférentiellement applicable par voie topique, en particulier un ingrédient ou composition cosmétique et/ou dermatologique, éventuellement formulé sous forme de composition topique, caractérisée en ce que la méthode comprend les étapes, préférentiellement in vitro, suivantes : i) avant l'application de l'ingrédient ou de la composition, éventuellement formulé sous forme de composition topique, sur au moins une zone cutanée à étudier présentant un microbiote dit de référence, à partir d'un premier prélèvement réalisé sur ladite zone cutanée, les analyses de la quantité de biomasse du microbiote cutané et, le cas échéant de sa biodiversité, en obtenant ainsi une première mesure dite de référence, de quantification de la biomasse et, le cas échéant, une première mesure dite de référence, de la biodiversité des bactéries dudit microbiote de référence ; et ii) après au moins une application de l'ingrédient ou de la composition, éventuellement formulé sous forme de composition topique, sur ladite zone cutanée, et après un temps prédéterminé d'action de l'ingrédient ou de la composition, à partir d'un deuxième prélèvement réalisé sur ladite zone cutanée, les analyses de la quantité de biomasse du microbiote cutané et, le cas échéant, de sa biodiversité, en obtenant ainsi une deuxième mesure dite d'activité de quantification de la biomasse et, le cas échéant, une deuxième mesure dite d'activité de la biodiversité ; et iii) à comparer la ou les deuxième(s) mesure(s) d'activité avec la ou les première(s) mesure(s) de référence, et à sélectionner l'ingrédient ou la composition, éventuellement formulé sous forme de composition topique, augmentant la biomasse du microbiote cutané, avantageusement sans diminuer sa biodiversité. Autrement dit, l'invention concerne une méthode, préférentiellement in vitro, de criblage ou sélection d'au moins un ingrédient ou composition, éventuellement formulé sous forme de composition topique, pour induire une augmentation de la biomasse d'un microbiote cutané, avantageusement sans réduire sa biodiversité comprenant les étapes suivantes: i) quantifier la biomasse du microbiote cutané et éventuellement mesurer la biodiversité d'un premier échantillon provenant d'une zone cutanée permettant d'obtenir ainsi une première mesure de quantification de la biomasse et le cas échéant une première mesure de la biodiversité d'un microbiote cutané dit de référence; et ii) quantifier la biomasse du microbiote cutané et éventuellement mesurer la biodiversité d'un deuxième échantillon provenant de ladite zone cutanée sur laquelle le au moins un ingrédient ou composition, éventuellement formulé sous forme de composition topique a été appliqué, permettant d'obtenir ainsi une deuxième mesure de quantification de la biomasse et le cas échéant une deuxième mesure de la biodiversité ; et iii) comparer la deuxième mesure de quantification de la biomasse avec la première mesure de quantification de la biomasse, et le cas échéant la deuxième mesure de la biodiversité avec la première mesure de la biodiversité et sélectionner le au moins un ingrédient ou composition, éventuellement formulé sous forme de composition topique, apte à augmenter la biomasse du microbiote cutané, le cas échéant sans diminuer sa biodiversité. Selon un mode de réalisation préféré, le temps prédéterminé d'action de l'ingrédient peut varier entre 1h et plusieurs jours, en particulier 31 jours, avantageusement 4 jours. Selon l'invention, un « microbiote cutané de référence » ou « microbiote cutané dit de référence » ou « microbiote de référence » ou « microbiote dit de référence » est par exemple un microbiote caractéristique d'une zone cutanée à étudier. Un microbiote de référence peut également être un milieu comprenant différentes bactéries dans diverses proportions qui correspond ou non à un microbiote existant dans la nature. Le microbiote de référence peut être obtenu notamment à partir d'un échantillon d'une zone cutanée d'un individu ou en préparant un milieu comprenant les différentes bactéries dans les proportions correspondant audit microbiote de référence. Par « milieu », on entend un milieu apte à la croissance des bactéries. Il peut s'agir d'un milieu solide ou liquide. Selon un mode de réalisation particulier, les niveaux de biomasse et de diversité d'un microbiote de référence caractéristique d'une zone cutanée est une moyenne des niveaux de biomasse et diversité et/ou de profil de diversité de microbiote mesurés sur des échantillons provenant de ladite zone de peau chez plusieurs individus. Selon un mode de réalisation préféré de l'invention, ledit microbiote de référence est celui d'une zone cutanée saine et/ou est appauvri, avantageusement suite à un stress ou une agression cutanée, plus avantageusement suite à l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Selon un mode de réalisation particulier de l'invention, on fournit une méthode de criblage ou de sélection ou d'évaluation de l'activité d'au moins un ingrédient préférentiellement applicable par voie topique, en particulier un ingrédient cosmétique et/ou dermatologique, éventuellement formulé sous forme de composition topique, caractérisée en ce que la méthode comprend : a) avant toute opération, la sélection d'une zone cutanée à étudier d'un ou plusieurs êtres humains, b) la réalisation d'au moins un premier prélèvement de matériel biologique ou génétique du microbiote présent sur cette zone sélectionnée, d'un ou plusieurs êtres-humains, constituant un microbiote dit de référence, suivi par l'analyse de la quantité de biomasse et de la biodiversité du microbiote cutané, en obtenant ainsi des premières mesures dites de référence respectivement de quantification de la biomasse du microbiote cutané et de biodiversité du microbiote cutané, c) l'application d'au moins un ingrédient ou une composition sur cette zone, de façon unique ou répétée, d) après une durée de 1 h à 31 jours, avantageusement 4 jours, après l'application unique ou les applications répétées, à nouveau la réalisation d'au moins un deuxième prélèvement de matériel biologique ou génétique du microbiote présent sur cette zone sélectionnée, d'un ou plusieurs êtres-humains, e) l'utilisation d'au moins une méthode permettant de quantifier la biomasse présente dans chacun des premiers et deuxièmes prélèvements précités en obtenant ainsi respectivement une première mesure, dite de référence, de quantification de la biomasse du microbiote cutané et une deuxième mesure, dite d'activité, de quantification de la biomasse du microbiote cutané, f) l'utilisation d'au moins une méthode permettant de mesurer la biodiversité microbienne présente dans chacun des premiers et deuxièmes prélèvements précités en obtenant ainsi respectivement une première mesure, dite de référence, de la biodiversité du microbiote cutané et une deuxième mesure, dite d'activité, de la biodiversité du microbiote cutané, g) l'évaluation de l'activité de l'ingrédient, ou éventuellement de la composition, appliqué par voie topique, sur la biomasse et la biodiversité du microbiote prélevé par comparaison de la biomasse et de biodiversité du microbiote entre lesdites premières mesures obtenues à partir du premier prélèvement et lesdites deuxièmes mesures obtenues à partir du deuxième prélèvement, et h) la sélection de l'ingrédient, éventuellement formulé sous forme de composition topique, qui augmente la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité. Selon un mode réalisation particulier, ledit ingrédient ou ladite composition à tester dans la méthode de sélection ou criblage selon l'invention est choisi parmi le groupe comprenant : a) au moins une source de carbone qui est susceptible d'être utilisée par les bactéries comme substrat de croissance et de multiplication, et préférentiellement choisie parmi : - les protéines hydrolysées partiellement ou totalement, issues de céréales (blé, maïs, orge, seigle, avoine), de légumineuses (soja, féverole, fève, pois), de lait (caséines, albumines), de levure, ainsi que les peptides et les acides aminés seuls ou en mélange ; - les oligosaccharides ou les polysaccharides à courte chaine, tels que les fructo oligosaccharides ou FOS ; les acides hyaluroniques, le pullulane, le chitosane, le dextran sulfate, le galactoarabinan, le carraghennane, l'alginate, la cellulose, avantageusement la cellulose microcristalline, l'amylose, l'amylopectine et leurs hydrolysats respectifs ; b) au moins un antioxydant qui présente une activité sur les espèces réactives de l'oxygène (ERO), avantageusement les radicaux libres, en particulier choisi dans le groupe constitué d'octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate ; 2,6-Bis(1,1- dimethylethyl)-4-methylphenol ; bis-ethylhexyl hydroxydimethoxy benzylmalonate ; tert-butylhydroquinone ; tetrabutyl ethylidenebisphenol ; thioglycolic acid ; thiotaurine ; thioctic acid ; dilauryl thiodipropionate ; sodium erythorbate ; sorbityl furfural ; erythorbic acid ; perillyl alcohol ; pyridyloxide t- butylnitrone ; ergothioneine ; melatonin ; acetyl cysteine ; cysteine ; lysine hydrochloride ; carnosic acid ; tyrosyl histidine HCl ; histidine hydrochloride ; pyridoxine serinate ; superoxide dismutase ; aminopropyl ascorbyl phosphate ; ascorbic acid ; ascorbyl dipalmitate ; ascorbyl glucoside ; ascorbyl linoleate ; ascorbyl methylsilanol pectinate ; ascorbyl palmitate ; ascorbyl tetraisopalmitate ; ascorbyl tocopheryl maleate ; trisodium ascorbyl palmitate phosphate ; disodium ascorbyl sulfate ; calcium ascorbate ; methylsilanol ascorbate ; sodium ascorbate ; sodium ascorbyl phosphate ; sodium ascorbyl/cholesteryl phosphate ; tetrahexyldecyl ascorbate ; magnesium ascorbyl phosphate ; tocopherol ; tocopheryl acetate ; tocopheryl linoleate ; tocopheryl oleate ; tocopheryl nicotinate ; tocopheryl retinoate ; sodium tocopheryl phosphate ; dioleyltocopheryl methylsilanol ; potassium ascorbyl tocopheryl 5 phosphate ; dodecyl gallate ; propyl gallate ; octyl gallate ; epigallocatechin gallate (EGCG) ; propyl gallate ; ethyl ferulate ; ethylhexyl ferulate ; chitosan ascorbate ; chitosan glycolate ; apigenin ; tiliroside ; alpha-arbutin ; arbutin ; baicalin ; quercetin ; quercetin caprylate ; isoquercetin ; diethylhexyl syringylidenemalonate ; dihydroxymethylchromone ; dimethoxy di-p-cresol ; dimethylmethoxy chromanol ; ethylbisiminomethylguaiacol manganese chloride ; hesperidin methyl chalcone ; kojic acid ; kojic dipalmitate ; madecassoside ; asiaticoside ; magnolol (5,5'- diallyl-2,2'-dihydroxybiphenyl) ; nordihydroguaiaretic acid ; phenylethyl resorcinol ; resveratrol ; troxerutin ; glucosylrutin,rutin (4H-1-benzopyran-4-one) ; disodium rutinyl disulfate ; tetrahydrobisdemethoxydiferuloylmethane ; tetrahydrodemethoxydiferuloylmethane ; tetrahydrodiferuloylmethane ; tococysteamide ; totarol ; hydroxydecyl ubiquinone ; ubiquinone ; caroténoïdes ; niacinamide ou l'un de ses dérivés ; créatine ; créatinine, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, et rhamnose ; plus particulièrement, dans le groupe comprenant tocophérol, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, rhamnose ; encore plus particulièrement la carnosine ; c) au moins un sel minéral ou organique topiquement acceptable, avantageusement choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, plus particulièrement le citrate de sodium. Selon un autre aspect, l'invention concerne également une méthode de préparation d'une composition, avantageusement topique, en particulier cosmétique ou dermatologique, préférentiellement écobiologique, caractérisée en ce qu'elle comprend (i) la mise en œuvre d'une méthode de sélection ou criblage selon l'invention (ii) la sélection d'au moins une composition, avantageusement topique, en particulier une composition cosmétique et/ou dermatologique, comprenant au moins un ingrédient sélectionné par la méthode de sélection ou criblage selon l'invention et apte à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité et (iii) le conditionnement de ladite composition. Selon un autre aspect, l'invention concerne une composition, avantageusement topique, préférentiellement écobiologique, en particulier cosmétique et/ou dermatologique, comprenant au moins un ingrédient ou une composition, sélectionné par la méthode de sélection ou criblage selon l'invention, pour son utilisation pour son utilisation pour maintenir et/ou rétablir un microbiote cutané sain. Avantageusement, la composition pour son utilisation selon l'invention comprend au moins une source de carbone, un antioxydant et un sel minéral ou organique, plus avantageusement ladite source de carbone, ledit antioxydant et/ou le dit sel étant sélectionné(s) par la méthode de sélection ou criblage selon l'invention. Selon un mode de réalisation préférentiel : a) la au moins une source de carbone est susceptible d'être utilisée par les bactéries comme substrat de croissance et de multiplication, et, préférentiellement, est choisi parmi : - les protéines hydrolysées partiellement ou totalement, issues de céréales (blé, maïs, orge, seigle, avoine), de légumineuses (soja, féverole, fève, pois), de lait (caséines, albumines), de levure, ainsi que les peptides et les acides aminés seuls ou en mélange ; - les oligosaccharides ou les polysaccharides à courte chaine, tels que les fructo oligosaccharides ou FOS ; les acides hyaluroniques, le pullulane, le chitosane, le dextran sulfate, le galactoarabinan, le carraghennane, l'alginate, la cellulose, avantageusement la cellulose microcristalline, l'amylose, l'amylopectine et leurs hydrolysats respectifs ; b) Le au moins un antioxydant présente une activité sur les espèces réactives de l'oxygène (ERO), avantageusement les radicaux libres, et, en particulier, est choisi dans le groupe constitué d'octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate ; 2,6-Bis(1,1- dimethylethyl)-4-methylphenol ; bis-ethylhexyl hydroxydimethoxy benzylmalonate ; tert-butylhydroquinone ; tetrabutyl ethylidenebisphenol ; thioglycolic acid ; thiotaurine ; thioctic acid ; dilauryl thiodipropionate ; sodium erythorbate ; sorbityl furfural ; erythorbic acid ; perillyl alcohol ; pyridyloxide t- butylnitrone ; ergothioneine ; melatonin ; acetyl cysteine ; cysteine ; lysine hydrochloride ; carnosic acid ; tyrosyl histidine HCl ; histidine hydrochloride ; pyridoxine serinate ; superoxide dismutase ; aminopropyl ascorbyl phosphate ; ascorbic acid ; ascorbyl dipalmitate ; ascorbyl glucoside ; ascorbyl linoleate ; ascorbyl methylsilanol pectinate ; ascorbyl palmitate ; ascorbyl tetraisopalmitate ; ascorbyl tocopheryl maleate ; trisodium ascorbyl palmitate phosphate ; disodium ascorbyl sulfate ; calcium ascorbate ; methylsilanol ascorbate ; sodium ascorbate ; sodium ascorbyl phosphate ; sodium ascorbyl/cholesteryl phosphate ; tetrahexyldecyl ascorbate ; magnesium ascorbyl phosphate ; tocopherol ; tocopheryl acetate ; tocopheryl linoleate ; tocopheryl oleate ; tocopheryl nicotinate ; tocopheryl retinoate ; sodium tocopheryl phosphate ; dioleyltocopheryl methylsilanol ; potassium ascorbyl tocopheryl 5 phosphate ; dodecyl gallate ; propyl gallate ; octyl gallate ; epigallocatechin gallate (EGCG) ; propyl gallate ; ethyl ferulate ; ethylhexyl ferulate ; chitosan ascorbate ; chitosan glycolate ; apigenin ; tiliroside ; alpha-arbutin ; arbutin ; baicalin ; quercetin ; quercetin caprylate ; isoquercetin ; diethylhexyl syringylidenemalonate ; dihydroxymethylchromone ; dimethoxy di-p-cresol ; dimethylmethoxy chromanol ; ethylbisiminomethylguaiacol manganese chloride ; hesperidin methyl chalcone ; kojic acid ; kojic dipalmitate ; madecassoside ; asiaticoside ; magnolol (5,5'- diallyl-2,2'-dihydroxybiphenyl) ; nordihydroguaiaretic acid ; phenylethyl resorcinol ; resveratrol ; troxerutin ; glucosylrutin,rutin (4H-1-benzopyran-4-one) ; disodium rutinyl disulfate ; tetrahydrobisdemethoxydiferuloylmethane ; tetrahydrodemethoxydiferuloylmethane ; tetrahydrodiferuloylmethane ; tococysteamide ; totarol ; hydroxydecyl ubiquinone ; ubiquinone ; caroténoïdes ; niacinamide ou l'un de ses dérivés ; créatine ; créatinine, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, et rhamnose ; plus particulièrement, dans le groupe comprenant tocophérol, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, rhamnose ; encore plus particulièrement, la carnosine ; c) Le au moins un sel minéral ou organique est topiquement acceptable et, avantageusement, est choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, plus particulièrement le citrate de sodium. Selon un mode de réalisation préféré, la composition pour son utilisation selon l'invention comprend du citrate de sodium et de la carnosine. Selon un mode de réalisation particulier, la composition pour son utilisation selon l'invention est une composition aqueuse, comprenant au moins 60% en poids de la composition totale d'eau, en particulier au moins 60%, mieux au moins 70%, ou au moins 80% ou au moins 90% ou au moins 95%. Préférentiellement, la phase aqueuse représente au moins 60% en poids de la composition totale, mieux au moins 70%, ou au moins 80% ou au moins 90% ou au moins 95%. Avantageusement, la phase aqueuse est caractérisée par : - une résistivité, à température ambiante, comprise entre 12,5 et 12500 Ohms.cm, en particulier entre 80 et 8000 Ohms.cm; et/ou - une pression osmotique comprise entre 70 et 2500 mOsmol/l et en particulier entre 100 et 800 mOsmol/l ainsi qu'un pH compris entre 5,0 et 9,0 et en particulier compris entre 6,0 et 8,0. Selon un mode de réalisation particulier, la composition pour son utilisation selon l'invention présente un pH compris entre 5,0 et 9,0 et en particulier compris entre 6,0 et 8,0. Selon un mode de réalisation particulier, la composition pour son utilisation selon l'invention est appliquée sur une zone cutanée saine, avant ou suite à une agression cutanée, avantageusement l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Selon un mode de réalisation particulier, ladite agression cutanée selon l'invention peut être unique ou répétée, préférentiellement répétée. Selon un mode de réalisation particulier, la composition pour son utilisation selon l'invention est pour son utilisation pour lutter contre une agression cutanée sur une peau saine, en stimulant la biomasse du microbiote présent au niveau cutané, ou présent sur une matière kératinique, pour augmenter la quantité de cette biomasse dudit microbiote afin d'éviter l'arrivée et/ou l'installation de nouvelles souches microbiennes liées à l'environnement. Selon un mode de réalisation particulier, la composition selon l'invention est pour son utilisation pour lutter contre une agression cutanée sur une peau saine en stimulant la biomasse du microbiote présent au niveau cutané, ou présent sur une matière kératinique, pour restaurer la quantité de cette biomasse dudit microbiote, fortement diminuée lors d'une agression cutanée et par exemple après l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Selon un autre aspect, l'invention concerne un procédé de traitement, avantageusement topique, en particulier cosmétique non thérapeutique ou dermatologique, préférentiellement écobiologique, pour maintenir et/ou rétablir un microbiote cutané sain, caractérisé par l'utilisation d'une composition telle que précédemment décrite permettant l'augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité., Selon un autre aspect, l'invention concerne l'utilisation d'au moins un ingrédient, ou au moins une substance, ou d'au moins une composition, sélectionnée par la méthode de sélection ou criblage selon l'invention pour la préparation d'une composition, préférentiellement topique, en particulier écobiologique, pour maintenir et/ou rétablir un microbiote cutané sain. Selon un autre aspect, l'invention concerne une méthode de soin topique, en particulier écobiologique, avantageusement cosmétique ou dermatologique, pour soigner la peau nécessitant un maintien ou une restauration de son microbiote sain, par l'augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité, caractérisée en ce qu'on sélectionne au moins une zone de la peau nécessitant ce soin, et on applique sur au moins cette zone de la peau un ingrédient, avantageusement formulé dans une composition, ou une composition sélectionnée à travers la méthode de sélection ou criblage décrite auparavant ou dans la description suivante. Selon un mode de réalisation particulier, l'invention concerne également une méthode de soin topique, de préférence écobiologique, caractérisée en ce qu'elle comprend (i) la mise en œuvre d'une méthode de sélection ou criblage définie auparavant ou dans la description suivante, et la sélection d'au moins un ingrédient ou une composition, en particulier un ingrédient ou une composition cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané décrite auparavant, en particulier sans réduire sa biodiversité (ii) la formulation des ingrédients ou compositions ainsi sélectionnés dans une formule à usage cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané sans réduire sa diversité (iii) le conditionnement de ladite formule (iv) l'application topique de ladite formulation sur une zone de peau ou une surface des annexes sélectionnées. Selon encore un autre aspect, l'invention concerne une méthode de soin, avantageusement topique, de préférence cosmétique ou dermatologique, en particulier écobiologique, pour lutter contre une agression cutanée sur une peau saine, caractérisée en ce qu'on sélectionne au moins une zone de la peau nécessitant ce soin et on réalise l'administration, préférentiellement par application topique, d'au moins un ingrédient ou composition, avantageusement applicable par voie topique, en particulier un ingrédient ou composition cosmétique et/ou dermatologique, en particulier formulé sous forme de composition topique, sélectionné par la méthode de criblage ou sélection décrite auparavant ou dans la description suivante, stimulant la biomasse du microbiote présent au niveau cutané, ou présent sur une matière kératinique, pour augmenter la quantité de cette biomasse dudit microbiote afin d'éviter l'arrivée de nouvelles souches microbiennes liées à l'environnement. Selon un autre aspect, l'invention concerne une méthode de soin, avantageusement topique, de préférence cosmétique ou dermatologique, en particulier écobiologique, pour lutter contre une agression cutanée sur une peau saine, caractérisée en ce qu'on sélectionne au moins une zone de la peau nécessitant ce soin et on réalise l'application, de préférence topique, d'au moins un ingrédient applicable par voie topique, en particulier un ingrédient cosmétique et/ou dermatologique, de préférence formulé sous forme de composition topique, sélectionné par la méthode de sélection ou criblage décrite auparavant ou dans la description suivante, stimulant la biomasse du microbiote présent au niveau cutané, ou présent sur une matière kératinique, pour restaurer la quantité de cette biomasse dudit microbiote contre sa diminution en quantité par une agression cutanée et par exemple après l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Sauf définition contraire, tous les termes techniques utilisés dans la présente invention ont la même signification que celle couramment comprise par un spécialiste ordinaire du domaine auquel se réfèrent les procédés et les compositions décrits. Les définitions générales peuvent être trouvées dans des ouvrages techniques relatifs au domaine de la biologie moléculaire, par exemple, Dictionary of Microbiology and Molecular Biology, 2e édition. (Singleton et al., 1994) ou The Harper Collins Dictionary of Biology (Hale et al., 1991). Tels qu'utilisés dans la présente invention, les termes et expressions suivants ont la signification qui leur est attribuée, sauf indication contraire. Par « matières kératiniques », on entend selon l'invention la peau, les semi- muqueuses (en particulier les lèvres), les muqueuses et les annexes dont les phanères. Par « méthode de quantification de la biomasse du microbiote », on entend selon l'invention, l'utilisation d'une méthode quantitative ou semi quantitative qui repose par exemple : - sur des techniques PCR quantitatives (ou qPCR), ou - sur de la cytométrie en flux, ou - sur des méthodes de dénombrement sur gélose, ou - sur toute méthode apparentée. Par « méthode de mesure de la biodiversité du microbiote », on entend selon l'invention, l'utilisation : - d'une méthode qui repose sur l'analyse d'une partie du gène hypervariable du ribosome 16S des bactéries après leur amplification ciblée, ou - d'une méthode utilisant la technique de shotgun métagénomique (sur ADN du microbiome), ou - d'une méthode dite de séquençage dit « shotgun » métatranscriptomique (sur ARN du microbiome), ou - de toute méthode apparentée, suivie par une méthode de calcul de la diversité microbienne à partir des données de la méthode précédente utilisée, telle que le calcul : - de l'indice de Shannon, - de l'indice de Simpson, - de l'indice de Inverse-Simpson, - du nombre d'espèces ou d'OTU, - de l'Evenness ou l'indice de Pielou, - de l'indice de Hurlbert, - de l'indice de Hill. Ces méthodes de calculs et indices sont bien décrits dans la littérature et connus par l'Homme de l'art. On les retrouvera par exemple dans le document de Eric Marcon, « Mesure de la Biodiversité, Kourou, France, ffcel-01205813v5ff ». Le prélèvement de matériel biologique ou génétique du microbiote peut se faire selon les méthodes connues de l'Homme de métier. Typiquement, le matériel biologique ou génétique est récupéré à l'aide d'un écouvillon qui permet de récolter les bactéries, en effectuant toujours la même manipulation pour standardiser le plus possible les quantités prélevées. A titre d'exemple l'écouvillon Sigma Trans Swab liquide Maies peut être utilisé. Dans un mode de réalisation préféré, le prélèvement est réalisé en effectuant 10 allers-retours en Z en exécutant une légère pression sur l'écouvillon. A ce stade, il est possible de congeler les écouvillons, qui peuvent être conservées à -80°C. A partir des écouvillons, les culots bactériens peuvent être récupérés par centrifugation. L'extraction d'ADN génomique (ADNg) peut être réalisée selon les méthodes connues à l'Homme de l'art comme par exemple avec le kit Nucleospin Microbial DNA kit (Macherey Nagel, France). Les prélèvements peuvent être effectués sur plusieurs zones du corps humain, comme par exemple, joue, front, avant-bras, afin d'étudier la réponse du microbiote des différentes zones biogéographiques aux ingrédients ou compositions applicables par voie topique. Dans un mode de réalisation préféré, le prélèvement est effectué à partir du visage ce qui permet de tester les ingrédients ou les compositions sur une zone réaliste et exposée à l'environnement, ou sur le dos ce qui permet de tester un ensemble de conditions différentes sur un microbiote relativement homogène (Grice et al., 2009). Selon un mode de réalisation préféré, l'aire intéressée peut être marquée par un cache, avantageusement délimitant une surface de prélèvement comprise entre 10 et 20 cm2, avantageusement de 15 cm2. Dans une mode de réalisation préféré, cette biomasse est évaluée en partant du matériel biologique ou génétique récupéré sur les matières kératiniques, mais elle peut être également quantifiée directement selon les méthodes connues à l'Homme du métier, par exemple par immunomarquage ou cytométrie de flux sur bactéries marquées. Par « standardisation » au sens de l'invention, on entend la normalisation de la quantité de biomasse prélevée. Par « amplification » au sens de l'invention on entend toute procédure in vitro qui produit des copies multiples d'une séquence d'acides nucléiques cible, ou de sa séquence complémentaire, ou des fragments de celle-ci (c'est-à-dire une séquence amplifiée contenant moins que l'acide nucléique cible complet). Avantageusement, cette amplification est effectuée par réaction de polymérisation en chaîne ou polymerase chain reaction (PCR), mais d'autres méthodes connues à l'Homme du métier, comme l'amplification isotherme, peuvent également être employées. Par « amplification quantitative » on entend au sens de l'invention toute procédure in vitro qui produit des copies multiples d'une séquence d'acides nucléiques cible et qui permet au même temps de déterminer la concentration de séquence cible initialement présente dans le mélange réactionnel. Plusieurs types de réactions d'amplification quantitatives sont décrits. On peut distinguer les amplifications quantitatives basées sur l'emploi d'un standard externe, les amplifications compétitives, utilisant un standard interne, et enfin les amplifications cinétiques, qui consistent à mesurer en temps réel, l'augmentation de la quantité de la séquence cible. Avantageusement, l'amplification quantitative est une amplification cinétique effectuée par PCR et correspond à la technique connue à l'Homme du métier sous le nom de PCR quantitative (qPCR) ou encore PCR en temps réel. Les réactions de PCR, que ce soit quantitative ou non-quantitative peuvent être réalisées avec le système « Light Cycler » (Roche Molecular Systems Inc.) et selon les procédures recommandées par le fournisseur. Le terme « séquence cible », tel qu'utilisé dans la présente invention, désigne la séquence nucléotidique particulière de l'acide nucléique cible qui doit être amplifiée et/ou détectée. La « séquence cible » comprend les séquences de complexation auxquelles des oligonucléotides (par exemple, les oligonucléotides d'amorçage et/ou les oligonucléotides promoteurs) se complexent au cours de processus d'amplification (par exemple, PCR). Lorsque l'acide nucléique cible est à l'origine simple brin, le terme « séquence cible » désignera également la séquence complémentaire de la « séquence cible » telle que présente dans l'acide nucléique cible. Lorsque l'acide nucléique cible est à l'origine double brin, le terme « séquence cible » désigne à la fois les brins sens (+) et antisens (-). Le terme « région », tel qu'utilisé dans la présente invention, désigne une portion d'un acide nucléique, ladite partie étant plus petite que l'acide nucléique entier. En particulier, avec « régions V1, V2, V3, V4 » on désigne des régions hypervariables de l'ADN ribosomal (ADNr) 16S bactérien définie par exemple dans Gray et al., 1984, qui constitue une portion de l'ADNg et qui permette l'identification des espèces auxquelles elles appartiennent lors du séquençage. Par « amplicon » ou « produit d'amplification », on entend une molécule d'acide nucléique générée dans une réaction d'amplification, que ce soit quantitative ou pas, d'acide nucléique et qui est dérivée d'un acide nucléique cible. Un amplicon ou un produit d'amplification contient une séquence d'acides nucléiques cible qui peut être du même sens ou d'un sens opposé à l'acide nucléique cible. Par « séquençage » au sens de l'invention on entend le décodage d'au moins un brin d'une séquence cible, et la lecture de la suite des nucléotides la composant dans leur ordre. Plusieurs méthodes de séquençages et assemblages des séquences lues sont connues à l'Homme du métier. Avantageusement le séquençage est effectué en utilisant la plateforme Illumina Miseq™. Par exemple, les documents WO03048387A2, WO2005024010A1 WO2006064199A1 et WO2007123744A1 ou encore WO2012096703A1 et les références citées dans ces documents décrivent des méthodes de préparation de échantillons et de séquençage adaptées à la présente invention. Le séquençage peut s'effectuer directement sur les amplicons, ou alternativement sur des banques d'amplicons. Selon un mode de réalisation préféré, le séquençage s'effectué à partir de banque d'amplicons, avantageusement les banques d'amplicons obtenues selon la solution Metabiote™ décrite dans Guillaume et al., 2014. Une « amorce » est un oligomère qui s'hybride à un acide nucléique et possède une extrémité 3' qui est étendue par polymérisation. Une amorce peut être éventuellement modifiée, par exemple en incluant une région 5' non complémentaire de la séquence cible. Une telle modification peut comprendre des additions fonctionnelles, telles que des étiquettes, des promoteurs ou d'autres séquences sans cible spécifique, utilisées ou utiles pour manipuler ou amplifier l'amorce ou l'oligonucléotide cible. Les amorces peuvent être « dégénérées ». Au sens de l'invention, « amorce dégénérée » désigne des amorces permettant incorporation des nucléotides différents à des positions déterminées. Les amorces dégénérées sont connues à l'Homme du métier ; le code suivant est utilisé universellement pour noter la dégénérescence des amorces à une position nucléotidique déterminée : I = Inosine (Nichols et al., 1994) ; R = G ou A ; K= G ou T ; S = G ou C ; W = A ou T ; M = A ou C ; Y = T ou C ; D = G ou A ou T ; V = G ou A ou C ; B = G ou T ou C ; H = A ou T ou C ; N = G ou A ou T ou C. Par « amorces universelles 16S » au sens de l'invention, on entend des oligonucléotides permettant l'amplification de régions hautement conservées de l'ADNr 16S. Selon une mode de réalisation particulier, il s'agit des amorces correspondant aux SEQ ID.18F (5'-AGAGTTTGATCCTGGCTCAG-3') and SEQ ID.21391R (5'-GACGGGCGGTGWGTRCA-3'). (Kong et al., 2012b). Selon un mode de réalisation alternatif, il s'agit des amorces correspondant aux SEQ ID.3533F (5'-GTGCCAGCAGCCGCGGTAA-3') et SEQ ID.4902R (5'- GTCAATTCITTTGAGTTTYARYC-3') (San Miguel et al., 2018). Par « amorces spécifiques 16S » au sens de l'invention on entend des oligonucléotides permettant l'amplification de régions hypervariables de l'ADNr 16S. Avantageusement, il s'agit des régions V1-V4 de l'ADNr 16S, de préférence les régions V1-V3 et/ou V3-V4, avantageusement les régions V3-V4. Avantageusement, les amorces spécifiques 16S permettent l'obtention d'amplicons de taille comprise entre 100 et 500 paires de base, avantageusement entre 200 et 400 paires de base. Selon un mode de réalisation, il s'agit des amorces pour les régions V1-V3 correspondant aux SEQ. ID 527F (5'- AGAGTTTGATCCTGGCTCAG-3') et SEQ. ID 6534R (5- ATTACCGCGGCTGCTGG-3) (San Miguel et al., 2018). Selon un mode de réalisation préféré, il s'agit des amorces pour les régions V3-V4 correspondant aux SEQ ID, 7338F (ACTCCTACGGGAGGCAGCAG) and SEQ ID.8806R (GGACTACHVGGGTWTCTAAT) (Li et al., 2020). Les expressions « banque(s) » ou « librairie(s) » seront utilisées indifféremment dans la présente description, ces deux expressions ayant le même sens. Au sens de l'invention, on entend par « banque » une collection de séquences nucléotidiques bactériennes issues de préférence de l'amplification de l'ADNr 16S et qui sont susceptibles d'être séquencées ou détectées de façon individuelle. La constitution de banque d'ADN peut se faire selon les méthodes connues à l'Homme du métier, par exemple en utilisant de kits comme le NEBNext™ Ultra™ DNA Library Prep Kit. Selon un mode de réalisation particulier, il est considéré dans le cadre de l'invention que l'ingrédient ou composition applicable par voie topique sélectionné a un effet significatif sur l'augmentation de la biomasse du microbiote cutané, si la biomasse augmente de façon statistiquement significative entre deux prélèvements, en particulier entre le premier prélèvement et le deuxième prélèvement précités, en particulier après de 1h à 31 jours de traitement. Selon l'invention, l'évaluation de l'activité de l'ingrédient ou composition applicable par voie topique sur la biomasse du microbiote prélevé peut être réalisée en comparant le matériel génétique bactérien total présent sur la zone prélevée avant et après le traitement avec l'ingrédient ou la composition par le biais de l'amplification quantitative avec les amorces universelle 16S, ce qui permet d'estimer le matériel génétique présent au moment du prélèvement avant et après le traitement. Dans un mode de réalisation particulier, l'au moins une amplification des quantités standardisées de matériel génétique est une amplification quantitative effectuée en utilisant les amorces universelles 16S. Pour cela, une lyse bactérienne suivie par une étape d'extraction d'ADN génomique avec le PureLink Mini kit (Life Technologies, Grand Island, NY) sont réalisées. La qPCR ciblée sur le gène de l'ARNr 16S est réalisée à l'aide d'amorces spécifiques (Bact-8F : AGAGTTTGATCCT GGCTCAG, Bacte338R : CTGCTGCCTCCCGTAGGAGT) avec une détection SYBR Green (Applied Biosystems, Foster City, CA) sur un équipement Mx300P (Agilent Technology). Les courbes standard sont préparées en amplifiant des quantités connues d'ADN d'E. coli. En comparant le nombre de cycles à la courbe standard il est possible de calculer le nombre de copies du gène ARN16S par microlitre d'échantillon. Dans un autre mode de réalisation particulier, l'évaluation de la biomasse du microbiote cutané est réalisée par quantification en cytométrie de flux. Pour cela, l'écouvillon est déchargé dans 1 mL de liquide Amies puis la suspension bactérienne est marquée avec du SYBRGreen, incubée 15 min à 37°C à l'abri de la lumière et analysée par un cytomètre de flux BD Accuri C6 (cytomètre BD Accuri, Belgique) équipé d'un laser de 50 mW émettant à une longueur d'onde fixe de 488 nm. L'intensité de la fluorescence est mesurée à Fl1 = 533 +/- 30 nm, Fl3> 670 nm. Les données sont traitées avec le logiciel BD Accuri CFlow. Un nombre de bactéries est ainsi obtenu par mL d'échantillon. Dans un autre mode de réalisation particulier, l'évaluation de la biomasse du microbiote cutané est réalisée par quantification par dénombrement sur gélose. Pour cela, après déchargement des écouvillons dans le liquide Amies, les suspensions bactériennes sont déposées sur gélose nutritive en boite de Pétri, et sont incubées pendant 48h à 35°C. Une numération des colonies bactériennes permet d'obtenir la concentration bactérienne en multipliant le nombre d'UFC par le facteur de dilution, et est exprimée en UFC/ml. Dans encore un autre mode de réalisation particulier, l'évaluation de la biomasse du microbiote cutané est réalisée par quantification par dénombrement direct au microscope. Pour cela les suspensions bactériennes sont ensemencées dans un bouillon nutritif tels que Mueller-Hinton ou Brain Heart Infusion. Après incubation, un dénombrement direct au microscope peut être réalisé sur une cellule de Thoma. Selon un mode de réalisation particulier de l'invention, Il est considéré qu'il n'y a pas eu de réduction de diversité bactérienne du microbiote cutané si les variations ne sont pas statistiquement significatives. Selon un mode de réalisation particulier de l'invention, l'évaluation de l'activité de l'ingrédient ou composition applicable par voie topique sur la diversité du microbiote prélevé peut être réalisée en comparant les espèces bactériennes présentes dans les différentes conditions expérimentales par le biais de l'amplification avec les amorces spécifiques 16S et le séquençage des amplicons obtenus. Plusieurs indices permettant d'estimer la diversité d'un matériel génétique constitué de mélanges d'espèces différentes sont connus à l'Homme du métier. Selon un mode de réalisation particulier, l'indice est celui dit de Shannon, qui est calculé en suivant la méthode décrite dans Lemos et al., 2011. L'indice de Shannon intègre deux paramètres distincts : le nombre d'unités taxonomiques opérationnelles (Operational Taxonomic Units ou OTUs) et la distribution de ces OTUs dans chaque échantillon. Ce dernier paramètre peut être calculé en prenant en compte l'abondance des 15 genres bactériens les plus représentés, ou alternativement, représentant le 90% des amplicons obtenus dans l'amplification avec les amorces spécifiques 16S. Les traitement bio-informatiques des séquences cibles peuvent être effectués selon les méthodes connues à l'Homme du métier, par exemple en utilisant un script PERL. Selon un autre mode de réalisation particulier, l'analyse par amplification avec les amorces spécifiques 16S puis, éventuellement, séquençage est effectuée seulement pour les échantillons de matériel génétique issu d'un prélèvement permettant l'obtention, lors de l'amplification quantitative avec les amorces universelles 16S dans un volume réactionnel de 4,38 µl, d'au moins 400 copies d'amplicon d'ADNr 16S/µL au bout de 30 cycles d'amplification et/ou d'au moins 500 copies d'amplicon d'ADNr 16S/µL au bout de 35 cycles d'amplification. Selon encore un mode de réalisation particulier, l'analyse de la diversité du microbiote cutané est réalisée par métagénomique de la façon suivante : après extraction de l'ADNg, la diversité microbienne est déterminée pour chaque échantillon par amplification ciblée d'une partie du gène ribosomal. Un fragment du gène d'ARNr 16S comprenant les régions hypervariables V3 à V4 (ou V1-V3) est amplifié avec les amorces 338F (ACTCCTACGGGAGGCAGCAG) et 806R (GGACTACHVGGGTWTCTAAT) (Munyaka et al., 2015). L'amplification est réalisée et les produits issus de PCR sont purifiés et normalisés en utilisant des billes magnétiques Agencourt AMPure XP (Beckman Coulter). Une fois purifié, les amplicons sont regroupés pour le séquençage Illumina, effectué en 2x300 pb sur une plate-forme Illumina MiSeq en utilisant MiSeq Reagent Kit v3 (600 cycles). Les séquences sont regroupées en unités taxonomiques opérationnelles (OTUs) avec un seuil d'identité de 97%. L'attribution taxonomique est réalisée en utilisant la classification Ribosomal Database Project (RDP) sur la base de données Greengenes V13_8. L'analyse est réalisée avec le logiciel R. L'indice de Shannon-Weaver, le nombre d'OTU observés et l'indice de dissimilarité de Bray- Curtis sont calculés avec les packages Phyloseq et Vegan. La significativité statistique est déterminée à l'aide du Test de Wilcoxon. Une valeur p <0,05 est considérée comme statistiquement significative. Selon un autre mode de réalisation, l'analyse de la diversité du microbiote cutané est réalisée par Shotgun : après extraction de l'ADNg, la diversité microbienne est déterminée pour chaque échantillon par Whole metagenome sequencing/WMS ou Shotgun metagenome sequencing c'est-à-dire un séquençage complet de tout le génome bactérien. Les librairies sont préparées en utilisant le kit de préparation NexteraXT (Illumina). Le séquençage est effectué au Penn Next Generation Sequencing Core sur la chimie Illumina MiSeq pour obtenir des lectures finales appariées de 150 pb. Les données de séquençage sont obtenues au format fastq. Les séquences dont la longueur est inférieure à 80 nucléotides sont retirées de l'analyse. DeconSeq est utilisé pour filtrer les fichiers fastq et l'un des reads appariés (SE1) est entré dans MetaPhlAn version 1.7.7 pour la classification taxonomique. L'une des extrémités appariées (SE1) de l'échantillon MCC est utilisé pour réaliser un BLAST contre une base de données personnalisée de génomes parmi les 20 espèces bactériennes attendues (blastn, max_target_seqs 1, e <10−10 ; alignement longueur> 50) afin de déterminer la composition des communautés bactériennes. L'indice de diversité alpha est ensuite calculé à l'aide du logiciel R et des packages Phyloseq et Vegan en utilisant l'indice de Shannon. Selon un autre mode de réalisation, l'analyse de la diversité du microbiote cutané est réalisée par Metatranscriptomic shotgun sequencing (MTS) : pour cela l'ARN total est extrait du prélèvement bactérien. Les ARNm polyadénylés sont séparés des non-polyadénylés (ARNr, ARN mitochondriaux, et ARNt) par affinité sur colonne poly-dT (Kit Dynabeads Oligo (dT), Dynal). L'ARNm enrichi est fragmenté mécaniquement à une gamme de taille de +/- 200 pb avec un ultrasonicator. La fragmentation de l'ARNm est évaluée à l'aide du kit Agilent RNA 6000 Pico sur un instrument 2100 Bioanalyzer (Agilent Technologies, Inc.). Les librairies de métatranscriptomes sont préparées en utilisant le NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs Inc). La qualité et la quantité de toutes les librairies finales sont analysées avec un kit Agilent DNA 1000 sur l'instrument 2100 Bioanalyzer et Qubit et sont quantifiées et validées par un dosage en qPCR en utilisant le kit de quantification de bibliothèque PerfeCTa NGS pour Illumina (Quanta Biosciences, Inc.) en utilisant le système de détection par PCR en temps réel CFX Connect (Rad Laboratories, Inc.). Le séquençage de l'une des bibliothèques MT est effectué sur un Illumina HiSeq 2000 à l'aide du réactif TruSeq SBS v3 pour une longueur de lecture de 100 paires (BGI Americas) (étiquetée HS100) et sur Illumina MiSeq à l'aide du kit de cycle v3-600 pour les paires 301 bases (étiquetées MS301). Un autre ensemble de douze bibliothèques est séquencé sur Illumina MiSeq en utilisant 151 « paired- end chemistry » (marquée MS151). Le protocole recommandé par le fabricant est utilisé pour effectuer la réaction de séquençage sur les plates-formes HiSeq et MiSeq. Pour les séquences ribosomales 16S amplifiées par PCR, des recherches BLASTN (paramètres par défaut, sauf taille de mot fixée à 7) sont effectuées contre la base de données de nucléotides GenBank nr (http : //www.ncbi.nlm.nih.gov/). Les séquences 16S sont ensuite attribuées à des phylums majeurs pour effectuer des analyses phylogénétiques (BioNJ et PhyM) en utilisant l'algorithme Best Blast Hit (BBH) et un ensemble de séquences de références pour chacun des différents phylums. Les analyses (alignements de séquence et analyses phylogénétiques) sont réalisées en utilisant SeaView. En utilisant les fichiers de sortie BLASTX, les ADNc sont taxonomiquement attribués à l'aide de MEGAN V 3.5 (www- ab.informatik.uni-tuebingen.de/software/megan). Selon encore un mode de réalisation particulier, la méthode selon l'invention comprend au moins deux prélèvements de matériel biologique ou génétique du microbiote : - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique dudit être humain, ledit prélèvement étant effectué dans une zone non-traitée avant le traitement, - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique dudit être humain ledit prélèvement étant effectué dans une zone traitée avec l'ingrédient ou composition, préférentiellement applicable par voie topique, après le traitement. Selon un autre mode de réalisation particulier, la méthode selon l'invention comprend au moins quatre prélèvements de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique, d'un ou plusieurs êtres-humains : - Un prélèvement effectué dans une zone à traiter avec l'ingrédient ou composition, préférentiellement applicable par voie topique, avant le traitement ; - Un prélèvement effectué dans une zone non-traitée avant le traitement ; - Un prélèvement effectué dans une zone traitée avec l'ingrédient ou composition, préférentiellement applicable par voie topique, après le traitement ; - Un prélèvement effectué dans une zone non-traitée après la période de traitement. Avantageusement, la « zone non-traitée » selon l'invention est une zone cutanée équivalente ou proche, fonctionnellement et/ou en composition microbienne, de ladite « zone à traiter ». Ainsi, selon un autre mode de réalisation particulier, l'invention concerne une méthode de sélection ou criblage ou d'évaluation de l'activité d'au moins un ingrédient ou composition, de préférence applicable par voie topique, en particulier un ingrédient cosmétique et/ou dermatologique, éventuellement formulé sous forme de composition topique, caractérisée en ce que la méthode comprend les étapes suivantes : - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, d'un ou plusieurs êtres-humains, ledit prélèvement étant effectué dans une zone à traiter avec l'ingrédient ou composition applicable par voie topique avant le traitement ; - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique, d'un ou plusieurs êtres- humains, ledit prélèvement étant effectué dans une zone non-traitée avant le traitement ; - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique, d'un ou plusieurs êtres humains ledit prélèvement étant effectué dans une zone traitée avec l'ingrédient ou composition applicable par voie topique après le traitement ; - un prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique, d'un ou plusieurs êtres- humains ledit prélèvement étant effectué dans une zone non-traitée après le traitement ; - une analyse de la biomasse, par exemple par amplification quantitative des quantités standardisées de matériel génétique effectuée en utilisant les amorces universelles 16S ; - une analyse de la biodiversité du microbiote prélevé, par exemple par amplification des quantités standardisées de matériel génétique prélevés, puis séquençage des amplicons ainsi produits en utilisant les amorces spécifiques 16S, puis calcul d'un indice de diversité permettant cette analyse. La durée du soin avec l'ingrédient ou la composition est variable. L'application du produit peut se faire quotidiennement ou plusieurs fois par jour ou sur la période de test. Selon un mode de réalisation, plusieurs prélèvements peuvent être effectués dans la zone traitée, à, par exemple, 5 minutes, 1h, 24h, 2j, 4j, 7j et 31j depuis le début du traitement. Selon un mode de réalisation particulier, le prélèvement à partir de la zone traitée est réalisé après 2 à 7 jours de traitement quotidien. Selon un mode de réalisation, l'application du produit se fait en raison d'un volume compris entre 0,1 et 100 µL/cm2, avantageusement entre 1 et 10 µl/cm2, en standardisant le plus possible l'application, par exemple en utilisant un bras robotique. Dans un mode de réalisation particulier, au moins un prélèvement est effectué sur plusieurs êtres humains. Cela offre l'avantage d'étayer la significativité statistique des données obtenues. Selon un mode de réalisation particulier, le groupe de sujets particulier à l'origine des prélèvements peuvent partager des caractéristiques (appartenance ethnique, âge, phototype, conditions dermatologiques chroniques, type de peau, diète, hygiène de vie, taux d'hydratation de la peau, géolocalisation et d'autres paramètres similaires) qui permettent la mise au point de méthode de soin, préférentiellement cosmétique et/ou dermatologique, ciblant spécifiquement lesdits groupes de sujets particuliers, également appelés « populations ». Selon un mode de réalisation avantageux et préféré, la population à l'origine des prélèvements est caractérisée par un microbiote cutané appauvri au moment des prélèvements. Par « microbiote cutané appauvri » au sens de l'invention on désigne tout déclin dans la biomasse du microbiote cutanée. En particulier, l'appauvrissement peut être lié à une agression cutanée selon l'invention, plus particulièrement à l'action de bactéricides ou de virucides ou de fongicides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. Dans un mode réalisation particulier, l'appauvrissement du microbiote cutané est causé par un traitement topique avec un agent antiseptique (c'est-à-dire fongicide, bactéricide et/ou virucide), avantageusement une composition comprenant un alcool C1-C8 choisi parmi l'éthanol, l'isopropanol, le propanol, le butanol, le pentanol, l'hexanol, l'heptanol, l'octanol, l'alcool benzylique et leur mélanges ; et/ou un agent antiseptique autre qu'un alcool, comme par exemple le chlorure de benzalkonium, le gluconate de chlorhexidine, l'hypochlorite de sodium, un savon réalisé par saponification d'un acide gras ; et/ou encore une composition comprenant un acide tel que l'acide lactique, malique, tartrique, glycolique ; dans un mode de réalisation alternatif, l'appauvrissement du microbiote cutané est causé par un traitement avec un agent virucide et/ou antifongique et/ou bactéricide et/ou antibiotique, administré par voie topique ou systémique. Selon un mode de réalisation particulier, le prélèvement de matériel biologique ou génétique du microbiote présent au niveau cutané, ou présent sur une matière kératinique, est effectué sur un seul être humain. Cette méthode permet de mettre au point des méthodes de soin, avantageusement cosmétique et/ou dermatologique, de préférence écobiologiques, personnalisées, basées sur les conditions, les spécificités et la réactivité au soin, du microbiote cutané de chaque individu. Ainsi, selon un autre mode de réalisation particulier, l'invention concerne encore une méthode personnalisée de sélection ou criblage ou d'évaluation, préférentiellement in vitro, de l'activité d'au moins un ingrédient ou composition, préférentiellement applicable par voie topique, en particulier un ingrédient cosmétique et/ou dermatologique, éventuellement formulé sous forme de composition, avantageusement topique, caractérisée en ce que la méthode comprend les étapes suivantes : - la sélection d'une zone cutanée ou des phanères à étudier, - au moins un prélèvement de matériel biologique ou génétique du microbiote présent sur cette zone, d'un seul être-humain, - l'application d'au moins un ingrédient ou une composition sur cette zone, de façon unique ou répétée, - après une durée de 1 h à 7 jours, au moins un prélèvement de matériel biologique ou génétique du microbiote présent sur cette zone, d'un seul être- humain, - au moins une méthode permettant de quantifier la biomasse présente dans les prélèvements, - au moins une méthode permettant de mesurer la diversité microbienne présente dans les prélèvements, - l'évaluation de l'activité de l'ingrédient ou composition applicable par voie topique sur la biomasse et la biodiversité du microbiote prélevé, - la sélection de l'ingrédient ou composition écobiologique applicable par voie topique permettant l'augmentation de la biomasse, en particulier sans réduire la diversité du microbiote cutané. Selon un mode de réalisation particulier, la méthode personnalisée selon l'invention peut être accompagnée par une analyse du profil dermatologique du seul être humain à l'origine de l'au moins un prélèvement sur la base de son appartenance ethnique, âge, phototype, conditions dermatologiques chroniques, type de peau, diète, hygiène de vie, taux d'hydratation de la peau, géolocalisation et d'autres paramètres similaires. Ces informations peuvent être récoltées par analyse directe ou par le biais d'un formulaire. Par exemple, les méthodes de pondération des caractéristiques cutanées, capillaires, ou unguéales et le programme d'ordinateur décrits dans le document FR 2983328 peuvent être combinées avec les résultats de la méthode de criblage personnalisée mentionnée auparavant pour sélectionner un traitement dermatologique écobiologique apte à induire l'augmentation de la biomasse du microbiote cutanée d'un individu, en particulier sans réduire sa diversité, et conforme aux besoins dermatologiques globaux dudit individu. L'invention concerne également une méthode de préparation d'une composition de préférence topique, caractérisée en ce qu'elle comprend (i) la mise en œuvre d'une méthode de criblage ou sélection définie auparavant et la sélection d'au moins un ingrédient ou une composition, en particulier un ingrédient ou une composition cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané en particulier sans diminuer sa biodiversité (ii) la formulation des ingrédients ou compositions ainsi sélectionnés dans une formule à usage cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans réduire sa biodiversité (iii) le conditionnement de ladite formule. L'invention concerne également une méthode de soin topique, préférentiellement cosmétique ou dermatologique, avantageusement écobiologique, caractérisée en ce qu'elle comprend (i) la mise en œuvre d'une méthode de criblage définie auparavant ou dans la description suivante et la sélection d'au moins un ingrédient ou une composition, en particulier un ingrédient ou une composition cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané décrite auparavant, en particulier sans diminuer sa biodiversité (ii) la formulation des ingrédients ou compositions ainsi sélectionnés dans une composition à usage cosmétique et/ou dermatologique, apte à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité (iii) le conditionnement de ladite formule (iv) l'application topique de ladite composition sur une zone de peau ou une surface des annexes sélectionnées. Comme mentionné auparavant pour la méthode de criblage ou sélection, la méthode de soin topique avantageusement écobiologique peut cibler des populations spécifiques, ou bien être personnalisée et adaptée aux microbiotes propres à chaque individu. Selon un mode de réalisation particulier, la population à soigner est caractérisée par un microbiote cutané appauvri. Selon un mode de réalisation alternatif, il s'agit d'une méthode personnalisée apte à soigner un individu caractérisé par un microbiote cutané appauvri. Ainsi, selon un autre aspect de l'invention, l'invention concerne une méthode de soin topique avantageusement cosmétique ou dermatologique, de préférence écobiologique, apte à induire l'augmentation de la biomasse du microbiote cutanée, en particulier sans impact ou diminution sur sa biodiversité, caractérisée en ce qu'on applique sur la peau une composition sélectionnée à travers la méthode de sélection ou criblage mentionnée auparavant ou décrite ci-après. Au sens de l'invention, par augmentation de la biomasse on désigne une augmentation statistiquement significative. A titre d'exemple, il peut s'agir d'une augmentation d'au moins 50% de la biomasse par rapport à une valeur de référence, c'est-à-dire avant application de l'ingrédient ou de la composition de l'invention. Dans un mode de réalisation alternatif, l'invention concerne une méthode de soin, de préférence cosmétique ou dermatologique, pour lutter contre une agression cutanée (par exemple contre une action bactéricide et/ou virucide et/ou fongicide), et/ou maintenir et/ou rétablir un microbiote sain, caractérisée en ce que la méthode comprend l'application topique d'au moins une composition topique sélectionnée dans la méthode de criblage ou sélection décrite auparavant ou dans la description suivante, ladite composition étant apte à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans réduire sa biodiversité, sur la peau ou sur une matière kératinique. Selon encore un mode de réalisation particulier, la composition sélectionnée à travers la méthode de criblage ou sélection mentionnée auparavant ou dans la description suivante et apte à induire l'augmentation de la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité, est associée à l'action de maintenir et/ou rétablir un microbiote cutané sain. Selon un autre mode de réalisation particulier, la composition à utiliser en particulier dans la mise en œuvre de la méthode de soin topique de préférence écobiologique apte à induire l'augmentation de la biomasse du microbiote cutanée, en particulier sans diminuer sa diversité, selon l'invention, est une composition aqueuse, comprenant en particulier au moins 60%, mieux au moins 70%, ou au moins 80%, ou au moins 90%, voire au moins 95% d'eau. Ainsi, l'invention concerne une méthode de soin topique de préférence écobiologique apte à induire l'augmentation de la biomasse du microbiote cutané en particulier sans réduire sa diversité, selon l'invention, avec une composition aqueuse, comprenant avantageusement au moins 60% d'eau, de préférence 70%, 80%, 90%, voire au moins 95% d'eau. Un mode de réalisation particulier, la composition aqueuse à utiliser en particulier dans la méthode de soin mentionnée auparavant ou dans la description suivante, comprend une eau qui est caractérisée par une résistivité comprise entre 12,5 et 12 500 Ohms.cm, en particulier entre 80 et 8000 Ohms.cm, à température ambiante De préférence, la composition à utiliser en particulier dans la méthode de soin mentionnée auparavant ou dans la description suivante présente une pression osmotique comprise entre 70 et 2500 mOsmol/l, et en particulier entre 100 et 800 mOsmol/l. Selon un mode de réalisation particulier de l'invention, la composition à utiliser en particulier dans la mise en œuvre de la méthode de soin topique de préférence écobiologique apte à induire l'augmentation de la biomasse du microbiote cutanée, en particulier sans diminuer sa biodiversité, comprend au moins un antioxydant et au moins un sel minéral ou organique, chacun ou au moins un de ces composants ou ingrédients ayant, avantageusement, été au préalable sélectionné par la méthode de criblage décrite auparavant ou dans la description suivante. Par « antioxydant », on entend un composé qui présente une activité antiradicalaire, en particulier sur les espèces réactives de l'oxygène (ERO), des carbonyles (RCS) ou de l'azote (RNS). Selon un mode de réalisation particulier, l'au moins un antioxydant est choisi dans le groupe comprenant l'octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate ; 2,6-Bis(1,1- dimethylethyl)-4-methylphenol ; bis-ethylhexyl hydroxydimethoxy benzylmalonate ; tert-butylhydroquinone ; tetrabutyl ethylidenebisphenol ; thioglycolic acid ; thiotaurine ; thioctic acid ; dilauryl thiodipropionate ; sodium erythorbate ; sorbityl furfural ; erythorbic acid ; perillyl alcohol ; pyridyloxide t- butylnitrone ; ergothioneine ; melatonin ; acetyl cysteine ; cysteine ; lysine hydrochloride ; carnosic acid ; tyrosyl histidine HCl ; histidine hydrochloride ; pyridoxine serinate ; superoxide dismutase ; aminopropyl ascorbyl phosphate ; ascorbic acid ; ascorbyl dipalmitate ; ascorbyl glucoside ; ascorbyl linoleate ; ascorbyl methylsilanol pectinate ; ascorbyl palmitate ; ascorbyl tetraisopalmitate ; ascorbyl tocopheryl maleate ; trisodium ascorbyl palmitate phosphate ; disodium ascorbyl sulfate ; calcium ascorbate ; methylsilanol ascorbate ; sodium ascorbate ; sodium ascorbyl phosphate ; sodium ascorbyl/cholesteryl phosphate ; tetrahexyldecyl ascorbate ; magnesium ascorbyl phosphate ; tocopherol ; tocopheryl acetate ; tocopheryl linoleate ; tocopheryl oleate ; tocopheryl nicotinate ; tocopheryl retinoate ; sodium tocopheryl phosphate ; dioleyltocopheryl methylsilanol ; potassium ascorbyl tocopheryl 5 phosphate ; dodecyl gallate ; propyl gallate ; octyl gallate ; epigallocatechin gallate (EGCG) ; propyl gallate ; ethyl ferulate ; ethylhexyl ferulate ; chitosan ascorbate ; chitosan glycolate ; apigenin ; tiliroside ; alpha-arbutin ; arbutin ; baicalin ; quercetin ; quercetin caprylate ; isoquercetin ; diethylhexyl syringylidenemalonate ; dihydroxymethylchromone ; dimethoxy di-p-cresol ; dimethylmethoxy chromanol ; ethylbisiminomethylguaiacol manganese chloride ; hesperidin methyl chalcone ; kojic acid ; kojic dipalmitate ; madecassoside ; asiaticoside ; magnolol (5,5'- diallyl-2,2'-dihydroxybiphenyl) ; nordihydroguaiaretic acid ; phenylethyl resorcinol ; resveratrol ; troxerutin ; glucosylrutin, rutin (4H-1-benzopyran-4-one) ; disodium rutinyl disulfate ; tetrahydrobisdemethoxydiferuloylmethane ; tetrahydrodemethoxydiferuloylmethane ; tetrahydrodiferuloylmethane ; tococysteamide ; totarol ; hydroxydecyl ubiquinone ; ubiquinone ; caroténoïdes ; niacinamide ou l'un de ses dérivés ; créatine ; créatinine, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, rhamnose. Plus particulièrement, l'au moins un antioxydant est choisi dans le groupe comprenant tocophérol, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, rhamnose. Selon un mode de réalisation encore plus particulier, l'au moins un antioxydant selon l'invention est la carnosine. Selon un mode de réalisation particulier, l'invention concerne une composition comprenant : - au moins un sel minéral ou organique choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, du chlorure de sodium, de l'hydrogénocarbonate de sodium, de l'hydrogénophosphate de sodium, du citrate de sodium, du gluconate de magnésium, - de la carnosine. Selon un autre mode de réalisation particulier, l'invention concerne une composition comprenant : - au moins un sel minéral ou organique choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, du chlorure de sodium, de l'hydrogénocarbonate de sodium, de l'hydrogénophosphate de sodium, du citrate de sodium, du gluconate de magnésium, - de la carnosine pour son utilisation pour maintenir et/ou rétablir un microbiote cutané sain. Selon un autre mode de réalisation particulier, l'invention concerne une composition comprenant : - du citrate de sodium, - de la carnosine. Selon un autre mode de réalisation particulier, l'invention concerne une composition comprenant : - du citrate de sodium, - de la carnosine pour son utilisation pour maintenir et/ou rétablir un microbiote cutané sain. Selon un mode de réalisation particulier, la composition selon l'invention, ou pour son utilisation selon l'invention, comprend en outre au moins une source de carbone. Par « source de carbone », on entend un composé susceptible d'être utilisé par les bactéries comme substrat de croissance et de multiplication, telles que : - les protéines hydrolysées partiellement ou totalement, issues de céréales (blé, maïs, orge, seigle, avoine), de légumineuses (soja, fèverole, fève, pois), de lait (caséines, albumines), de levure, ainsi que les peptides et les acides aminés et leurs dérivés seuls ou en mélange ; - les oligosaccharides ou les polysaccharides, avantageusement à courte chaine, tels que les fructooligosaccharides, l'acide hyaluronique, le pullulane, l'alginate, la cellulose, avantageusement la cellulose microcristalline, le carraghénane, le chitosane, le dextrane et le dextrane sulfate, le galactoarabinane, l'amylose, l'amylopectine, les maltodextrines, les pectines, et leurs hydrolysats respectifs, ainsi que les sucres, oses, osides et leurs dérivés tels que glucose, galactose, fructose, xylose, mannose, glucosamine, lactose, ribose, rhamnose et leurs dérivés phosphates et acétylés en particulier. Les fructo oligosaccharides ou FOS peuvent être issu de matières végétales comme la betterave, l'artichaut ou le maïs. La matière première correspondant à la désignation INCI fructooligosaccharides peut être utilisée dans le cadre de la présente invention comme par exemple le produit commercialisé par la société Shaanxi Bolin Biotechnology Co sous le nom de FRUCTOOLIGOSACCHARIDES. Avantageusement, les FOS représentent de 0,001% à 20 % en poids total de la composition, avantageusement de 0,01% à 10 %, encore plus avantageusement de 0,1% à 5 %. Selon un mode de réalisation particulier, l'invention concerne une composition comprenant : - au moins un sel minéral ou organique choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, du chlorure de sodium, de l'hydrogénocarbonate de sodium, de l'hydrogénophosphate de sodium, du citrate de sodium, du gluconate de magnésium, - de la carnosine - au moins une source de carbone choisi parmi l'acide hyaluronique, et la cellulose, avantageusement la cellulose microcristalline. Selon un mode de réalisation particulier, l'invention concerne une composition comprenant : - au moins un sel minéral ou organique choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, du chlorure de sodium, de l'hydrogénocarbonate de sodium, de l'hydrogénophosphate de sodium, du citrate de sodium, du gluconate de magnésium, - de la carnosine ; - au moins une source de carbone choisi parmi l'acide hyaluronique, et la cellulose, avantageusement la cellulose microcristalline pour son utilisation pour maintenir et/ou rétablir un microbiote cutané sain. Selon un autre mode de réalisation particulier, l'invention concerne une composition comprenant : - du citrate de sodium, - de la carnosine. - au moins une source de carbone choisi parmi l'acide hyaluronique, et la cellulose, avantageusement la cellulose microcristalline Selon un autre mode de réalisation particulier, l'invention concerne une composition comprenant : - du citrate de sodium, - de la carnosine - au moins une source de carbone choisi parmi l'acide hyaluronique, et la cellulose, avantageusement la cellulose microcristalline pour son utilisation pour maintenir et/ou rétablir un microbiote cutané sain. Selon un mode de réalisation particulier, on utilise une forme pure ou hautement purifiée de carnosine dans la composition selon l'invention. Selon une variante, la carnosine est obtenue par synthèse chimique. A titre d'exemple, on peut citer la matière cosmétique L-Carnosine correspondant à la dénomination INCI carnosine et commercialisée par l'entreprise MERCK KgaA. Selon un autre mode de réalisation de l'invention, la carnosine représente entre 0,0001% et 5% en poids total de la composition, avantageusement entre 0,001% et 2%. En pratique, dans la composition selon l'invention on peut utiliser une forme pure ou hautement purifiée d'hypotaurine. Selon une variante, l'hypotaurine est obtenue par synthèse chimique. A titre d'exemple, on peut citer la matière première cosmétique hypotaurine correspondant à la dénomination INCI aminoethanesulfinic acid et commercialisée par l'entreprise DKSH. Selon un mode de réalisation particulier de l'invention, l'hypotaurine représente entre 0,0001% et 5% en poids total de la composition, avantageusement entre 0,001% et 2%. Selon un autre mode de réalisation particulier, le tocophérol utilisé est un mélange de tocophérols naturels purifiés, en particulier d'α-tocophérol, de β-tocophérol, de γ-tocophérol, et de δ-tocophérol ; ce mélange peut être utilisé notamment dans une huile choisie parmi les huiles végétales, minérales, siliconées, et de préférence végétales. Comme mélange de tocophérols naturels utilisable selon l'invention, on peut citer le mélange vendu par la Société Nisshin OilliO Group, Ltd sous la dénomination Tocopherol 80 et correspondant à la dénomination INCI « Tocopherol ». Selon un autre mode de réalisation particulier de l'invention, le tocophérol représente entre 0,0001% et 5% en poids total de la composition, avantageusement entre 0,001% et 2%. La bétaïne est un osmolyte et un antioxydant qu'on retrouve notamment dans le règne végétal. La bétaïne peut être d'origine végétale ou obtenue par biotechnologie. Avantageusement une forme hautement purifiée de bétaïne, d'une pureté d'au moins 95%, est utilisée dans les compositions selon l'invention. Comme source de bétaïne utilisable selon l'invention, on peut citer la matière première vendue par la société Evonik Nutrition & Care GmbH sous la dénomination de Tego Natural Bétaïne (Evonik Nutrition & Care GmbH) et correspondant à la dénomination INCI « Betaine ». Selon un autre mode de réalisation particulier de l'invention, la bétaïne représente entre 0,0001% et 5% en poids total de la composition, avantageusement entre 0,001% et 2%. Le xylitol peut être du xylitol en poudre, par exemple d'origine végétale, notamment obtenu à partir du bois par hydrogénation des xyloses, présentant en particulier une pureté supérieure à 95%. A titre d'exemple, la société Orient Star LCC commercialise le produit OriStar XLT correspondant à la dénomination INCI « Xylitol » pouvant être mis en œuvre dans le cadre de l'invention. De manière adaptée, le xylitol représente de 0,001% à 5% en poids total de la composition, avantageusement de 0,1% à 2%. Le mannitol peut par exemple être du mannitol en poudre, d'origine végétale (maïs, pomme de terre, blé), de préférence présentant une pureté supérieure à 98%. A titre d'exemple, la société American Biochemicals, Inc. commercialise le produit D-mannitol correspondant à la dénomination INCI mannitol pouvant être mis en œuvre dans le cadre de l'invention. De manière adaptée, le mannitol représente de 0,001% à 5% en poids total de la composition, avantageusement de 0,1% à 2%. L'inositol est un composé cyclique dérivé du cyclohexane et porteur de six fonctions alcool secondaire. Conformément à un mode de réalisation particulier de l'invention, on utilise de l'inositol d'origine végétale, synthétisé naturellement à partir du glucose-6-phosphate (G6P), issu de la photosynthèse ou de l'hydrolyse des réserves carbonées de la plante. De préférence, on utilise de l'inositol extrait du riz par les techniques connues de l'Homme du métier. On peut également utiliser de l'inositol préparé par synthèse chimique ou enzymatique. A titre d'exemple, la société Dupont Industrial Biosciences commercialise le produit Genencare OSMS MI correspondant à la dénomination INCI inositol pouvant être mis en œuvre dans le cadre de l'invention. De manière adaptée, l'inositol représente de 0,001% à 5% en poids total de la composition, avantageusement de 0,1% à 2%. Tous les sels minéraux ou organiques cosmétiquement ou pharmaceutiquement acceptables sont adaptés à l'utilisation dans la composition selon l'invention ou pour son utilisation selon l'invention. Avantageusement, il s'agit d'au moins un sel minéral ou organique choisi dans le groupe constitué par un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc, et leurs mélanges, plus particulièrement le citrate de sodium. Selon un mode de réalisation particulier de l'invention, l'au moins un sel minéral ou organique est choisi dans le groupe comprenant le chlorure de calcium ; le chlorure de potassium ; le phosphate de potassium ; le sulfate de magnésium ; le chlorure de sodium ; l'hydrogénocarbonate de sodium ; l'hydrogénophosphate de sodium ; le citrate de sodium ; le gluconate de magnésium. Ces sels minéraux sont disponibles auprès plusieurs fournisseurs de matières premières cosmétiques comme par exemple, COOPER INDUSTRIE ou BRENNTAG FRANCE. Dans un mode de réalisation particulier, la composition selon l'invention comprend au moins l'un des sels minéraux suivants, en pourcentage en poids : - De 0,000001% à 0,1% en poids total de la composition de chlorure de calcium, en particulier de 0,0001% à 0,1% ; - De 0,0001% à 0,1% en poids total de la composition de chlorure de potassium, en particulier de 0,001% à 0,1% ; - De 0,00001% à 0,1% en poids total de la composition de phosphate de potassium, en particulier de 0,0001% à 0,1% ; - De 0,0001% à 0,1% en poids total de la composition de sulfate de magnésium, en particulier de 0,001% à 0,1% ; - De 0,001 à 4% en poids total de la composition de chlorure de sodium, en particulier de 0,1 à 4% ; - De 0,0001 à 0,1% en poids total de la composition d'hydrogénocarbonate de sodium ; - De 0,0001% à 0,1% en poids total de la composition d'hydrogénophosphate de sodium ; - De 0,0001% à 1% en poids total de la composition d'acide citrique, en particulier de 0,001% à 1% ; - De 0,001% à 4% en poids total de la composition de citrate de sodium ; - De 0,001% à 4% en poids total de la composition de gluconate de magnésium. Selon un autre mode de réalisation particulier de l'invention, le poids total des sels minéraux et/ou organiques est inférieur ou égal à 1% en poids, en particulier est inférieur ou égal à 0,1% en poids du poids total de la composition. En particulier les sels minéraux et organiques sont utilisés à une concentration qui permet de se situer dans la fourchette de pression osmotique recherchée. Selon un mode de réalisation particulier, le citrate de sodium représente de 0,001 à 4% en poids total de la composition et la carnosine représente de 0,0001% à 5% en poids total de la composition, avantageusement de 0,001% à 2%. Plusieurs sources cosmétiques d'acide hyaluronique sont connues. Selon un mode de réalisation particulier de l'invention, l'acide hyaluronique a un poids moléculaire compris entre compris entre 20 et 50 kDa. A titre d'exemple, les matières premières PRIMALHYAL™ 50 ou HyaCare 50 commercialisées respectivement par les sociétés SOLIANCE et EVONIK et correspondantes à l'acide hyaluronique de poids moléculaire (Mw) compris entre 20 et 50 kDa (INCI : hyaluronic acid) peuvent être utilisées dans le cadre de la présente invention. Selon un mode de réalisation particulier, l'acide hyaluronique représente de 0,001 à 4% en poids total de la composition, avantageusement de 0,001% à 2%. Toute source commerciale de cellulose microcristalline adaptée à l'application topique et cosmétique peut être mis en œuvre dans les compositions selon l'invention. A titre d'exemple on peut citer Avicel PH-101 NF commercialisé par la société FMC Corporation et correspondant à a désignation INCI microcrystalline cellulose. Selon un mode de réalisation particulier, la cellulose microcristalline représente de 0,001 à 5% en poids total de la composition, avantageusement de 0,001% à 3%. Selon un autre mode de réalisation particulier, la composition selon l'invention ou pour son utilisation selon l'invention ou à utiliser en particulier dans la méthode de soin mentionnée auparavant ou dans la description suivante est destinée aux populations caractérisées par un microbiote cutané appauvri. La manière dont l'invention peut être réalisée et les avantages qui en découlent ressortiront mieux des exemples de réalisation qui suivent, donnés à titre indicatif et non limitatif, à l'appui des figures annexées. Dans les exemples les pourcentages sont donnés en poids, la pression est la pression atmosphérique et la température est la température ambiante habituellement comprise entre environ 18°C et environ 30°C, sauf indication contraire. [Fig.1] et [Fig.2] montrent l'effet des ingrédients purs vaseline (Fig 1) et du silicone (Fig 2) après 4 jours de contact versus zone non traitée (ZNT), sur la biomasse bactérienne du microbiote cutané, mesurée par le nombre de copies d'ADNg 16S chez les volontaires pris en compte dans l'analyse. La significativité statistique est déterminée par un test de Wilcoxon. (ns, non significatif : p>0.05 ; significatif * : p≤0.05 ; significatif ** : p≤0.01 ; significatif *** : p≤0.001). [Fig.3] et [Fig.4] montrent l'effet de la formule 1 (fle 1 ; Fig 3) et de la formule 2 (fle 2 ; Fig 4) après 4 jours de contact versus zone non traitée (ZNT), sur la biomasse bactérienne du microbiote cutané, mesurée par le nombre de copies d'ADNg 16S chez les volontaires pris en compte dans l'analyse. La significativité statistique est déterminée par un test de Wilcoxon. (ns, non significatif : p>0.05 ; significatif * : p≤0.05 ; significatif ** : p≤0.01 ; significatif *** : p≤0.001). [Fig.5] et [Fig.6] montrent l'effet des ingrédients purs vaseline [Fig.5] et silicone [Fig.6] après 4 jours de contact versus zone non traitée (ZNT), sur la diversité bactérienne du microbiote cutané, mesurée par l'indice de Shannon, chez les volontaires pris en compte pour l'analyse. La significativité statistique est déterminée par un test apparié Wilcoxon. (ns, non significatif : p>0.05 ; significatif * : p≤0.05 ; significatif ** : p≤0.01 ; significatif *** : p≤0.001). [Fig.7] et [Fig.8] montrent l'effet de la formule 1 (fle 1 ; Fig.7) et de la formule 2 (fle 2 ; Fig.8) après 4 jours de contact versus zone non traitée (ZNT), sur la diversité bactérienne du microbiote cutané, mesurée par l'indice de Shannon, chez les volontaires pris en compte pour l'analyse. La significativité statistique est déterminée par un test apparié Wilcoxon. (ns, non significatif : p>0.05 ; significatif * : p≤0.05 ; significatif ** : p≤0.01 ; significatif *** : p≤0.001). Exemples 1 de réalisation de l'Evaluation de l'effet de deux compositions cosmétiques et de deux ingrédients purs sur le microbiote cutané de volontaires sains [Description] METHOD FOR SELECTING TOPICAL ECOBIOLOGICAL INGREDIENTS AND COMPOSITIONS SUITABLE FOR RESTORING OR STRENGTHENING THE SKIN MICROBIOTA Technical Field of the Invention The present invention relates to the study of the skin microbiota and its use for the restoration and/or maintenance of healthy skin microbiota. STATE OF THE ART The skin, the largest human organ, is colonized by billions of microorganisms such as bacteria, yeasts, fungi or viruses collectively called microbiota or microflora. These microorganisms inhabit the epidermis, and are mainly found in the upper layers of the stratum corneum as well as in the ducts of sweat glands and pilosebaceous follicles (Kong et al., 2012a). Historically, the study of the human microbiota has focused first on the intestinal microflora, whose crucial role in maintaining human physiological balance is now indisputably recognized. The cutaneous microbiota, which is much less abundant, had to wait for the development and fine-tuning of metagenomic methods allowing its detailed characterization. These techniques, now available, allow the amplification and sequencing of the genetic material of the microbial communities of the skin. To date, the composition of the skin flora is only partially known. Only the bacterial composition of the skin is studied in detail. The number of bacteria present on the skin, whether on its surface, at the level of the glands or on the appendages, can reach nearly 1 billion per cm2 (Kong et al., 2012a). The human skin flora can be subdivided into two groups: - The resident flora is made up of commensal germs, ie living at the expense of their host, without causing them any harm. The resident human flora is dominated by Proteobacteria, including about ten types of Gram-positive aerobic bacteria such as Staphylococcus epidermidis and Staphylococcus hominis (Otto, 2010). The resident flora is also composed of Gram-positive anaerobic bacteria belonging to the Actinobacteria division (genera Cutibacterium, Corynebacterium, Dermabacter and Micrococcus) (Kligman et al., 1976). Among the microorganisms of the other kingdoms, Malassezia is the fungal species most frequently found on the skin. Finally, the mites Demodex folliculorum and Demodex brevis, or "eyelash mites", are also considered members of the resident human flora. - Transient flora is composed of mostly harmless fungi, viruses and bacteria, called saprophytes, that is to say, which feed on decomposing organic matter from the environment. This flora can also consist of opportunistic pathogenic bacteria that can cause disease in the host. The transient flora does not establish itself permanently on the surface of the skin, varying during the day, according to the activities carried out and the surrounding conditions. However, it can persist for hours or even days. Its density is low on dry areas and particularly high on hairy areas, folds and areas subject to perspiration. The most common transient species are Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and several species of the genus Bacillus. Several studies have shown that the skin microbiota has considerable differences between genders, ethnic groups and even individuals and that the distribution of microbial residents in different areas of the body is also highly variable (Grice et al., 2009). Despite these marked interpersonal and biogeographical differences, there is strong evidence that the entire resident microflora plays an important role in the health of the skin, particularly in terms of its immune defenses and its resistance to colonization by pathogenic microorganisms, also involving imbalances in the microbiota and the appearance or aggravation of dermatological conditions such as atopic dermatitis or psoriasis (Nakatsuji and al., 2017; Gao et al., 2008; Kong et al., 2012b). Indeed, just as in large-scale ecosystems, a loss of diversity of the microbiota is accompanied by a sometimes irreversible deregulation of its balances, in which a limited number of opportunistic species proliferate in an uncontrolled manner at the expense of the others. In the case of the skin microbiota, it is for example possible to cite in particular the species responsible for serious infections, often of nosocomial origin, Staphylococcus aureus and Pseudomonas aeruginosa. It is difficult to treat the skin infections caused by these strains which have sometimes acquired during their evolution, one or more resistances to conventional antibiotics, in particular beta-lactams. For example, methicillin-resistant Staphylococcus aureus (MRSA) represents a concrete public health risk to date, having in 2020 been responsible for more than a third of all community and hospital infections associated with mortality and morbidity high. The management of MRSA requires the use of new antibiotics, which can however further destabilize the entire human microbiota by slowing down the recovery of its diversity. Several indices making it possible to evaluate the diversity of the microbiota are developed, among which one can quote the indices of Shannon or Simpson, the most used being the index of Shannon which makes it possible at the same time to evaluate the number of species of this environment (specific richness) and the distribution of individuals within these species (specific equitability; Lemos et al., 2011). It is known that the use of topical products can have a significant impact on the composition and balances of cutaneous microbial communities (Kong, 2011; Kong et al., 2012a). Antiseptic products (i.e. fungicides, bactericides and/or virucides) in particular, including ethanol in particular, impoverish not only the quantity, but also the diversity of the skin microbiota and this potentially more durably (San Miguel et al., 2018). The application of biocides to the surface of the skin was relatively rare and limited to the hospital setting until 2020. However, since the global pandemic of COVID-19, the use of this category of products has become commonplace and widespread. to levels never before achieved. For alcohol-based products, the indiscriminate inhibition of the growth of all of the microbiota is added to its effect of destruction of the hydrolipidic film, which leads to an alteration of its barrier effect, with increased penetration of allergens and irritants. The reduction in the quality of the skin microbiota, caused in particular by biocidal products and antibiotic treatments, requires the development of a new class of ecobiological topical skincare products to maintain and/or quickly restore a healthy and diversified skin microbiota. There therefore remains an obvious need to develop methods allowing the selection of ingredients, or even topical compositions, capable of maintaining and/or restoring, preferably rapidly, a healthy microbiota. Description of the invention The present invention is based on the demonstration of the interest, under non-pathological conditions, of promoting the increase in the biomass of the cutaneous microbiota (multiplication of the microbiota), without reducing its overall diversity (conservation or increase in the diversity of said microbiota). The healthy skin microbiota and its balance are then reinforced and/or reconditioned to better resist potential opportunistic dysregulation and colonization. Surprisingly, the Applicant has thus found a way that is simple, effective, affordable and usable on an industrial scale, in particular in cosmetics or dermatology, for selecting ingredient(s) and/or composition(s) suitable to restore and/or strengthen the healthy skin microbiota, by seeking to increase the biomass of the skin microbiota, in particular without reducing its overall diversity. For this, in one embodiment of the invention, two methods are combined: a method for measuring the biomass on a skin area and a method for measuring the microbial diversity on this same area. According to the invention, the “microbiota” concerns all of the microbial flora, including microbes, bacteria, yeasts, fungi and viruses. According to the invention, by “cutaneous microbiota” or “keratinous material(s)”, is meant the microbiota of the skin, of the semi-mucous membranes (in particular the lips), of the mucous membranes and of the appendages including appendages. According to the invention, the “microbiome” relates to all the genes or genomes present in the microbiota. According to the invention, by “biodiversity” or “diversity” or “diversity of the microbiota” or “diversity of the microbiome” or “bacterial diversity”, one refers to all bacterial populations or species present in the skin microbiota. This biodiversity can be characterized in particular by the number of different bacterial genetic sequences within said microbiome (also denoted OTU, for “Operational Taxonomic Unit”) and/or their relative abundance. Several indices making it possible to evaluate the diversity of the microbiota have been developed and are well known to those skilled in the art, among which mention may be made of the Shannon or Simpson indices. According to the invention, by “biomass of the skin microbiota” or “biomass” or “bacterial load”, is meant the number or quantity of bacteria on a skin area. This biomass can be characterized in particular by the amount of bacterial genetic material, in particular 16S rDNA, from said microbiota. The biomass and diversity levels of a microbiota can be measured by any appropriate technique well known to those skilled in the art, such as quantitative amplification and 16S rDNA sequencing. According to the invention, “healthy” is understood to mean non-pathological. According to the invention, by “healthy skin area”, is meant a skin area not exhibiting dermatosis. According to the invention, by “healthy skin microbiota” or “healthy skin microbiota”, is meant a stable and diversified skin microbiota on a given skin area, such as on the same healthy skin area not subjected to stress conditions or particular aggressions (such as, but not limited to, treatments or care or non-ecobiological environments, in particular dermatological, cosmetics, hygiene, pollution, etc.). The microbiota of healthy skin protects in particular against invasive pathogens, but also against drying out of the skin. According to the invention, by "stress" or "aggression" is meant any event, of any nature whatsoever, single or repeated, in particular of external origin, which may be the cause of destabilization or modifying a system. At the cutaneous level, such an attack can in particular lead to a reduction in the biomass and/or the diversity of the cutaneous microbiota. In particular, it may be linked to non-ecobiological treatments or care or environments, in particular dermatological, cosmetic, hygiene, pollution, etc., and, more particularly, to the action of bactericides or virucides or fungicides or soaps or hygiene or care products that are aggressive towards the microbiota. By “ecobiological”, we mean respectful of people, their interactions with the world and the planet. The present invention relates to a method for selecting or screening ingredients and compositions, preferably ecobiological, cosmetic or dermopharmaceutical or dermatological, advantageously applicable topically, according to their effect on the cutaneous microbiota, in particular their ability to induce the increasing the biomass of the cutaneous microbiota, in particular without reducing its biodiversity. The present invention also relates to a method for preparing a composition, preferably topical, capable of inducing an increase in the biomass of the cutaneous microbiota, as well as its use for the preparation of care compositions, preferably topical, advantageously ecobiological and ecobiological care methods, preferably topically, based on the use of the aforementioned compositions or ingredients. The present invention also relates to a composition for its use for maintaining and/or restoring a healthy skin microbiota via an increase in the biomass of the skin microbiota, in particular without reducing its biodiversity. In other words, the invention aims for the first time to analyze and quantify the biomass of the cutaneous microbiota, as well as, where appropriate, its biodiversity and to select the substances and compositions which make it possible to increase the biomass of the cutaneous microbiota, in particular without diminishing its biodiversity. The present invention therefore relates to a method for selecting or screening ingredients, substances or topical compositions making it possible to maintain and/or restore, preferably rapidly, a healthy microbiota. The present invention relates in particular to a screening method whose objective is to select ingredients, preferably topically acceptable, individually and/or compositions comprising a mixture of several aforementioned ingredients, optionally combined with excipients, preferably topically acceptable, capable of increase the biomass of the cutaneous microbiota, in particular without reducing its biodiversity. The present invention also relates to a composition, preferably topical, selected by the screening method. The present invention also relates to one or more ingredients, optionally in the form of a composition, preferably topical, to be used for the manufacture of a composition, preferably topical, for the care of the skin, mucous membranes or appendages capable of increase the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, to maintain and/or restore the healthy nature of the skin, mucous membranes or appendages. The present invention finally relates to a care method, preferably by topical route, using one or more ingredients, advantageously topically acceptable, capable of increasing the biomass of the cutaneous microbiota, by particular without reducing its biodiversity, to maintain and/or restore the healthy character of the skin, mucous membranes or appendages. Thus, according to a first aspect, the invention relates to a method, preferably in vitro, for selecting an ingredient or composition, preferably ecobiological, advantageously applicable topically, in particular cosmetic or dermatological, useful in maintaining or/and the restoration of a healthy skin microbiota, comprising the steps consisting in: a) bringing an ingredient or composition to be tested into contact with bacteria of a reference microbiota, b) analyzing the quantity of biomass and, advantageously the biodiversity of the microbiota after bringing into contact, and c) selecting as an ingredient or composition useful in maintaining and/or restoring a healthy skin microbiota, an ingredient or a composition capable of increasing the biomass of the skin microbiota, advantageously without reduce its biodiversity, compared to the quantification of the biomass and, where applicable, the biodiversity of the reference microbiota. According to a particular embodiment, the method according to the invention is personalized, based on the conditions, specificities and reactivity of the skin microbiota of a given individual. According to a particular embodiment, step a is carried out in a single or repeated manner and/or step b is carried out from 1 hour to 31 days, preferably 4 days, after step a. According to a variant embodiment, the invention relates to a method, preferably in vitro, for screening or evaluating or selecting the activity of at least one ingredient or composition, preferably applicable topically, in particular an ingredient or cosmetic composition and/or dermatological agent, optionally formulated in the form of a topical composition, characterized in that the method comprises the following steps, preferably in vitro: i) before the application of the ingredient or of the composition, optionally formulated in the form of a topical composition, on at least one skin area to be studied presenting a so-called reference microbiota, from a first sample taken from said skin area, the analyzes of the amount of biomass of the skin microbiota and, where appropriate, of its biodiversity, thus obtaining a first so-called reference measurement, for quantifying the biomass and, where appropriate, a first so-called reference measurement, of the biodiversity of the bacteria of said reference microbiota; and ii) after at least one application of the ingredient or of the composition, optionally formulated in the form of a topical composition, on the said skin area, and after a predetermined time of action of the ingredient or of the composition, from a second sample taken from said skin area, the analyzes of the amount of biomass of the skin microbiota and, where appropriate, of its biodiversity, thus obtaining a second so-called measure of biomass quantification activity and, where appropriate , a second so-called biodiversity activity measure; and iii) comparing the second activity measurement(s) with the first reference measurement(s), and selecting the ingredient or composition, optionally formulated as a composition topical, increasing the biomass of the cutaneous microbiota, advantageously without reducing its biodiversity. In other words, the invention relates to a method, preferably in vitro, of screening or selecting at least one ingredient or composition, optionally formulated in the form of a topical composition, to induce an increase in the biomass of a cutaneous microbiota, advantageously without reduce its biodiversity including the following steps: i) quantifying the biomass of the skin microbiota and possibly measuring the biodiversity of a first sample from a skin area, thus making it possible to obtain a first quantification measurement of the biomass and, where appropriate, a first measurement of the biodiversity of a so-called reference skin microbiota; and ii) quantifying the biomass of the skin microbiota and optionally measuring the biodiversity of a second sample from said skin area to which the at least one ingredient or composition, optionally formulated in the form of a topical composition, has been applied, thus making it possible to obtain a second biomass quantification measurement and, if necessary, a second biodiversity measurement; and iii) comparing the second biomass quantification measurement with the first biomass quantification measurement, and where appropriate the second biodiversity measurement with the first biodiversity measurement and selecting the at least one ingredient or composition, optionally formulated in the form of a topical composition, capable of increasing the biomass of the cutaneous microbiota, if necessary without reducing its biodiversity. According to a preferred embodiment, the predetermined action time of the ingredient can vary between 1 hour and several days, in particular 31 days, advantageously 4 days. According to the invention, a “reference skin microbiota” or “so-called reference skin microbiota” or “reference microbiota” or “so-called reference microbiota” is for example a microbiota characteristic of a skin zone to be studied. A reference microbiota can also be a medium comprising different bacteria in various proportions which may or may not correspond to a microbiota existing in nature. The reference microbiota can be obtained in particular from a sample of a skin zone of an individual or by preparing a medium comprising the different bacteria in the proportions corresponding to said reference microbiota. By "medium" is meant a medium suitable for the growth of bacteria. It can be a solid or liquid medium. According to a particular embodiment, the levels of biomass and diversity of a reference microbiota characteristic of a skin area is an average of the levels of biomass and diversity and/or of microbiota diversity profile measured on samples from said area of skin in several individuals. According to a preferred embodiment of the invention, said reference microbiota is that of a healthy skin area and/or is depleted, advantageously following stress or cutaneous aggression, more advantageously following the action of bactericides or virucides or soaps or hygiene or care products that are aggressive towards the microbiota. According to a particular embodiment of the invention, a method is provided for screening or selecting or evaluating the activity of at least one ingredient which is preferentially applicable topically, in particular a cosmetic and/or dermatological ingredient, optionally formulated in the form of a topical composition, characterized in that the method comprises: a) before any operation, the selection of a skin area to be studied from one or more human beings, b) the taking of at least a first sample of biological or genetic material of the microbiota present on this selected area, of a or several human beings, constituting a so-called reference microbiota, followed by the analysis of the quantity of biomass and the biodiversity of the skin microbiota, thus obtaining first so-called reference measurements respectively for quantification of the biomass of the skin microbiota and of biodiversity of the cutaneous microbiota, c) the application of at least one ingredient or a composition to this area, in a single or repeated manner, d) after a period of 1 hour to 31 days, advantageously 4 days, after the application single or repeated applications, again the taking of at least a second sample of biological or genetic material from the microbiota present in this selected area, from one or more human beings, e) the use of at least one method making it possible to quantify the biomass present in each of the aforementioned first and second samples by thus obtaining respectively a first measurement, called reference, of quantification of the biomass of the cutaneous microbiota and a second measurement, called activity, of quantification of the biomass of the cutaneous microbiota, f) the use of at least one method making it possible to measure the microbial biodiversity present in each of the aforementioned first and second samples, thus obtaining respectively a first measurement, called a reference, of the biodiversity of the cutaneous microbiota and a second measurement, called activity, of the biodiversity of the cutaneous microbiota, g) the evaluation of the activity of the ingredient, or possibly of the composition, applied topically, on the biomass and the biodiversity of the microbiota sampled by comparison of the biomass and biodiversity of the microbiota between said first measurements obtained from the first p reading and said second measurements obtained from the second sample, and h) selecting the ingredient, optionally formulated as a composition topical, which increases the biomass of the cutaneous microbiota, in particular without reducing its biodiversity. According to a particular embodiment, said ingredient or said composition to be tested in the selection or screening method according to the invention is chosen from the group comprising: a) at least one carbon source which is capable of being used by bacteria such as substrate for growth and multiplication, and preferably chosen from: - partially or totally hydrolyzed proteins, from cereals (wheat, corn, barley, rye, oats), legumes (soya, fava beans, broad beans, peas), milk ( caseins, albumins), yeast, as well as peptides and amino acids alone or as a mixture; - short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) at least one antioxidant which has an activity on reactive oxygen species (ROS), advantageously free radicals, in particular chosen from the group consisting of octadecyl di-t-butyl-4-hydroxyhydrocinnamate; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t-butylnitrone; ergothioneine; melatonin; acetyl cysteine; cysteine; lysine hydrochloride; carnosic acid; tyrosyl histidine HCl; histidine hydrochloride; pyridoxine serinate; superoxide dismutase; aminopropyl ascorbyl phosphate; ascorbic acid; ascorbyl dipalmitate; ascorbyl glucoside; ascorbyl linoleate; ascorbyl methylsilanol pectinate; ascorbyl palmitate; ascorbyl tetraisopalmitate; ascorbyl tocopheryl maleate; trisodium ascorbyl palmitate phosphate; disodium ascorbyl sulphate; calcium ascorbate; methylsilanol ascorbate; sodium ascorbate; sodium ascorbyl phosphate; sodium ascorbyl/cholesteryl phosphate; tetrahexyldecyl ascorbate; magnesium ascorbyl phosphate; tocopherol; tocopheryl acetate; tocopheryl linoleate; tocopheryl oleate; tocopheryl nicotinate; tocopheryl retinoate; sodium tocopheryl phosphate; dioleyltocopheryl methylsilanol; potassium ascorbyl tocopheryl 5 phosphate; dodecyl gallate; propyl gallate; octyl gallate; epigallocatechin gallate (EGCG); propyl gallate; ethylferulate; ethylhexyl ferulate; chitosan ascorbate; chitosan glycolate; apigenin; tiliroside; alpha-arbutin; arbutin; baicalin; quercetine; quercetin caprylate; isoquercetin; diethylhexyl syringylidenemalonate; dihydroxymethylchromone; dimethoxy di-p-cresol; dimethylmethoxy chromanol; ethylbisiminomethylguaiacol manganese chloride; hesperidin methyl chalcone; kojic acid; kojic dipalmitate; madecassoside; asiaticoside; magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl); nordihydroguaiaretic acid; phenylethyl resorcinol; resveratrol; troxerutin; glucosylrutin, rutin (4H-1-benzopyran-4-one); disodium rutinyl disulfate; tetrahydrobisdemethoxydiferuloylmethane; tetrahydrodemethoxydiferuloylmethane; tetrahydrodiferuloylmethane; tococysteamide; totarol; hydroxydecyl ubiquinone; ubiquinone; carotenoids; niacinamide or one of its derivatives; creatine; creatinine, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, and rhamnose; more particularly, in the group comprising tocopherol, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose; even more particularly carnosine; c) at least one topically acceptable inorganic or organic salt, advantageously chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular the sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium, copper or zinc gluconate, more particularly sodium citrate. According to another aspect, the invention also relates to a method for preparing a composition, advantageously topical, in particular cosmetic or dermatological, preferably ecobiological, characterized in that it comprises (i) the implementation of a method of selection or screening according to the invention (ii) the selection of at least one composition, advantageously topical, in particular a cosmetic and/or dermatological composition, comprising at least one ingredient selected by the selection or screening method according to the invention and capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity and (iii) the conditioning of the said composition. According to another aspect, the invention relates to a composition, advantageously topical, preferentially ecobiological, in particular cosmetic and/or dermatological, comprising at least one ingredient or a composition, selected by the selection or screening method according to the invention, for its use for its use in maintaining and/or restoring a healthy skin microbiota. Advantageously, the composition for its use according to the invention comprises at least one carbon source, an antioxidant and an inorganic or organic salt, more advantageously said carbon source, said antioxidant and/or said salt being selected by selection or screening method according to the invention. According to a preferred embodiment: a) the at least one carbon source is capable of being used by bacteria as a substrate for growth and multiplication, and, preferentially, is chosen from: - partially or totally hydrolyzed proteins, from cereals (wheat, corn, barley, rye, oats), legumes (soya, fava beans, beans, peas), milk (caseins, albumins), yeast, as well as peptides and amino acids alone or as a mixture; - short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) The at least one antioxidant exhibits activity on reactive oxygen species (ROS), advantageously free radicals, and, in particular, is chosen from the group consisting of octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t-butylnitrone; ergothioneine; melatonin; acetyl cysteine; cysteine; lysine hydrochloride; carnosic acid; tyrosyl histidine HCl; histidine hydrochloride; pyridoxine serinate; superoxide dismutase; aminopropyl ascorbyl phosphate; ascorbic acid; ascorbyl dipalmitate; ascorbyl glucoside; ascorbyl linoleate; ascorbyl methylsilanol pectinate; ascorbyl palmitate; ascorbyl tetraisopalmitate; ascorbyl tocopheryl maleate; trisodium ascorbyl palmitate phosphate; disodium ascorbyl sulphate; calcium ascorbate; methylsilanol ascorbate; sodium ascorbate; sodium ascorbyl phosphate; sodium ascorbyl/cholesteryl phosphate; tetrahexyldecyl ascorbate; magnesium ascorbyl phosphate; tocopherol; tocopheryl acetate; tocopheryl linoleate; tocopheryl oleate; tocopheryl nicotinate; tocopheryl retinoate; sodium tocopheryl phosphate; dioleyltocopheryl methylsilanol; potassium ascorbyl tocopheryl 5 phosphate; dodecyl gallate; propyl gallate; octyl gallate; epigallocatechin gallate (EGCG); propyl gallate; ethylferulate; ethylhexyl ferulate; chitosan ascorbate; chitosan glycolate; apigenin; tiliroside; alpha-arbutin; arbutin; baicalin; quercetine; quercetin caprylate; isoquercetin; diethylhexyl syringylidenemalonate; dihydroxymethylchromone; dimethoxy di-p-cresol; dimethylmethoxy chromanol; ethylbisiminomethylguaiacol manganese chloride; hesperidin methyl chalcone; kojic acid; kojic dipalmitate; madecassoside; asiaticoside; magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl); nordihydroguaiaretic acid; phenylethyl resorcinol; resveratrol; troxerutin; glucosylrutin, rutin (4H-1-benzopyran-4-one); disodium rutinyl disulfate; tetrahydrobisdemethoxydiferuloylmethane; tetrahydrodemethoxydiferuloylmethane; tetrahydrodiferuloylmethane; tococysteamide; totarol; hydroxydecyl ubiquinone; ubiquinone; carotenoids; niacinamide or one of its derivatives; creatine; creatinine, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, and rhamnose; more particularly, in the group comprising tocopherol, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose; even more particularly, carnosine; c) The at least one inorganic or organic salt is topically acceptable and, advantageously, is chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, more particularly sodium citrate. According to a preferred embodiment, the composition for its use according to the invention comprises sodium citrate and carnosine. According to a particular embodiment, the composition for its use according to the invention is an aqueous composition, comprising at least 60% by weight of the total composition of water, in particular at least 60%, better still at least 70%, or at least 80% or at least 90% or at least 95%. Preferably, the aqueous phase represents at least 60% by weight of the total composition, better still at least 70%, or at least 80% or at least 90% or at least 95%. Advantageously, the aqueous phase is characterized by: - a resistivity, at ambient temperature, of between 12.5 and 12500 Ohms.cm, in particular between 80 and 8000 Ohms.cm; and/or - an osmotic pressure between 70 and 2500 mOsmol/l and in particular between 100 and 800 mOsmol/l as well as a pH between 5.0 and 9.0 and in particular between 6.0 and 8, 0. According to a particular embodiment, the composition for its use according to the invention has a pH comprised between 5.0 and 9.0 and in particular comprised between 6.0 and 8.0. According to a particular embodiment, the composition for its use according to the invention is applied to a healthy skin area, before or following a skin attack, advantageously the action of bactericides or virucides or soaps or hygiene products or aggressive treatments for the microbiota. According to a particular embodiment, said cutaneous aggression according to the invention can be single or repeated, preferentially repeated. According to a particular embodiment, the composition for its use according to the invention is for its use for combating cutaneous aggression on healthy skin, by stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, for increase the amount of this biomass of said microbiota in order to avoid the arrival and/or installation of new microbial strains linked to the environment. According to a particular embodiment, the composition according to the invention is for its use in combating cutaneous aggression on healthy skin by stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, to restore the quantity of this biomass of said microbiota, greatly reduced during a skin attack and for example after the action of bactericides or virucides or soaps or hygiene or care products that are aggressive towards the microbiota. According to another aspect, the invention relates to a method of treatment, advantageously topical, in particular non-therapeutic or dermatological cosmetic, preferably ecobiological, for maintaining and/or restoring a healthy skin microbiota, characterized by the use of a composition such as previously described, allowing the biomass of the skin microbiota to be increased, in particular without reducing its biodiversity. According to another aspect, the invention relates to the use of at least one ingredient, or at least one substance, or at least one at least one composition, selected by the selection or screening method according to the invention for the preparation of a composition, preferably topical, in particular ecobiological, for maintaining and/or restoring a healthy skin microbiota. According to another aspect, the invention relates to a method of topical care, in particular ecobiological, advantageously cosmetic or dermatological, for treating the skin requiring maintenance or restoration of its healthy microbiota, by increasing the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, characterized in that at least one area of the skin requiring this care is selected, and one applies to at least this area of the skin an ingredient, advantageously formulated in a composition, or a composition selected through the selection or screening method described previously or in the following description. According to a particular embodiment, the invention also relates to a topical care method, preferably ecobiological, characterized in that it comprises (i) the implementation of a selection or screening method defined previously or in the description following, and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing the increase in the biomass of the cutaneous microbiota described previously, in particular without reducing its biodiversity ( ii) the formulation of the ingredients or compositions thus selected in a formula for cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota without reducing its diversity (iii) the packaging of said formula (iv) the topical application of said formulation to an area of skin or a surface of the selected appendages. According to yet another aspect, the invention relates to a treatment method, advantageously topical, preferably cosmetic or dermatological, in particular ecobiological, for combating cutaneous aggression on healthy skin, characterized in that at least one zone is selected of the skin requiring this care and the administration is carried out, preferably by topical application, of at least one ingredient or composition, advantageously applicable topically, in particular a cosmetic and/or dermatological ingredient or composition, in particular formulated in the form of topical composition, selected by the screening or selection method described previously or in the following description, stimulating the biomass of the microbiota present in the skin, or present on a keratin material, to increase the quantity of this biomass of said microbiota in order to avoid the arrival of new microbial strains linked to the environment. According to another aspect, the invention relates to a treatment method, advantageously topical, preferably cosmetic or dermatological, in particular ecobiological, for combating cutaneous aggression on healthy skin, characterized in that at least one area of the skin requiring this care and the application is carried out, preferably topically, of at least one ingredient which can be applied topically, in particular a cosmetic and/or dermatological ingredient, preferably formulated in the form of a topical composition, selected by the method of selection or screening described previously or in the following description, stimulating the biomass of the microbiota present at the cutaneous level, or present on a keratin material, to restore the quantity of this biomass of said microbiota against its reduction in quantity by a cutaneous aggression and for example after the action of bactericides or virucides or soaps or aggressive hygiene or care products vis-à-vis the mic robot. Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the field to which the disclosed methods and compositions relate. General definitions can be found in technical books relating to the field of molecular biology, for example, Dictionary of Microbiology and Molecular Biology, 2nd edition. (Singleton et al., 1994) or The Harper Collins Dictionary of Biology (Hale et al., 1991). As used in the present invention, the following terms and expressions have the meaning attributed to them, unless otherwise indicated. By “keratinous materials”, is meant according to the invention the skin, the semi-mucous membranes (in particular the lips), the mucous membranes and the appendages including the skin appendages. By "method for quantifying the biomass of the microbiota", we mean according to the invention, the use of a quantitative or semi-quantitative method which is based for example: - on quantitative PCR (or qPCR) techniques, or - on flow cytometry, or - on agar counting methods, or - on any related method. By "method for measuring the biodiversity of the microbiota", is meant according to the invention, the use: - of a method which is based on the analysis of part of the hypervariable gene of the 16S ribosome of bacteria after their targeted amplification , or - a method using the metagenomic shotgun technique (on microbiome DNA), or - a so-called metatranscriptomic shotgun sequencing method (on microbiome RNA), or - any related method, followed by a method for calculating the microbial diversity from the data of the previous method used, such as the calculation: - of the Shannon index, - of the Simpson index, - of the Inverse-Simpson index, - the number of species or OTU, - the Evenness or the Pielou index, - the Hurlbert index, - the Hill index. These calculation methods and indices are well described in the literature and known to those skilled in the art. They can be found, for example, in the document by Eric Marcon, “Measurement of Biodiversity, Kourou, France, ffcel-01205813v5ff”. The sampling of biological or genetic material from the microbiota can be done according to the methods known to those skilled in the art. Typically, the biological or genetic material is recovered using a swab which allows the bacteria to be collected, always performing the same manipulation to standardize the quantities taken as much as possible. As an example the Sigma Trans Swab liquid Maies swab can be used. In a preferred embodiment, the sample is taken by carrying out 10 return trips in Z by executing a slight pressure on the swab. At this stage, it is possible to freeze the swabs, which can be stored at -80°C. From the swabs, the bacterial pellets can be recovered by centrifugation. The extraction of genomic DNA (gDNA) can be carried out according to methods known to those skilled in the art, for example with the Nucleospin Microbial DNA kit (Macherey Nagel, France). The samples can be taken from several areas of the human body, such as, for example, the cheek, forehead, forearm, in order to study the response of the microbiota of the different biogeographical areas to the ingredients or compositions applied topically. In a preferred embodiment, the sample is taken from the face, which makes it possible to test the ingredients or compositions on a realistic area exposed to the environment, or on the back, which makes it possible to test a set of different conditions. on a relatively homogeneous microbiota (Grice et al., 2009). According to a preferred embodiment, the area concerned can be marked by a mask, advantageously delimiting a sampling surface of between 10 and 20 cm2, advantageously 15 cm2. In a preferred embodiment, this biomass is evaluated starting from the biological or genetic material recovered from the keratin materials, but it can also be quantified directly according to the methods known to those skilled in the art, for example by immunolabeling or flow cytometry on labeled bacteria. By “standardization” within the meaning of the invention, is meant the standardization of the quantity of biomass sampled. By "amplification" within the meaning of the invention is meant any in vitro procedure which produces multiple copies of a target nucleic acid sequence, or of its complementary sequence, or of fragments thereof (i.e. i.e. an amplified sequence containing less than the complete target nucleic acid). Advantageously, this amplification is carried out by polymerase chain reaction or polymerase chain reaction (PCR), but other methods known to those skilled in the art, such as isothermal amplification, can also be used. By “quantitative amplification” is meant within the meaning of the invention any in vitro procedure which produces multiple copies of a target nucleic acid sequence and which at the same time makes it possible to determine the concentration of target sequence initially present in the reaction mixture. . Several types of quantitative amplification reactions are described. We can distinguish quantitative amplifications based on the use of an external standard, competitive amplifications, using an internal standard, and finally kinetic amplifications, which consist in measuring in real time, the increase in the quantity of the target sequence . Advantageously, the quantitative amplification is a kinetic amplification carried out by PCR and corresponds to the technique known to those skilled in the art under the name of quantitative PCR (qPCR) or even real-time PCR. The PCR reactions, whether quantitative or non-quantitative, can be carried out with the “Light Cycler” system (Roche Molecular Systems Inc.) and according to the procedures recommended by the supplier. The term "target sequence", as used in the present invention, refers to the particular nucleotide sequence of the target nucleic acid which is to be amplified and/or detected. "Target sequence" includes complexing sequences to which oligonucleotides (eg, primer oligonucleotides and/or promoter oligonucleotides) complex during amplification processes (eg, PCR). When the target nucleic acid is originally single-stranded, the term “target sequence” will also designate the sequence complementary to the “target sequence” as present in the target nucleic acid. When the target nucleic acid is originally double-stranded, the term "target sequence" refers to both sense (+) and antisense (-) strands. The term "region", as used in the present invention, means a portion of a nucleic acid, said part being smaller than the entire nucleic acid. In particular, with “regions V1, V2, V3, V4” is meant hypervariable regions of the bacterial 16S ribosomal DNA (rDNA) defined for example in Gray et al., 1984, which constitutes a portion of the gDNA and which allows the identification of the species to which they belong during sequencing. By "amplicon" or "amplification product" is meant a nucleic acid molecule generated in an amplification reaction, whether quantitative or not, of nucleic acid and which is derived from a target nucleic acid. An amplicon or amplification product contains a target nucleic acid sequence which may be of the same or opposite sense to the target nucleic acid. By “sequencing” within the meaning of the invention is meant the decoding of at least one strand of a target sequence, and the reading of the sequence of nucleotides composing it in their order. Several methods for sequencing and assembling the sequences read are known to those skilled in the art. Advantageously, the sequencing is carried out using the Illumina Miseq™ platform. For example, the documents WO03048387A2, WO2005024010A1 WO2006064199A1 and WO2007123744A1 or also WO2012096703A1 and the references cited in these documents describe sample preparation and sequencing methods suitable for the present invention. The sequencing can be carried out directly on the amplicons, or alternatively on banks of amplicons. According to a preferred embodiment, the sequencing is carried out using a library of amplicons, advantageously the libraries of amplicons obtained according to the Metabiote™ solution described in Guillaume et al., 2014. A “primer” is an oligomer which is 'hybridizes to a nucleic acid and has a 3' end which is extended by polymerization. A primer can optionally be modified, for example by including a 5' region that is not complementary to the target sequence. Such modification may include functional additions, such as tags, promoters or other non-specific target sequences, used or useful to manipulate or amplify the primer or target oligonucleotide. Primers can be "degenerate". Within the meaning of the invention, “degenerate primer” denotes primers allowing incorporation of different nucleotides at determined positions. Degenerate primers are known to those skilled in the art; the following code is universally used to note the degeneracy of primers at a given nucleotide position: I=Inosine (Nichols et al., 1994); R = G or A; K= G or T; S = G or C; W = A or T; M = A or C; Y = T or C; D = G or A or T; V = G or A or C; B = G or T or C; H = A or T or C; N=G or A or T or C. By “universal 16S primers” within the meaning of the invention, is meant oligonucleotides allowing the amplification of highly conserved regions of the 16S rDNA. According to a particular embodiment, these are the primers corresponding to SEQ ID.18F (5'-AGAGTTTGATCCTGGCTCAG-3') and SEQ ID.21391R (5'-GACGGGCGGTGWGTRCA-3'). (Kong et al., 2012b). According to an alternative embodiment, these are the primers corresponding to SEQ ID.3533F (5'-GTGCCAGCAGCCGCGGTAA-3') and SEQ ID.4902R (5'-GTCAATTCITTTGAGTTTYARYC-3') (San Miguel et al., 2018 ). By “16S specific primers” within the meaning of the invention is meant oligonucleotides allowing the amplification of hypervariable regions of 16S rDNA. Advantageously, these are the V1-V4 regions of 16S rDNA, preferably the V1-V3 and/or V3-V4 regions, advantageously the V3-V4 regions. Advantageously, the 16S specific primers make it possible to obtain amplicons of size between 100 and 500 base pairs, advantageously between 200 and 400 base pairs. According to one embodiment, these are the primers for the V1-V3 regions corresponding to SEQ. ID 527F (5'-AGAGTTTGATCCTGGCTCAG-3') and SEQ. ID 6534R (5-ATTACCGCGGCTGCTGG-3) (San Miguel et al., 2018). According to a preferred embodiment, these are the primers for the V3-V4 regions corresponding to SEQ ID, 7338F (ACTCCTACGGGAGGCAGCAG) and SEQ ID.8806R (GGACTACHVGGGTWTCTAAT) (Li et al., 2020). The expressions “bank(s)” or “library(s)” will be used interchangeably in the present description, these two expressions having the same meaning. Within the meaning of the invention, the term “bank” means a collection of bacterial nucleotide sequences preferably resulting from the amplification of 16S rDNA and which are capable of being sequenced or detected individually. The constitution of a DNA library can be done according to the methods known to those skilled in the art, for example by using kits such as the NEBNext™ Ultra™ DNA Library Prep Kit. According to a particular embodiment, it is considered within the scope of the invention that the selected ingredient or composition applied topically has a significant effect on the increase in the biomass of the cutaneous microbiota, if the biomass increases statistically significant between two samples, in particular between the first sample and the second sample mentioned above, in particular after 1 hour to 31 days of treatment. According to the invention, the evaluation of the activity of the ingredient or composition applied topically on the biomass of the sampled microbiota can be carried out by comparing the total bacterial genetic material present on the sampled area before and after the treatment with the ingredient or composition through quantitative amplification with universal 16S primers, which makes it possible to estimate the genetic material present at the time of sampling before and after treatment. In a particular embodiment, the at least one amplification of the standardized amounts of genetic material is a quantitative amplification carried out using the universal 16S primers. For this, bacterial lysis followed by a genomic DNA extraction step with the PureLink Mini kit (Life Technologies, Grand Island, NY) is carried out. 16S rRNA gene-targeted qPCR is performed using specific primers (Bact-8F: AGAGTTTGATCCT GGCTCAG, Bacte338R: CTGCTGCCTCCCGTAGGAGT) with SYBR Green detection (Applied Biosystems, Foster City, CA) on equipment Mx300P (Agilent Technology). Standard curves are prepared by amplifying known amounts of DNA from E. coli. By comparing the number of cycles to the standard curve it is possible to calculate the number of copies of the RNA16S gene per microliter of sample. In another particular embodiment, the evaluation of the biomass of the cutaneous microbiota is carried out by quantification in flow cytometry. For it, the swab is discharged into 1 mL of Amies liquid then the bacterial suspension is labeled with SYBRGreen, incubated for 15 min at 37°C protected from light and analyzed by a BD Accuri C6 flow cytometer (BD Accuri cytometer, Belgium) equipped with a 50 mW laser emitting at a fixed wavelength of 488 nm. Fluorescence intensity is measured at Fl1 = 533 +/- 30 nm, Fl3 > 670 nm. Data are processed with BD Accuri CFlow software. A number of bacteria is thus obtained per mL of sample. In another particular embodiment, the evaluation of the biomass of the cutaneous microbiota is carried out by quantification by counting on agar. For this, after unloading the swabs in the Amies liquid, the bacterial suspensions are deposited on nutrient agar in a Petri dish, and are incubated for 48 hours at 35°C. A count of the bacterial colonies makes it possible to obtain the bacterial concentration by multiplying the number of CFU by the dilution factor, and is expressed in CFU/ml. In yet another particular embodiment, the evaluation of the biomass of the cutaneous microbiota is carried out by quantification by direct counting under a microscope. For this, the bacterial suspensions are inoculated in a nutrient broth such as Mueller-Hinton or Brain Heart Infusion. After incubation, a direct enumeration under the microscope can be carried out on a Thoma cell. According to a particular embodiment of the invention, it is considered that there has been no reduction in the bacterial diversity of the cutaneous microbiota if the variations are not statistically significant. According to a particular embodiment of the invention, the evaluation of the activity of the ingredient or composition applied topically on the diversity of the sampled microbiota can be performed by comparing the bacterial species present in the different experimental conditions through amplification with specific 16S primers and sequencing of the amplicons obtained. Several indices making it possible to estimate the diversity of a genetic material consisting of mixtures of different species are known to those skilled in the art. According to a particular embodiment, the index is the so-called Shannon index, which is calculated by following the method described in Lemos et al., 2011. The Shannon index integrates two distinct parameters: the number of operational taxonomic units ( Operational Taxonomic Units or OTUs) and the distribution of these OTUs in each sample. This last parameter can be calculated by taking into account the abundance of the 15 most represented bacterial genera, or alternatively, representing the 90% of the amplicons obtained in the amplification with the specific 16S primers. The bioinformatics processing of the target sequences can be carried out according to the methods known to those skilled in the art, for example by using a PERL script. According to another particular embodiment, the analysis by amplification with the specific 16S primers then, optionally, sequencing is carried out only for the samples of genetic material from a sample making it possible to obtain, during the quantitative amplification with the 16S universal primers in a reaction volume of 4.38 µl, at least 400 copies of 16S rDNA amplicon/µL after 30 cycles of amplification and/or at least 500 copies of amplicon of 16S/µL rDNA after 35 cycles of amplification. According to yet another particular embodiment, the analysis of the diversity of the cutaneous microbiota is carried out by metagenomics in the following way: after extraction of the gDNA, the microbial diversity is determined for each sample by targeted amplification of part of the gene ribosomal. A fragment of the 16S rRNA gene comprising the hypervariable regions V3 to V4 (or V1-V3) is amplified with primers 338F (ACTCCTACGGGAGGCAGCAG) and 806R (GGACTACHVGGGTWTCTAAT) (Munyaka et al., 2015). The amplification is carried out and the products resulting from PCR are purified and normalized using Agencourt AMPure XP magnetic beads (Beckman Coulter). Once purified, amplicons are pooled for Illumina sequencing, performed in 2x300 bp on an Illumina MiSeq platform using MiSeq Reagent Kit v3 (600 cycles). The sequences are grouped into operational taxonomic units (OTUs) with an identity threshold of 97%. Taxonomic assignment is performed using the Ribosomal Database Project (RDP) classification on the Greengenes V13_8 database. The analysis is carried out with the R software. The Shannon-Weaver index, the number of OTUs observed and the Bray-Curtis dissimilarity index are calculated with the Phyloseq and Vegan packages. Statistical significance is determined using the Wilcoxon test. A p-value <0.05 is considered statistically significant. According to another embodiment, the analysis of the diversity of the cutaneous microbiota is carried out by Shotgun: after extraction of the gDNA, the microbial diversity is determined for each sample by Whole metagenome sequencing/WMS or Shotgun metagenome sequencing, i.e. i.e. complete sequencing of the entire bacterial genome. The libraries are prepared using the NexteraXT preparation kit (Illumina). Sequencing is performed at the Penn Next Generation Sequencing Core on Illumina MiSeq chemistry to obtain final paired reads of 150 bp. Sequencing data is obtained in fastq format. Sequences shorter than 80 nucleotides are removed from the analysis. DeconSeq is used to filter fastq files and one of the paired reads (SE1) is entered into MetaPhlAn version 1.7.7 for taxonomic classification. One of the matched ends (SE1) of the MCC sample is used to perform a BLAST against a personalized database of genomes among the 20 expected bacterial species (blastn, max_target_seqs 1, e <10−10; alignment length > 50) to determine the composition of bacterial communities. The alpha diversity index is then calculated using the R software and the Phyloseq and Vegan packages using the Shannon index. According to another embodiment, the analysis of the diversity of the cutaneous microbiota is carried out by Metatranscriptomic shotgun sequencing (MTS): for this the total RNA is extracted from the bacterial sample. The polyadenylated mRNAs are separated from the non-polyadenylated ones (rRNA, mitochondrial RNA, and tRNA) by affinity on a poly-dT column (Dynabeads Oligo (dT) kit, Dynal). The enriched mRNA is mechanically fragmented to a size range of +/- 200 bp with an ultrasonicator. mRNA fragmentation is assessed using the Agilent RNA 6000 Pico Kit on a 2100 Bioanalyzer instrument (Agilent Technologies, Inc.). Metatranscriptome libraries are prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs Inc). The quality and quantity of all final libraries are analyzed with an Agilent DNA 1000 kit on the 2100 Bioanalyzer and Qubit instrument and are quantified and validated by a qPCR assay using the PerfeCTa NGS library quantification kit for Illumina (Quanta Biosciences, Inc.) using the CFX Connect real-time PCR detection system (Rad Laboratories, Inc.). Sequencing of one of the MT libraries is performed on an Illumina HiSeq 2000 using TruSeq SBS v3 reagent for 100 pair read length (BGI Americas) (labeled HS100) and on Illumina MiSeq using the v3-600 cycle kit for 301 base pairs (labeled MS301). Another set of twelve libraries is sequenced on Illumina MiSeq using 151 “paired-end chemistry” (marked MS151). The manufacturer's recommended protocol is used to perform the sequencing reaction on the HiSeq and MiSeq platforms. For PCR-amplified 16S ribosomal sequences, BLASTN searches (default settings, except word size set to 7) are performed against the GenBank nr nucleotide database (http://www.ncbi.nlm.nih.gov /). The 16S sequences are then assigned to major phyla to perform phylogenetic analyzes (BioNJ and PhyM) using the Best Blast Hit (BBH) algorithm and a set of reference sequences for each of the different phyla. Analyzes (sequence alignments and phylogenetic analyses) are performed using SeaView. Using BLASTX output files, cDNAs are taxonomically assigned using MEGAN V 3.5 (www-ab.informatik.uni-tuebingen.de/software/megan). According to yet another particular embodiment, the method according to the invention comprises at least two samples of biological or genetic material from the microbiota: - a sample of biological or genetic material from the microbiota present in the skin, or present on a keratin material of said being human, said sample being taken from an untreated area before the treatment, - a sample of biological or genetic material from the microbiota present in the skin, or present on a keratin material of said human being, said sample being taken from an area treated with the ingredient or composition, preferably applicable topically, after the treatment. According to another particular embodiment, the method according to the invention comprises at least four samples of biological or genetic material of the microbiota present in the skin, or present on a keratin material, from one or more human beings: - A sample carried out in an area to be treated with the ingredient or composition, preferably applied topically, before the treatment; - A sample taken from an untreated area before treatment; - A sample taken from an area treated with the ingredient or composition, preferably applicable topically, after the treatment; - A sample taken in an untreated area after the treatment period. Advantageously, the “untreated zone” according to the invention is a skin zone equivalent or close, functionally and/or in microbial composition, to said “zone to be treated”. Thus, according to another particular embodiment, the invention relates to a method for selecting or screening or evaluating the activity of at least one ingredient or composition, preferably applicable topically, in particular a cosmetic ingredient and /or dermatological, optionally formulated in the form of a topical composition, characterized in that the method comprises the following steps: - a sample of biological or genetic material of the microbiota present in the skin, of one or more human beings, said sample being performed in an area to be treated with the topically applicable ingredient or composition prior to treatment; - a sample of biological or genetic material of the microbiota present in the skin, or present on a keratinous material, of one or more human beings, said sample being taken in an untreated area before the treatment; - a sample of biological or genetic material of the microbiota present on the skin, or present on a keratin material, of one or more human beings, said sample being taken from an area treated with the ingredient or composition applicable topically after the treatment ; - a sample of biological or genetic material of the microbiota present in the skin, or present on a keratinous material, of one or more human beings, said sample being taken in an untreated area after the treatment; - an analysis of the biomass, for example by quantitative amplification of the standardized quantities of genetic material carried out using the universal primers 16S; - an analysis of the biodiversity of the sampled microbiota, for example by amplification of the standardized quantities of genetic material sampled, then sequencing of the amplicons thus produced using the specific 16S primers, then calculation of a diversity index allowing this analysis. The duration of the treatment with the ingredient or the composition is variable. The application of the product can be done daily or several times a day or over the test period. According to one embodiment, several samples can be taken from the treated zone, at, for example, 5 minutes, 1 hour, 24 hours, 2 days, 4 days, 7 days and 31 days from the start of the treatment. According to a particular embodiment, the sample is taken from the treated zone after 2 to 7 days of daily treatment. According to one embodiment, the application of the product is done due to a volume of between 0.1 and 100 μL/cm2, advantageously between 1 and 10 μl/cm2, standardizing the application as much as possible, for example by using a robotic arm. In a particular embodiment, at least one sample is taken from several human beings. This offers the advantage of supporting the statistical significance of the data obtained. According to a particular embodiment, the particular group of subjects at the origin of the samples can share characteristics (ethnicity, age, phototype, chronic dermatological conditions, type of skin, diet, lifestyle, rate of hydration of the skin, geolocation and other similar parameters) which allow the development of treatment methods, preferably cosmetic and/or dermatological, specifically targeting said groups of particular subjects, also called "populations". According to an advantageous and preferred embodiment, the population at the origin of the samples is characterized by an impoverished skin microbiota at the time of the samples. By “depleted skin microbiota” within the meaning of the invention is meant any decline in the biomass of the skin microbiota. In particular, the depletion can be linked to a skin aggression according to the invention, more particularly to the action of bactericides or virucides or fungicides or soaps or aggressive hygiene or care products vis-à-vis of the microbiota. In a particular embodiment, the depletion of the cutaneous microbiota is caused by a topical treatment with an antiseptic agent (that is to say fungicide, bactericide and/or virucide), advantageously a composition comprising a C1-C8 alcohol chosen from ethanol, isopropanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, benzyl alcohol and mixtures thereof; and/or an antiseptic agent other than an alcohol, such as for example benzalkonium chloride, chlorhexidine gluconate, sodium hypochlorite, a soap produced by saponification of a fatty acid; and/or also a composition comprising an acid such as lactic, malic, tartaric or glycolic acid; in an alternative embodiment, the depletion of the skin microbiota is caused by treatment with a virucidal and/or antifungal and/or bactericidal and/or antibiotic agent, administered topically or systemically. According to a particular embodiment, the sampling of biological or genetic material of the microbiota present in the skin, or present on a keratinous material, is carried out on a single human being. This method makes it possible to develop care methods, advantageously cosmetic and/or dermatological, preferably ecobiological, personalized, based on the conditions, specificities and responsiveness to care, of the skin microbiota of each individual. Thus, according to another particular embodiment, the invention also relates to a personalized method for selecting or screening or evaluating, preferably in vitro, the activity of at least one ingredient or composition, preferably applicable topically, in particular a cosmetic and/or dermatological ingredient, optionally formulated in the form of a composition, advantageously topical, characterized in that the method comprises the following steps: - the selection of a skin area or skin appendages to be studied, - at least one sample of biological or genetic material of the microbiota present in this area, of a single human being, - the application of at least one ingredient or a composition to this area, in a single or repeated manner, - after a period of 1 hour at 7 days, at least one sample of biological or genetic material from the microbiota present in this area, from a single human being, - at least one method for quantifying the biomass present in the samples, - at least one method making it possible to measure the microbial diversity present in the samples, - the evaluation of the activity of the ingredient or composition applied topically on the biomass and the biodiversity of the microbiota sampled, - the selection of the ingredient or ecobiological composition applicable topically allowing the increase of the biomass, in particular without reducing the diversity of the cutaneous microbiota. According to a particular embodiment, the personalized method according to the invention can be accompanied by an analysis of the dermatological profile of the single human being at the origin of the at least one sample on the basis of his ethnicity, age, phototype, chronic dermatological conditions, skin type, diet, lifestyle, skin hydration level, geolocation and other similar parameters. This information can be collected by direct analysis or through a form. For example, the methods for weighting cutaneous, capillary, or nail characteristics and the computer program described in document FR 2983328 can be combined with the results of the personalized screening method mentioned above to select an ecobiological dermatological treatment capable of inducing the increase in the biomass of the cutaneous microbiota of an individual, in particular without reducing its diversity, and in accordance with the overall dermatological needs of said individual. The invention also relates to a method for preparing a composition, preferably topical, characterized in that it comprises (i) the implementation of a screening or selection method defined previously and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing an increase in the biomass of the cutaneous microbiota in particular without reducing its biodiversity (ii) the formulation of the ingredients or compositions thus selected in a formula with cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity (iii) the packaging of the said formula. The invention also relates to a topical care method, preferably cosmetic or dermatological, advantageously ecobiological, characterized in that it comprises (i) the implementation of a screening method defined previously or in the following description and the selection of at least one ingredient or a composition, in particular an ingredient or a cosmetic and/or dermatological composition, capable of inducing the increase in the biomass of the cutaneous microbiota described above, in particular without reducing its biodiversity (ii) the formulation of the ingredients or compositions thus selected in a composition for cosmetic and/or dermatological use, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity (iii) the packaging of said formula (iv) the topical application of said composition to an area of skin or a surface of the selected appendages. As mentioned before for the screening or selection method, the advantageously ecobiological topical care method can target specific populations, or be personalized and adapted to the microbiota specific to each individual. According to a particular embodiment, the population to be treated is characterized by an impoverished skin microbiota. According to an alternative embodiment, it is a personalized method capable of treating an individual characterized by an impoverished cutaneous microbiota. Thus, according to another aspect of the invention, the invention relates to an advantageously cosmetic or dermatological topical care method, preferably ecobiological, capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without impact or reduction on its biodiversity, characterized in that a composition selected through the selection or screening method mentioned above or described below is applied to the skin. Within the meaning of the invention, by increase in the biomass is meant a statistically significant increase. By way of example, it may be an increase of at least 50% in the biomass compared to a reference value, that is to say before application of the ingredient or of the composition of the invention. In an alternative embodiment, the invention relates to a care method, preferably cosmetic or dermatological, for combating a skin attack (for example against a bactericidal and/or virucidal and/or fungicidal action), and/or maintaining and /or restore a healthy microbiota, characterized in that the method comprises the topical application of at least one topical composition selected in the screening or selection method described previously or in the following description, said composition being capable of inducing the increase of the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, on the skin or on a keratin material. According to yet another particular embodiment, the composition selected through the screening or selection method mentioned above or in the following description and capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity, is associated with the action of maintaining and/or restoring a healthy skin microbiota. According to another particular embodiment, the composition to be used in particular in the implementation of the preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its diversity, according to the The invention is an aqueous composition, comprising in particular at least 60%, better still at least 70%, or at least 80%, or at least 90%, or even at least 95% of water. Thus, the invention relates to a preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota in particular without reducing its diversity, according to the invention, with an aqueous composition, advantageously comprising at least 60% of water, preferably 70%, 80%, 90%, or even at least 95% water. A particular embodiment, the aqueous composition to be used in particular in the care method mentioned above or in the following description, comprises water which is characterized by a resistivity of between 12.5 and 12,500 Ohms.cm, in particular between 80 and 8000 Ohms.cm, at room temperature Preferably, the composition to be used in particular in the care method mentioned above or in the following description has an osmotic pressure of between 70 and 2500 mOsmol/l, and in particular between 100 and 800 mOsmol/l. According to a particular embodiment of the invention, the composition to be used in particular in the implementation of the preferably ecobiological topical care method capable of inducing an increase in the biomass of the cutaneous microbiota, in particular without reducing its biodiversity , comprises at least one antioxidant and at least one inorganic or organic salt, each or at least one of these components or ingredients having, advantageously, been selected beforehand by the screening method described above or in the following description. By “antioxidant”, is meant a compound which exhibits an antiradical activity, in particular on reactive oxygen (ROS), carbonyl (RCS) or nitrogen (RNS) species. According to a particular embodiment, the at least one antioxidant is chosen from the group comprising octadecyl di-t-butyl-4-hydroxyhydrocinnamate; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t- butylnitrone; ergothioneine; melatonin; acetyl cysteine; cysteine; lysine hydrochloride; carnosic acid; tyrosyl histidine HCl; histidine hydrochloride; pyridoxine serinate; superoxide dismutase; aminopropyl ascorbyl phosphate; ascorbic acid; ascorbyl dipalmitate; ascorbyl glucoside; ascorbyl linoleate; ascorbyl methylsilanol pectinate; ascorbyl palmitate; ascorbyl tetraisopalmitate; ascorbyl tocopheryl maleate; trisodium ascorbyl palmitate phosphate; disodium ascorbyl sulphate; calcium ascorbate; methylsilanol ascorbate; sodium ascorbate; sodium ascorbyl phosphate; sodium ascorbyl/cholesteryl phosphate; tetrahexyldecyl ascorbate; magnesium ascorbyl phosphate; tocopherol; tocopheryl acetate; tocopheryl linoleate; tocopheryl oleate; tocopheryl nicotinate; tocopheryl retinoate; sodium tocopheryl phosphate; dioleyltocopheryl methylsilanol; potassium ascorbyl tocopheryl 5 phosphate; dodecyl gallate; propyl gallate; octyl gallate; epigallocatechin gallate (EGCG); propyl gallate; ethylferulate; ethylhexyl ferulate; chitosan ascorbate; chitosan glycolate; apigenin; tiliroside; alpha-arbutin; arbutin; baicalin; quercetine; quercetin caprylate; isoquercetin; diethylhexyl syringylidenemalonate; dihydroxymethylchromone; dimethoxy di-p-cresol; dimethylmethoxy chromanol; ethylbisiminomethylguaiacol manganese chloride; hesperidin methyl chalcone; kojic acid; kojic dipalmitate; madecassoside; asiaticoside; magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl); nordihydroguaiaretic acid; phenylethyl resorcinol; resveratrol; troxerutin; glucosylrutin, rutin (4H-1-benzopyran-4-one); disodium rutinyl disulfate; tetrahydrobisdemethoxydiferuloylmethane; tetrahydrodemethoxydiferuloylmethane; tetrahydrodiferuloylmethane; tococysteamide; totarol; hydroxydecyl ubiquinone; ubiquinone; carotenoids; niacinamide or one of its derivatives; creatine; creatinine, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose. More particularly, the at least one antioxidant is chosen from the group comprising tocopherol, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose. According to an even more particular embodiment, the at least one antioxidant according to the invention is carnosine. According to a particular embodiment, the invention relates to a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine. According to another particular embodiment, the invention relates to a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, chloride of sodium, sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine for its use in maintaining and/or restoring a healthy skin microbiota. According to another particular embodiment, the invention relates to a composition comprising: - sodium citrate, - carnosine. According to another particular embodiment, the invention relates to a composition comprising: - sodium citrate, - carnosine for its use in maintaining and/or restoring a healthy skin microbiota. According to one particular embodiment, the composition according to the invention, or for its use according to the invention, also comprises at least one carbon source. "Carbon source" means a compound likely to be used by bacteria as a substrate for growth and multiplication, such as: - partially or totally hydrolyzed proteins from cereals (wheat, corn, barley, rye, oats), legumes (soya, broad bean, broad bean, pea), milk (caseins, albumins), yeast, as well as peptides and amino acids and their derivatives alone or in mixtures; - oligosaccharides or polysaccharides, advantageously short-chain, such as fructooligosaccharides, hyaluronic acid, pullulan, alginate, cellulose, advantageously microcrystalline cellulose, carrageenan, chitosan, dextran and dextran sulphate, galactoarabinan, amylose, amylopectin, maltodextrins, pectins, and their respective hydrolysates, as well as sugars, oses, osides and their derivatives such as glucose, galactose, fructose, xylose, mannose, glucosamine, lactose, ribose , rhamnose and their phosphate and acetylated derivatives in particular. Fructo oligosaccharides or FOS can be derived from plant materials such as beets, artichokes or corn. The raw material corresponding to the INCI designation fructooligosaccharides can be used in the context of the present invention, for example the product marketed by the company Shaanxi Bolin Biotechnology Co under the name of FRUCTOOLIGOSACCHARIDES. Advantageously, the FOS represent from 0.001% to 20% by total weight of the composition, advantageously from 0.01% to 10%, even more advantageously from 0.1% to 5%. According to a particular embodiment, the invention relates to a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously cellulose microcrystalline. According to a particular embodiment, the invention relates to a composition comprising: - at least one inorganic or organic salt chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc , cobalt and mixtures thereof, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate, sodium chloride , sodium hydrogen carbonate, sodium hydrogen phosphate, sodium citrate, magnesium gluconate, - carnosine; - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose for its use in maintaining and/or restoring a healthy skin microbiota. According to another particular embodiment, the invention relates to a composition comprising: - sodium citrate, - carnosine. - at least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose According to another particular embodiment, the invention relates to a composition comprising: - sodium citrate, - carnosine - au least one carbon source chosen from hyaluronic acid, and cellulose, advantageously microcrystalline cellulose for its use in maintaining and/or restoring a healthy skin microbiota. According to a particular embodiment, a pure or highly purified form of carnosine is used in the composition according to the invention. According to a variant, the carnosine is obtained by chemical synthesis. By way of example, mention may be made of the cosmetic material L-Carnosine corresponding to the INCI name carnosine and marketed by the company MERCK KgaA. According to another embodiment of the invention, the carnosine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%. In practice, in the composition according to the invention, a pure or highly purified form of hypotaurine can be used. According to a variant, the hypotaurine is obtained by chemical synthesis. By way of example, mention may be made of the cosmetic raw material hypotaurine corresponding to the INCI name aminoethanesulfinic acid and marketed by the company DKSH. According to a particular embodiment of the invention, the hypotaurine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%. According to another particular embodiment, the tocopherol used is a mixture of purified natural tocopherols, in particular α-tocopherol, β-tocopherol, γ-tocopherol, and δ-tocopherol; this mixture can be used in particular in an oil chosen from vegetable, mineral, silicone, and preferably vegetable oils. As a mixture of natural tocopherols that can be used according to the invention, mention may be made of the mixture sold by the company Nisshin OilliO Group, Ltd under the name Tocopherol 80 and corresponding to the INCI name “Tocopherol”. According to another particular embodiment of the invention, the tocopherol represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%. Betaine is an osmolyte and an antioxidant found in particular in the plant kingdom. Betaine can be of vegetable origin or obtained by biotechnology. Advantageously a highly purified form of betaine, with a purity of at least 95%, is used in the compositions according to the invention. As a source of betaine that can be used according to the invention, mention may be made of the raw material sold by the company Evonik Nutrition & Care GmbH under the name of Tego Natural Betaine (Evonik Nutrition & Care GmbH) and corresponding to the INCI name “Betaine”. According to another particular embodiment of the invention, the betaine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 2%. The xylitol can be powdered xylitol, for example of vegetable origin, in particular obtained from wood by hydrogenation of xyloses, having in particular a purity greater than 95%. By way of example, the company Orient Star LCC markets the product OriStar XLT corresponding to the INCI name “Xylitol” which can be implemented within the framework of the invention. Suitably, the xylitol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%. The mannitol can for example be powdered mannitol, of plant origin (corn, potato, wheat), preferably having a purity greater than 98%. By way of example, the company American Biochemicals, Inc. markets the product D-mannitol corresponding to the INCI name mannitol which can be used in the context of the invention. Suitably, the mannitol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%. Inositol is a cyclic compound derived from cyclohexane and carrying six secondary alcohol functions. In accordance with a particular embodiment of the invention, inositol of plant origin is used, synthesized naturally from glucose-6-phosphate (G6P), resulting from photosynthesis or from the hydrolysis of the carbon reserves of the plant. Preferably, inositol extracted from rice by techniques known to those skilled in the art is used. It is also possible to use inositol prepared by chemical or enzymatic synthesis. By way of example, the company Dupont Industrial Biosciences markets the product Genencare OSMS MI corresponding to the INCI name inositol which can be used in the context of the invention. Suitably, the inositol represents from 0.001% to 5% by total weight of the composition, advantageously from 0.1% to 2%. All cosmetically or pharmaceutically acceptable inorganic or organic salts are suitable for use in the composition according to the invention or for its use according to the invention. Advantageously, it is at least one inorganic or organic salt chosen from the group consisting of a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulfate, potassium or sodium phosphate, copper or zinc gluconate, and mixtures thereof, more particularly sodium citrate. According to a particular embodiment of the invention, the at least one inorganic or organic salt is chosen from the group comprising calcium chloride; potassium chloride; potassium phosphate; magnesium sulfate; sodium chloride; sodium hydrogen carbonate; sodium hydrogen phosphate; sodium citrate; magnesium gluconate. These mineral salts are available from several suppliers of cosmetic raw materials such as, for example, COOPER INDUSTRIE or BRENNTAG FRANCE. In a particular embodiment, the composition according to the invention comprises at least one of the following mineral salts, in percentage by weight: - From 0.000001% to 0.1% by total weight of the composition of calcium chloride , in particular from 0.0001% to 0.1%; - From 0.0001% to 0.1% by total weight of the potassium chloride composition, in particular from 0.001% to 0.1%; - From 0.00001% to 0.1% by total weight of the phosphate composition of potassium, in particular from 0.0001% to 0.1%; - From 0.0001% to 0.1% by total weight of the magnesium sulphate composition, in particular from 0.001% to 0.1%; - From 0.001 to 4% by total weight of the composition of sodium chloride, in particular from 0.1 to 4%; - From 0.0001 to 0.1% by total weight of the sodium hydrogencarbonate composition; - From 0.0001% to 0.1% by total weight of the sodium hydrogen phosphate composition; - From 0.0001% to 1% by total weight of the citric acid composition, in particular from 0.001% to 1%; - From 0.001% to 4% by total weight of the sodium citrate composition; - From 0.001% to 4% by total weight of the magnesium gluconate composition. According to another particular embodiment of the invention, the total weight of the mineral and/or organic salts is less than or equal to 1% by weight, in particular is less than or equal to 0.1% by weight of the total weight of the composition. In particular, the inorganic and organic salts are used at a concentration which makes it possible to be within the range of osmotic pressure sought. According to a particular embodiment, the sodium citrate represents from 0.001% to 4% by total weight of the composition and the carnosine represents from 0.0001% to 5% by total weight of the composition, advantageously from 0.001% to 2%. Several cosmetic sources of hyaluronic acid are known. According to a particular embodiment of the invention, the hyaluronic acid has a molecular weight comprised between comprised between 20 and 50 kDa. By way of example, the raw materials PRIMALHYAL™ 50 or HyaCare 50 marketed respectively by the companies SOLIANCE and EVONIK and corresponding to hyaluronic acid with a molecular weight (Mw) of between 20 and 50 kDa (INCI: hyaluronic acid) can be used in the context of the present invention. According to a particular embodiment, the hyaluronic acid represents from 0.001 to 4% by total weight of the composition, advantageously from 0.001% to 2%. Any commercial source of microcrystalline cellulose suitable for topical and cosmetic application can be used in the compositions according to the invention. By way of example, mention may be made of Avicel PH-101 NF marketed by the company FMC Corporation and corresponding to the INCI designation microcrystalline cellulose. According to a particular embodiment, the microcrystalline cellulose represents from 0.001 to 5% by total weight of the composition, advantageously from 0.001% to 3%. According to another particular embodiment, the composition according to the invention or for its use according to the invention or to be used in particular in the care method mentioned above or in the following description is intended for populations characterized by an impoverished cutaneous microbiota. The way in which the invention can be implemented and the resulting advantages will emerge better from the examples of embodiment which follow, given by way of indication and not limitation, in support of the appended figures. In the examples the percentages are given by weight, the pressure is atmospheric pressure and the temperature is room temperature usually between about 18° C. and about 30° C., unless otherwise indicated. [Fig.1] and [Fig.2] show the effect of the pure ingredients petrolatum (Fig 1) and silicone (Fig 2) after 4 days of contact versus untreated area (ZNT), on the bacterial biomass of the skin microbiota, measured by the number of 16S gDNA copies in the volunteers taken into account in the analysis. Statistical significance is determined by a Wilcoxon test. (ns, not significant: p>0.05; significant*: p≤0.05; significant**: p≤0.01; significant***: p≤0.001). [Fig.3] and [Fig.4] show the effect of formula 1 (fle 1; Fig 3) and formula 2 (fle 2; Fig 4) after 4 days of contact versus untreated area (ZNT) , on the bacterial biomass of the skin microbiota, measured by the number of 16S gDNA copies in the volunteers taken into account in the analysis. Statistical significance is determined by a Wilcoxon test. (ns, not significant: p>0.05; significant*: p≤0.05; significant**: p≤0.01; significant***: p≤0.001). [Fig.5] and [Fig.6] show the effect of the pure ingredients petrolatum [Fig.5] and silicone [Fig.6] after 4 days of contact versus untreated area (ZNT), on the bacterial diversity of the microbiota cutaneous, measured by the Shannon index, in the volunteers taken into account for the analysis. Statistical significance is determined by a paired Wilcoxon test. (ns, not significant: p>0.05; significant*: p≤0.05; significant**: p≤0.01; significant***: p≤0.001). [Fig.7] and [Fig.8] show the effect of formula 1 (fle 1; Fig.7) and formula 2 (fle 2; Fig.8) after 4 days of contact versus untreated area ( ZNT), on the bacterial diversity of the skin microbiota, measured by the Shannon index, in the volunteers taken into account for the analysis. Statistical significance is determined by a paired Wilcoxon test. (ns, not significant: p>0.05; significant*: p≤0.05; significant**: p≤0.01; significant***: p≤0.001). Examples 1 of the Evaluation of the Effect of Two Cosmetic Compositions and Two Pure Ingredients on the Skin Microbiota of Healthy Volunteers
L'objectif de l'étude est d'évaluer l'effet à 4 jours de 2 ingrédients cosmétiquesThe objective of the study is to evaluate the effect at 4 days of 2 cosmetic ingredients
(vaseline et silicone) et de 2 produits finis (formule 1 et formule 2) sur le microbiote cutané en examinant leur impact sur la biomasse prélevée ainsi que sur la diversité du microbiote cutané. (vaseline and silicone) and 2 finished products (formula 1 and formula 2) on the skin microbiota by examining their impact on the biomass sampled as well as on the diversity of the skin microbiota.
Les produits suivants sont testés simultanément dans l'étude clinique : The following products are tested simultaneously in the clinical study:
[Tableau 1] - produits testés [Table 1] - products tested
[Tableau 2] - Formule 1 (fle 1) - émulsion H/E [Table 2] - Formula 1 (fle 1) - O/W emulsion
[Tableau 3] - Formule 2 (fle 2) - lotion aqueuse Une application standardisée (Ί ,5 μL/cm2) de chacun des produits à tester est réalisée sur le dos de 19 volontaires pendant 4 jours à raison de 1 application par jour. Le jour du prélèvement, les volontaires ont pour consigne de ne pas se doucher au minimum 12 heures avant les prélèvements. Le prélèvement est réalisé à l'aide d'un écouvillon (Sigma Trans Swab liquide Amies réf MW176S d'Elitech) en réalisant 10 allers-retours en Z sur une surface de 15 cm2 et en exerçant une légère pression sur l' écouvillon. L' écouvillon est ensuite maintenu dans la glace et analysé. Afin de mesurer l'effet de l'ingrédient ou de la composition (produit fini) sur la biomasse et sur la diversité du microbiote cutané, l'approche méthodologique repose sur un procédé constitué des étapes suivantes: a) Extraction de l'ADNg (ADN génomique) à partir des écouvillons b) Analyse de la quantité de biomasse par une approche qPCR c) L'analyse de la diversité microbienne qui s'appuie sur : [Table 3] - Formula 2 (fle 2) - aqueous lotion A standardized application (Ί.5 μL/cm 2 ) of each of the products to be tested is carried out on the backs of 19 volunteers for 4 days at the rate of 1 application per day. On the day of the sample, the volunteers are instructed not to shower at least 12 hours before the samples. The sample is taken using a swab (Sigma Trans Swab liquid Amies ref MW176S from Elitech) by carrying out 10 round trips in Z over a surface of 15 cm 2 and by exerting slight pressure on the swab. The swab is then kept on ice and analyzed. In order to measure the effect of the ingredient or the composition (finished product) on the biomass and on the diversity of the skin microbiota, the methodological approach is based on a process consisting of the following steps: a) Extraction of the gDNA ( genomic DNA) from swabs b) Analysis of the quantity of biomass using a qPCR approach c) Analysis of microbial diversity based on:
- la préparation des librairies d'amplicons selon la solution Metabiote®, limitant les biais d'amplifications entre échantillons et incluant un contrôle positif (communauté bactérienne artificielle « ABC control », ainsi qu'un contrôle négatif (bruit de fond de PCR du processus total de préparation des librairies).- the preparation of amplicon libraries according to the Metabiote® solution, limiting amplification biases between samples and including a positive control (artificial bacterial community "ABC control", as well as a negative control (PCR background noise of the process library prep total).
- le séquençage des librairies d'amplicons sur un « run » unique (ou cycle unique) Illumina MiSeq « paired-end » en chimie 2x300 bases et le tri des séquences par échantillon et le réassemblage des lectures « paired-end » permettant d'obtenir des lectures ADNr 16S (ADN ribosomique 16S) pleine longueur. - the sequencing of amplicon libraries on a single "run" (or single cycle) Illumina MiSeq "paired-end" in 2x300 base chemistry and the sorting of sequences by sample and the reassembly of "paired-end" reads allowing obtain full-length 16S rDNA (16S ribosomal DNA) reads.
En détail, l'étape a) d'extraction de l'ADN génomique à partir des écouvillons consiste en une étape de récupération optimisée des bactéries (consistant en un essorage de l'écouvillon et une récupération des culots bactériens par centrifugation) qui est réalisée avant l'étape d'extraction. L'extraction d'ADN génomique a ensuite été réalisée avec le kit Nucleospin Microbial DNA kit (Macherey Nagel, France). Les ADNg extraits ne sont pas « dosables », une quantification de la biomasse et un contrôle de qualité a donc été réalisé par qPCR 16S. En détail, l'étape b) d'analyse de la biomasse et de contrôle qualité par qPCR16S consiste en la réalisation d'une gamme standard dans les conditions expérimentales standardisées. Sur l'ensemble des points de dilution de la gamme, l'utilisation de 4 points permet d'obtenir une efficacité proche de 94%, sachant qu'il est communément considéré que l'efficacité de qPCR est optimale entre 90 et 110 %. Il est considéré que cette zone de nombre de copies 16S de la communauté artificielle présentait les caractéristiques nécessaires pour être utilisée en gamme standard de quantification. Les échantillons d'ADN extraits n'étant pas quantifiables car trop faiblement concentrés, les qPCR sont réalisées, en triplicat, à partir d'un volume fixe de 4,38 μL correspondant au volume maximum qu'il est possible d'intégrer dans le volume réactionnel de la qPCR.In detail, step a) of extracting the genomic DNA from the swabs consists of an optimized bacteria recovery step (consisting of wiping the swab and recovering the bacterial pellets by centrifugation) which is carried out before the extraction step. The genomic DNA extraction was then carried out with the Nucleospin Microbial DNA kit (Macherey Nagel, France). The extracted gDNAs are not “assayable”, a quantification of the biomass and a quality control was therefore carried out by 16S qPCR. In detail, step b) of analysis of the biomass and quality control by qPCR16S consists of carrying out a standard range under standardized experimental conditions. On all the dilution points of the range, the use of 4 points makes it possible to obtain an efficiency close to 94%, knowing that it is commonly considered that the efficiency of qPCR is optimal between 90 and 110%. It is considered that this 16S copy number area of the artificial community presented the necessary characteristics to be used in standard range of quantification. Since the extracted DNA samples cannot be quantified because they are too weakly concentrated, the qPCRs are carried out, in triplicate, from a fixed volume of 4.38 μL corresponding to the maximum volume that it is possible to integrate into the qPCR reaction volume.
Le témoin négatif nommé « Tneg » est le témoin négatif de qPCR réalisé avec de l'eau stérile. The negative control called “Tneg” is the negative control for qPCR carried out with sterile water.
À partir des courbes standard et de leur équation sur la zone d'efficacité, il est possible de calculer le nombre de copies ADNr 16S bactérien contenu au sein de chaque échantillon. From the standard curves and their equation on the efficiency zone, it is possible to calculate the number of bacterial 16S rDNA copies contained within each sample.
Les amplifications quantitatives (qPCR) sont effectuées avec des amorces universelles 16S [SEQ ID. 1 8F (5 AGAGTTT G ATCCT GGCT CAG-3 ') and SEQ ID. 2 139 IR (5 '-GACGGGCGGTGWGTRCA-3 ')] (Kong et al. 2012b). Les conditions de l'amplification et équipement était conforme à ce qui est décrit dans SanMiguel et al. 2018. The quantitative amplifications (qPCR) are carried out with 16S universal primers [SEQ ID. 18F (5 AGAGTTT G ATCCT GGCT CAG-3') and SEQ ID. 2,139 IR (5'-GACGGGCGGTGWGTRCA-3')] (Kong et al. 2012b). The amplification conditions and equipment were as described in SanMiguel et al. 2018.
En détail, et concernant l'étape c) d'analyse de la diversité du microbiome cutané, il est à noter que pour ce type d'échantillons à biomasse rare, une quantité maximale de matériel est utilisée pour la création des librairies spécifiques Illumina MiSeq. L'étape d'amplification est effectué avec les amorces pour les régions V1-V3 correspondant aux SEQ. ID 5 27F (5'- AGAGTTTGATCCT GGCT CAG-3' ) et SEQ. ID 6 534R (5- ATTACCGCGGCTGCTGG-3) conformément à San Miguel et al. 2018, avec les modifications qui suivent. Deux contrôles supplémentaires sont intégrés au cours de la préparation des librairies Metabiote®. Un contrôle interne positif, nommé « ABCv2 » pour « Artificial Bacterial Community ». Il s'agit d'une communauté artificielle réalisée à partir d'un mélange d'ADNg de différentes souches bactériennes. Le second contrôle est un témoin négatif appelé « BF », celui-ci est également intégré au processus de préparation et permet d'évaluer le bruit de fond inhérent aux réactifs utilisés après l'étape d'extraction. Le contrôle quantitatif des librairies finales est réalisé par une méthode fluorométrique. Ensuite, la communauté contrôle ABC dont la composition est connue (17 bactéries dont 2 archaebactéries) est introduite dans le séquençage ; elle sert de contrôle positif au cours de notre procédé MetaBiote®, mais également de contrôle positif lors de la réalisation de l'affiliation taxonomique des séquences. Le témoin négatif de création des librairies (BF.V1V3) sert à identifier ce qui peut être obtenu comme profil si l'on n'introduit pas de biomasse initiale et que l'on pousse le procédé de préparation MetaBiote®. Ce témoin permet de distinguer les échantillons à exclure des comparaisons. In detail, and concerning step c) of analysis of the diversity of the cutaneous microbiome, it should be noted that for this type of rare biomass samples, a maximum quantity of material is used for the creation of specific Illumina MiSeq libraries . The amplification step is carried out with the primers for the V1-V3 regions corresponding to SEQ. ID 5 27F (5'-AGAGTTTTGATCCT GGCT CAG-3') and SEQ. ID 6534R (5-ATTACCGCGGCTGCTGTGG-3) according to San Miguel et al. 2018, with the following modifications. Two additional controls are integrated during the preparation of the Metabiote® libraries. A positive internal control, named “ABCv2” for “Artificial Bacterial Community”. It is an artificial community made from a mixture of gDNA from different bacterial strains. The second control is a negative control called "BF", this one is also integrated into the preparation process and makes it possible to evaluate the background noise inherent to the reagents used after the extraction step. The quantitative control of the final libraries is carried out by a fluorometric method. Then, the ABC control community whose composition is known (17 bacteria including 2 archaebacteria) is introduced into the sequencing; it serves as a positive control during our MetaBiote® process, but also as a positive control when performing the taxonomic affiliation of the sequences. The negative library creation control (BF.V1V3) is used to identify what profile can be obtained if no initial biomass is introduced and the MetaBiote® preparation process is pushed. This control makes it possible to distinguish the samples to be excluded from the comparisons.
Les indices de diversité alpha des échantillons ainsi que les mesures de raréfaction sont définis, permettant ainsi d'observer la diversité taxonomique (en nombre d'OTUs) des échantillons. The alpha diversity indices of the samples as well as the measures of rarefaction are defined, thus making it possible to observe the taxonomic diversity (in number of OTUs) of the samples.
L'analyse bio-informatique effectuée a permis d'éliminer les échantillons qui ont présenté un séquençage de mauvaise qualité. Résultats : The bioinformatics analysis carried out made it possible to eliminate the samples which presented poor quality sequencing. Results :
Les résultats sont présentés dans les figures 1 à 4. L'analyse sur la biomasse, réalisée par le biais des amplifications quantitatives avec les amorces universelles 16S, montre que la vaseline et la silicone induisent une légère diminution de la biomasse après 4 jours d'application journalière (respectivement, une diminution moyenne de 4,5% et 2% ; figures 1 et 2). The results are presented in figures 1 to 4. The analysis on the biomass, carried out by means of quantitative amplifications with the universal primers 16S, shows that petroleum jelly and silicone induce a slight reduction in the biomass after 4 days of daily application (respectively, an average reduction of 4.5% and 2%; figures 1 and 2).
Ces deux ingrédients ne sont donc pas adaptés, du moins sous forme pure, à la mise en œuvre d'une composition pour son utilisation selon l'invention. Ils sont donc écartés. En revanche, la formule 1 (fle 1) induit une augmentation moyenne de la biomasse de 134% au bout de 4 jours d'application (figure 3) chez les volontaires pris en compte pour l'analyse, tandis que formule 2 (fle 2) induit une augmentation moyenne de 150% (figure 4). La différence entre la zone non-traitée (ZNT) et la zone traitée (ZT) est significative dans les deux cas (test deThese two ingredients are therefore not suitable, at least in pure form, for the implementation of a composition for its use according to the invention. They are therefore discarded. On the other hand, formula 1 (fle 1) induces an average increase in biomass of 134% after 4 days of application (figure 3) in the volunteers taken into account for the analysis, while formula 2 (fle 2 ) induces an average increase of 150% (Figure 4). The difference between the untreated zone (ZNT) and the treated zone (ZT) is significant in both cases (test of
Wilcoxon). Les formules, sélectionnées grâce au procédé de criblage décrit dans la présente demande, sont donc adaptées pour la mise en œuvre d'une composition pour son utilisation selon l'invention, à savoir pour induire une augmentation de la biomasse du microbiote cutanée. Wilcoxon). The formulas, selected using the screening method described in the present application, are therefore suitable for the implementation of a composition for its use according to the invention, namely to induce an increase in the biomass of the cutaneous microbiota.
L'amplification avec les amorces spécifiques 16S puis le séquençage des amplicons obtenus montre que la vaseline et la silicone n'ont, en pratique, aucun effet sur la diversité du microbiote cutané (figures 5 et 6 ; p>0,05 dans les deux cas donc différence non significative ; voir les tableaux 4 et 5). Cette observation est conforme avec les résultats de l'analyse de la biomasse et indique que ces ingrédients sont substantiellement inertes vis-à-vis du microbiote cutané. Amplification with specific 16S primers followed by sequencing of the amplicons obtained shows that petroleum jelly and silicone have, in practice, no effect on the diversity of the skin microbiota (figures 5 and 6; p>0.05 in both case therefore difference not significant; see tables 4 and 5). This observation is consistent with the results of the biomass analysis and indicates that these ingredients are substantially inert with respect to the skin microbiota.
[Tableau 4] - Indice Shannon et OTUs et pourcentages relatifs au traitement avec la vaseline [Table 4] - Shannon index and OTUs and percentages relating to treatment with petroleum jelly
[Tableau 5] - Indice Shannon et OTUs et pourcentages relatifs au traitement avec la silicone [Table 5] - Shannon index and OTUs and percentages relating to treatment with silicone
L'étude des formules 1 et 2 (fle 1 et fle 2) montre que dans les deux cas les variations observées dans l'indice de Shannon en comparant la zone non-traité aux zones traitées avec les compositions à l'étude est non-significative (p = 0,054 et p= 0,76 ; voir également tableaux 6 et 7 pour la variation de l'indice de Shannon et le pourcentage OTUs). The study of formulas 1 and 2 (fle 1 and fle 2) shows that in both cases the variations observed in the Shannon index by comparing the untreated zone to the zones treated with the compositions under study is non- significant (p = 0.054 and p = 0.76; see also tables 6 and 7 for the variation of the Shannon index and the percentage OTUs).
[Tableau 6] - Indice Shannon et OTUs et pourcentages relatifs au traitement avec la fle 1 [Table 6] - Shannon index and OTUs and percentages relating to treatment with fle 1
[Tableau 7] - Indice Shannon et OTUs et pourcentages relatifs au traitement avec la fle 2 [Table 7] - Shannon index and OTUs and percentages relating to treatment with fle 2
Dans les deux cas, il peut être observé que : In both cases, it can be observed that:
- l'indice de Shannon ne varie pas significativement, - le nombre d'OTUs ne varie pas significativement. - the Shannon index does not vary significantly, - the number of OTUs does not vary significantly.
L'impact négligeable des formules fle 1 et fle 2 sur les équilibres du microbiote cutanée est confirmée également par l'analyse fine de modification de l'abondance relative de 15 genres bactériens plus représentés entre ZNT et ZT - fle 1 et - fle 2 (tableaux 8 et 9). The negligible impact of the fle 1 and fle 2 formulas on the balance of the cutaneous microbiota is also confirmed by the detailed analysis of the modification of the relative abundance of 15 more represented bacterial genera between ZNT and ZT - fle 1 and - fle 2 ( tables 8 and 9).
[Tableau 8] - Comparaison de l'abondance relative de 15 genres bactériens plus représentés entre ZNT et ZT avec la fle 1 [Table 8] - Comparison of the relative abundance of 15 more represented bacterial genera between ZNT and ZT with fle 1
[Tableau 9] - Comparaison de l'abondance relative de 15 genres bactériens plus représentés entre ZNT et ZT avec la fle 2 [Table 9] - Comparison of the relative abundance of 15 more represented bacterial genera between ZNT and ZT with fle 2
Il est possible de décrire les résultats des essais réalisés dans l'exemple 1, sous forme d'un tableau de synthèse (Voir le tableau 10) : [Tableau 10] - Présentation des résultats de façon synthétique It is possible to describe the results of the tests carried out in example 1, in the form of a summary table (See table 10): [Table 10] - Presentation of the results in a synthetic way
Légende : pas de variation significative (=), augmentation significative (+), diminution significative (-) Il peut donc être conclu que l'augmentation de la biomasse du microbiote cutanée observée en réponse à l'application des deux formules fle 1 et fle 2, qui contiennent un sel organique (citrate de sodium), un antioxydant (carnosine) et deux sources de carbone selon l'invention (acide hyaluronique et cellulose microcristalline) n'a pas été accompagnée par une diminution de diversité des genres bactériens, notamment les plus représentés. Les deux formules, sélectionnées grâce à la méthode de criblage décrite auparavant, sont adaptées pour l'utilisation, notamment dans le contexte d'un traitement visant à augmenter la biomasse globale du microbiote cutané, tout en respectant les proportions entre ses constituants. Legend: no significant change (=), significant increase (+), significant decrease (-) It can therefore be concluded that the increase in the biomass of the cutaneous microbiota observed in response to the application of the two formulas fle 1 and fle 2, which contain an organic salt (sodium citrate), an antioxidant (carnosine) and two sources of carbon according to the invention (hyaluronic acid and microcrystalline cellulose) was not accompanied by a reduction in the diversity of the bacterial genera, in particular the most represented. The two formulas, selected using the screening method described above, are suitable for use, particularly in the context of a treatment aimed at increasing the overall biomass of the cutaneous microbiota, while respecting the proportions between its constituents.
Exemple 2 - Etude d'ingrédients et de compositions Example 2 - Study of ingredients and compositions
Les techniques décrites dans l'exemple 1 sont reproduites à l'identique. Les ingrédients listés au Tableau 11 sont placés à un pourcentage indiqué en gramme pour 100 grammes (qsp 100 g d'eau), les compositions sont appliquées pures.The techniques described in Example 1 are reproduced identically. The ingredients listed in Table 11 are placed at a percentage indicated in grams per 100 grams (qsp 100 g of water), the compositions are applied neat.
Leur pH est ajusté entre 5 et 9, et leur pression osmotique est ajustée pour obtenir une valeur comprise entre 100 et 800 mOsm.l-1. Their pH is adjusted between 5 and 9, and their osmotic pressure is adjusted to obtain a value of between 100 and 800 mOsm.l -1 .
Les produits sont appliqués dans le dos de séries de 10 volontaires. Les applications sont quotidiennes pendant 4 jours et les prélèvements de matériel biologique cutané sont effectués avant la première application et après le 4ème jour. L'analyse de la biomasse est effectuée par qPCR (amorces universelles 16S) et l'analyse de la diversité microbienne par amplification avec des amorces spécifiques 16S, puis analyse des résultats par mesure de l'indice de Shannon. Les résultats synthétiques sont regroupés dans le tableau 11 suivant : The products are applied to the back of series of 10 volunteers. Applications are daily for 4 days and samples of biological material from the skin are taken before the first application and after the 4th day. The biomass analysis is carried out by qPCR (16S universal primers) and the analysis of the microbial diversity by amplification with specific 16S primers, then analysis of the results by measuring the Shannon index. The summary results are grouped in the following table 11:
On note « = » l'absence de variation significative, « + » une augmentation significative et « - » une réduction significative. We note “=” the absence of significant variation, “+” a significant increase and “-” a significant reduction.
[Tableau 11] - Synthèse [Table 11] - Summary
Légende : pas de variation significative (=), augmentation significative (+), diminution significative (-) Exemple 3 : Etude sur une zone cutanée caractérisée par une biomasse du microbiote cutané réduite après l'action d'une solution éthanoliqueLegend: no significant variation (=), significant increase (+), significant decrease (-) Example 3: Study on a cutaneous area characterized by a reduced cutaneous microbiota biomass after the action of an ethanolic solution
Composition testée [Tableau 12] Composition tested [Table 12]
L'étude est réalisée sur le front de 5 volontaires. Un prélèvement de la biomasse est réalisé à T0 sur le front des volontaires. Puis, une désinfection de la totalité du front avec une compresse imbibée de 5 mL d'alcool 70° (alcool modifié de Cooper, lot 013199) est réalisée en frottant pendant 30 sec et en laissant poser 3 min supplémentaires. Après 5 min de séchage, 2 prélèvements sont réalisés : T5min à gauche (G) et T5min à droite (D). Puis une compresse imbibée de 5 mL de la composition ci-dessus est déposée sur la moitié droite du front pendant 3 min. Après 2h, une brumisation de 2 sec de la composition ci-dessus est réalisée sur la moitié droite du front. Après 2h supplémentaires, 2 prélèvements sont réalisés : T4h côté alcool et T4h côté composition à tester. The study is carried out on the forehead of 5 volunteers. A biomass sample is taken at T0 on the forehead of the volunteers. Then, disinfection of the entire forehead with a compress soaked in 5 mL of 70° alcohol (modified alcohol from Cooper, batch 013199) is carried out by rubbing for 30 seconds and leaving for an additional 3 minutes. After 5 min of drying, 2 samples are taken: T5 min on the left (G) and T5 min on the right (D). Then a compress soaked in 5 mL of the above composition is placed on the right half of the forehead for 3 min. After 2 hours, a 2 sec mist of the above composition is applied to the right half of the forehead. After 2 additional hours, 2 samples are taken: T4h on the alcohol side and T4h on the composition to be tested side.
Les prélèvements sont réalisés à l'aide d'écouvillons (Sigma Trans Swab liquide Amies ref MW176S d'Elitech) aux temps T0, T5min, T2h et T4h, en réalisant 10 allers-retours en Z sur une surface de 3,75 cm2 et en exerçant une légère pression sur l'écouvillon de manière standardisée. L'écouvillon est ensuite maintenu dans la glace. Effet sur la biomasse : The samples are taken using swabs (Sigma Trans Swab liquid Amies ref MW176S from Elitech) at times T0, T5min, T2h and T4h, by performing 10 round trips in Z over an area of 3.75 cm 2 and exerting light pressure on the swab in a standardized manner. The swab is then kept on ice. Effect on biomass:
Cette suspension est ensuite diluée au 1/2, 1/10 et 1/50 dans du bouillon nutritif. Un ensemencement de 1 mL de chaque dilution est ensuite réalisé sur gélose TSA en boite de Pétri et incubé à 30-35°C pendant 48h. Après 48h d'incubation les colonies (UFC) sont comptées en totalité sur les boites. La biomasse bactérienne est obtenue en multipliant le nombre d'UFC comptées par son facteur de dilution et est exprimée en UFC/mL. This suspension is then diluted 1/2, 1/10 and 1/50 in nutrient broth. A seeding of 1 mL of each dilution is then carried out on TSA agar in a Petri dish and incubated at 30-35° C. for 48 hours. After 48 hours of incubation the colonies (CFU) are counted in full on the dishes. The bacterial biomass is obtained by multiplying the number of CFU counted by its dilution factor and is expressed in CFU/mL.
La désinfection avec l'alcool 70° a induit une forte diminution de la quantité de biomasse des bactéries cultivables de 99,8 %, 5 min après application de l'alcool. L'application du produit à tester après 4h de contact sur le microbiote appauvri augmente cette biomasse de 160% par rapport au témoin alcool 70° seul. Disinfection with 70° alcohol induced a sharp decrease in the amount of biomass of cultivable bacteria by 99.8%, 5 min after application of the alcohol. Application of the product to be tested after 4 hours of contact on the depleted microbiota increases this biomass by 160% compared to the 70° alcohol control alone.
Effet sur la diversité : Les tests réalisés selon le protocole décrit dans l'exemple 1 montrent que la diversité du microbiome cutané n'est pas réduite après application du produit évalué. On note « = » l'absence de variation significative, « + » une augmentation significative et « - » une réduction significative. Les résultats sont regroupés dans le tableau 13 suivant. Effect on diversity: The tests carried out according to the protocol described in example 1 show that the diversity of the cutaneous microbiome is not reduced after application of the product evaluated. We note “=” the absence of significant variation, “+” a significant increase and “-” a significant reduction. The results are collated in Table 13 below.
[Tableau 13] - Synthèse [Table 13] - Summary
Légende : pas de variation significative (=), augmentation significative (+), diminution significative (-) Legend: no significant change (=), significant increase (+), significant decrease (-)
Exemple 4 : Etude sur une zone cutanée caractérisée par une biomasse du microbiote cutané réduite après l'action d'un produit d'hygiène à base de savon de Marseille Example 4: Study on a skin area characterized by a reduced skin microbiota biomass after the action of a hygiene product based on Marseille soap
Composition testée : [Tableau 14] Composition tested: [Table 14]
L'étude est réalisée sur le front de 5 volontaires. Un prélèvement de la biomasse est réalisé à T0 sur le front des volontaires. Puis, un lavage de la totalité du front avec une compresse imbibée de 5 mL d'une solution de savon de Marseille à 2% en poids (savon de Marseille, Persavon) est réalisé en frottant pendant 30 sec. Deux rinçages sont ensuite réalisés avec des compresses imbibées d'eau. Après 5 min de séchage, un prélèvement est réalisé : T5min. Puis une compresse imbibée de 5 mL de la composition ci-dessus est déposée sur la moitié droite du front pendant 3 min. Après 2h, une brumisation de 2 sec de la composition ci-dessus est réalisée sur la moitié droite du front. Après 2h supplémentaires, 2 prélèvements sont réalisés : T4h côté savon de Marseille et T4h côté composition à tester. Les prélèvements sont réalisés à l'aide d'écouvillons (Sigma Trans Swab liquide Amies ref MW176S d'Elitech) aux temps T0, T5min, T2h et T4h, en réalisant 10 allers-retours en Z sur une surface de 3,75 cm2 et en exerçant une légère pression sur l'écouvillon de manière standardisée. L'écouvillon est ensuite maintenu dans la glace. The study is carried out on the forehead of 5 volunteers. A biomass sample is taken at T0 on the forehead of the volunteers. Then, washing the entire forehead with a compress soaked in 5 mL of a 2% by weight Marseille soap solution (Marseille soap, Persavon) is carried out by rubbing for 30 seconds. Two rinses are then carried out with compresses soaked in water. After 5 min of drying, a sample is taken: T5min. Then a compress soaked in 5 mL of the above composition is placed on the right half of the forehead for 3 min. After 2 hours, a 2 sec mist of the above composition is applied to the right half of the forehead. After 2 additional hours, 2 samples are taken: T4h on the Marseille soap side and T4h on the composition to be tested side. The samples are taken using swabs (Sigma Trans Swab liquid Amies ref MW176S from Elitech) at times T0, T5min, T2h and T4h, by performing 10 round trips in Z over an area of 3.75 cm 2 and exerting light pressure on the swab in a standardized manner. The swab is then kept on ice.
Effet sur la biomasse : Effect on biomass:
Cette suspension est ensuite diluée au 1/2, 1/10 et 1/50 dans du bouillon nutritif. Un ensemencement de 1 mL de chaque dilution est ensuite réalisé sur gélose TSA en boite de Pétri et incubé à 30-35°C pendant 48h. Après 48h d'incubation les colonies (UFC) sont comptées en totalité sur les boites. La biomasse bactérienne est obtenue en multipliant le nombre d'UFC comptées par son facteur de dilution et est exprimée en UFC/ml. This suspension is then diluted 1/2, 1/10 and 1/50 in nutrient broth. A seeding of 1 mL of each dilution is then carried out on TSA agar in a Petri dish and incubated at 30-35° C. for 48 hours. After 48 hours of incubation, the colonies (CFU) are counted in their entirety on the dishes. The bacterial biomass is obtained by multiplying the number of CFU counted by its dilution factor and is expressed in CFU/ml.
Le nettoyage avec la solution de savon de Marseille a induit une forte diminution de la quantité de biomasse des bactéries cultivables de 78%, 5 min après application de la solution de savon de Marseille. L'application du produit à tester après 4h de contact sur le microbiote appauvri augmente cette biomasse de 3 fois par rapport au témoin savon de Marseille. Cleaning with the Marseille soap solution induced a strong decrease in the amount of biomass of cultivable bacteria by 78%, 5 min after application of the Marseille soap solution. The application of the product to be tested after 4 hours of contact on the depleted microbiota increases this biomass by 3 times compared to the Marseille soap control.
Effet sur la diversité : Effect on diversity:
Les tests réalisés selon le protocole décrit dans l'exemple 1 montrent que la diversité du microbiome cutané n'est pas réduite après application du produit évalué. On note « = » l'absence de variation significative, « + » une augmentation significative et « - » une réduction significative. The tests carried out according to the protocol described in example 1 show that the diversity of the cutaneous microbiome is not reduced after application of the product evaluated. We note “=” the absence of significant variation, “+” a significant increase and “-” a significant reduction.
Les résultats sont regroupés dans le tableau 15 suivant. [Tableau 15] - Synthèse The results are collated in Table 15 below. [Table 15] - Summary
Légende : pas de variation significative (=), augmentation significative (+), diminution significative (-) Bibliographie Legend: no significant change (=), significant increase (+), significant decrease (-) Bibliography
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Claims

Revendications Claims
1. Méthode de sélection d'un ingrédient ou composition, de préférence écobiologique, avantageusement applicable par voie topique, en particulier cosmétique ou dermatologique, utile dans le maintien ou/et la restauration d'un microbiote cutané sain, comprenant les étapes consistant à : a) mettre un ingrédient ou composition à tester en contact avec des bactéries d'un microbiote de référence, b) analyser la quantité de biomasse et, avantageusement la biodiversité du microbiote après la mise en contact, et c) sélectionner en tant qu'ingrédient ou composition utile dans le maintien et/ou la restauration d'un microbiote cutané sain, un ingrédient ou une composition capable d'augmenter la biomasse du microbiote cutané, avantageusement sans en diminuer la biodiversité, par rapport à la quantification de la biomasse et, le cas échéant, la biodiversité du microbiote de référence. 1. Method for selecting an ingredient or composition, preferably ecobiological, advantageously applicable topically, in particular cosmetic or dermatological, useful in maintaining and/or restoring a healthy skin microbiota, comprising the steps consisting of: a) bringing an ingredient or composition to be tested into contact with bacteria of a reference microbiota, b) analyzing the quantity of biomass and, advantageously, the biodiversity of the microbiota after contacting, and c) selecting as an ingredient or composition useful in maintaining and/or restoring a healthy skin microbiota, an ingredient or a composition capable of increasing the biomass of the skin microbiota, advantageously without reducing its biodiversity, with respect to the quantification of the biomass and, where appropriate, the biodiversity of the reference microbiota.
2. Méthode de sélection selon la revendication 1, comprenant les étapes consistant à : a) mettre un ingrédient ou composition à tester en contact avec des bactéries d'un microbiote de référence, b) analyser la quantité de biomasse et la biodiversité du microbiote après la mise en contact, et c) sélectionner en tant qu'ingrédient ou composition utile dans le maintien et/ou la restauration d'un microbiote cutané sain, un ingrédient ou une composition capable d'augmenter la biomasse du microbiote cutané sans en diminuer la biodiversité, par rapport à la quantification de la biomasse et la biodiversité du microbiote de référence. 2. Selection method according to claim 1, comprising the steps consisting in: a) bringing an ingredient or composition to be tested into contact with bacteria of a reference microbiota, b) analyzing the quantity of biomass and the biodiversity of the microbiota after bringing into contact, and c) selecting, as an ingredient or composition useful in maintaining and/or restoring a healthy skin microbiota, an ingredient or a composition capable of increasing the biomass of the skin microbiota without reducing its biodiversity, compared to the quantification of the biomass and the biodiversity of the reference microbiota.
3. Méthode de sélection selon la revendication 1 ou 2, caractérisée en ce que ledit microbiote de référence est celui d'une zone cutanée saine. 3. Selection method according to claim 1 or 2, characterized in that said reference microbiota is that of a healthy skin area.
4. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce que ledit microbiote de référence est appauvri, avantageusement suite à une agression cutanée, plus avantageusement suite à l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à-vis du microbiote. 4. Selection method according to one of the preceding claims, characterized in that said reference microbiota is depleted, advantageously following a skin attack, more advantageously following the action of bactericides or virucides or soaps or products of hygiene or aggressive care vis-à-vis the microbiota.
5. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce que l'étape b est réalisée de 1 h à 31 jours, préférentiellement 4 jours, après l'étape a. 5. Selection method according to one of the preceding claims, characterized in that step b is carried out from 1 hour to 31 days, preferably 4 days, after step a.
6. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce qu'elle comprend les étapes in vitro suivantes : i) avant l'application dudit ingrédient ou composition, éventuellement formulé sous forme de composition topique, sur au moins une zone cutanée à étudier, à partir d'un premier prélèvement réalisé sur ladite zone cutanée, les analyses de la quantité de biomasse du microbiote cutané, ainsi que de sa biodiversité, en obtenant ainsi une première mesure dite de référence de quantification de la biomasse et une première mesure dite de référence de la biodiversité des bactéries dudit microbiote de référence ; et ii) après au moins une application dudit ingrédient ou de ladite composition, éventuellement formulé sous forme de composition topique, sur ladite zone cutanée, et après un temps prédéterminé d'action de l'ingrédient ou de la composition, à partir d'un deuxième prélèvement réalisé sur ladite zone cutanée, les analyses de la quantité de biomasse du microbiote cutané, ainsi que de sa biodiversité, en obtenant ainsi une deuxième mesure dite d'activité de quantification de la biomasse et une deuxième mesure dite d'activité de la biodiversité ; et iii) à comparer les deuxièmes mesures d'activité avec les premières mesures de référence, et à sélectionner l'ingrédient ou la composition, éventuellement formulé sous forme de composition topique, augmentant la biomasse du microbiote cutané, en particulier sans diminuer sa biodiversité. 6. Selection method according to one of the preceding claims, characterized in that it comprises the following in vitro steps: i) before the application of the said ingredient or composition, optionally formulated in the form of a topical composition, on at least one area skin to be studied, from a first sample taken from said skin area, the analyzes of the amount of biomass of the skin microbiota, as well as its biodiversity, thus obtaining a first so-called reference measurement for the quantification of the biomass and a first so-called reference measurement of the biodiversity of the bacteria of said reference microbiota; and ii) after at least one application of said ingredient or of said composition, optionally formulated in the form of a topical composition, on said skin area, and after a predetermined time of action of the ingredient or of the composition, from a second sample taken from said cutaneous area, the analyzes of the quantity of biomass of the cutaneous microbiota, as well as of its biodiversity, thus obtaining a second measurement called the activity of quantification of the biomass and a second measurement called the activity of the biodiversity; and iii) comparing the second activity measurements with the first reference measurements, and selecting the ingredient or composition, optionally formulated in the form of a topical composition, increasing the biomass of the cutaneous microbiota, in particular without reducing its biodiversity.
7. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce qu'on utilise au moins une méthode permettant de quantifier la biomasse du microbiote choisie parmi les méthodes quantitative ou semi quantitative suivantes 7. Selection method according to one of the preceding claims, characterized in that at least one method is used making it possible to quantify the biomass of the microbiota chosen from the following quantitative or semi-quantitative methods
- PCR quantitatives (ou qPCR), - Quantitative PCR (or qPCR),
- cytométrie en flux, - méthodes de dénombrement sur gélose. - flow cytometry, - enumeration methods on agar.
8. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce qu'on utilise au moins une méthode permettant de mesurer la biodiversité du microbiote choisie parmi les méthodes suivantes : - analyse d'une partie du gène hypervariable du ribosome 16S des bactéries après leur amplification ciblée, 8. Selection method according to one of the preceding claims, characterized in that at least one method is used making it possible to measure the biodiversity of the microbiota chosen from the following methods: - analysis of part of the hypervariable gene of the 16S ribosome of bacteria after their targeted amplification,
- technique de shot gun métagénomique, - metagenomic shot gun technique,
- technique de séquençage shotgun métatranscriptomique, suivie par une méthode de calcul de la diversité microbienne à partir des données de la méthode précédente utilisée, telle que le calcul : - metatranscriptomic shotgun sequencing technique, followed by a method for calculating the microbial diversity from the data of the previous method used, such as the calculation:
- de l'indice de Shannon, - the Shannon index,
- de l'indice de Simpson, - Simpson's index,
- de l'indice de Inverse-Simpson, - the Inverse-Simpson index,
- du nombre d'espèces ou d'OTU, - de l'Evenness ou l'indice de Pielou, - the number of species or OTUs, - the Evenness or the Pielou index,
- de l'indice de Hurlbert, - the Hurlbert index,
- de l'indice de Hill. - the Hill index.
9. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce qu'elle comprend au moins deux étapes d'amplification des quantités standardisée de matériel génétique : 9. Selection method according to one of the preceding claims, characterized in that it comprises at least two steps of amplifying the quantities standardized genetic material:
- une amplification quantitative effectuée en utilisant les amorces universelles 16S afin de mesurer la biomasse, - a quantitative amplification carried out using universal 16S primers in order to measure the biomass,
- une amplification effectuée en utilisant les amorces spécifiques 16S afin de mesurer la biodiversité du microbiote, - an amplification carried out using specific 16S primers in order to measure the biodiversity of the microbiota,
- la mesure de l'indice de Shannon. - measurement of the Shannon index.
10. Méthode de sélection selon l'une des revendications précédentes, caractérisée en ce que ledit ingrédient ou ladite composition à tester est choisi parmi le groupe comprenant : a) au moins une source de carbone qui est susceptible d'être utilisée par les bactéries comme substrat de croissance et de multiplication, et préférentiellement choisie parmi : 10. Selection method according to one of the preceding claims, characterized in that said ingredient or said composition to be tested is chosen from the group comprising: a) at least one carbon source which is capable of being used by bacteria such as growth and multiplication substrate, and preferably chosen from:
- les protéines hydrolysées partiellement ou totalement, issues de céréales (blé, maïs, orge, seigle, avoine), de légumineuses (soja, féverolle, fève, pois), de lait- partially or totally hydrolysed proteins, from cereals (wheat, corn, barley, rye, oats), legumes (soya, faba bean, broad bean, pea), milk
(caséines, albumines), de levure, ainsi que les peptides et les acides aminés seuls ou en mélange ; (caseins, albumins), yeast, as well as peptides and amino acids alone or as a mixture;
- les oligosaccharides ou les polysaccharides à courte chaîne, tels que les fructo oligosaccharides ou FOS ; les acides hyaluroniques, le pullulan, le chitosane, le dextran sulfate, le galactoarabinan, le carraghennane, l'alginate, la cellulose, avantageusement la cellulose microcristalline, l'amylose, l'amylopectine et leurs hydrolysats respectifs ; b) au moins un antioxydant qui présente une activité sur les espèces réactives de l'oxygène (ERO), avantageusement les radicaux libres, en particulier choisi dans le groupe constitué d'octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate ; 2,6-Bis(1,1- dimethylethyl)-4-methylphenol ; bis-ethylhexyl hydroxydimethoxy benzylmalonate ; tert-butylhydroquinone ; tetrabutyl ethylidenebisphenol ; thioglycolic acid ; thiotaurine ; thioctic acid ; dilauryl thiodipropionate ; sodium erythorbate ; sorbityl furfural ; erythorbic acid ; perillyl alcohol ; pyridyloxide t- butylnitrone ; ergothioneine ; melatonin ; acetyl cysteine ; cysteine ; lysine hydrochloride ; camosic acid ; tyrosyl histidine HC1 ; histidine hydrochloride ; pyridoxine serinate ; superoxide dismutase ; aminopropyl ascorbyl phosphate ; ascorbic acid ; ascorbyl dipalmitate ; ascorbyl glucoside ; ascorbyl linoleate ; ascorbyl methylsilanol pectinate ; ascorbyl palmitate ; ascorbyl tetraisopalmitate ; ascorbyl tocopheryl maleate ; trisodium ascorbyl palmitate phosphate ; disodium ascorbyl sulfate ; calcium ascorbate ; methylsilanol ascorbate ; sodium ascorbate ; sodium ascorbyl phosphate ; sodium ascorbyl/cholesteryl phosphate ; tetrahexyldecyl ascorbate ; magnésium ascorbyl phosphate ; tocopherol ; tocopheryl acetate ; tocopheryl linoleate ; tocopheryl oleate ; tocopheryl nicotinate ; tocopheryl retinoate ; sodium tocopheryl phosphate ; dioleyltocopheryl methylsilanol ; potassium ascorbyl tocopheryl 5 phosphate ; dodecyl gallate ; propyl gallate ; octyl gallate ; epigallocatechin gallate (EGCG) ; propyl gallate ; ethyl ferulate ; ethylhexyl ferulate ; chitosan ascorbate ; chitosan glycolate ; apigenin ; tiliroside ; alpha-arbutin ; arbutin ; baicalin ; quercetin ; quercetin caprylate ; isoquercetin ; diethylhexyl syringylidenemalonate ; dihydroxymethylchromone ; dimethoxy di-p-cresol ; dimethylmethoxy chromanol ; ethylbisiminomethylguaiacol manganèse chloride ; hesperidin methyl chalcone ; kojic acid ; kojic dipalmitate ; madecassoside ; asiaticoside ; magnolol (5,5'- diallyl-2,2'-dihydroxybiphenyl) ; nordihydroguaiaretic acid ; phenyl ethyl resorcinol ; resveratrol ; troxerutin ; glucosylrutin,rutin (4H-l-benzopyran-4-one)- short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) at least one antioxidant which has an activity on reactive oxygen species (ROS), advantageously free radicals, in particular chosen from the group consisting of octadecyl di-t-butyl-4-hydroxyhydrocinnamate; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t- butylnitrone; ergothioneine; melatonin; acetyl cysteine; cysteine; lysine hydrochloride; camosic acid; tyrosyl histidine HC1; histidine hydrochloride; pyridoxine serinate; superoxide dismutase; aminopropyl ascorbyl phosphate; ascorbic acid; ascorbyl dipalmitate; ascorbyl glucoside; ascorbyl linoleate; ascorbyl methylsilanol pectinate; ascorbyl palmitate; ascorbyl tetraisopalmitate; ascorbyl tocopheryl maleate; trisodium ascorbyl palmitate phosphate; disodium ascorbyl sulphate; calcium ascorbate; methylsilanol ascorbate; sodium ascorbate; sodium ascorbyl phosphate; sodium ascorbyl/cholesteryl phosphate; tetrahexyldecyl ascorbate; magnesium ascorbyl phosphate; tocopherol; tocopheryl acetate; tocopheryl linoleate; tocopheryl oleate; tocopheryl nicotinate; tocopheryl retinoate; sodium tocopheryl phosphate; dioleyltocopheryl methylsilanol; potassium ascorbyl tocopheryl 5 phosphate; dodecyl gallate; propyl gallate; octyl gallate; epigallocatechin gallate (EGCG); propyl gallate; ethylferulate; ethylhexyl ferulate; chitosan ascorbate; chitosan glycolate; apigenin; tiliroside; alpha-arbutin; arbutin; baicalin; quercetine; quercetin caprylate; isoquercetin; diethylhexyl syringylidenemalonate; dihydroxymethylchromone; dimethoxy di-p-cresol; dimethylmethoxy chromanol; ethylbisiminomethylguaiacol manganese chloride; hesperidin methyl chalcone; kojic acid; kojic dipalmitate; madecassoside; asiaticoside; magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl); nordihydroguaiaretic acid; phenyl ethyl resorcinol; resveratrol; troxerutin; glucosylrutin,rutin (4H-l-benzopyran-4-one)
; disodium rutinyl disulfate ; tetrahydrobisdemethoxydiferuloylmethane ; tetrahydrodemethoxydiferuloylmethane ; tetrahydrodiferuloylmethane ; tococysteamide ; totarol ; hydroxydecyl ubiquinone ; ubiquinone ; caroténoïdes ; niacinamide ou l'un de ses dérivés ; créatine ; créatinine, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, et rhamnose ; c) au moins un sel minéral ou organique topiquement acceptable, avantageusement choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc. ; disodium rutinyl disulfate; tetrahydrobisdemethoxydiferuloylmethane; tetrahydrodemethoxydiferuloylmethane; tetrahydrodiferuloylmethane; tococysteamide; totarol; hydroxydecyl ubiquinone; ubiquinone; carotenoids; niacinamide or one of its derivatives; creatine; creatinine, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, and rhamnose; c) at least one topically acceptable inorganic or organic salt, advantageously chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular the sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate.
11. Composition , avantageusement écobiologique, en particulier cosmétique et/ou dermatologique, comprenant au moins un ingrédient ou une composition, sélectionné par la méthode de sélection selon l'une des revendications précédentes, pour son utilisation pour maintenir et/ou restaurer un microbiote cutané sain. 11. Composition, advantageously ecobiological, in particular cosmetic and/or dermatological, comprising at least one ingredient or composition, selected by the selection method according to one of the preceding claims, for its use to maintain and/or restore a skin microbiota healthy.
12. Composition pour son utilisation selon la revendication 11, caractérisée en ce qu'elle comprend au moins une source de carbone, un antioxydant et un sel minéral ou organique, avantageusement ladite source de carbone, ledit antioxydant et/ou le dit sel étant sélectionné par la méthode de sélection selon l'une des revendications 1 à 10. 12. Composition for its use according to claim 11, characterized in that it comprises at least one carbon source, an antioxidant and an inorganic or organic salt, advantageously said carbon source, said antioxidant and/or said salt being selected by the selection method according to one of claims 1 to 10.
13. Composition pour son utilisation selon l'une des revendications 11 ou 12, caractérisée en ce qu'il s'agit d'une composition aqueuse, comprenant au moins 60% en poids de la composition totale d'eau, en particulier au moins 60%, mieux au moins 70%, ou au moins 80% ou au moins 90% ou au moins 95%. 13. Composition for its use according to one of claims 11 or 12, characterized in that it is an aqueous composition, comprising at least 60% by weight of the total composition of water, in particular at least 60%, better at least 70%, or at least 80% or at least 90% or at least 95%.
14. Composition pour son utilisation selon l'une des revendications 11 à 13, caractérisée en ce qu'elle comprend une phase aqueuse qui est caractérisée par une résistivité, à température ambiante, comprise entre 12,5 et 12 500 Ohms. cm, en particulier entre 80 et 8 000 Ohms.cm. 14. Composition for its use according to one of claims 11 to 13, characterized in that it comprises an aqueous phase which is characterized by a resistivity, at ambient temperature, of between 12.5 and 12,500 Ohms. cm, in particular between 80 and 8000 Ohms.cm.
15. Composition pour son utilisation, selon l'une des revendications 11 à 14, caractérisée en ce que la phase aqueuse présente une pression osmotique comprise entre 70 et 2500 mOsmol/1 et en particulier entre 100 et 800 mOsmol/1 ainsi qu'un pH compris entre 5,0 et 9,0 et en particulier compris entre 6,0 et 8,0. 15. Composition for its use, according to one of claims 11 to 14, characterized in that the aqueous phase has an osmotic pressure of between 70 and 2500 mOsmol / 1 and in particular between 100 and 800 mOsmol / 1 as well as a pH between 5.0 and 9.0 and in particular between 6.0 and 8.0.
16. Composition pour son utilisation selon l'une des revendications 11 à 15, caractérisée en ce que la composition présente un pH compris entre 5,0 et 9,0 et en particulier compris entre 6,0 et 8,0. 16. Composition for its use according to one of claims 11 to 15, characterized in that the composition has a pH of between 5.0 and 9.0 and in particular between 6.0 and 8.0.
17. Composition pour son utilisation selon l'une des revendications 12 à 16, caractérisée en ce que : a) la au moins une source de carbone est susceptible d'être utilisée par les bactéries comme substrat de croissance et de multiplication, et, préférentiellement, est choisie parmi : - les protéines hydrolysées partiellement ou totalement, issues de céréales (blé, maïs, orge, seigle, avoine), de légumineuses (soja, féverolle, fève, pois), de lait (caséines, albumines), de levure, ainsi que les peptides et les acides aminés seuls ou en mélange ; 17. Composition for its use according to one of claims 12 to 16, characterized in that: a) the at least one carbon source is capable of being used by bacteria as a growth and multiplication substrate, and, preferentially , is chosen from: - partially or totally hydrolyzed proteins, derived from cereals (wheat, corn, barley, rye, oats), legumes (soya, fava beans, broad beans, peas), milk (caseins, albumins), yeast , as well as peptides and amino acids alone or as a mixture;
- les oligosaccharides ou les polysaccharides à courte chaîne, tels que les fructo oligosaccharides ou FOS ; les acides hyaluroniques, le pullulan, le chitosane, le dextran sulfate, le galactoarabinan, le carraghennane, l'alginate, la cellulose, avantageusement la cellulose microcristalline, l'amylose, l'amylopectine et leurs hydrolysats respectifs ; b) Le au moins un antioxydant présente une activité sur les espèces réactives de l'oxygène (ERO), avantageusement les radicaux libres, et, en particulier, est choisi dans le groupe constitué d'octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate ; 2,6-Bis(1,1- dimethylethyl)-4-methylphenol ; bis-ethylhexyl hydroxydimethoxy benzylmalonate ; tert-butylhydroquinone ; tetrabutyl ethylidenebisphenol ; thioglycolic acid ; thiotaurine ; thioctic acid ; dilauryl thiodipropionate ; sodium erythorbate ; sorbityl furfural ; erythorbic acid ; perillyl alcohol ; pyridyloxide t- butylnitrone ; ergothioneine ; melatonin ; acetyl cysteine ; cysteine ; lysine hydrochloride ; camosic acid ; tyrosyl histidine HCl ; histidine hydrochloride ; pyridoxine serinate ; superoxide dismutase ; aminopropyl ascorbyl phosphate ; ascorbic acid ; ascorbyl dipalmitate ; ascorbyl glucoside ; ascorbyl linoleate ; ascorbyl methylsilanol pectinate ; ascorbyl palmitate ; ascorbyl tetraisopalmitate ; ascorbyl tocopheryl maleate ; trisodium ascorbyl palmitate phosphate ; disodium ascorbyl sulfate ; calcium ascorbate ; methylsilanol ascorbate ; sodium ascorbate ; sodium ascorbyl phosphate ; sodium ascorbyl/cholesteryl phosphate ; tetrahexyldecyl ascorbate ; magnésium ascorbyl phosphate ; tocopherol ; tocopheryl acetate ; tocopheryl linoleate ; tocopheryl oleate ; tocopheryl nicotinate ; tocopheryl retinoate ; sodium tocopheryl phosphate ; dioleyltocopheryl methylsilanol ; potassium ascorbyl tocopheryl 5 phosphate ; dodecyl gallate ; propyl gallate ; octyl gallate ; epigallocatechin gallate (EGCG) ; propyl gallate ; ethyl ferulate ; ethylhexyl ferulate ; chitosan ascorbate ; chitosan glycolate ; apigenin ; tiliroside ; alpha-arbutin ; arbutin ; baicalin ; quercetin ; quercetin caprylate ; isoquercetin ; diethylhexyl syringylidenemalonate ; dihydroxymethylchromone ; dimethoxy di-p-cresol ; dimethylmethoxy chromanol ; ethylbisiminomethylguaiacol manganèse chloride ; hesperidin methyl chalcone ; kojic acid ; kojic dipalmitate ; madecassoside ; asiaticoside ; magnolol (5,5'- diallyl-2,2'-dihydroxybiphenyl) ; nordihydroguaiaretic acid ; phenyl ethyl resorcinol ; resveratrol ; troxerutin ; glucosylrutin,rutin (4H-l-benzopyran-4-one)- short-chain oligosaccharides or polysaccharides, such as fructo-oligosaccharides or FOS; hyaluronic acids, pullulan, chitosan, dextran sulphate, galactoarabinan, carrageenan, alginate, cellulose, advantageously microcrystalline cellulose, amylose, amylopectin and their respective hydrolysates; b) The at least one antioxidant exhibits activity on reactive oxygen species (ROS), advantageously free radicals, and, in particular, is chosen from the group consisting of octadecyl di-t-butyl-4-hydroxyhydrocinnamate ; pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate; 2,6-Bis(1,1-dimethylethyl)-4-methylphenol; bis-ethylhexyl hydroxydimethoxy benzylmalonate; tert-butylhydroquinone; tetrabutyl ethylidenebisphenol; thioglycolic acid; thiotaurine; thioctic acid; dilauryl thiodipropionate; sodium erythorbate; sorbityl furfural; erythorbic acid; perillyl alcohol; pyridyloxide t-butylnitrone; ergothioneine; melatonin; acetyl cysteine; cysteine; lysine hydrochloride; camosic acid; tyrosyl histidine HCl; histidine hydrochloride; pyridoxine serinate; superoxide dismutase; aminopropyl ascorbyl phosphate; ascorbic acid; ascorbyl dipalmitate; ascorbyl glucoside; ascorbyl linoleate; ascorbyl methylsilanol pectinate; ascorbyl palmitate; ascorbyl tetraisopalmitate; ascorbyl tocopheryl maleate; trisodium ascorbyl palmitate phosphate; disodium ascorbyl sulphate; calcium ascorbate; methylsilanol ascorbate; sodium ascorbate; sodium ascorbyl phosphate; sodium ascorbyl/cholesteryl phosphate; tetrahexyldecyl ascorbate; magnesium ascorbyl phosphate; tocopherol; tocopheryl acetate; tocopheryl linoleate; tocopheryl oleate; tocopheryl nicotinate; tocopheryl retinoate; sodium tocopheryl phosphate; dioleyltocopheryl methylsilanol; potassium ascorbyl tocopheryl 5 phosphate; dodecyl gallate; propyl gallate; octyl gallate; epigallocatechin gallate (EGCG); propyl gallate; ethylferulate; ethylhexyl ferulate; chitosan ascorbate; chitosan glycolate; apigenin; tiliroside; alpha-arbutin; arbutin; baicalin; quercetine; quercetin caprylate; isoquercetin; diethylhexyl syringylidenemalonate; dihydroxymethylchromone; dimethoxy di-p-cresol; dimethylmethoxy chromanol; ethylbisiminomethylguaiacol manganese chloride; hesperidin methyl chalcone; kojic acid; kojic dipalmitate; madecassoside; asiaticoside; magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl); nordihydroguaiaretic acid; phenyl ethyl resorcinol; resveratrol; troxerutin; glucosylrutin,rutin (4H-l-benzopyran-4-one)
; disodium rutinyl disulfate ; tetrahydrobisdemethoxydiferuloylmethane ; tetrahydrodemethoxydiferuloylmethane ; tetrahydrodiferuloylmethane ; tococysteamide ; totarol ; hydroxydecyl ubiquinone ; ubiquinone ; caroténoïdes ; niacinamide ou l'un de ses dérivés ; créatine ; créatinine, carnosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, et rhamnose ; c) Le au moins un sel minéral ou organique est topiquement acceptable et, avantageusement, est choisi dans le groupe comprenant un sel de sodium, de magnésium, de calcium, de potassium, de phosphore, de cuivre, de zinc, de cobalt et leurs mélanges, en particulier le citrate de sodium, le lactate de sodium, le sulfate de magnésium, de cuivre ou de zinc, le phosphate de potassium ou de sodium, le gluconate de cuivre ou de zinc. ; disodium rutinyl disulfate; tetrahydrobisdemethoxydiferuloylmethane; tetrahydrodemethoxydiferuloylmethane; tetrahydrodiferuloylmethane; tococysteamide; totarol; hydroxydecyl ubiquinone; ubiquinone; carotenoids; niacinamide or one of its derivatives; creatine; creatinine, carnosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, and rhamnose; c) The at least one inorganic or organic salt is topically acceptable and, advantageously, is chosen from the group comprising a salt of sodium, magnesium, calcium, potassium, phosphorus, copper, zinc, cobalt and their mixtures, in particular sodium citrate, sodium lactate, magnesium, copper or zinc sulphate, potassium or sodium phosphate, copper or zinc gluconate.
18. Composition pour son utilisation selon l'une des revendications 12 à 17, caractérisée en ce ledit au moins un antioxydant est choisi dans le groupe comprenant tocophérol, camosine, hypotaurine, inositol, tréhalose, bétaïne, xylitol, mannitol, rhamnose. 18. Composition for its use according to one of claims 12 to 17, characterized in that said at least one antioxidant is chosen from the group comprising tocopherol, camosine, hypotaurine, inositol, trehalose, betaine, xylitol, mannitol, rhamnose.
19. Composition pour son utilisation selon l'une des revendications 12 à 18, caractérisée en ce qu'elle comprend du citrate de sodium et de la camosine. 19. Composition for its use according to one of claims 12 to 18, characterized in that it comprises sodium citrate and camosine.
20. Composition pour son utilisation selon l'une des revendications 12 à 19, caractérisée en ce que la composition est appliquée sur une zone cutanée saine, avant ou suite à une agression cutanée, avantageusement l'action de bactéricides ou de virucides ou de savons ou de produits d'hygiène ou de soin agressifs vis-à- vis du microbiote. 20. Composition for its use according to one of claims 12 to 19, characterized in that the composition is applied to a healthy skin area, before or following a skin attack, advantageously the action of bactericides or virucides or soaps or hygiene or care products that are aggressive towards the microbiota.
EP22723166.9A 2021-04-20 2022-04-20 Method for selecting ecobiological topical compositions and ingredients capable of restoring or reinforcing the skin microbiota Pending EP4326892A1 (en)

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