EP3250707A1 - Détection sélective de bactéries lactiques et/ou acétiques ou de champignons - Google Patents

Détection sélective de bactéries lactiques et/ou acétiques ou de champignons

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Publication number
EP3250707A1
EP3250707A1 EP16707831.0A EP16707831A EP3250707A1 EP 3250707 A1 EP3250707 A1 EP 3250707A1 EP 16707831 A EP16707831 A EP 16707831A EP 3250707 A1 EP3250707 A1 EP 3250707A1
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EP
European Patent Office
Prior art keywords
flora
fungal
matrix
adenosine triphosphate
bacterial
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.)
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Application number
EP16707831.0A
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German (de)
English (en)
French (fr)
Inventor
Charles Cervin
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Individual
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Individual
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Publication of EP3250707A1 publication Critical patent/EP3250707A1/fr
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/008—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions for determining co-enzymes or co-factors, e.g. NAD, ATP
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/66—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • G01N21/763—Bioluminescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food
    • G01N33/14—Beverages
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2304/00—Chemical means of detecting microorganisms
    • C12Q2304/60—Chemiluminescent detection using ATP-luciferin-luciferase system

Definitions

  • the present application relates to the selective detection of bacteria or the selective detection of fungi in a mixture of these bacteria and fungi.
  • the present application thus relates to the detection, measurement or estimation of the presence of lactic or acetic bacteria in a mixture comprising these lactic acid bacteria or these acetic bacteria and comprising fungi, in particular yeasts or molds.
  • Such a mixture is found in drinks such as beer, wine, in the sugar industry when pressing sugar beet juice or sugar cane and sugary carbonated beverages such as lemonade, or fruit juices when their manufacture.
  • the present application also relates to the detection, measurement or estimation of the presence of fungi, in a mixture comprising lactic acid bacteria or acetic bacteria and comprising said fungi, in particular yeasts or molds.
  • the present application relates even more particularly to the estimation, in a medium containing fungi, of the presence of lactic or acetic bacteria, by measuring the adenosine triphosphate or ATP of these bacteria, or ATPmetria, in particular using luciferin. , luciferase and a measure of bioluminescence.
  • the present application also relates to said estimation by the addition of a luminescence reagent, such as luminol or a viability marker, enabling viable microorganisms to emit measurable or detectable luminescence, with or without excitation, by any optical means.
  • a luminescence reagent such as luminol or a viability marker
  • An acetic bacterium refers below to a Gram-negative aerobic bacterium capable of converting ethanol to acetic acid. Acetic bacteria are present in wine and cider and rarely in beer saturated with carbon dioxide. The development of acetic bacteria is undesirable in wine after alcoholic and / or malolactic fermentation, as well as in beer, and in carbonated and sugary beverages.
  • a lactic acid bacterium refers below to a gram-positive bacterium, optional anaerobic or microaerophilic, in particular in the form of shells or rods or tetrads and capable of fermenting sugars lactic acid.
  • Lactic acid bacteria are present in wine, cider, sugar beet juice and sugar cane, and can be found in carbonated beverages such as soda and beverage juices. sweet.
  • the development of lactic acid bacteria is undesirable after wine fermentation and sulphiting, and in cider without malo-lactic fermentation because they can alter the organoleptic qualities of wine and cider.
  • Lactic acid bacteria are undesirable in beer, except for certain lambics or Hopkins Weisse type lactic acid bacteria, which are added during fermentation.
  • certain lactic acid bacteria such as "leuconostoc mesenteroides" consume sucrose in sugar beet juice causing filtration problems and financial losses.
  • lactic acid bacteria or acetic bacteria should be understood throughout the application as meaning either lactic acid bacteria or acetic bacteria or lactic acid bacteria and acetic bacteria.
  • bacteria or "bacterial flora” are to be included in the application as synonymous with “lactic or acetic bacteria” unless otherwise stated.
  • muscles or "fungal flora” must be included in the application as being synonymous with “molds or yeasts”,
  • composition of matter that includes bacteria and fungi.
  • Such a mixture may be solid, especially when it is dried as a filtrate or after centrifugation, or wet if it is dispersed in a liquid, or gaseous, for example a sample of air.
  • agri-food matrix must be understood as a solid or liquid or gaseous support containing in volume or on its surface a microbial flora found in the agri-food industry, including agriculture, or during production homemade food or drink.
  • application in the liquid phase of a product A to a product B mean in the application an operation the result of which is a liquid containing A and B in solution or in dispersion.
  • An aqueous phase application means a liquid phase application for a liquid which is water or a liquid compatible with the survival of the bacteria in the solution obtained by the application.
  • ATP or adenosine triphosphate denote in the application, the molecule which, in the biochemistry of all known living organisms, provides by hydrolysis, the energy necessary for the chemical reactions of the metabolism.
  • free ATP mean ⁇ out of any cell, whether or not this ATP is available for a bioluminescence reaction.
  • Luminescence words refer to any non-naturally occurring luminescence in the sample analyzed, but caused by the addition of one or more reagents essential for this luminescence reaction, said luminescence being detectable in viable microbial cells of the sample. sample in particular by any optical system with or without excitation of this luminescence.
  • bacteria refers to a treatment having the effect of reducing a bacterial population of at least three decimal logarithms of colony forming units / ml upon an agar count of a sample after treatment.
  • Ergosterol refers to a component of the membrane or wall of fungi.
  • a “sterilized precursor of ergosterol” refers to a molecule having the chemical structure of a sterol, namely a lipid having a sterane nucleus whose carbon 3 carries a hydroxyl group produced during the biosynthesis of ergosterol in the fungal cell.
  • the words "beta-glucans” refer to polysaccharides consisting of beta-linked D-glucose which may be ⁇ (1-3), ⁇ (1-4) or ⁇ (1-6), with the numbers indicating D-glucose concerned.
  • MRS refers to either broth or agar of Man, Rogosa and Sharpe.
  • Luminol denotes a molecule of chemical formulation C8H7N302 which produces a chemiluminescence with a characteristic blue luster, when it is mixed with certain oxidants. This luminescence reaction with luminol can also be catalyzed by addition of a catalyst, such as menadione. A number of studies show a correlation between the luminol luminescence signal and the number of bacterial cells in the sample.
  • viability marker refers to any marker used either to check the cell wall status of dead, damaged or intact cells such as propidium iodide, or ethidium bromide, or to detect intracellular enzyme activity such as for example the esterase activity of a viable microbial cell often measured by introducing a non-luminescent molecule that becomes luminescent after enzymatic conversion in the viable cell, or to simultaneously verify the integrity of the cell wall and the intracellular enzyme activity of the viable cell.
  • the labeling is nonspecific to a particular species of microorganism and does not include nucleic probes, primers Adn or Arn. This viability marker is detectable by luminescence. It can be used with a counter-marker to enhance the luminescence detection of labeled cells.
  • the prior art knows the technique of ATPmetry which allows a measurement of free dans in a solution to estimate the biomass present in the solution.
  • This technique firstly involves making available, by release into the solution, ⁇ biomass, for example by a lysing agent that causes the death of living organisms and releases their ATP in the solution, then assumes to perform a luminescence measurement proportional to the amount of free ATP in the solution.
  • a known bioluminescence reaction using the luciferin-luciferase enzyme-substrate complex is used.
  • the bioluminescence reaction occurs through prior activation of luciferin by the hydrolysis of adenosine triphosphate.
  • the light intensity produced can be made proportional to the amount of free ATP in the mixture and it is thus possible, thanks to a luminometer, to obtain optically an evaluation of the amount of ATP in a sample.
  • lysing agent means a product capable of opening the membrane of bacteria or fungi. However, most lysing agents only provide access to the total amount of intracellular ATP of the total biomass present in a solution.
  • the selective measurement by ATPmetry of lactic or acetic bacterial biomass in a mixture of a bacterial flora and a fungal flora is therefore for the prior art a difficult problem. It is the same for obtaining the selective measurement of fungal biomass in such a mixture, except to neglect the bacterial biomass.
  • the invention relates to a method for the detection of lactic acid bacteria and / or acetic acid in an agro-alimentary matrix comprising a microbial flora, the microbial flora comprising a lactic and / or acetic bacterial flora and comprising a fungal flora, the flora bacterium containing a bacterial adenosine triphosphate, the fungal flora containing a fungal adenosine triphosphate, which comprises the following steps:
  • an antifungal agent having a lethal antifungal action at a first expiry, on the fungal flora, and a non-lethal antibiotic action at a second expiry after the first expiry, on the bacterial flora,
  • the antifungal agent is an ergosterol inhibitor.
  • the antifungal agent is an inhibitor of a sterol precursor of ergosterol.
  • the antifungal agent has a glutarimide functional group and is an inhibitor of the translocation step of the elongation phase of the eukaryotic protein synthesis.
  • the ergosterol inhibitor is a molecule of the polyene family.
  • the inhibitor of a sterilized precursor of ergosterol is a molecule of the family of morpholines.
  • the steroid precursor inhibitor of ergosterol is a molecule of the azole family.
  • the antifungal agent is an inhibitor of the synthesis of beta-glucans.
  • the invention also relates to a device which comprises an antifungal having, after application, an antifungal activity which is lethal at a first time, on a fungal flora, and a non-lethal antibiotic activity at a second time after the first expiry, on a lactic bacterial flora and or acetic, and which comprises a means for detecting a microbial flora comprising fungal flora and lactic and / or acetic bacterial flora.
  • the means for detecting the microbial flora comprises a lysing agent.
  • the invention also relates to an application of the above method to the separate estimation of adenosine triphosphate of fungal origin and of adenosine triphosphate of bacterial origin in an agri-food matrix comprising a lactic bacterial flora and / or acetic acid and fungal flora, characterized in that it comprises the following steps:
  • the invention also relates to a method for the detection of fungi in an agro-alimentary matrix comprising a microbial flora, the microbial flora comprising a lactic and / or acetic bacterial flora and comprising a fungal flora, the flora bacterium containing a bacterial adenosine triphosphate, the fungal flora containing a fungal adenosine triphosphate, which comprises the following steps:
  • an antifungal agent having a lethal antifungal action at a first expiry, on the fungal flora, and a non-lethal antibiotic action at a second expiry after the first expiry, on the bacterial flora,
  • an antifungal of the family of polyenes is selected, it is the antifungal named:
  • Molecule A1 Amphotericin B
  • This polyene family targets the ergosterol membrane of fungi.
  • the polyene family includes amphotericin B, nystatin, natamycin.
  • the toxicity results for yeasts and bacteria are presented in the table below showing the exposure time of the fungal flora to the A1 molecule and the associated fluorescence measurement: The lysis of the yeasts after exposure to the fungicide was confirmed by microscopic examination and staining with methylene blue. The words "contact time” denote below the exposure time of the microorganisms to the fungicidal product, here the A1 molecule.
  • Molecule A1 dosage 400 mg / L (milligram / liter) contact time 4 hours
  • the concentration threshold which makes it possible to obtain the effect of the invention of being non-bactericidal for lactic and acetic bacteria while being antifungal for the yeast populations is between 50 mg / L or 50mg / l and 100 mg / l.
  • the skilled person may constitute a first list of antifungal products and a second list of non-bactericidal products for bacteria and select the products appearing in the first and second list for which fungicidia intervenes before the bactericide.
  • the application of the method is then made during the rearing of post-sulphiting wine to monitor the abnormal growth of lactic and acetic bacteria whose number must decrease as the various withdrawals, or brewery during the phase alcoholic fermentation of brew musts, after and before filtration of the beer, for the undesirable growth monitoring of lactic acid bacteria for the purpose of a quality control.
  • the initial mixture with an antifungal in this case A1 is exposed to a concentration of, for example, 100 mg / l, by application in the aqueous phase, for example, for 8 hours.
  • a concentration of, for example, 100 mg / l for example, 100 mg / l
  • the antifungal and the initial mixture are each in aqueous phase, they will be mixed; if only one of them is powdered, it will be mixed in the aqueous phase of the other and if both are in powder, they may be mixed with an aqueous buffer.
  • the fungal cells After exposure of the initial mixture to the antifungal, the fungal cells have released their ATP in the aqueous phase and it is possible to remove this fungal ATP from the aqueous phase by various known means (filtration, ATPase, etc.).
  • Bacteria are the only viable microorganisms remaining at this stage and are larger than ATP molecules, which allows, for example, filtration.
  • the method of the invention may therefore, after removal of fungalteil, use any method of counting the remaining bacteria, as long as this method is insensitive to the presence in the aqueous phase of dead cell membranes of fungal cells.
  • Any method of counting living bacteria for example, solid and flow cytometry, solid-phase cytometry and enumeration, impedancemetry, flow cytometry, fountain flow cytometry, viability marker, luminol, luminescence reagent .
  • a pore diameter filter of the order of 0.2 micron or 0.45 micron, that is to say of a size greater than the molecules of ATP but less to the size of the bacteria, to obtain filter residues on the filter containing dead fungal cell membranes, lacking ATP and living bacteria containing ⁇ .
  • any means for removing free ATP in the aqueous phase at this stage is an equivalent means for the invention.
  • a method of choice for the counting of bacteria, it will then be used with the invention a method combining total lysis and ATPmetry. For this, it will cause cell death by exposure to a lysing agent in the liquid phase, generally aqueous, but especially capable of releasing ATP cells, their survival no longer useful at this stage.
  • a lysing agent in the liquid phase, generally aqueous, but especially capable of releasing ATP cells, their survival no longer useful at this stage.
  • Other liquids than water, are therefore possible for this application of a lysing agent provided that these liquids do not substantially inhibit the enzymatic reaction of bioluminescence of ATP
  • the liquid phase application of the lysing agent is, for example, non-specific to bacteria, it will cause the lysis of all the microbial cells present in the lysing agent.
  • aqueous phase therefore a priori lactic and acetic bacterial cells mainly present in the initial mixture, releasing their bacterial ATP in the liquid phase. If other bacteria are present, they will be a systematic error and the invention is thus particularly applicable to beverages in the fermentation phase (including beer, wine, cider) in which the presence of other bacteria that lactic and acetic is weakly possible, because of the pH of these drinks or the presence of carbon dioxide in the beer.
  • the luciferin / luciferase couple and the bioluminescence reaction with the ATP of this pair will be conveniently used to optically determine the bacterial ATP and deduce an estimate of the biomass of bacterial origin in the initial mixture.
  • ATPmetry was verified for the destruction of the fungal flora and the safety of the antifungal treatment on the lactic or acetic bacterial flora for a contact time of 4 hours or 8 hours and on different strains.
  • Lum-1 ATPmetry device model analogous to the New Horizons Diagnostics Profile-1 biolumometer
  • bioluminescence reagents from Biothema Ab, namely the "ATP Eliminating Buffer", "Extractant B / S” reagents "ATP Reagent HS” and Filtravette TM porosity 0.45 ⁇ filter cuvettes from New Horizons Diagnostics. Description of the initial protocol:
  • Gluconobacter oxydans 1.10 7 CFU / 500 ⁇ 4 ; 1 .10 6 CFU / 500 ⁇
  • Bioluminescence values before and after treatment demonstrate treatment efficacy Saccharomyces diastaticus before and after fungicide treatment.
  • the above method is applicable with all embodiments of the invention, i.e. with any selective antifungal for lactic or acetic bacteria.
  • the molecule A1 is therefore adapted to the invention.
  • an antifungal of the family Morpholine is selected, it is the antifungal named:
  • Molecule A2 Amorolfine This family of morpholines targets a sterol precursor of the ergosterol membrane of fungi.
  • Yeast and bacterial toxicity results are presented in the table below showing the exposure time of the fungal flora to the A2 molecule and are obtained by counting on MRS agar.
  • Molecule A2 Dosage: 200 mg / L contact time 4 hours
  • the concentration threshold which makes it possible to obtain the effect of the invention of being non-bactericidal for lactic and acetic bacteria while being antifungal for yeast populations is between 100 and 100. mg / l and 200 mg / l.
  • the same methods as in the first embodiment are applicable for determining bacterial or fungal biomass.
  • an antifungal of the family Azolés is selected, it is the antifungal named:
  • Molecule A3 voriconazole.
  • This family of azoles targets a sterol precursor of the ergosterol membrane of fungi.
  • the toxicity results for yeasts and bacteria are presented in the table below showing the exposure time of the fungal flora to the molecule A3.
  • the molecule A3 is therefore suitable for the invention at a concentration of 400 mg / l.
  • Molecule A3 Dosage: 200 mg / L contact time 18 hours
  • the concentration threshold for obtaining the effect of the invention is between 200 mg / l and 300 mg / l.
  • the molecule A3 is therefore suitable for the invention at a concentration of 200 mg / l.
  • an antifungal agent which has a glutarimide functional group and is an inhibitor of the phase translocation step
  • Molecule A4 cycloheximide
  • This family also includes the following compound: lacimidomycin.
  • Yeast and bacterial toxicity results are presented in the table below showing the duration of exposure of the fungal flora to the A4 molecule.
  • the concentration threshold for obtaining the effect of the invention is greater than 300 mg / l.
  • the molecule A4 is suitable for the invention.
  • the A4 molecule is particularly suitable for wine in breeding phase after sulphiting, targeting wild yeasts present in and around lees.
  • an antifungal of the echinocandin family, inhibitors of the synthesis of beta-glucans of fungal cells it is the antifungal named:
  • Molecule A5 Caspofungin This family of compounds inhibits the synthesis of beta-glucans in the fungal cell by a noncompetitive inhibition of the enzyme 1, 3- ⁇ "glucan synthase” and comprises at least the following compounds: pneumocandins, 1 echinocandin B, cilofungin, caspofungin (or casponfungin), micafungin or micafungin, anidulafungin or anidulafungin.
  • the concentration threshold which makes it possible to obtain the effect of the invention of being non-bactericidal for acetic bacteria while being antifungal for the yeast populations is between 100 mg / l. and 200 mg / l.
  • the invention thus makes it possible, by applying certain antifungals, to a beverage or a juice containing a lactic and / or acetic flora and containing fungi, to measure the bacterial biomass reliably and independently of the fungal biomass.
  • the invention also makes it possible to determine the fungal biomass by subtracting the bacterial biomass from the total biomass.
  • the invention thus makes it possible in a simple manner, to the precision of the measurement of bacterial ATP performed or to the precision of any method of counting bacteria used as a replacement for the ATPmetry applied to the remaining bacteria after the application of the antifungal non-bactericidal, to detect or estimate the value of bacterial biomass in a mixture of a fungal flora and a bacterial, lactic and / or acetic flora.
  • a fungal membrane component such as mannans or chitins
  • mannans or chitins the antifungal inhibitors of this component or of one of its precursors in the path of biosynthesis of said component as follows:
  • the selection may generally relate to an inhibitor of a biosynthetic reaction specific to the fungal kingdom, that is to say on a branch of the path of biosynthesis of the component.
  • the selection is thus made on the specific branch from lanosterol.
  • Members of the Allylamine family are, for example, not part of the sterol precursor inhibitors of ergosterol biosynthesis. A molecule of this family was thus tested without a yeast or bactericidal effect is found in less than twenty four hours of contact time.
  • the selection may relate to any other luminescence reagent, such as the use of luminol, or any cell viability marker, made detectable by any optical means.
  • the invention also relates to its teaching and the definition of "antifungal product with bactericidal action posterior to a lethal antifungal action", an inhibitor of a biosynthesis reaction of a fungal membrane component, c that is, this inhibitor inhibits a reaction occurring in the biosynthetic pathway of the component.
  • the invention is particularly susceptible of industrial application in the production of wine, beer, cider, soda and carbonated drinks and in the production of sugar from beet juice or sugar cane juice.
  • the invention can be applied in all its embodiments in the form of a method for the selective detection of fungi in an agro-alimentary matrix comprising a microbial flora, the microbial flora comprising a lactic and / or acetic bacterial flora and comprising a fungal flora, the bacterial flora containing a bacterial adenosine triphosphate, the fungal flora containing a fungal adenosine triphosphate, by following the steps of: applying to the matrix, an antifungal agent having an antifungal action lethal to a first maturity, on the fungal flora, and a non-lethal antibiotic action at a second time after the first deadline, on the bacterial flora, said lethal action releasing adenosine triphosphate of fungal origin for the first time; measure, between the first expiry and the second expiry, the adenosine triphosphate of fungal origin released
  • an antifungal according to the invention can be used for selective measurement of bacteria in a mixture of these two flora by measuring, in particular by ATPmetry, ⁇ bacteria remaining after the lethal action or ⁇ fungi, in a time window during which the fungi are dead and the bacteria are alive.

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EP16707831.0A 2015-01-28 2016-01-25 Détection sélective de bactéries lactiques et/ou acétiques ou de champignons Withdrawn EP3250707A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR1500163A FR3031988B1 (fr) 2015-01-28 2015-01-28 Detection selective de bacteries lactiques et/ou acetiques ou de champignons
PCT/FR2016/000011 WO2016120534A1 (fr) 2015-01-28 2016-01-25 Détection sélective de bactéries lactiques et/ou acétiques ou de champignons

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EP3250707A1 true EP3250707A1 (fr) 2017-12-06

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US (1) US10415074B2 (zh)
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CN (1) CN107406870A (zh)
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WO (1) WO2016120534A1 (zh)

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CN113444766B (zh) * 2021-06-04 2024-05-24 海天醋业集团有限公司 用于发酵酒陈酿过程中变质菌的富集培养基及的检测方法
CN115369148A (zh) * 2022-08-22 2022-11-22 武汉新华扬生物股份有限公司 一种微生态产品中乳酸菌的计数方法

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CA1273308A (en) * 1985-07-08 1990-08-28 John E. Hill Dried medium culture film solution and method for making medium lactobacillacea selective
JP3547882B2 (ja) * 1995-12-28 2004-07-28 キッコーマン株式会社 Ａｔｐ消去剤、ａｔｐ消去法、それを用いた生物細胞測定試薬及び生物細胞測定法
CN100547388C (zh) * 2006-08-16 2009-10-07 中国科学院电子学研究所 一种抗干扰的食品细菌总数快速检测方法及试剂
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US10415074B2 (en) 2019-09-17
FR3031988B1 (fr) 2018-10-19
FR3031988A1 (fr) 2016-07-29
WO2016120534A1 (fr) 2016-08-04
CN107406870A (zh) 2017-11-28

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