CN113677434B - Device for determining the presence of bacterial contamination in a fluid - Google Patents
Device for determining the presence of bacterial contamination in a fluid Download PDFInfo
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- CN113677434B CN113677434B CN202080027540.2A CN202080027540A CN113677434B CN 113677434 B CN113677434 B CN 113677434B CN 202080027540 A CN202080027540 A CN 202080027540A CN 113677434 B CN113677434 B CN 113677434B
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
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- B01D63/087—Single membrane modules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/10—Supported membranes; Membrane supports
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/56—Labware specially adapted for transferring fluids
- B01L3/563—Joints or fittings ; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
- B01L3/5635—Joints or fittings ; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors connecting two containers face to face, e.g. comprising a filter
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0668—Trapping microscopic beads
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0681—Filter
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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- B01L2300/0803—Disc shape
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
- G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
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- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
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- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a device (1) for determining contamination of a fluid by microorganisms, the device (1) having: a housing (2) having an interior volume; a cover (3) that closes the housing; a fluid inlet (11); at least one filtering member (4); at least one nutritive layer (5) comprising a composition of microbial culture medium, characterized in that the device comprises a fluid outlet (11) and in that the cover (3) has, in the inner volume, an inner surface extending radially around the fluid inlet (11) up to the peripheral edge of the cover, which inner surface is inclined and converging towards the fluid inlet (11), and in that the bottom of the housing (2) has a surface extending radially around the fluid outlet (12) up to the side wall of the housing, which inner surface is inclined and converging towards the fluid outlet (12).
Description
Technical Field
The invention belongs to the technical field of microbiological analysis. More particularly, the present invention relates to a microbiological test device for testing a liquid to be analyzed, which liquid is susceptible to containing at least one microorganism.
Background
The invention can be applied in the field of industrial, food, pharmaceutical, cosmetic or veterinary microbiological testing, for terrestrial or space use.
The invention was developed following work supported by the french national space research Center (CNES) and the company INITIAL.
Background
There are many instances where a test liquid is present to determine the presence of (typically to determine the absence of) at least one microorganism. Of course, the liquid to be analyzed may be a biological fluid (whole blood, serum, plasma, urine, cerebrospinal fluid, organ secretions, etc.). However, the liquid may also be an industrial liquid, in particular a food liquid (water, other beverages, in particular fruit juices, milk, soft drinks, etc.) or a pharmaceutical or cosmetic or veterinary liquid (milk from sick animals).
A number of laboratory techniques are known for filtering a liquid to be analyzed, to collect any microorganisms contained in the liquid, to culture these microorganisms in order to subsequently detect, count, characterize and/or identify these microorganisms. These techniques require a certain number of actions and specific infrastructure (filter ramps, laboratory benches, incubators) well known to laboratory assistants.
These techniques generally require the use of a filtration device comprising an enclosed interior space defined by a housing designed to receive the liquid to be analyzed. Such a technique is particularly called "membrane filtration". A microbial filtration means, such as a filtration membrane, is disposed in the enclosed interior space and separates a first compartment from a second compartment of the enclosed interior space in the enclosed interior space. The device (filter cartridge) has an inlet for the liquid to be analysed, which opens into a first compartment enclosing an inner space.
In some known filtration devices, the device is opened to retrieve the filtration means, and then the filtration means is transferred to a culture device to culture the microorganisms. Such techniques are easily performed in the laboratory.
In contrast, such techniques are difficult to implement in operating environments, on the ground, in international space stations, and in industrial environments where liquids are produced, packaged, distributed, or used.
From document WO2018189478 A1 a microbiological test device and method for testing a liquid to be analyzed are known, which enable particularly simplified test operations that can be used or performed remotely from a microbiological laboratory (including in an industrial environment). This document describes a microbiological test device for testing a liquid to be analyzed, which liquid is liable to contain at least one microorganism. The test device includes: a closed housing designed to receive a liquid to be analyzed; a microbial filtration means disposed in the enclosed interior space, separating a first compartment from a second compartment of the enclosed interior space in the enclosed interior space; an inlet for a liquid to be analysed leading to a first compartment enclosing an interior space; and a nutrient layer comprising a microbial culture medium. The main disadvantage of this device is that the liquid remains in the device after filtration, cannot be recycled, and the liquid remaining in the device may leach out the PAD, dilute the nutritive layer and generate false negatives. Furthermore, the manufacture of the device requires the housing to be placed in a vacuum, which is a significant limitation.
Disclosure of Invention
The present invention is an improvement over the aforementioned devices, being more compact and more practical.
To this end, the invention relates to a device for determining microbial contamination of a fluid, the device comprising:
-a housing having an interior volume bounded by at least one sidewall and a bottom;
-a cover closing the housing and positioned opposite the bottom;
-a fluid inlet arranged on the cover, open to the inner volume of the housing, the fluid inlet extending along an axis substantially secant and preferably perpendicular to the cover;
-at least one filter member arranged in the inner volume;
-at least one nutrient layer comprising a microbial culture medium;
it is characterized in that the device comprises:
-a fluid outlet arranged on the housing, open to the inner volume of said housing, and the cover having, in the inner volume, an inner surface extending radially around the fluid inlet up to the peripheral edge of the cover (3), said inner surface being inclined or curved and converging towards the fluid inlet;
-a support grid configured to support a nutrition layer;
and in that the bottom of the housing has a surface extending radially around the fluid outlet up to the side wall of the housing, said surface being inclined and converging towards the fluid outlet.
Advantageously, in the device according to the invention, the fluid passes through the device and does not stagnate in the bottom of the casing and on the filter, which ensures that the liquid passes through the device quickly (which requires less force to do so), prevents leaching of the nutritive layer, and ensures that the microorganisms are collected and cultured under optimal conditions. Furthermore, the inclination of the inner surface of the cover helps to improve the distribution of the fluid in the device and ensures that all the fluid is distributed uniformly over the filter, to ensure a uniform distribution of the microorganisms over the filter, thus preventing any formation of microbial nodules on the filter, which might generate false negatives.
Advantageously, the device is designed to quantify and detect two fecal contaminants, e.g., E.coli and enterococcus.
In the present invention, the term "port" means a fluid channel having a central orifice longitudinally through the channel.
The device according to the invention is preferably used vertically, i.e. the fluid inlet and the fluid outlet are positioned vertically.
In the present application, a vertically positioned object is an object arranged in a direction parallel to the direction of gravity.
According to a feature of the invention, the fluid is a liquid, such as water, or the fluid is a gas, such as air.
According to another characteristic of the invention, the inclination of the inner surface of the cover is strictly greater than +4 °. Preferably, the inclination of the inner surface of the cap is preferably between +5 ° and +15 °, and more preferably substantially 10 °.
According to another characteristic of the invention, the inclination of the surface of the bottom is strictly less than 0 °. Preferably, the inclination of the surface of the bottom is preferably between-5 ° and-10 °, and more preferably between-6.5 ° and-7.5 °.
According to another feature of the invention, the fluid inlet projects into the interior volume of the housing relative to the inner surface of the cover.
Preferably, the fluid inlet is arranged substantially in the centre of the lid.
According to another feature of the invention, the inlet has a first end extending outside the cap, the first end being designed to cooperate with the plug.
According to another feature of the present invention, the fluid inlet has a second end extending within the interior volume, the second end having at least one lateral aperture opening into the interior volume.
According to another feature of the invention, the at least one lateral orifice is oriented to eject the fluid at least partially towards the inner surface of the cover.
According to another feature of the invention, the inlet has at least two lateral orifices, preferably arranged opposite each other.
According to another feature of the invention, the fluid outlet has a first end extending outside the housing, the first end being designed to cooperate with the plug.
According to another feature of the invention, the fluid inlet has a second end flush with the bottom of the housing.
According to a feature of the invention, the filter member is arranged in the housing at a given distance h from the inlet. Advantageously, the given distance h is strictly greater than 1mm, and preferably between 1.5mm and 5mm, and more preferably between 2.5mm and 3.5 mm. The filter member must be at a given distance h so that the microorganisms can grow and form readily visible colonies when the device is incubated. The given distance depends on the minimum volume of air required to grow the microorganisms.
According to another characteristic of the invention, the device according to the invention is fluid-tight when the inlet and the fluid outlet are blocked by the plugs.
According to a feature of the invention, the lid is transparent or translucent, which enables the microbial colonies to be observed after incubation.
According to a feature of the invention, the housing is substantially cylindrical or polygonal.
According to a feature of the invention, the housing has a first circumferential bearing surface shaped to cooperate complementarily with the edge of the cover. Advantageously, the cover and the housing are ultrasonically welded together.
According to a feature of the invention, the housing has at least one second bearing surface extending in the internal volume and shaped to receive a peripheral portion of the filter member, said second bearing surface being shaped to cooperate with a portion of the cover such that the peripheral portion of the filter member is (preferably uniformly) clamped between the cover and the housing such that the filter member is stable and stationary as fluid passes through the device to ensure that there is no leakage and all fluid passes through the filter member.
According to another feature of the invention, the bottom of the housing has support ribs arranged radially with respect to the fluid outlet.
According to a feature of the invention, the ribs are designed to hold the support grid and to direct the fluid towards the fluid outlet.
According to a feature of the invention, the support ribs project from a surface of the base and extend substantially perpendicular to said surface of the base.
According to a feature of the invention, the internal pressure of the casing is substantially equal to the pressure outside the casing under ground conditions, i.e. when no depression is applied before sealing the cover on the casing.
According to a feature of the invention, the support grid is designed to support the nutritive layer.
According to a feature of the invention, the support grid is also designed to support the filtering member.
According to a feature of the invention, a support grid is arranged between the nutrition layer and the bottom of the housing.
According to a feature of the invention, the support grid is generally cylindrical or polygonal.
According to a feature of the invention, the support grid has a plurality of through holes distributed over the grid, which helps to distribute the fluid that has passed through the nutritive layer over the entire surface of the bottom, in order to drain said liquid more quickly and to prevent the liquid from stagnating in the device.
According to a feature of the invention, the support grid preferably rests on support ribs arranged on the bottom of the housing.
According to one feature of the invention, the support grid is preferably of the same size and shape as the overlying nutritive layer, so as to prevent any deformation of the nutritive layer during the passage of the liquid.
According to one feature of the invention, the perimeter of the support grid is shaped to fit the shape of the housing, so that the grid is retained by friction in the housing and in the internal volume.
According to one feature of the invention, the housing has a third bearing surface 25 designed to receive the support grid, and more particularly the peripheral edge of the support grid.
According to one feature of the invention, the third bearing surface extends within the inner volume of the housing and is an external shoulder of the housing.
According to one feature of the invention, the filter membrane is permeable to fluids and in particular to gases or to liquids of viscosity enabling microbial filtration without any solid particles.
According to another feature of the invention, the filter member is a microorganism-impermeable filter member, preferably a bacteria-impermeable filter member, which enables microorganisms and bacteria to be retained on the surface of the at least one filter member.
According to a feature of the invention, the filtering means is distinct from the nutritive layer.
According to a feature of the invention, the filter membrane is permeable to the nutrients and any additives comprised in the nutrient layer.
According to a feature of the invention, the filtering member is a porous membrane and may be made, for example, of one or more materials or derivatives of these materials, such as latex, polytetrafluoroethylene, poly (vinylidene fluoride), polycarbonate, polystyrene, polyamide, polysulfone, polyethersulfone, cellulose or a mixture of cellulose and nitrocellulose.
Advantageously, the filtering means has a surface above the surface of the nutritive layer, so that the surface of said nutritive layer is completely covered by the filtering means.
Preferably, the filter member is white or similarly colored, which helps to optimize the differentiation of colored colonies on its surface. Alternatively, the filter member may be dark to facilitate visibility of white or cream-colored colonies.
Advantageously, the filtering capacity and hydrophilicity of the filter member serve to enable and optimize passage of nutrients and any additives in the nutrient layer to the upper surface of the filter member after rehydration thereof, while preventing or limiting migration of filtered bacteria, yeasts and the like to the upper surface of the filter member in the opposite direction.
Advantageously, the filtering member has pores with a diameter comprised between 0.01 μm and 0.8 μm, preferably between 0.2 μm and 0.6 μm, in order to retain bacteria, yeasts and moulds on its surface. According to a specific embodiment, the filtering means have pores with a diameter between 0.25 μm and 0.6 μm, for example between 0.3 μm and 0.6 μm or between 0.4 μm and 0.6 μm. Alternatively, a layer without measurable pores may be used, such as a dialysis membrane. For example, the filter member may be "Fisherbrand" sold by Fisher Scientific Company L.L.C. of Industrial road 300, pittsburgh, pa., code 15275 TM Typically a membrane filter "series, or a membrane in the" Nitocellulose membrane filter "series manufactured by Zefon International, inc, of tokala, ohara, florida, usa (zip code: 34474) zeofen International No. 5350.
According to a feature of the invention, the nutritive layer is arranged below the filtering means and preferably in contact with said filtering means.
According to a feature of the invention, the nutrient layer comprises a carrier comprising a microbiological culture medium.
According to a feature of the invention, the carrier can be made of various absorbent compounds, preferably with very high water retention capacity, such as rayon, cotton, natural or chemically modified cellulose fibers (for example carboxymethyl cellulose), absorbent or superabsorbent chemical polymers (for example polyacrylates, acrylate/acrylamide copolymers).
According to another characteristic of the invention, the support can be impregnated with the microbial culture medium in liquid form.
Advantageously, the microbial culture medium may advantageously be dehydrated, i.e. have a water activity (Aw) incompatible with microbial development. Alternatively, the carrier may be covered with the microorganism culture medium in powder form or a component thereof or dry impregnated. Alternatively, liquid impregnation may be performed by adding the powder after dehydration.
By microbial culture medium is meant a culture medium comprising nutrients required for the survival and/or growth of the microorganism, in particular one or more of the following nutrients: carbohydrates including sugars, peptones, growth promoters, mineral salts and/or vitamins, etc. In practice, the skilled person will select the microbial culture medium according to the target microorganism, based on well-known criteria available to said skilled person. The nutritional layer may comprise additives such as:
-one or more selective agents, such as inhibitors or antibiotics, which promote the growth and development of one species/strain of a particular organism but not another;
-buffer, colorant.
Typically, the nutritive layer may also comprise a substrate for detecting an enzyme or metabolic activity in the target microorganism using a directly or indirectly detectable signal. For direct detection, the matrix may be attached to a moiety that serves as a fluorescent or chromogenic label. For indirect detection, the nutrition layer according to the invention may also comprise a pH indicator, which is sensitive to pH changes caused by consumption of the matrix, which reveals the growth of the target microorganism. The pH indicator may be a chromophore or a fluorophore. Neutral red, aniline blue, and bromocresol blue are exemplary chromophores. 4-methylumbelliferone, hydroxycoumarin derivatives and resorufin derivatives are all fluorophores. Thus, a preferred PC-PLC fluorescent matrix for carrying out the method according to the invention is 4-methyl-umbelliferyl-phosphocholine (4 MU-CP).
According to a feature of the invention, the microbial culture medium of the nutritive layer is dehydrated in the delivery configuration of the device according to the invention before use. In this case, after the carrier of the nutrition layer is impregnated by the dehydrated microorganism culture medium dry method, the nutrition layer may be subjected to a calendering operation. The pressure and heat generated by calendering enables the dehydrated microbial culture medium in the carrier of the nutritive layer to be retained and remain stable over time, which ensures the retention of nutrients and any additives in the nutritive layer. Calendering of the nutritive layer also helps ensure that the surface of the nutritive layer is flat and smooth. Calendering also helps to accelerate rehydration of the nutritive layer relative to an uncalendered nutritive layer due to the resulting compression of the nutritive layer. If the carrier is made of fibres, this compression, associated with the presence of dehydrated medium in the nutritive layer, generates a significant increase in the capillary capacity of the nutritive layer, which causes it to rehydrate almost immediately. This may also lead to a suction phenomenon of the individual microbial filtration members arranged against the carrier surface. Thus, the microbial filtration member may be pressed against the nutritive layer, thereby eliminating or reducing the space between the two and ensuring optimal microbial growth and/or survival on the surface of the microbial filtration means. This helps to avoid the need for a bonding means between the microbial filtration member and the nutritive layer when they are separate (e.g. to avoid the need for a bonding layer). This represents a significant advantage as such a means of attachment will slow the passage of nutrients and any additives from the rehydrated nutrient layer to the microorganisms present on the microbial filtration means, thereby reducing the chances of growth and/or survival of these microorganisms.
Preferably, the nutritive layer may be a microbial culture device described in patent application FR19/03751 filed on 8.4.2019.
According to a feature of the invention, the nutrition layer is a microbial culture device comprising some or all of the dehydrated microbial culture medium in powder form, the microbial culture medium having at least two portions made of absorbent hydrophilic material having an at least substantially flat upper surface, wherein the dehydrated microbial culture medium in powder form is arranged between two successive portions and the microbial culture medium comprises at least one gelling agent in powder form.
Many absorbent, hydrophilic and water-insoluble materials may be used to make the absorbent material portion of the microbial culture device according to the present invention. These materials are selected primarily for their absorption capacity, their ability to retain aqueous solutions and their ability to allow aqueous solutions to pass in all directions.
According to a feature of the invention, the absorbent material is made in part from a matrix of short nonwoven fibers that form an assembly having structural integrity and mechanical consistency. Particularly suitable substrates are made from natural cellulose fibers (e.g. cotton) or synthetic cellulose fibers (e.g. rayon), modified cellulose fibers (e.g. carboxymethyl cellulose and nitrocellulose) and absorbent chemical polymer fibers (e.g. polyacrylates and acrylate/acrylamide copolymers). Advantageously, the absorbent material is partly made of a nonwoven fabric made of cellulose fibres.
In the present invention, the absorbent hydrophilic material portions of a given device may have the same mass density or different mass densities. Similarly, the absorbent hydrophilic material portions of a given microbial culture device according to the present invention may have the same thickness or different thicknesses.
According to one characteristic of the invention, the portion of absorbent hydrophilic material has a density of 0.045g/cm 3 To 0.10g/cm 3 Preferably 0.05g/cm 3 To 0.07g/cm 3 Mass density of (d) in between.
The absorbent hydrophilic material portion of a given microbial cultivation device according to the present invention may have a thickness of between 0.5mm and 2 mm. Preference is given toThe absorbent hydrophilic material portion has a thickness of 0.8mm to 1.8mm, more preferably between 1mm to 1.5 mm. The surface of the absorbent hydrophilic material portion is 1cm 2 To 40cm 2 Preferably between 10cm 2 To 30cm 2 More preferably between 15cm 2 To 25cm 2 In the meantime.
According to a feature of the invention, the absorbent hydrophilic material portion is capable of retaining a volume of water greater than 2mL, preferably greater than 3 mL. Thus, it had a thickness of 25cm after calendering 2 And a porous support of 1mm thickness will be able to hold 3mL of water.
According to a feature of the invention, the microbial culture apparatus has undergone a calendaring operation. The pressure and heat generated by the calendering enables the dehydration reaction medium in the microbial culture device, including the gelling agent or agents, to be retained and remain stable over time, which ensures that different elements (e.g., nutrients) are retained between the absorbent hydrophilic material portions.
Preferably, the calendering is performed at a temperature higher than ambient temperature, preferably between 30 ℃ and 60 ℃. Temperatures below 60 ℃ ensure that the thermolabile compounds in the medium do not denature.
Calendering also ensures that the reaction medium is retained within the microorganism culture apparatus, thereby facilitating its handling.
According to another characteristic of the invention, the culture medium also comprises at least one gelling agent, also in powder form. Once activated and rehydrated by the liquid to be analyzed, the gelling agent or agents help create an ensemble having a degree of structural integrity and mechanical consistency. In addition, by contacting the culture medium, the gelling agent or agents provide a gelling consistency to the microorganism culture device that promotes establishment of the microorganism and enables elements (e.g., nutrients or active agents) that make up the culture medium to be as close as possible to the microorganism.
According to another characteristic of the invention, the gelling agent is one of the following: xanthan gum, alginate, gellan gum, galactomannan gum, locust bean gum, starch or a mixture thereof.
Preferably, the cultivation of the device according to the inventionThe base comprises 0.0030g/cm 3 To 0.020g/cm 3 At least one gelling agent.
The invention also relates to an assembly comprising at least one fluid supply and at least one device according to the invention, said device being connected to the fluid supply via a fluid inlet.
Advantageously, the assembly further comprises a second device according to the invention, mounted in parallel with the first device according to the invention.
According to another characteristic of the invention, the assembly comprises a plurality of devices according to the invention mounted parallel to each other.
Advantageously, the plurality of devices may be identical or dedicated to different microorganism types.
According to another feature of the invention, the fluid outlet may be connected to a receiver or otherwise.
According to a feature of the invention, one or more devices according to the invention may be located in a compartment in an industrial pipeline or other structure.
According to a feature of the invention, the device has a capacity to enable analysis of 100mL of liquid.
Drawings
The invention will be better understood from the following description of an embodiment of the invention, given as a non-limiting example and explained with reference to the accompanying schematic drawings. The schematic drawings are listed below:
FIG. 1 is a perspective view of a device for determining contamination according to the present invention;
FIG. 2 is a middle cross-sectional view of the device of FIG. 1;
FIG. 3 is a detailed view of FIG. 2;
FIG. 4 is a partial perspective view of the apparatus according to the present invention showing the support grid located in the housing;
FIG. 5 is a perspective bottom view of a cover of the device according to the present invention;
FIG. 6 is a perspective view of the interior of the housing of the device according to the present invention;
FIG. 7 is a cross-sectional view of a cover and filter member of the device according to the invention;
FIG. 8 is a cross-sectional view of the housing of the device according to the present invention;
fig. 9 is a schematic view of a first step of a first mode of use of the device according to the invention;
fig. 10 is a schematic view of a second step of the first mode of use of the device according to the invention;
fig. 11 is a schematic view of a third step of the first mode of use of the device according to the invention;
fig. 12 is a schematic view of a first step of a second mode of use of the device according to the invention;
fig. 13 is a schematic view of a second step of a second mode of use of the device according to the invention; and
fig. 14 is a schematic view of a third step of the second mode of use of the device according to the invention.
Detailed Description
The device 1 for determining the contamination of a fluid by microorganisms according to the present invention comprises a housing 2, a cover 3, a filter member 4, a nutrition layer 5 and a grid 6, as particularly shown in fig. 2.
As shown in fig. 1, only the cover 3 and the housing 2 are visible from the outside. Advantageously, the cover 3 and the housing 2 are sealed together by ultrasound. According to the cross-section shown in fig. 2, the filtering member 4 is arranged in the inner volume of the housing 2, the nutrient layer 5 is arranged below, preferably in contact with, the filtering member 3, and the support grid 6 is arranged between the nutrient layer 5 and the bottom 22 of the housing 2.
According to the invention, the filtering membrane 4 is permeable to fluids and in particular to gases or to liquids whose viscosity enables a microbiological filtration free from any solid particles.
The device 1 further comprises a fluid inlet 11 and a fluid outlet 12, as shown in fig. 1. The fluid inlet 11 is located on the cover 3 and the fluid outlet 12 is located on the housing 2, preferably on the bottom of the housing 2. The fluid inlet 11 and the fluid outlet 12 open into the inner volume of the housing 2, as shown in fig. 2. The device according to the invention is fluid tight when the inlet and the fluid outlet are blocked by the plug.
As shown in fig. 2, for example, the fluid inlet 11 protrudes into the interior volume of the housing 2 relative to the inner surface 31 of the cover 3. Preferably, the fluid inlet 11 is arranged substantially in the centre of the lid 3. The fluid inlet has: a first end 11a, extending outside the lid 3, designed to cooperate with a plug (not shown); and a second end 11b, which extends inside the internal volume, the second end being provided with at least one lateral orifice 11c, which is open to the internal volume and is oriented to eject the fluid at least partially towards the internal surface 31 of the cover 3.
As shown in fig. 2, for example, the fluid outlet 12 has: a first end 12a, extending outside the housing 2, designed to cooperate with a plug; and a second end 12b, which is flush with the bottom 22 of the housing 2.
The fluid inlet 11 and the fluid outlet 12 extend along an axis substantially secant and preferably perpendicular to the bottom of the cover 3 or the housing 2, respectively. In the example shown in fig. 2, the fluid inlet 11 and the fluid outlet 12 are aligned and arranged opposite to each other.
The housing 2 of the device 1 is described in more detail below with reference to fig. 1, 2, 6 and 8. As shown in fig. 1, the housing 2 is substantially cylindrical.
As shown in fig. 2, the housing 2 has an interior volume bounded by at least one side wall 21 and a bottom 22.
Fig. 2 and 8 show a cross-section of the housing 2 showing the first circumferential bearing surface 23 shaped to complementarily cooperate with the edge 33 of the cover 3. Furthermore, according to the invention, the casing 2 has at least one second bearing surface 24 extending in the internal volume and shaped to receive a peripheral portion 41 of the filter member 4, said second bearing surface 24 being shaped to cooperate with the portion 34 of the cover 3 so that the peripheral portion 41 of the filter member 4 is clamped between the cover 3 and the casing 2, as shown in detail in fig. 3. Furthermore, the casing 2 has a third bearing surface 25 designed to receive the support grid 6 and, more specifically, the peripheral edge 65 of the support grid 6. The third bearing surface 25 extends within the inner volume of the housing 2 and is an outer shoulder 26 of the housing 2, as shown in fig. 8.
As shown in fig. 8, the respective bearing surfaces 23, 24, 25 have different diameters and advantageously the first bearing portion 23 has a larger diameter than the second bearing portion 24, the second bearing portion 24 in turn having a larger diameter than the third bearing portion 25.
As best shown in fig. 6, the bottom 22 of the housing 2 has a plurality of support ribs 27 designed to support the support grid 6 and direct fluid toward the fluid outlet 12. The support ribs 27 are arranged radially with respect to the fluid outlet 12. The support ribs 27 project from the surface of the base 22 and extend generally perpendicular to the surface of the base 22, as shown in cross-section in fig. 8.
As shown in particular in fig. 8, the bottom 22 of the casing 2 has a surface 28 extending radially around the fluid outlet 12 as far as the lateral wall 21 of the casing 2, said inner surface 28 being inclined and converging towards the fluid outlet 12 to facilitate the discharge of the fluid. In the example shown in fig. 2 and 8, the inclination β of the surface 28 of the bottom 22 is strictly less than 0 ° and between-5 ° and-10 °. The applicant has carried out tests which demonstrate that if the bottom is flat, the fluid is only partially expelled or not expelled at all.
The cover 3 of the device 1 is described in more detail below with reference to fig. 1, 2, 5 and 7.
As shown in fig. 1, the cap 3 has a generally cylindrical base with a fluid inlet 11 at the top. The cover 3 has an inner surface 31 that closes the inner volume of the housing 2. The inner surface of the cover 3 extends radially around the fluid inlet 11 as far as the peripheral edge 32 of the cover 3, said inner surface 31 being inclined and converging towards the fluid inlet 11. Thus, the inner surface 31 has a general frustoconical shape extending and widening from the fluid inlet 11 up to the peripheral edge 32 of the cover 3. In a variant not shown, the inner surface may be curved.
In the example shown in fig. 2 and 7, the inclination α of the inner surface 31 of the cover 3 is strictly greater than +4 ° and between +5 ° and +15 °. The applicant has carried out tests which demonstrate that if the inner surface of the cover has an inclination equal to or less than 4 °, the fluid is distributed unevenly on the filtering member 4.
As shown in fig. 7, the filter member 4 is disposed in the housing 2 at a given distance h from the fluid inlet 11. Advantageously, the given distance h is strictly greater than 1mm, preferably between 1.5mm and 5 mm. In fact, tests carried out by the applicant have demonstrated that the filtering means 4 must be at a given distance h to enable the growth of microorganisms when the device 1 according to the invention is cultivated. The given distance h depends on the minimum air volume required for growing the microorganisms. In the examples shown in fig. 1, 2, 5 and 7, the cover 3 is transparent or translucent, which enables the growth of microorganisms to be observed.
According to a feature of the invention, the support grid is designed to support the nutritive layer and the filtering member.
The support grid 6 of the device 1 is described in more detail below with reference to fig. 2 and 4.
As shown in fig. 2, the support grid 6 is arranged between the nutrition layer 5 and the bottom 22 of the housing 2. The support grid 6 has an overall cylindrical shape and is the same size as the nutrition layer 5.
As shown in fig. 4, the support grid 6 has a plurality of through holes 61 distributed over the whole grid 6, which helps to distribute the fluid that has passed through the nutritive layer 5 over the whole surface 28 of the bottom 22, so as to drain said fluid as quickly as possible.
As shown in fig. 2, the support grid 6 rests on the support ribs 27 of the housing 2. The support grid 6 has a peripheral edge 65 which rests on the third bearing surface 25 of the housing 2, as shown in fig. 2 and 3.
In a variant not shown, the grid rests only on the third bearing surface 25 of the housing, and the support ribs are optional.
Two possible uses of the device according to the invention are described below with reference to fig. 9 to 14.
According to a first use illustrated in fig. 9 to 11, a syringe 100 containing the fluid to be analyzed or any other element comprising a piston is connected to the fluid inlet 11 of the device 1 and a collector 101 or another syringe is connected to the fluid outlet 12 of the device 1, as shown in fig. 9. A three-way valve 102 is located between the device 1 and the injector 100, the injector 100 being indirectly connected to the fluid inlet 11 of the device 1. First, as shown in fig. 9, the first inlet 102a of the valve 102 is connected to the injector 100 and closed, the second inlet 102b is opened and connected to the air inlet, and the outlet 102c is also opened and connected to the fluid inlet 11 of the device. This enables a vacuum to be created before the device is used and before the fluid to be analysed/filtered is introduced.
Subsequently, as shown in fig. 10, the first inlet 102a of the valve 102 is inserted and the fluid is injected into the device 1, the piston of the syringe 100 enabling the fluid to be pushed into the device 1, then the second inlet 102b of the valve is closed and the outlet 102c is opened to enable the fluid to flow through the device 1.
Once all the fluid to be analyzed has been injected through the device 1, the fluid is recovered in the collector 101, as shown in fig. 11.
According to a second use, illustrated in fig. 12 to 14, a canister 103 containing the fluid to be analyzed is connected to the fluid inlet 11 of the device 1, and a syringe 104, or any other element comprising a piston and enabling the aspiration of the fluid, is connected to the fluid outlet 12 of the device 1, as shown in fig. 12. A three-way valve 105 is located between the device 1 and the injector 100, the injector 100 being indirectly connected to the fluid inlet 11 of the device 1. First, as shown in fig. 12, the first inlet 105a of the valve 102 is connected to the collector 103 and closed, the second inlet 105b is opened and connected to the air inlet, and the outlet 105c is also opened and connected to the fluid inlet 11 of the device 1.
Subsequently, as shown in fig. 13, the first inlet 105a of the valve 105 is opened and the fluid contained in the collector 103 is aspirated by the syringe 104 located downstream of the device 1, then the second inlet 105b of the valve is closed and the outlet 105c is opened to enable the fluid to flow through the device 1.
Once all the fluid to be analysed has been aspirated through the device 1, the fluid is recovered in the syringe 104, as shown in figure 14. Of course, the capacity of the syringe and the canister or collector is adapted to the volume of fluid to be analyzed.
Regardless of the use of the device described above, once the fluid has been injected (fig. 11 and 14), it is advantageous to inject sterile air via the air inlet 110 to eliminate any stagnant water from the surface of the filtering means 4 of the device 1.
Indeed, even a small amount of fluid present on the surface of the filtering member may result in the spread of bacterial colonies, false negatives or difficulty in counting the colonies after the incubation step. For this purpose, sterile air is drawn by syringe 100 or canister 103 through valves 102, 105 and then injected into device 1 in order to dry the surface of filter member 4.
The advantages of these uses are that these use techniques are simple, they do not require effort (especially the first use), the workflow lasts less than one minute in total, no laboratory infrastructure is required to perform these operations, no microbiologically qualified personnel are required, and the operations are very reproducible.
The invention is of course not limited to the embodiments described and/or illustrated in the drawings. The invention may be modified, particularly in respect of the construction of different elements or by substitution of technical equivalents, without thereby departing from the scope of protection of the invention.
Claims (13)
1. A device (1) for determining contamination of a fluid by a microorganism, the device (1) comprising:
-a casing (2) having an internal volume delimited by at least one side wall (21) and a bottom (22);
-a cover (3) closing the casing (2) and positioned opposite the bottom (22);
-a fluid inlet (11) arranged on the cover (3) open to the inner volume of the housing (2), the fluid inlet (11) extending along an axis perpendicular to the cover (3);
-at least one filtering member (4) arranged in said internal volume;
-at least one nutrient layer (5) comprising a microbial culture medium;
characterized in that said device (1) comprises:
-a fluid outlet (12) arranged on the housing (2), open to the inner volume of the housing (2), and in that the cover (3) has an inner surface (31) in the inner volume extending radially around the fluid inlet (11) up to a peripheral edge of the cover (3), which inner surface (31) is inclined or curved and converges towards the fluid inlet (11);
-a support grid configured to support the nutritive layer (5);
and in that the bottom (22) of the housing (2) has a surface (28) extending radially around the fluid outlet (12) as far as the side wall (21) of the housing (2), which surface (28) is inclined and converging towards the fluid outlet (12).
2. Device according to claim 1, characterized in that the inclination of the inner surface (31) of the cover (3) is greater than +4 °.
3. Device according to claim 1, characterized in that the inclination of the surface (28) of the bottom (22) is less than 0 °.
4. The device according to claim 1, wherein the fluid inlet (11) protrudes into the inner volume of the housing (2) with respect to the inner surface (31) of the cover (3).
5. Device according to claim 1, characterized in that said filtering means (4) are arranged in said casing (2) at a given distance h from said inlet (11), said given distance h being greater than 1 mm.
6. Device according to claim 1, characterized in that the housing (2) has a first circumferential bearing surface (23) shaped to cooperate complementarily with an edge (33) of the cover (3).
7. The device according to any one of claims 1 to 6, characterized in that said casing (2) has at least one second bearing surface (24) extending in said internal volume and shaped to receive a peripheral portion (41) of said filtering member (4), said second bearing surface (24) being shaped to cooperate with a portion (34) of said cover (3) so that said peripheral portion (41) of said filtering member (4) is clamped between said cover (3) and said casing (2).
8. The device according to any one of claims 1 to 6, characterized in that the bottom (22) of the housing (2) has support ribs (27) arranged radially with respect to the fluid outlet (12), the support ribs (27) being designed to hold the support grid (6) and to direct the fluid towards the fluid outlet (12).
9. Device according to claim 2, characterized in that said inclination of said inner surface (31) of said cover (3) is comprised between +5 ° and +15 °.
10. Device according to claim 9, characterized in that said inclination of said inner surface (31) of said cover (3) is 10 °.
11. The device according to claim 3, characterized in that said inclination of said surface of said bottom is between-5 ° and-10 °.
12. The device according to claim 11, characterized in that said inclination of said surface of said bottom is comprised between-6.5 ° and-7.5 °.
13. Assembly comprising at least one fluid supply and at least one device (1) according to any one of claims 1-12, the device (1) being connected to the fluid supply by means of the fluid inlet (11).
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1903807 | 2019-04-09 | ||
| FR1903807A FR3094986B1 (en) | 2019-04-09 | 2019-04-09 | Device for determining bacteriological contamination in a fluid |
| FR1911739 | 2019-10-21 | ||
| FR1911739A FR3094987B1 (en) | 2019-04-09 | 2019-10-21 | Device for determining bacteriological contamination in a fluid |
| PCT/FR2020/000076 WO2020208309A1 (en) | 2019-04-09 | 2020-03-31 | Device for determining the presence of a bacteriological contamination in a fluid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113677434A CN113677434A (en) | 2021-11-19 |
| CN113677434B true CN113677434B (en) | 2023-04-11 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202080027540.2A Active CN113677434B (en) | 2019-04-09 | 2020-03-31 | Device for determining the presence of bacterial contamination in a fluid |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN113677434B (en) |
| FR (2) | FR3094986B1 (en) |
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Also Published As
| Publication number | Publication date |
|---|---|
| FR3094987B1 (en) | 2021-05-21 |
| FR3094987A1 (en) | 2020-10-16 |
| FR3094986A1 (en) | 2020-10-16 |
| CN113677434A (en) | 2021-11-19 |
| FR3094986B1 (en) | 2021-05-21 |
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