Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
  • C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
  • C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
• • 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
  • C12Q1/06—Quantitative determination
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D2201/00—Details relating to filtering apparatus
  • B01D2201/20—Pressure-related systems for filters
  • B01D2201/202—Systems for applying pressure to filters
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/02—Devices for withdrawing samples
  • G01N1/10—Devices for withdrawing samples in the liquid or fluent state
  • G01N1/14—Suction devices, e.g. pumps; Ejector devices
  • G01N2001/1418—Depression, aspiration
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/02—Devices for withdrawing samples
  • G01N1/10—Devices for withdrawing samples in the liquid or fluent state
  • G01N1/20—Devices for withdrawing samples in the liquid or fluent state for flowing or falling materials
  • G01N1/2035—Devices for withdrawing samples in the liquid or fluent state for flowing or falling materials by deviating part of a fluid stream, e.g. by drawing-off or tapping
  • G01N2001/2071—Removable sample bottle
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

• the invention relates to a device for collecting micro-organisms from a liquid in a filter, a method for collecting micro-organisms from a liquid and a method for identifying and quantifying micro-organisms.
• Such a device is generally known, such as for instance at different certified or non- certified agencies which are employed in the Netherlands, on the basis of the Drinking Water Decree, to determine the quantity of micro-organisms, and legionella bacteria in particular, in a determined quantity of liquid, in particular water.
• Companies and organizations legally obliged to prevent legionella must have their water supply system tested periodically by an external service provider.
• This service provider takes on-site samples of the water which are subsequently propagated in the laboratory using a bacterial culture and then analysed by means of counting. In this way it takes a minimum of a week before the result of such a test is known. Many litres of water must moreover be transported, this entailing high energy and transport costs.
• the object of the invention is to design a device for efficient collection of microorganisms such as the legionella bacteria from water, wherein the collected micro-organisms take up the smallest possible volume in order to minimize transport costs to the testing location.
• This device has to be user-friendly and environmentally-friendly and it must be possible to use it many times. It must further be possible to analyse in reliable manner the micro-organisms collected in this way.
• the present invention has for its object to provide a device of the type stated in the preamble, comprising
• a filter holder suitable for placing at least during use on the balloon holder (3) and comprising a filter (8), the pore size of which is such that the micro-organisms are stopped;
• the gas supply to the device is preferably provided by a gas bottle such as a gas cartridge.
• the device according to the invention more preferably comprises a filter holder having on the underside a lower filter shell part (5) with thickened portions fitting into the recesses of the balloon holder, the filter (8) with filter plate (6), and on the upper side an upper filter shell part (9).
• the different components of the filter holder can also be attached to each other, this being the case particularly for the filter plate (6) and the upper filter shell part (9). All components of the filter holder are preferably manufactured from a polymer, more preferably from ABS (acrylonitrile butadiene styrene) polymer.
• the balloon holder (3) comprises a ring with recesses (3') and is provided with a first edge (3") lying substantially perpendicularly of the ring and having a dimension such that it fits into the upper side of the bottle and wherein the inner ring of the ring is reinforced with a second edge (3 "') with a dimension such that the lower filter shell part (5) fits therein.
• the first edge is particularly provided with an O-ring (2).
• the balloon holder is preferably a synthetic polymer, more preferably of nylon. Any gas can in principle be used, although C0 2 is recommended since use thereof is simple and it is freely available on the market.
• the volume of the bottle preferably amounts to 1000-2000 ml, more preferably 1500-1800 ml, in particular 1650-1700 ml, more particularly 1680 ml.
• the volume of the balloon preferably amounts to 400-600 ml, more preferably 450-550 ml, in particular 520 ml.
• the ratio of the volume of the balloon and the volume of the bottle must however always be such here that sufficient pressure can be developed. This ratio preferably lies between 0.20 and 0.60, more preferably between 0.25 and 0.35, and is in particular 0.30. It will be evident to a person with ordinary skill in the art that the neck of the balloon may not be so narrow that it is squeezed together by the pressure to be developed.
• the filter holder particularly has a thickness of 10 mm to 25 mm, preferably of 15 mm to 20 mm.
• the filter plate (6) preferably has a pore size of 0.1 mm to 5 mm.
• the filter (8) preferably has a pore size of 1 ⁇ - 3 ⁇ , in particular about 2 ⁇ .
• the maximum thickness of the filter holder is particularly determined by the opening of the letterbox which can receive a package for mailing.
• the pore size of the filter must be such that the micro-organisms for analysis are stopped.
• an insert part (17) with punch (18) and press-through pin suitable for causing the gas to flow via the cover (19) out of a gas bottle such as a gas cartridge.
• the material from which the press- through pin, the punch and the insert part are made is not generally of critical importance. Suitable materials are alloys; hardened steel for instance for the press-through pin and the punch and brass for instance for the insert part.
• Preferably arranged on the filter (8) in a determined quantity is a standard microorganism with which the application of the device according to the invention can be controlled in determining a quantity of micro-organisms in a liquid.
• the present invention also provides a method for collecting micro-organisms from a liquid while applying the device according to the invention as described above, wherein
• the filter holder with filter plate with the collected micro-organisms is removed and the micro-organisms isolated.
• the gas is supplied particularly using a gas cartridge.
• the micro-organisms legionella bacteria are preferably collected in this way.
• This separate filter holder provides the great advantage that it can be removed without the filter plate or filter having to be touched.
• the pressure developed amounts in this method to preferably 2-3.5 bar, more preferably 2.5-3 bar. It is further possible in an embodiment of the invention, instead of arranging a standard micro-organism on the filter as described above, to add to the liquid a standard micro- organism with which the application of the device according to the invention can be controlled in determining a quantity of micro-organisms in a liquid.
• Another aspect of the invention is formed by a method for identifying and quantifying micro-organisms by applying a device according to the invention, wherein nucleic acid such as DNA from the collected micro-organisms is preferably amplified while applying nucleic acid amplification techniques, such as for instance the polymerase chain reaction, and analysed using for instance sequencing.
• nucleic acid such as DNA from the collected micro-organisms
• nucleic acid amplification techniques such as for instance the polymerase chain reaction
• the device according to the invention comprises a hose (22) which is suctioned centrally into the balloon. It is hereby generally easier to empty the whole balloon.
• the gas supply in the device according to the invention is preferably controllable, for instance by applying a gas pressure controller at the connection point (20) for gas supply and/or by applying a valve (23) when a gas bottle such as a gas cartridge is used.
• the device according to the invention is preferably portable and provides the option of automatically filtering a quantity of liquid at a location under all conditions with the purpose of collecting the micro-organisms present in the liquid in a filter, following which they can be identified and/or quantified. Envisaged in the first instance here is the detection and quantifying of legionella bacteria for the purpose of legionella prevention. Samples of the water are generally taken on site by a service provider and subsequently analysed using a bacterial culture in the laboratory. The device according to the invention makes it possible for companies to draw off and filter water themselves.
• the filter which is situated in a closed filter holder, is generally then sent by post to the laboratory, after which the company is informed in the not too distant future of the result.
• the device provides a financially favourable solution for all other companies and consumers wishing to monitor their draw-off points for safety purposes and legionella prevention.
• the device according to the invention is unique in that it enables companies and consumers, at a reasonable price, to monitor the water in their water supply for the presence of the legionella bacteria.
• an external company has usually been brought in to take water samples which then sends these directly to a laboratory for analysis.
• the method according to the invention ensures that companies and consumers can themselves take the water samples in safe manner, whereby no costs need be incurred for an external intermediary plus transport.
• the water is after all immediately processed in the device according to the invention in uniform manner, and the filter plate with filter and collected micro-organisms is sent directly to the laboratory.
• the filter holder is sent unopened and preferably by post to the laboratory. If the culture method is applied, the bacteria must remain alive, wherein the filter with bacteria must be transported in cooled state.
• the use of the device further prevents many errors in monitoring. This is because there can be no more in the balloon than the fixed quantity of water necessary for the analysis.
• the use of an internal monitoring enables later evaluation of the filtration, transport and analysis in the laboratory, this significantly increasing the quality of the result. All this contributes toward a standardization of the whole legionella monitoring process. Because water draw-off, monitoring and analysis are standardized, the margin of error in the whole process will decrease and the quality of the whole monitoring system will thus be increased.
• the device can be reused frequently, although a new balloon and filter holder with a filter therein will have to be placed for each sample.
• Gas cartridges, ready-to-use filter holders and balloons must always be replaced as non-durable items and preferably in the form of disposables.
• FR French patent application
• the detection of bacteria preferably takes place on the basis of DNA testing. This method is quicker and more sensitive than the culture method generally used and, since it is based on the detection of DNA, it does not matter whether the bacteria survive transport or not.
• Another detection method such as the culture method followed by counting, can of course also be applied and this will generally take place in a laboratory. Whichever detection method is used, the result will answer the question of whether the legionella bacteria are present in the filtered water and in what quantity.
• the present invention is further elucidated on the basis of the following drawings of two embodiments of the invention for collecting micro-organisms from a fluid in a filter according to the invention and the balloon holder applied in this device.
• Figure 1 shows an oblique front view of the device according to the invention comprising a bottle (1) with balloon holder (3) and an O-ring (2) therebetween, a balloon (4); a filter holder comprising a filter (8) with filter plate (6), on the underside a lower filter shell part (5), an O-ring (7) and on the upper side an upper filter shell part (9); a cover (19) comprising a pressure-relief valve (14) with venting spring (11) and venting closure (12) and venting O-ring (13), an O-ring (10) between upper filter shell part (9) and cover (19), a part for opening the gas cartridge comprising a punch (18), insert part (17) and a press-through pin (16) with O-ring (15), and outlet for the liquid (21) and a connection point (20) for a gas cartridge.
• Figure 2 shows balloon holder (3) comprising a ring with recesses (3') and provided with a first edge (3") lying substantially perpendicularly of ring (3') and wherein the inner ring of the ring is reinforced with a second edge (3'").
• Figure 3 is an oblique front view of the device according to the invention as shown schematically in Figure 1 but with the addition of hose (22) and a gas cartridge provided with a valve (23).
• FIG. 1 shows in detail an oblique front view of the device according to the invention.
• the device consists of a bottle (1) in which, in a balloon holder (3), a balloon (4), preferably of latex, can be placed.
• the bottle is preferably of plastic, but can also be of any other suitable material such as tin, aluminium or glass.
• the balloon is preferably of latex, but can also be of any other suitable material, such as polyester.
• the water sample is received in the balloon, and added to the water as control is for instance an internal standard with which the quality of the filtration, transport and laboratory analysis can be demonstrated and which therefore significantly increases the reliability of the result.
• a filter holder with a filter (8) therein for instance of polyester, with a pore size such that micro-organisms, in particular bacteria such as for instance legionella bacteria, are stopped, and a filter plate (6).
• a cover (19) is then screwed onto the bottle (1) so that the filter holder clamps onto the balloon.
• Pressure is developed between the wall of bottle (1) and balloon (4) by the outflowing gas, whereby the water is pressed through the filter.
• Pressure- relief valve (14) ensures that the pressure in the bottle does not become too high.
• Figure 2 shows balloon holder (3) for a balloon (4) as shown in Figure 1 , which is an advantageous element in the device according to the invention.
• the recesses serve to create a fixed system in which nothing can be changed, whereby the draw-off of water is standardized.
• first edge (3") then drops into balloon (3).
• the lower filter shell part (5) shown in Figure 1 fits into the second edge (3"').
• Figure 3 shows the device according to Figure 1 , but now with an additional hose (22) for the purpose of allowing more chance of success in suctioning in the solutions with microorganisms for analysis, emptying the whole balloon and allowing the micro-organisms to be collected wholly on the filter.
• the gas supply is preferably also controllable, for instance by applying a valve (23) during use of a gas cartridge.

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• Life Sciences & Earth Sciences (AREA)
• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Engineering & Computer Science (AREA)
• Organic Chemistry (AREA)
• Zoology (AREA)
• Wood Science & Technology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Biotechnology (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• Microbiology (AREA)
• General Health & Medical Sciences (AREA)
• Genetics & Genomics (AREA)
• Molecular Biology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Biomedical Technology (AREA)
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• Biophysics (AREA)
• Analytical Chemistry (AREA)
• Physics & Mathematics (AREA)
• Toxicology (AREA)
• Apparatus Associated With Microorganisms And Enzymes (AREA)

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• 2011
  • 2011-09-11 NL NL2007389A patent/NL2007389C2/nl not_active IP Right Cessation
• 2012
  • 2012-09-07 EP EP12758925.7A patent/EP2753406A1/en not_active Withdrawn
  • 2012-09-07 WO PCT/NL2012/050632 patent/WO2013036127A1/en active Application Filing

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