WO1999041354A1 - Protocol for simulated natural biofilm formation - Google Patents
Protocol for simulated natural biofilm formation Download PDFInfo
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- WO1999041354A1 WO1999041354A1 PCT/US1999/003123 US9903123W WO9941354A1 WO 1999041354 A1 WO1999041354 A1 WO 1999041354A1 US 9903123 W US9903123 W US 9903123W WO 9941354 A1 WO9941354 A1 WO 9941354A1
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- Prior art keywords
- biofilm
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- slide
- reservoir
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- 230000032770 biofilm formation Effects 0.000 title claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 49
- 230000001580 bacterial effect Effects 0.000 claims abstract description 28
- 230000000694 effects Effects 0.000 claims abstract description 19
- 238000012360 testing method Methods 0.000 claims abstract description 19
- 239000002054 inoculum Substances 0.000 claims abstract description 15
- 235000015097 nutrients Nutrition 0.000 claims description 26
- 230000001332 colony forming effect Effects 0.000 claims description 8
- 239000010935 stainless steel Substances 0.000 claims description 7
- 229910001220 stainless steel Inorganic materials 0.000 claims description 7
- 238000013019 agitation Methods 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 239000007900 aqueous suspension Substances 0.000 claims description 4
- 239000002028 Biomass Substances 0.000 claims description 3
- 230000003115 biocidal effect Effects 0.000 claims description 3
- 239000003139 biocide Substances 0.000 claims description 3
- 239000004615 ingredient Substances 0.000 claims description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 2
- 239000011707 mineral Substances 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 239000001888 Peptone Substances 0.000 claims 1
- 108010080698 Peptones Proteins 0.000 claims 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 235000019319 peptone Nutrition 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 239000000047 product Substances 0.000 description 19
- 239000002609 medium Substances 0.000 description 18
- 239000000523 sample Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 229910001868 water Inorganic materials 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000011521 glass Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000009472 formulation Methods 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 230000002906 microbiologic effect Effects 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000001974 tryptic soy broth Substances 0.000 description 3
- 108010050327 trypticase-soy broth Proteins 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229920002444 Exopolysaccharide Polymers 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 241000191940 Staphylococcus Species 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229920000515 polycarbonate Polymers 0.000 description 2
- 239000004417 polycarbonate Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000588986 Alcaligenes Species 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000002064 Dental Plaque Diseases 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000589565 Flavobacterium Species 0.000 description 1
- 206010070245 Foreign body Diseases 0.000 description 1
- 229920002306 Glycocalyx Polymers 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 210000004517 glycocalyx Anatomy 0.000 description 1
- 210000002768 hair cell Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000018343 nutrient deficiency Nutrition 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 239000008257 shaving cream Substances 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000001635 urinary tract Anatomy 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
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- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/04—Flat or tray type, drawers
-
- 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
- C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- 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
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/97—Test strip or test slide
Definitions
- This invention generally relates to methodology for production of experimental biofilm matrices on selected surfaces. More specifically this invention relates to the production of simulated natural bio films for testing the activities of formulated products for inhibition or removal of the simulated biofilms from a support surface. BACKGROUND ART
- Biofilm science is reviewed in publications such as Food Reviews International, 8 (4), 573 (1992); Microbial Ecology in Health and Disease, 8, 305 (1995); Annu. Rev. Microbial, 49, 711 (1995); Applied and Environmental Microbiology, 4014 (Nov. 1996); and "Biofilms” by W. G. Characklis and K. C. Marshall (John Wiley & Sons, Inc., New York, 1989).
- a biofilm consists of cells immobilized on a substratum and embedded in an organic polymer matrix of microbial origin.
- a biofilm is a surface accumulation, which is not necessarily uniform in time or space.
- a biofilm may be composed of a significant fraction of inorganic or abiotic substances held cohesively by the biotic matrix.
- a biofilm is a protective matrix for bacteria, with the essential purpose of survival in an environment of limited nutrient supply.
- Biofilms consist of both host microbes and their extracellular products, usually exopolysaccharides. Microbes have a tendency to form these protective exopolysaccharide matrices after they have adhered to a surface. The formation of biofilm complexes requires only humid conditions and/or water systems and contact with a support surface. With respect to nutrients, a nutrient deficiency in fact may increase the biofilm formation capacity of microbes, as reported in Adv. Appl. Microbiol., 29, 93 (1983). -2-
- Biofilms generally can be produced by almost all microbes under suitable conditions.
- the most common biofilm producers belong to the genera Pseudomonas, Enter obacter, Flavobacterium, Alcaligenes, Staphylococcus, and Bacillus.
- biofilms can cause energy losses and blockages in condenser and heat exchange tubes, interfere with water and waste water systems, and form drag-inducing encrustations on ship hulls.
- a biofilm formed by bacteria such as a Pseudomonas species can be the systemic causation of diseases of the lungs or the gastrointestinal and urinary tracts. Additionally, a biofilm formed by bacteria such as Staphylococcus species can be a serious contamination problem in foreign-body instruments such as cardiac pacemakers, catheters, prostheses, artificial valves, and the like. Dental plaque is also a typical form of biofilm.
- biofilms One of the main purposes of natural biofilm formation is for the protection of the host microbes from a hostile environment. As a consequence, there is a combative interaction between microbes in biofilms and biocidal vehicles such as preservatives, disinfectants and antibiotics. Further, the sessile mode of bacterial growth in biofilms differs from that of the same bacteria species that are present as planktonic cells in a circulating aqueous medium which interfaces with the biofilm. Biofilms also act as a trap for nutrient acquisition, which is an important factor when bacteria grow on surfaces and the nutrient supply is oligotrophic.
- biofilm formation Because of the manifold ramifications of biofilm formation, there is a serious commitment to biofilm research in a broad range of scientific investigations.
- Methods of studying biofilm formation include microbiological, physical, and chemical methods. When microbes from extreme natural environments are cultured, standard plate counts usually do not provide accurate estimates. Thus, the classical evaluation methods relying on microbiological plating have questionable value with respect to the laboratory study of biofilms which are intended to achieve authentic correspondence with natural biofilms which exist in the biosphere.
- formation of a natural type biofilm in the laboratory environment is difficult, mainly because there are no standardized methodologies currently available. -3-
- One or more objects of the present invention are accomplished by the provision of a method for controlled biofilm formation in accordance with a monitored protocol which comprises (1) arranging at least one removable slide within the internal reservoir of a reactor; (2) introducing a bacterial inoculum and aqueous nutrient medium into the reservoir; (3) providing the aqueous suspension medium with agitation for an incubation period sufficient to initiate bacterial attachment to the slide surface; (4) draining the suspension medium from the reservoir; (5) recharging the reservoir with aqueous nutrient medium to maintain bacterial growth for an elapsed time period sufficient to establish a biofilm of prescribed thickness on the slide surface; and (6) retrieving the slide from the reservoir, and optionally determining the population of bacterial Colony Forming Units per square centimeter of the biofilm on the slide surface.
- a suitable laboratory scale reactor can comprise an annular reactor which has dimensions between about 2-30 centimeters in diameter and 5-50 centimeters in height.
- a preferred type of reactor is one constructed of polycarbonate, with a rotating inner drum which is adapted to secure stainless steel type retrievable slides.
- this invention provides a method for controlled biofilm formation in accordance with a monitored protocol which comprises (1) arranging removable slides within the internal reservoir of a reactor; (2) introducing a bacterial inoculum and aqueous nutrient medium into the reservoir; (3) providing the aqueous suspension medium with agitation for an incubation period sufficient to initiate bacterial attachment to the slide surfaces; (4) draining the suspension medium from the reservoir; (5) recharging the reservoir with aqueous nutrient medium at least once every 24 hours of elapsed time period; (6) repeating the step (5) regimen for an extended period sufficient to establish a biofilm of prescribed thickness on the slide surfaces; and (7) retrieving the slides from the reservoir, and optionally determining the population of bacterial Colony Forming Units per square centimeter of the biofilm on the slide surfaces.
- a standardized Mineral Salts nutrient medium can be employed for the biofilm bacterial growth cycles. Suitable nutrient media for biofilm formation are described in technical publications such as Biotechnol. Bioeng., 5 (5), 459 (1997); incorporated herein by reference.
- a typical nutrient medium contains sources of carbon, nitrogen, phosphate and trace nutrients.
- the bacterial inoculum is derived from a natural biofilm. This is illustrated by a bacterial inoculum which is extracted from a household drain biofilm.
- the monitored protocol in the reactor is designed to provide essential correspondence with simulated household drain conditions, thereby promoting the formation of a simulated biofilm in the reactor which has close correspondence with a household drain natural biofilm.
- simulated biofilm refers to a derivative biofilm which has essential phenotypic correspondence with the bacterial consortia of its sampled natural biofilm.
- natural biofilm refers to a biospheric surface- mediated bacterial consortium which is in a dynamic relationship with environmental parameters. MODES OF CARRYING OUT THE INVENTION
- the present invention methodology has particular advantage for the preparation of simulated biofilms which are a convenient and reliable vehicle for testing the activities of formulated products, where said products are intended for inhibition or removal of natural biofilms having a derivative relationship with the simulated biofilms respectively.
- the present invention methodology can be illustrated by reference to a simulated household drain biofilm which is derived from an inoculum of a natural household drain biofilm.
- the inoculum and aqueous nutrient medium are introduced into an annular reactor which holds a set of stainless steel slides.
- the reactor contents are incubated for between about 4-60 hours at a temperature between about 15 ° -45 ° C.
- the reactor reservoir is drained and recharged with aqueous nutrient medium at least once every 48 hours, over an elapsed time period of at least about 72 hours.
- the reservoir is drained and recharged at least twice every 24 hours, over an elapsed time period between about 2-45 days.
- Simulated biofilm growth on the slide surfaces in the annular reactor is continued until a desired biofilm thickness is achieved, e.g., between about 10-500 microns.
- the biotic population in a simulated biofilm typically will be between about 10 5 -10 15 bacterial Colony Forming Units per square centimeter (CFU/cm 2 ) on the slide surfaces.
- the methodology of the present invention has general utility for the production of simulated biofilms which are intended for application in a further protocol for testing the activity of an experimental product or a commercial product.
- the further protocol can include testing the activity of products such as biocides and cleaners, and products for opening conduits which are clogged with biomass, and the like.
- EXAMPLE I This Example illustrates invention methodology for growing a simulated biofilm, as derived from a household drain natural biofilm inoculum, in an annular reactor under simulated household drain conditions.
- An annular reactor with a rotatable inner drum is utilized (Biosurface
- the reactor has a 23 cm height and a 13 cm diameter, and is constructed of autoclavable polycarbonate.
- the reactor is adapted to secure the 12 stainless steel slides which are employed in the protocol.
- a biofilm sample is extracted from a household drain, and used as the protocol inoculum.
- aqueous nutrient medium which has the following composition: 0.2% yeast extract 0.2% Trypticase soy broth 0.1% kaolin solution 7 g/L K 2 HPO 4
- the reactor contents are incubated for two days at room temperature; (6) the reactor liquid volume is drained, and the reactor is refilled with nutrient medium and shaving residue;
- step (6) drain and refill procedure is repeated two times per 24 hours to simulate household drain usage, and the procedure is continued for a time period between about 4-6 weeks sufficient to develop a 40-50 micron thick biofilm on the stainless steel slides (10 9 -10 12 Colony Forming Units per square centimeter);
- biofilm-coated slides are utilized in a further protocol to test the activities of laboratory formulations and commercial products.
- EXAMPLE II This Example illustrates a method for determining the Colony Forming Units per square centimeter (CFU/cm 2 ) on biofilm-coated slides.
- a stainless slide retrieved from the annular reactor in Example I is placed in a beaker with 10 ml of PBS nutrient medium.
- the slide is scraped from top to bottom using a sterile cell scraper to provide a biofilm sample.
- the sample is homogenized one minute with a tissumizer operating at moderate speed.
- One ml of the homogenized biofilm sample is diluted in 9 ml of PBS medium.
- the resulting 10 ml sample is vortexed, and then the sample is diluted serially.
- a 0.1 ml quantity of a diluted sample is plated on R2A agar. Each sample dilution is plated in triplicate.
- the dilution sample on each plate is spread with a plate spreader, and the plates are incubated at room temperature for five days.
- a plated sample which has approximately 30-300 colonies is counted.
- the colony counts, dilution, and the scraped area of the slide is used to determine CFU/cm 2 . -8-
- EXAMPLE III This Example illustrates a method for testing the activities of household drain care products on laboratory-grown biofilms which simulate natural household drain biofilms.
- a sink and drain system is assembled from three 6" straight sections of 1.25"
- the drain system is flushed with one gallon of water (38 ° C).
- One biofilm-coated slide in accordance with the Example I protocol is secured to the top of the horizontal glass pipe section after the P-trap, and another biofilm- coated slide is secured to the bottom of the horizontal glass pipe section after the P-trap.
- a household drain-care product is dispensed into the drain system, and the product is maintained in the drain system for 8 hours.
- the drain system then is gently rinsed with one gallon of water (38 ° C).
- the treated slides are removed and placed into wells of a slotted tray.
- the slides are submerged in 5% sodium thiosulfate solution for two minutes, and then transferred to a second tray with deionized water.
- the CFU/cm 2 are determined, and compared with the initial count of control samples.
- the comparative data indicate that Formulation DC-X effectively treats a greater biofilm surface area than the PS Liquid Plumr product.
- Formulation DC-X removes more biofilm than Plumr on the top slide in the drain system, as determined by the remaining CFU/cm 2 count on the slide. With respect to the bottom slide in the drain system, DC-X and Plumr are equally effective.
- Top and bottom slides treated with DC-X have similar reduction in the amount of biofilm, while the top slide treated with Plumr has less reduction than the bottom slide treated with Plumr.
- a method for producing a biofilm useful for testing cleaning products and other formulated products intended to inhibit, remove, or otherwise affect such films is useful, for example, in the development of commercial cleaning products.
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK99906994T DK1053299T3 (en) | 1998-02-13 | 1999-02-12 | Protocol for forming a simulated natural biofilm |
AU26771/99A AU2677199A (en) | 1998-02-13 | 1999-02-12 | Protocol for simulated natural biofilm formation |
CA002321258A CA2321258C (en) | 1998-02-13 | 1999-02-12 | Protocol for simulated natural biofilm formation |
DE69900632T DE69900632T2 (en) | 1998-02-13 | 1999-02-12 | PROTOCOL FOR THE PRODUCTION OF A SIMULATED, NATURAL ORGANIC SURFACE |
EP99906994A EP1053299B1 (en) | 1998-02-13 | 1999-02-12 | Protocol for simulated natural biofilm formation |
AT99906994T ATE211165T1 (en) | 1998-02-13 | 1999-02-12 | PROTOCOL FOR PRODUCING A SIMULATED NATURAL BIOLOGICAL COVER |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/023,520 US5928889A (en) | 1998-02-13 | 1998-02-13 | Protocol for simulated natural biofilm formation |
US09/023,520 | 1998-02-13 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1999041354A1 true WO1999041354A1 (en) | 1999-08-19 |
Family
ID=21815603
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1999/003123 WO1999041354A1 (en) | 1998-02-13 | 1999-02-12 | Protocol for simulated natural biofilm formation |
Country Status (10)
Country | Link |
---|---|
US (1) | US5928889A (en) |
EP (1) | EP1053299B1 (en) |
AT (1) | ATE211165T1 (en) |
AU (1) | AU2677199A (en) |
CA (1) | CA2321258C (en) |
DE (1) | DE69900632T2 (en) |
DK (1) | DK1053299T3 (en) |
ES (1) | ES2165228T3 (en) |
PT (1) | PT1053299E (en) |
WO (1) | WO1999041354A1 (en) |
Cited By (3)
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EP2229959A2 (en) | 2009-03-18 | 2010-09-22 | EMPA Eidgenössische Materialprüfungs- und Forschungsanstalt | Standarized production of mature biofilms |
EP3027727A4 (en) * | 2013-07-29 | 2017-03-08 | UTI Limited Partnership | Annular culture dish |
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US6599714B1 (en) | 1996-03-13 | 2003-07-29 | University Technologies International Inc. | Method of growing and analyzing a biofilm |
US20060166358A1 (en) * | 1997-03-12 | 2006-07-27 | University Technologies International, Inc. | Biofilm incubation |
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US6207405B1 (en) * | 1998-06-29 | 2001-03-27 | S. C. Johnson & Son, Inc. | Pilot drain system for rapid biofilm formation |
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US20030032079A1 (en) * | 2000-04-17 | 2003-02-13 | Howard Ceri | Apparatus and method for testing effects of materials and surface coating on the formation of biofilms |
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US7192615B2 (en) | 2001-02-28 | 2007-03-20 | J&J Consumer Companies, Inc. | Compositions containing legume products |
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US6793880B2 (en) * | 2001-07-13 | 2004-09-21 | Minntech Corporation | Apparatus and method for monitoring biofilm cleaning efficacy |
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- 1999-02-12 AU AU26771/99A patent/AU2677199A/en not_active Abandoned
- 1999-02-12 AT AT99906994T patent/ATE211165T1/en not_active IP Right Cessation
- 1999-02-12 ES ES99906994T patent/ES2165228T3/en not_active Expired - Lifetime
- 1999-02-12 CA CA002321258A patent/CA2321258C/en not_active Expired - Fee Related
- 1999-02-12 PT PT99906994T patent/PT1053299E/en unknown
- 1999-02-12 DE DE69900632T patent/DE69900632T2/en not_active Expired - Fee Related
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Also Published As
Publication number | Publication date |
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DE69900632T2 (en) | 2002-06-13 |
CA2321258C (en) | 2005-02-08 |
EP1053299B1 (en) | 2001-12-19 |
PT1053299E (en) | 2002-05-31 |
AU2677199A (en) | 1999-08-30 |
ATE211165T1 (en) | 2002-01-15 |
DK1053299T3 (en) | 2002-01-28 |
CA2321258A1 (en) | 1999-08-19 |
ES2165228T3 (en) | 2002-03-01 |
DE69900632D1 (en) | 2002-01-31 |
EP1053299A1 (en) | 2000-11-22 |
US5928889A (en) | 1999-07-27 |
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