CA2248808A1 - Biofilm incubation - Google Patents
Biofilm incubationInfo
- Publication number
- CA2248808A1 CA2248808A1 CA002248808A CA2248808A CA2248808A1 CA 2248808 A1 CA2248808 A1 CA 2248808A1 CA 002248808 A CA002248808 A CA 002248808A CA 2248808 A CA2248808 A CA 2248808A CA 2248808 A1 CA2248808 A1 CA 2248808A1
- Authority
- CA
- Canada
- Prior art keywords
- biofilm
- plural
- adherent sites
- sites
- growth medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000011534 incubation Methods 0.000 title claims 2
- 239000007788 liquid Substances 0.000 claims abstract 25
- 239000001963 growth medium Substances 0.000 claims abstract 23
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract 19
- 230000000845 anti-microbial effect Effects 0.000 claims abstract 18
- 238000003556 assay Methods 0.000 claims abstract 6
- 230000035945 sensitivity Effects 0.000 claims abstract 3
- 230000001464 adherent effect Effects 0.000 claims 56
- 238000000034 method Methods 0.000 claims 37
- 238000000576 coating method Methods 0.000 claims 4
- 239000004599 antimicrobial Substances 0.000 claims 2
- 239000011248 coating agent Substances 0.000 claims 2
- 239000000463 material Substances 0.000 claims 2
- 238000011109 contamination Methods 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract 1
- 239000011435 rock Substances 0.000 abstract 1
Classifications
-
- 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
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
- B01L3/50853—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates with covers or lids
-
- 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
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/06—Plates; Walls; Drawers; Multilayer plates
-
- 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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/16—Vibrating; Shaking; Tilting
-
- 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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
Abstract
Biofilm forming organisms are incubated to form a biofilm on projections by providing a flow of liquid growth medium across projections, the direction of the flow of liquid being repeatedly changed, and an assay made of the resulting biofilm. Biofilm forming organisms are incubated to form a biofilm on projections arranged in rows, with several projections in each row, while providing a flow of liquid growth medium across each row of projections, and an assay made of the resulting biofilm. Sensitivity of the biofilm to antimicrobial reagent may be determined by treating the projections with antimicrobial reagent before carrying out the assay, by treating each row of projections with a different antimicrobial reagent, and each of the projections in a row with a different concentration of antibacterial reagent. A biofilm assay device includes a vessel including at least one channel for flow of liquid growth medium, projections arranged in at least one row and having a support for supporting the projections within the channel, and a tilt table to flow liquid growth medium along each channel in different directions across the projections. A further biofilm assay device includes a vessel including channels for flow of liquid growth medium, projections arranged in rows and having a support for supporting the projections within the channels, and a rocking table to rock the vessel and flow liquid growth medium along each channel across the projections.
Claims (45)
1. A method of growing a biofilm, the method comprising the steps of:
incubating biofilm forming organisms on plural biofilm adherent sites while providing a flow of liquid growth medium across the plural biofilm adherent sites, the direction of the flow of liquid being repeatedly changed, to thereby create a biofilm at the plural biofilm adherent sites.
incubating biofilm forming organisms on plural biofilm adherent sites while providing a flow of liquid growth medium across the plural biofilm adherent sites, the direction of the flow of liquid being repeatedly changed, to thereby create a biofilm at the plural biofilm adherent sites.
2. The method of claim 1 further comprising the step of assaying the biofilm at the plural biofilm adherent sites to analyze a characteristic of the biofilm.
3. The method of claim 2 in which the characteristic of the biofilm is the sensitivity of the biofilm to antimicrobial reagent and the method further comprises the step of, before assaying the biofilm, treating the biofilm adherent sites with antimicrobial reagent.
4. The method of claim 3 further comprising:
after treating the biofilm adherent sites with antimicrobial reagent, dislodging the biofilm from the biofilm adherent sites before performing the assay.
after treating the biofilm adherent sites with antimicrobial reagent, dislodging the biofilm from the biofilm adherent sites before performing the assay.
5. The method of claim 4 further including, after dislodging the biofilm from the biofilm adherent sites, further incubating the biofilm.
6. The method of claim 4 in which dislodging the biofilm from the biofilm adherent sites includes dislodging the biofilm from each biofilm adherent site into a separate well of a well plate.
7. The method of claim 3 in which the biofilm adherent sites are formed in rows, and treating the biofilm adherent sites with an antimicrobial reagent includes:
treating each row of biofilm adherent sites with a different antimicrobial reagent.
treating each row of biofilm adherent sites with a different antimicrobial reagent.
8. The method of claim 7 further including treating each biofilm adherent site in a row with a different concentration of antimicrobial reagent.
9. The method of claim 3, 7 or 8 in which flow direction of the liquid growth medium is repeatedly reversed.
10. The method of claim 9 in which the liquid growth medium flows in channels of a vessel, and the direction of flow of the liquid growth medium is reversed by rocking of the vessel.
11. The method of claim 10 in which the biofilm adherent sites are projections from a lid and incubating the biofilm includes:
suspending the projections in liquid growth medium in the channels while rocking the vessel so as to provide shear forces on the biofilm during growth of the biofilm.
suspending the projections in liquid growth medium in the channels while rocking the vessel so as to provide shear forces on the biofilm during growth of the biofilm.
12. The method of claim 2 further including treating each of the biofilm adherent sites with a different concentration of antimicrobial reagent.
13. The method of claim 12 further including:
before incubating biofilm forming organisms on plural biofilm adherent sites, applying a biofilm resistant coating to the plural biofilm adherent sites.
before incubating biofilm forming organisms on plural biofilm adherent sites, applying a biofilm resistant coating to the plural biofilm adherent sites.
14. The method of claim 13 further including applying different coatings to different biofilm adherent sites.
15. The method of claim 1 further including:
incubating the biofilm forming organisms in the presence of host material in the liquid growth medium.
incubating the biofilm forming organisms in the presence of host material in the liquid growth medium.
16. The method of claim 1 further including the step of, after initially incubating the biofilm forming organisms on plural biofilm adherent sites to create a biofilm, subsequently incubating the biofilm forming organisms in separate micro-reactors.
17. The method of claim 3 in which different biofilm adherent sites are treated with different antimicrobial reagents.
18. Apparatus for analyzing biofilms, the apparatus comprising:
a vessel including at least one channel for flow of liquid growth medium;
plural biofilm adherent sites having a support for supporting the plural biofilm adherent sites within the channel; and means to flow liquid growth medium along each channel in different directions across the plural biofilm adherent sites.
a vessel including at least one channel for flow of liquid growth medium;
plural biofilm adherent sites having a support for supporting the plural biofilm adherent sites within the channel; and means to flow liquid growth medium along each channel in different directions across the plural biofilm adherent sites.
19. The apparatus of claim 18 in which:
the plural biofilm adherent sites are formed in plural rows, with more than one biofilm adherent site in each row;
the vessel includes plural channels having substantially the same shape.
the plural biofilm adherent sites are formed in plural rows, with more than one biofilm adherent site in each row;
the vessel includes plural channels having substantially the same shape.
20. The apparatus of claim 19 in which the biofilm adherent sites have substantially the same shape.
21. The apparatus of claim 20 in which the support for the plural biofilm adherent sites forms a lid for the vessel.
22. The apparatus of claims 18, 19 or 20 in which the means to flow liquid growth medium across the plural biofilm adherent sites is a tilt table.
23. A method of growing a biofilm, the method comprising the steps of:
incubating biofilm forming organisms on plural biofilm adherent sites arranged in plural rows, with plural biofilm adherent sites in each row, while providing a flow of liquid growth medium across each of the plural biofilm adherent sites, to thereby create a biofilm at each of the plural biofilm adherent sites.
incubating biofilm forming organisms on plural biofilm adherent sites arranged in plural rows, with plural biofilm adherent sites in each row, while providing a flow of liquid growth medium across each of the plural biofilm adherent sites, to thereby create a biofilm at each of the plural biofilm adherent sites.
24. The method of claim 23 further comprising the step of assaying the biofilm at the plural biofilm adherent sites to analyze a characteristic of the biofilm.
25. The method of claim 24 in which providing a flow of liquid growth medium includes flowing liquid growth medium backwards and forwards along uniform channels of a
26 vessel to create a uniform biofilm at each biofilm adherent site.
26. The method of claim 24 in which the characteristic of the biofilm is the sensitivity of the biofilm to antimicrobial reagent and the method further comprises the step of, before assaying the biofilm, treating the biofilm adherent sites with antimicrobial reagent.
26. The method of claim 24 in which the characteristic of the biofilm is the sensitivity of the biofilm to antimicrobial reagent and the method further comprises the step of, before assaying the biofilm, treating the biofilm adherent sites with antimicrobial reagent.
27. The method of claim 26 further comprising:
after treating the biofilm adherent sites with antimicrobial reagent, dislodging the biofilm from the biofilm adherent sites before assaying the biofilm.
after treating the biofilm adherent sites with antimicrobial reagent, dislodging the biofilm from the biofilm adherent sites before assaying the biofilm.
28. The method of claim 27 further including, after dislodging the biofilm, further incubating the biofilm.
29. The method of claim 27 in which dislodging the biofilm from the biofilm adherent sites includes:
dislodging the biofilm from each biofilm adherent site into a separate well of a well plate.
dislodging the biofilm from each biofilm adherent site into a separate well of a well plate.
30. The method of claim 29 in which treating the biofilm adherent sites with an antimicrobial reagent includes treating each row of biofilm adherent sites with a different antimicrobial reagent.
31. The method of claim 30 further including:
treating each biofilm adherent site in a row with a different concentration of antimicrobial reagent.
treating each biofilm adherent site in a row with a different concentration of antimicrobial reagent.
32. The method of claim 23 in which the biofilm adherent sites are projections from a lid and incubating the biofilm includes:
suspending the projections in liquid growth medium in uniform channels of a vessel while rocking the vessel so as to provide uniform shear forces on the projections during growth of the biofilm.
suspending the projections in liquid growth medium in uniform channels of a vessel while rocking the vessel so as to provide uniform shear forces on the projections during growth of the biofilm.
33. The method of claim 24 in which assaying the biofilm comprises applying a vital dye to the biofilm and automatically measuring the intensity of light given off by the dyed biofilm.
34. The method of claim 23 further including the step of, after initially incubating the biofilm forming organisms on plural biofilm adherent sites to create a biofilm, subsequently incubating the biofilm forming organisms in separate micro-reactors.
35. The method of claim 23 further including:
before incubating biofilm forming organisms on the plural biofilm adherent sites, applying a biofilm resistant coating to the plural biofilm adherent sites.
before incubating biofilm forming organisms on the plural biofilm adherent sites, applying a biofilm resistant coating to the plural biofilm adherent sites.
36. The method of claim 35 further including applying different coatings to different biofilm adherent sites.
37. The method of claim 23 further including incubating the biofilm forming organisms in the presence of host material in the liquid growth medium.
38. The method of claim 26 in which different biofilm adherent sites are treated with different antimicrobial reagents.
39. Apparatus for growing biofilms, the apparatus comprising:
a vessel including plural channels for flow of liquid growth medium;
plural biofilm adherent sites arranged in plural rows and having a support for supporting the plural biofilm adherent sites within the channels; and means to avoid contamination of the inside of the vessel during incubation.
a vessel including plural channels for flow of liquid growth medium;
plural biofilm adherent sites arranged in plural rows and having a support for supporting the plural biofilm adherent sites within the channels; and means to avoid contamination of the inside of the vessel during incubation.
40. The apparatus of claim 39 in which each channel has substantially the same shape.
41. The apparatus of claim 40 in which each biofilm adherent site has substantially the same shape.
42. The apparatus of claim 41 further including:
means to flow liquid growth medium along each channel across plural biofilm adherent sites supported within the channel.
means to flow liquid growth medium along each channel across plural biofilm adherent sites supported within the channel.
43. The apparatus of claims 41 or 42 in which:
the support for the plural biofilm adherent sites forms a lid for the vessel.
the support for the plural biofilm adherent sites forms a lid for the vessel.
44. The apparatus of claim 43 in which the plural biofilm adherent sites are projections from the lid.
45. The apparatus of claim 42 in which the means to flow liquid growth medium across the plural biofilm adherent sites is a tilt table.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/614,593 | 1996-03-13 | ||
| US08/614,593 US6599714B1 (en) | 1996-03-13 | 1996-03-13 | Method of growing and analyzing a biofilm |
| PCT/CA1997/000171 WO1997033972A1 (en) | 1996-03-13 | 1997-03-12 | Biofilm incubation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2248808A1 true CA2248808A1 (en) | 1997-09-18 |
| CA2248808C CA2248808C (en) | 2010-01-05 |
Family
ID=24461943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002248808A Expired - Fee Related CA2248808C (en) | 1996-03-13 | 1997-03-12 | Biofilm incubation |
Country Status (8)
| Country | Link |
|---|---|
| US (5) | US6599714B1 (en) |
| EP (1) | EP0904347B1 (en) |
| JP (1) | JP3886152B2 (en) |
| AT (1) | ATE301183T1 (en) |
| AU (1) | AU724282B2 (en) |
| CA (1) | CA2248808C (en) |
| DE (1) | DE69733888T2 (en) |
| WO (1) | WO1997033972A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005098023A1 (en) * | 2004-04-12 | 2005-10-20 | University Technologies International Inc. | Microbial growth assay |
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| US6599714B1 (en) * | 1996-03-13 | 2003-07-29 | University Technologies International Inc. | Method of growing and analyzing a biofilm |
| US5928889A (en) | 1998-02-13 | 1999-07-27 | S.C. Johnson & Son, Inc. | Protocol for simulated natural biofilm formation |
| CH692583A5 (en) | 1998-03-03 | 2002-08-15 | Weidmann H Ag | Culture vessel. |
-
1996
- 1996-03-13 US US08/614,593 patent/US6599714B1/en not_active Expired - Fee Related
-
1997
- 1997-03-12 DE DE69733888T patent/DE69733888T2/en not_active Expired - Lifetime
- 1997-03-12 JP JP53214097A patent/JP3886152B2/en not_active Expired - Fee Related
- 1997-03-12 US US09/380,714 patent/US6410256B1/en not_active Expired - Lifetime
- 1997-03-12 AT AT97906060T patent/ATE301183T1/en not_active IP Right Cessation
- 1997-03-12 WO PCT/CA1997/000171 patent/WO1997033972A1/en active IP Right Grant
- 1997-03-12 CA CA002248808A patent/CA2248808C/en not_active Expired - Fee Related
- 1997-03-12 AU AU20887/97A patent/AU724282B2/en not_active Ceased
- 1997-03-12 EP EP97906060A patent/EP0904347B1/en not_active Expired - Lifetime
-
1998
- 1998-07-31 US US09/126,957 patent/US6051423A/en not_active Expired - Lifetime
-
2000
- 2000-01-18 US US09/484,527 patent/US6326190B1/en not_active Expired - Lifetime
-
2002
- 2002-03-27 US US10/106,115 patent/US7041470B2/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005098023A1 (en) * | 2004-04-12 | 2005-10-20 | University Technologies International Inc. | Microbial growth assay |
Also Published As
| Publication number | Publication date |
|---|---|
| US7041470B2 (en) | 2006-05-09 |
| US6326190B1 (en) | 2001-12-04 |
| JP3886152B2 (en) | 2007-02-28 |
| WO1997033972A1 (en) | 1997-09-18 |
| ATE301183T1 (en) | 2005-08-15 |
| US6410256B1 (en) | 2002-06-25 |
| DE69733888D1 (en) | 2005-09-08 |
| US20020146809A1 (en) | 2002-10-10 |
| AU724282B2 (en) | 2000-09-14 |
| AU2088797A (en) | 1997-10-01 |
| US6599714B1 (en) | 2003-07-29 |
| DE69733888T2 (en) | 2006-04-06 |
| EP0904347A1 (en) | 1999-03-31 |
| EP0904347B1 (en) | 2005-08-03 |
| JP2000506021A (en) | 2000-05-23 |
| US6051423A (en) | 2000-04-18 |
| CA2248808C (en) | 2010-01-05 |
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