WO2004042082A1 - Procede de controle de la presence de micro-organismes nocifs dans l'industrie du papier - Google Patents

Procede de controle de la presence de micro-organismes nocifs dans l'industrie du papier Download PDF

Info

Publication number
WO2004042082A1
WO2004042082A1 PCT/FI2003/000839 FI0300839W WO2004042082A1 WO 2004042082 A1 WO2004042082 A1 WO 2004042082A1 FI 0300839 W FI0300839 W FI 0300839W WO 2004042082 A1 WO2004042082 A1 WO 2004042082A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
biofilm
ccc
microorganism
probe
Prior art date
Application number
PCT/FI2003/000839
Other languages
English (en)
Other versions
WO2004042082A9 (fr
Inventor
German Jurgens
Marko Kolari
Frederick Rainey
Mirja Salkinoja-Salonen
Hanna Laatikainen
Päivi TAMMELA
Pia Vuorela
Pentti VÄÄTÄNEN
Original Assignee
Kemira Oyj
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority claimed from FI20021986A external-priority patent/FI116030B/fi
Priority claimed from FI20021987A external-priority patent/FI115502B/fi
Application filed by Kemira Oyj filed Critical Kemira Oyj
Priority to AU2003276306A priority Critical patent/AU2003276306A1/en
Publication of WO2004042082A1 publication Critical patent/WO2004042082A1/fr
Publication of WO2004042082A9 publication Critical patent/WO2004042082A9/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/06—Coniferophyta [gymnosperms], e.g. cypress
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A01N65/16—Ericaceae [Heath or Blueberry family], e.g. rhododendron, arbutus, pieris, cranberry or bilberry
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A01N65/22—Lamiaceae or Labiatae [Mint family], e.g. thyme, rosemary, skullcap, selfheal, lavender, perilla, pennyroyal, peppermint or spearmint
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A01N65/34—Rosaceae [Rose family], e.g. strawberry, hawthorn, plum, cherry, peach, apricot or almond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
    • A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H21/00—Non-fibrous material added to the pulp, characterised by its function, form or properties; Paper-impregnating or coating material, characterised by its function, form or properties
    • D21H21/02—Agents for preventing deposition on the paper mill equipment, e.g. pitch or slime control
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H21/00—Non-fibrous material added to the pulp, characterised by its function, form or properties; Paper-impregnating or coating material, characterised by its function, form or properties
    • D21H21/02—Agents for preventing deposition on the paper mill equipment, e.g. pitch or slime control
    • D21H21/04—Slime-control agents
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/02—Non-contaminated water, e.g. for industrial water supply
    • C02F2103/023—Water in cooling circuits
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/26—Nature of the water, waste water, sewage or sludge to be treated from the processing of plants or parts thereof
    • C02F2103/28—Nature of the water, waste water, sewage or sludge to be treated from the processing of plants or parts thereof from the paper or cellulose industry
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H17/00—Non-fibrous material added to the pulp, characterised by its constitution; Paper-impregnating material characterised by its constitution
    • D21H17/02—Material of vegetable origin
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H17/00—Non-fibrous material added to the pulp, characterised by its constitution; Paper-impregnating material characterised by its constitution
    • D21H17/03—Non-macromolecular organic compounds
    • D21H17/05—Non-macromolecular organic compounds containing elements other than carbon and hydrogen only
    • D21H17/06—Alcohols; Phenols; Ethers; Aldehydes; Ketones; Acetals; Ketals

Definitions

  • the present invention relates to a method for monitoring the presence of harmful microorganisms in paper and board making industry and to the use of the method for determining the need for an anti-biofilm agent in the process. Furthermore, oligonucleotide probes as well as a kit assembly containing such probes are also provided.
  • the paper and board making processes contain warm process waters (e.g. 45-60°C) that are rich in biodegradable nutrients and have a beneficial pH (e.g. pH 4-9) thus providing a good environment for the growth of microorganisms.
  • the microbes in the process show biofilm-forming, i.e. surface-attached, growth and free-swimming, i.e. planktonic, growth.
  • biofilms i.e. slime layers, which develop on the surfaces of the process equipments and can rip off from the surfaces.
  • Biofouling of the machine surfaces and the loose biofilm/agglomerates can cause severe process disturbances: reduce water flow; block filters, wires, etc.; deteriorate the end-product quality, e.g. by causing holes or colored spots in the end product; or break the whole paper web.
  • the biofilms are difficult to remove from the surfaces of the process equipment and often require the use of very strong chemicals.
  • biocides For controlling the microbe growth e.g. biocides have been added to the process waters. Planktonic microbes have been efficiently controlled by the biocides; however, the use of biocides has not solved all biofilm problems in the paper or board machines. Several reasons exist for that, e.g. a wide variety of microbes are growing in the papermaking process and it is now also known that bacteria growing in a biofilm are generally more resistant to biocides than the planktonic microbes.
  • the primary biofilm-formers seem to be a prerequisite for successful surface colonization of several other microbes in paper and board machines.
  • E.g. Deinococcus geothermalis is a primary biofilm-former and strains of several Bacillus species adhere to the primary biofilms of this bacterium.
  • Thermomonas such as T. haemolytica or T. hydrothermalis.
  • the primary biofilm-formers are typically moderately fhermophilic, some with maximum growth temperatures as high as 67°C, and most of the strains showing the fastest growth at temperatures of 45 to 55°C. To overcome the problem with the biofilms in paper industry was, however, not discussed in said thesis.
  • the efficient use of biocides would require an effective monitoring method.
  • the typical monitoring method for the microbe growth in paper industry is the cultivation of a sample of process water in laboratory. Such cultivation methods are, however, time-consuming and complicated, i.e. they take several days and require i.a. the use of various growth media for isolation and identification of different types of microbes in a sample.
  • the microbe growth in laboratory is culture-medium-dependent, thus part of the microbes present in a sample are often unable to grow under the laboratory conditions.
  • the monitoring is typically focused on the free-swimming microbes. Efficient methods for monitoring the effects of anti-biofilm agents on the biofilm-forming microbes have been lacking. Therefore, in the prior art biocides are conventionally used for non-selective inhibition of all microbes in the process, whereby high amounts of biocides are often required.
  • Identification and quantification methods based on the detection of nucleic acid sequences of organisms are well known in the field of biosciences. These methods include hybridization methods, wherein a probe of an oligonucleotide labelled with a detectable marker is developed which is complementary to a region of a DNA or RNA, e.g. rPvNA, molecule of the organism of interest.
  • a probe of an oligonucleotide labelled with a detectable marker is developed which is complementary to a region of a DNA or RNA, e.g. rPvNA, molecule of the organism of interest.
  • Such an in situ hybridization technique also known as FISH (Fluorescence-In-Situ-Hybridization) is a well- established technique in the studies of microbial ecology (e.g. Amann, R. et al, Current Opinion in Biotechnology, 12, 2001, p.231-236).
  • WO 0185340, WO 02103043 and WO 02102824 describe in situ hybridization method and/or a kit for certain industrial applications.
  • the disclosed industrial applications relate to "pure” systems or systems with “specific microbes”, wherein practically no microbe growth or only minor or specific microbe growth occurs, such as the use of in situ hybridization method in pharmaceutical industry or for detecting harmful microbes from drinking water or beer.
  • the object of the present invention is to provide a further method for the paper industry, which enables an efficient and timesaving means for monitoring the microbiological state of a paper or board making process, and, moreover, enables the recognition of harmful microbes before process failures emerge.
  • a further object of the invention is to provide a method, which can be used for specific monitoring of microbes that are relevant for biofouling in paper industry, to determine the need for an anti-biofilm agent in the process, and the effects of such an anti-biofilm agent.
  • Another object of the invention is to provide new oligonucleotide probes, which can be used for a reliable and fast monitoring of the harmful microbes, and for recognition of the causative bacteria in the case of slime outbreaks.
  • a still further object of the invention is to provide a kit assembly, which is very feasible for carrying out the method of the invention on the process site.
  • Figure 1A and Figure IB present the epifluorescence microscope image and phase contrast light microscope image of the specific binding of the oligonucleotide probe shown in SEQ ID NO. 1 with a sample of the hybridization example 1.
  • Figure 2 presents epifluorescence microscope image of a paper machine biofilm of a red-gray slime sample after in situ hybridization with the probe shown in SEQ ID NO. 1 of the hybridization example 2.
  • Figure 3 presents epifluorescence microscope image of a paper machine biofilm of a red slime sample after in situ hybridization with the probe shown in SEQ ID NO. 1 of the hybridization example 2.
  • Figure 4 presents epifluorescence microscope image of a board machine biofilm on a sample support after in situ hybridization with the probe shown in SEQ ID NO. 1 of the hybridization example 2.
  • Figure 5 presents epifluorescence microscope image of Meiothermus ruber DSM1279 cells after in situ hybridization with the oligonucleotide probe shown in SEQ ID No. 14 of the hybridization example 3.
  • the invention is based on the idea that by controlling the growth of the biofilm- formers, and especially the primary biofilm-formers, during the paper or board making process the problems with slime formation could be prevented efficiently. Furthermore, the inventors found that by monitoring the biofilm-formers during the process the development of the more problematic slime in the whole biomass, which actually causes the biofouling, could be foreseen and/or predicted and preventive actions be started at an early stage, whereby the detrimental disturbances in the process could be decreased or fully avoided. However, the time consuming prior art methods used in paper industry are not suitable for monitoring quick changes in the process.
  • the present idea can unexpectedly be realized with in situ hybridization technique which was found to be surprisingly utilizable in a highly demanding microbial environment of the paper or board making process for monitoring selectively the presence of harmful microorganisms, especially the biofilm-formers and, among these, particularly the most harmful primary biofilm-formers. Moreover, it was found that the in situ hybridization method can be applied directly to samples taken from the process, i.e. without any prior cultivation or isolation steps and without any dilution/concentration steps, and that the targeted harmful microbe can be detected selectively in the presence of the versatile microbe population of the process.
  • the in situ hybridization method of the invention is very fast to carry out, i.e. samples taken from the process can be analyzed in three to four hours, whereby it is possible to take samples continuously from one or several locations of the process line, and to react quickly to the changes in the microbe growth within the process, i.e. to start of preventive countermeasures before process failures emerge, e.g. by dosing an anti-biofilm agent and by monitoring its effects by the said method.
  • the present method makes it possible to monitor more reliably and quickly the actual occurrence of the relevant microbe to be detected compared to the cultivation-based methods in current use.
  • the method is also very sensitive, as single active cells of harmful microbes can be detected, and thereby the process personnel can be alerted before any visible process failures emerge.
  • Paper or board making process and “process line” as used herein covers the process lines of wet manufacturing processes for raw materials of paper and pulp industry, pulping process, the pulp drying process, the actual paper or board making process, and the equipment thereof including the paper and board machines.
  • the paper and board machines are understood to comprise both the wet-end with wire section and the drying section.
  • the present invention provides a method for monitoring harmful microorganisms in paper and board making processes, wherein the presence or absence of the microorganism of interest in a sample is detected by in situ hybridization method using an oligonucleotide probe hybridizable with a region of a nucleic acid molecule of said microorganism.
  • the oligonucleotide probe is hybridizable with a nucleotide sequence of a nucleic acid molecule present in the harmful microorganism desired to be detected (referred herein as "target microorganism” or “target microbe” or “microbe of interest”).
  • target microorganism or "target microbe” or “microbe of interest”
  • the probe is hybridizable with an agglomerate- and/or a biofilm-forming microorganism, more preferably with a primary biofilm-forming microorganism.
  • oligonucleotide probes can be designed for a chosen group of microbes or to a specific species.
  • the nucleotide sequence of the probe is targeted to a region of a nucleic acid molecule of the microbe of interest, which region is specific for that biofilm-forming genus or species.
  • one or more probes can be used to hybridize with one or more target microbe species present in the sample, whereby each probe may be targeted to one microbe species.
  • the oligonucleotide probe is hybridizable with a region of a DNA or RNA molecule, preferably of an RNA molecule, and more preferably of an rRNA molecule, of the harmful target microorganism. It is known that rRNA molecules are good target molecules for the in situ hybridization, as they exist in all living cells in detectable levels.
  • the oligonucleotide probe hybridizes with a region, i.e. a nucleic acid sequence, present in the 16S or 23S rRNA molecule of the target microorganism.
  • the probe hybridizes with a region of the 16S rRNA of the desired microbe.
  • sequence of the probe is complementary to a specific region of the DNA or RNA molecule of the target microbe, whereby “complementary” means herein that the nucleotide sequence of the probe is complementary with at least part of the nucleotides within the length of the specific region of the DNA or RNA molecule of the target microbe, so that the probe hybridizes to the target region under suitable hybridization conditions.
  • complementary means herein that the nucleotide sequence of the probe is complementary with at least part of the nucleotides within the length of the specific region of the DNA or RNA molecule of the target microbe, so that the probe hybridizes to the target region under suitable hybridization conditions.
  • the sequence for the oligonucleotide probe specific to a target microorganism can be designed and prepared by methods well known in the field of bioscience using the nucleic acid sequences, e.g. rRNA gene sequences, of the target microorganism.
  • the nucleic acid sequences of the target microorganisms may be found from the literature and public databases, or be identified e.g. by using the known PCR (polymerase chain reaction) and sequencing techniques.
  • the probes designed can be compared for their specificity with the sequence information contained in the public databases such as the EMBL Nucleotide Sequence Database or the RDP II.
  • the length of a probe can be between 12 and 1000 nucleotides.
  • the probe is typically provided with a detectable marker to enable the detection thereof after the hybridization.
  • the marker is preferably attached to the 5 '-end of the sequence of the probe in a manner known in the art.
  • markers may be commercially available and include a fluorescent marker, radioactive marker, enzymatically active group etc.
  • One suitable marker is a fluorescent label, such as commercially available Alexa, Cy3, FITC or TRITC-labels.
  • the sample can be taken from any part of the process line including the wet end part of the paper or board machines and the dry end of the process line. Furthermore the samples can be taken from any process media including process waters, pulp suspensions, circulating and waste waters, from any surfaces of the machines including liquid containing and splash areas of the machines, from wires, from filters, from raw materials, reagents, intermediate products and end products etc.
  • the splash area in the process system includes splash areas of the paper or board machines, e.g. splash areas in the wire sections and process water containers.
  • the sample is from i) slimes collected from the machine surfaces, ii) raw materials of the process, iii) end products of the process, iv) process water, or v) biofilms grown on surface of sampler plates during immersion in the process water e.g. using a flow-through biofilm reactor.
  • the fresh sample is subjected to the support, e.g. an objective slide or a multi-well objective slide known in the art, for the fixing step.
  • the support e.g. an objective slide or a multi-well objective slide known in the art
  • the sample e.g. slime
  • the fixation step e.g. in a suitable nutrient medium such as yeast extract, e.g. 30 min to 2 h, such as 1 h, between the ambient temperature and 65°C, preferably close to the temperature of the sampling site, e.g. at 35-65°C, e.g. at 40-60°C, to increase the rRNA content of the cells.
  • a suitable nutrient medium such as yeast extract, e.g. 30 min to 2 h, such as 1 h, between the ambient temperature and 65°C, preferably close to the temperature of the sampling site, e.g. at 35-65°C, e.g. at 40-60°C, to increase the rRNA content of the cells.
  • the "fixing step” means the well-known permeabilization and/or immobilization of the microorganism present in the sample to a support to enable the hybridization of the probe with the desired DNA or RNA region of the target microbe.
  • dehydrating the sample by air-drying and/or treating with ethanol could be mentioned.
  • step (c) the hybridization is effected by incubating the fixed sample together with the oligonucleotide probe labeled with a detectable marker in conditions enabling the hybridization of the probe with the harmful target microbe, preferably with an agglomerate- and/or biofilm-forming microorganism, more preferably with a target primary biofilm-former, and then removing any unbound probe.
  • the specificity of the probe to the target microbe is controlled by the hybridization conditions.
  • the hybridization can be effected in moderate or stringent conditions, preferably in stringent conditions to enable only the specific binding to the target RNA or DNA, such as rRNA, of the microorganism of interest.
  • the stringency can be adjusted in the known manner e.g.
  • the concentration of the hybridization buffer, of NaCl and/or of formamide is changed by changing the concentration of the hybridization buffer, of NaCl and/or of formamide. Only as an example, e.g. 20 min to 3 h, preferably 2 h, at 40 to 55°C, preferably at 46°C, in a hybridization buffer such as 20 mM Tris-HCl (pH 7.2) including a detergent, preferably sodium dodecyl sulphate (SDS) 0.01%, sodium chloride, preferably 0.9 M, and formamide, preferably 20-45%, could be mentioned for achieving the stringent conditions.
  • a hybridization buffer such as 20 mM Tris-HCl (pH 7.2) including a detergent, preferably sodium dodecyl sulphate (SDS) 0.01%, sodium chloride, preferably 0.9 M, and formamide, preferably 20-45%, could be mentioned for achieving the stringent conditions.
  • SDS sodium dodecyl sulphate
  • the fixed sample Before the actual hybridization step (c) the fixed sample may optionally be pretreated with an unlabelled non-specific oligonucleotide probe(s) for blocking any unspecific binding sites in a manner known in the art.
  • the unbound probe is removed in a known manner by washing with a buffer solution.
  • a buffer solution e.g. Tris-HCl buffer (pH 7.2) including SDS, e.g. 0.01% SDS, EDTA, e.g. 5 mM EDTA, and NaCl, e.g. 10-300 mM NaCl, at 40-60°C, e.g. at 48°C, for 20-30 min, could be mentioned.
  • Hybridized probe is then detected (d) qualitatively or quantitatively in a manner known in the art, e.g. a probe with a fluorescent label can be detected microscopically using an epifluorescence microscope.
  • the hybridized probes with the detectable marker can be separated in a known manner from the sample using e.g. solvents and quantified with a detection device such as a fluorometer.
  • the standard in situ hybridization protocols, i.e. fixing, hybridizing and detection steps and the design of the probe have been widely described in the handbooks in the field and the principles are within the skills of an artisan.
  • the present method can be used for detecting the relevant microbes from a biofilm formed in situ in the process on the surface of a sampler device and that the fixation and hybridization can unexpectedly be effected with said biofilm on the surface of said test sampler.
  • the sample is a biofilm formed by biofilm-formers on a surface of a sampler support immersed in the process water or placed in a splash area within the process system for a period of time, e.g. for 4 h to 7 d, preferably 12 h to 48 h.
  • microorganisms of the biofilm adhered to the surface of the sampler support do preferably not require fixing for immobilization on said support, only for permeabilization,
  • a harmful target microbe preferably a primary biofilm-former, present in the fixed biofilm is then hybridized on said sampler support with a labelled probe hybridizable therewith.
  • the sampler support is a test plate, e.g. a coupon or slide, preferably a slide of a stainless steel, and the biofilm is grown in situ in the process on the surface of the said test plate, (b) the fixing of the sample is effected on said plate, and (c) the in situ hybridization is performed on the surface of said plate.
  • the hybridization is effected in a disposable hybridization chamber attached, e.g. glued, on the test plate after the fixing step.
  • disposable chambers are known from the bioscience and commercially available.
  • the sampler support device can be brought in the splash area or immersed in the liquid streams for enabling the biofilm-formers to form the biofilm on the surface of the sampler support.
  • the sampler support may comprise means for fixing it e.g. on the surface of the paper making equipment, e.g. paper machine.
  • the sample taking points of the process line may be provided with a mounting means, wherein the sampler device can be fixed for a period of time to enable the biofilm formation on the surface thereof.
  • the time required for forming the biofilm on the surface of the device in the process depends on the place in the process line and can be chosen accordingly. E.g. when the device is in a wire water line of a neutral paper or board machine can some primary biofilm-formers be detected akeady after a six-hours exposure.
  • the oligonucleotide probe is hybridizable with a region of a DNA or RNA molecule, preferably of a rRNA molecule, of a microorganism selected from a group comprising species Deinococcus geothermalis, genus Meiothermus, particularly species M. silvanus or M. ruber, genus Azospirillum, genus Burkholderia, particularly B. multivorans or B. cepacia, genus Porphyrobacter, particularly P. cryptus, genus "Rubellimicrobium " and genus Thermomonas, particularly T. haemolytica or T. hydrothermalis.
  • a microorganism selected from a group comprising species Deinococcus geothermalis, genus Meiothermus, particularly species M. silvanus or M. ruber, genus Azospirillum, genus Burkholderia, particularly B. multivorans or B. cepac
  • the probe is specific to the microorganism to be detected.
  • the invention further provides oligonucleotide probes, which were designed for the specific detection of the above listed microbes as given above, especially specific for a region of the rRNA molecule or its gene, especially of the 16S rRNA molecule, of said microbes.
  • These probes provide an advantageous means for the method of the invention.
  • Said probes include new specific oligonucleotide probes which comprise a nucleotide sequence selected from the listed sequences below, which sequences are also presented in the Sequence Listing as a part of this application with SEQ ID NO's indicated below. Accordingly, the probe of the invention comprises a nucleotide sequence selected: (from 5' — » 3' direction)
  • nucleotide sequences which are particularly suitable for the detection of Deinococcus geothermalis in a process sample, and preferably targeted for a region of the rRNA or its gene of said microbe:
  • nucleotide sequences which are particularly suitable for the detection of genus Meiothermus, and preferably targeted for a region of the rRNA or its gene of said microbe:
  • nucleotide sequences which are particularly suitable for the detection of Porphyrobacter cryptus, and preferably targeted for a region of the rRNA or its gene of said microbe:
  • nucleotide sequences which are particularly suitable for the detection of genus "Rubellimicrobium ", and preferably targeted for a region of the rRNA or its gene of said microbe:
  • one or more probes preferably one or more new probes as specified above, can be used.
  • “Functional variant thereof means that the probe sequence may have variations compared to the nucleic acid sequence listed above, but the variant probe is still capable to hybridize to the target microbe in suitable hybridization conditions.
  • the sequence of the probe may have 60 % or more, e.g. 80 % or 95 %, similarity with the corresponding above listed sequence. Thus mismatches are allowed in certain degree as mentioned above for the term “complementary”.
  • “Functional variant thereof means also peptide nucleic acid molecules that could substitute for the oligonucleotides.
  • the method of the invention may preferably be used for monitoring the presence of the harmful microorganisms, especially the primary biofilm-forming microorgamsms, in the paper and board machines, in pulp-drying machines, and in the wet manufacturing processes for raw materials of paper and pulp industry.
  • the presence of the biofilm-forming microorganisms, especially the primary biofilm-formers is monitored for determining the need of an anti-biofilm agent in the paper or board making process and/or for determining the effects of such an anti-biofilm agent.
  • the invention further provides a method for inhibiting the formation of agglomerates and/or a biofilm by thermophilic microbes capable of adhering to the surfaces of the paper and board machines and/or for removing such agglomerates and/or biofilms from said surfaces, wherein at least one anti-biofilm agent is added in an amount effective against said microbes to the circulation water of said paper or board making machines, whereby the need of said anti-biofilm agent is monitored with the detection method of the invention.
  • the process can be monitored continuously, whereby the need of an anti-biofilm agent, the timing of the addition and/or of any further addition as well as the doses of the anti-biofilm agent can be optimized over to the prior art.
  • Anti-biofilm agent means herein generally any substance suitable to paper industry for the inhibition or prevention of the growth of harmful microbes, preferably of biofilm- and agglomerates-forming microorganisms, and, particularly, for the inhibition or prevention of the formation of a biofilm and/or removal of the formed biofilm from the surfaces of the process equipment.
  • substances include any compounds, mixtures of compounds and extracts, i.a. chemicals and biocides conventionally used in the paper industry, such as 2,2-dibromo-3- nitrilopropionamide (DBNPA) and methylene bisthiocyanate (MBT), or effective agents of plant origin, compounds and extracts, such as those mentioned below.
  • the anti-biofilm agent used for inhibiting the biofilm-formation in the present method is a pure substance isolated from a plant or a plant extract, or a mixture thereof.
  • agent originating from a plant means a natural agent isolated from a plant or a synthetic equivalent or derivative thereof, which has an ability to inhibit the formation of the biofilm by thermophilic microbes and/or to remove such biofilms from a surface.
  • the reduction achieved in the biofilm-formation should be at least 50 %, preferably at least 70 % and more preferably at least 90 %.
  • the agents of plant origin can reduce the adhering of the biofilm-formers, some of them up to 90 % or more.
  • the plant extracts can prevent effectively, 91-99.7 %, the agglomerate forming D. geothermalis of adhering on the surface of the paper machine steel (AISI 316).
  • the prevention achieved e.g. with betuline and flavonols varies between 84-99.95 % depending on the used agent.
  • the "agents originating from the plant” can be used as raw extracts or as more effective components isolated from said extracts.
  • the "agent originating from a plant” or “pure compound” or “plant extract” effective against biofilm- formation may originate e.g. from Japanese rose (Rosa rugosa), rosebay willow herb (Epilobium angustifolium), meadowsweet (Filipendula ulmarid), or salvia (Salvia officinalis), and against agglomeration e.g.
  • the extract can be obtained by extracting the plant or a part of it with a solvent or a mixture of solvents.
  • a solvent is methanol.
  • suitable solvents for the extraction include cold water, acetone, ethanol, hexane and chloroform.
  • phenolic compounds such as an ester of a phenolic acid.
  • a preferable ester of a phenolic acid is an alkyl ester of a gallic acid, preferably octyl gallate or lauryl gallate or a mixture thereof, and against agglomeration betuline and flavonols, such as pentahydroxyflavone and trihydroxyflavone, or a mixture thereof.
  • agents originating from a plant such as “pure compound” or “extract” may be added e.g. to circulating waters of a paper or board machine to a product concentration of 1-1000 ppm, preferably 5-200 ppm, and more preferably 10- 100 ppm, calculated from the dry weight of the "agent originating from a plant”.
  • the invention further provides an assembly kit for monitoring the presence of agglomerate- and/or biofilm-forming microorganisms in a paper or board making process.
  • Said kit comprises one or more oligonucleotide probes hybridizable with primary biofilm-forming microorganism(s) occurring in paper and board industry and labelled with a detectable marker.
  • the assembly kit according to the invention comprises one or more new specific oligonucleotide probe(s) as defined above.
  • the reagents for fixing the sample and the hybridizing and washing buffers are in general known to a skilled person and may be chosen to be suitable for the present microorganisms and probe(s). Suitable examples are described above and can be generalized from the examples below.
  • the support for receiving the sample is a sampler support as defined above, e.g. a test plate, to be placed in the process for enabling the biofilm-formers to form a biofilm on the surface thereof in situ in the process, to be used as a support for the fixation, i.e. permeabilization, of the microorganisms present in the biofilm formed on the surface thereof and for the hybridization of the oligonucleotide probe(s) with the target microorganism(s) present in the biofilm formed on the surface thereof, and optionally for the detection step.
  • the kit may further comprises miniature hybridization chamber(s), e.g. those known in the field of bioscience and commercially available, which can be attached, e.g. glued, on the support for retaining the hybridization buffer on the support, e.g. on the test plate.
  • the assembly kit can also be provided with a detection device for the label used for the probe.
  • a detection device for the label used for the probe E.g. for a fluorescent label an epifluorescent microscope can be used.
  • the hybridization method and the assembly kit of the invention can be used for detecting the efficacy of an anti-biofilm agent against biofilm-formers in a process, whereby the specific hybridization for the biofilm- formers is performed after the sample has been exposed to an anti-biofilm agent.
  • Samples in the experiments were 1) bacterial pure cultures in active growth phase in liquid growth media, 2) slime samples freshly collected from surfaces of paper and board machines, and 3) biofilms grown on stainless steel test plates which had been immersed in the process water of paper or board machine for 2 d.
  • Sample pretreatment Bacterial cultures (a 8 ⁇ l) or slime samples were immobilized by air-drying to the surface of microscopy slides (Teflon printed glass slides, 10- well, coated with gelatin) and then dehydrated by successive passages through 50%, 80%, 96% and abs. ethanol (3 min each). Biofilms grown on stainless steel test plates were treated similarly, except that miniature hybridization chambers (Secure- SealTM hybridization chamber, Molecular Probes, Eugene, OR, USA) were glued on the stainless steel test plates after the dehydration step. These disposable plastic chambers were used in order to prevent the hybridization buffer from running off the steel surface during hybridization.
  • Successful in situ hybridization requires stringent conditions that allow binding of the probe to the completely matching target rRNA sequences only. This is achieved by adjusting the formamide concentration of the hybridization buffer (X 1 ) and the NaCl concentration of the washing buffer (X ). The stringent conditions for each probe were experimentally screened.
  • SEQ ID NO.l An oligonucleotide probe shown in SEQ ID NO.l (5'-CCC ATC CGG GGC CTT TCG- 3') was designed for specific detection of the primary biofilm-former Deinococcus geothermalis.
  • the sequence of SEQ ID NO.l -probe was compared to the prokaryotic 16S rRNA gene sequences (ca. 72000 sequences at 26.6.2003) in the RDP-II database (Ribosomal Database Project at http://rdp.cme.msu.edu/html/). Only four strains with completely matching target 16S rRNA sequences were found, covering all strains representing the species D.
  • geothermalis in the database strains AG-3a T , AG-5a and RSPS-2a from hot springs and E50053 from paper machine.
  • the probe was separated from the next nearest hit in the RDP database (Brochothrix thermosphacta ATCC11509 T ) by three mismatching bases indicating that SEQ ID No.l -probe is specific for D. geothermalis.
  • Figure 1 shows specific binding of the probe shown in SEQ ID NO.l to the cells of primary biofilm-former Deinococcus geothermalis.
  • the specimen was a mixture of broth cultures of eight different species: D. geothermalis E50053, Bacillus amyloliquefaciens TSP55, B. licheniformis D50141, B. megaterium HAMBI696, B. pumilus TSP66, D. radiodurans DSM20539 , Micrococcus luteus HAMBI1399 and Staphylococcus aureus HAMBI66.
  • the epifluorescence microscope image in figure 1A shows that only cells of D. geothermalis E50053 were labeled by the fluorescently tagged SEQ ID NO.l -probe, whereas the phase contrast image in Figure IB shows cells of all eight species in the specimen.
  • Figure 1 demonstrates that cells of the primary biofilm-forming species D. geothermalis can be specifically hybridized in situ with the probe shown in SEQ ID NO.l and thereafter microscopically detected despite of the presence of other bacteria in the same specimen.
  • the red-gray slime was sampled from the vicinity of the first press cylinder of a magazine paper machine (52°C, pH 7).
  • Image shows a brightly fluorescent microcolony (arrow) of coccoidal cells that were labeled with SEQ ID NO.l -probe indicating that they are D. geothermalis cells.
  • the red slime was sampled from the splash area of wire section of a newsprint machine (52°C, pH 5). Almost all cells visible in the image were labeled by the SEQ ID NO.l -probe (intensive orange fluorescence) indicating that they were D. geothermalis cells. Based on microscopic cell counting D. geothermalis constituted ca. 10% of the all bacteria in this particular slime specimen.
  • the biofilm was formed on a stainless steel test plate during a two-day immersion in the cloudy-filtrate container (52°C, pH 7). Image shows only a few cells labeled with the SEQ ID NO.l -probe.
  • D. geothermalis constituted less than 1% of the all bacteria in this particular slime specimen indicating that in this site of the process the major primary biofilm-former was other species than D. geothermalis.
  • Figures 2, 3 and 4 demonstrate that the presence of the biofilm-forming D. geothermalis in process samples can be monitored using in situ hybridization and specific probes. This approach is faster than the traditional cultivation-based approaches.
  • the cells of D. geothermalis were abundant, whereas in the board machine specimen ( Figure 4) this adverse bacterium was present only in low amounts.
  • SEQ ID NO.14 An oligonucleotide probe shown in SEQ ID NO.14 was designed for specific detection of the primary biofilm-forming bacteria of the genus Meiothermus.
  • the sequence of SEQ ID NO.14 -probe was compared to the prokaryotic 16S rRNA gene sequences in the RDP-II database. Twelve completely matching target 16S rRNA sequences were found, all originating from strains belonging to the genus Meiothermus.
  • the probe was separated from the two next nearest hits in the RDP database (Lactobacillus vaginalis and Paenibacillus pabul ⁇ ) by four mismatching bases indicating that SEQ ID NO.14 -probe is specific for bacteria of the genus Meiothermus.
  • Figure 5 demonstrates that primary biofilm-forming bacteria of the genus Meiothermus can be specifically hybridized with the SEQ ID NO.14 -probe.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Agronomy & Crop Science (AREA)
  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Hydrology & Water Resources (AREA)
  • Environmental & Geological Engineering (AREA)
  • Water Supply & Treatment (AREA)
  • Organic Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de contrôle de micro-organismes nocifs dans l'industrie du papier et du carton. Selon ce procédé, la présence ou l'absence du micro-organisme cible nocif est détectée par un procédé d'hybridation in situ au moyen d'une sonde oligonucléotidique pouvant être hybridée avec une zone d'une molécule d'acide nucléique dudit micro-organisme. Plus particulièrement, ce micro-organisme peut être choisi dans un groupe comprenant l'espèce Deinococcus geothermalis et les genres Meiothermus, Azospirillum, Burkholderia, Porhyrobacter, « Rubellimicrobium » et Thermomonas.
PCT/FI2003/000839 2002-11-06 2003-11-06 Procede de controle de la presence de micro-organismes nocifs dans l'industrie du papier WO2004042082A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003276306A AU2003276306A1 (en) 2002-11-06 2003-11-06 A method for monitoring the presence of harmful microorganisms in paper industry

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
FI20021987 2002-11-06
FI20021986A FI116030B (fi) 2002-11-06 2002-11-06 Paperi- ja kartonkikoneiden termofiilisten mikrobien biofilminmuodostuksen esto
FI20021986 2002-11-06
FI20021987A FI115502B (fi) 2002-11-06 2002-11-06 Bakteerien aiheuttaman kertymänmuodostuksen estäminen paperi- ja kartonkikoneiden pinnoille

Publications (2)

Publication Number Publication Date
WO2004042082A1 true WO2004042082A1 (fr) 2004-05-21
WO2004042082A9 WO2004042082A9 (fr) 2005-05-26

Family

ID=32313914

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FI2003/000839 WO2004042082A1 (fr) 2002-11-06 2003-11-06 Procede de controle de la presence de micro-organismes nocifs dans l'industrie du papier

Country Status (2)

Country Link
AU (1) AU2003276306A1 (fr)
WO (1) WO2004042082A1 (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1939352A1 (fr) * 2002-09-30 2008-07-02 Nalco Company Procédé pour la détection de matériaux améliorant la performance dans un cylindre à crêper
US8613837B2 (en) 2012-01-24 2013-12-24 Nalco Company Detection and quantification of nucleic acid to assess microbial biomass in paper defects and machine felts
WO2014015044A1 (fr) * 2012-07-17 2014-01-23 Nalco Company Procédé de construction d'indices de diversité et de viabilité de micro-organismes dans des échantillons de traitement
US20140141443A1 (en) * 2012-01-24 2014-05-22 Nalco Company Detection and quantification of nucleic acid to assess microbial biomass in paper defects and machine felts
US20150000853A1 (en) * 2012-01-20 2015-01-01 Kemira Oyj Device and method for monitoring biocide dosing in a machine
WO2015025083A1 (fr) * 2013-08-19 2015-02-26 Oy Separation Research Ab Composition d'hydrophobisation, procédé d'hydrophobisation et substrat
WO2015100123A1 (fr) * 2013-12-23 2015-07-02 Nalco Company Détection et quantification d'acides nucléiques pour évaluer la biomasse microbienne dans des défauts de papier et feutres pour machine à papier
US9295745B2 (en) 2012-01-24 2016-03-29 Nalco Company Method for constructing a diversity index and a viability index of microorganisms in process samples
WO2016071805A1 (fr) * 2014-11-07 2016-05-12 Stora Enso Oyj Procédé amélioré pour la détermination de micro-organismes
US9908796B2 (en) 2012-10-23 2018-03-06 Ecolab Usa Inc. Use of oxidizing and non-oxidizing biocides for control of bacteria tolerant to stabilized-oxidant treatment
EP3476928A1 (fr) * 2017-10-27 2019-05-01 Envall Consulting AB Détection de la croissance bactérienne
US11608516B2 (en) 2015-04-15 2023-03-21 Ecolab Usa Inc. Method for determination of diversity and viability thresholds used to assess microorganisms in process samples

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001007649A2 (fr) * 1999-07-22 2001-02-01 Vermicon Ag Procede de detection de micro-organismes
WO2001036673A2 (fr) * 1999-11-17 2001-05-25 Creatogen Ag Test pour micro-organismes
EP1130115A2 (fr) * 2000-03-03 2001-09-05 GSF - Forschungszentrum für Umwelt und Gesundheit, GmbH Sondes oligonucléotidiques pour la détection spécifique de genre et d'espèce de bactéries qui stimulent la croissance des plantes.
WO2002054865A1 (fr) * 2001-01-09 2002-07-18 Kurita Water Industries Ltd. Procede de selection d'un agent antimicrobien et procede d'utilisation de ce dernier

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001007649A2 (fr) * 1999-07-22 2001-02-01 Vermicon Ag Procede de detection de micro-organismes
WO2001036673A2 (fr) * 1999-11-17 2001-05-25 Creatogen Ag Test pour micro-organismes
EP1130115A2 (fr) * 2000-03-03 2001-09-05 GSF - Forschungszentrum für Umwelt und Gesundheit, GmbH Sondes oligonucléotidiques pour la détection spécifique de genre et d'espèce de bactéries qui stimulent la croissance des plantes.
WO2002054865A1 (fr) * 2001-01-09 2002-07-18 Kurita Water Industries Ltd. Procede de selection d'un agent antimicrobien et procede d'utilisation de ce dernier

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DAIMS H ET AL: "THE DOMAIN-SPECIFIC PROBE EUB338 IS INSUFFICIENT FOR THE DETECTION OF ALL BACTERIA: DEVELOPMENT AND EVALUATION OF A MORE COMPREHENSIVE PROBE SET", SYSTEMATIC AND APPLIED MICROBIOLOGY, XX, XX, vol. 22, no. 3, September 1999 (1999-09-01), pages 434 - 444, XP009012835, ISSN: 0723-2020 *
DATABASE WPI Section Ch Week 200273, Derwent World Patents Index; Class B04, AN 2002-682647, XP002272494 *
LEMKE MICHAEL J L ET AL: "Comparison of methods for the concentration of bacterioplankton for in situ hybridization", JOURNAL OF MICROBIOLOGICAL METHODS, vol. 29, no. 1, 1997, pages 23 - 29, XP002272492, ISSN: 0167-7012 *
PALLUD CELINE ET AL: "Combined use of a specific probe and PCAT medium to study Burkholderia in soil", JOURNAL OF MICROBIOLOGICAL METHODS, vol. 47, no. 1, October 2001 (2001-10-01), pages 25 - 34, XP002272491, ISSN: 0167-7012 *
STOFFELS MARION ET AL: "Bacterial community dynamics during start-up of a trickle-bed bioreactor degrading aromatic compounds", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 64, no. 3, March 1998 (1998-03-01), pages 930 - 939, XP002272493, ISSN: 0099-2240 *

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1939352A1 (fr) * 2002-09-30 2008-07-02 Nalco Company Procédé pour la détection de matériaux améliorant la performance dans un cylindre à crêper
US20150000853A1 (en) * 2012-01-20 2015-01-01 Kemira Oyj Device and method for monitoring biocide dosing in a machine
US9341560B2 (en) * 2012-01-20 2016-05-17 Kemira Oyj Device and method for monitoring biocide dosing in a machine
US9295745B2 (en) 2012-01-24 2016-03-29 Nalco Company Method for constructing a diversity index and a viability index of microorganisms in process samples
US20140141443A1 (en) * 2012-01-24 2014-05-22 Nalco Company Detection and quantification of nucleic acid to assess microbial biomass in paper defects and machine felts
KR102037510B1 (ko) 2012-01-24 2019-10-28 날코 컴퍼니 종이 결함 및 기계 펠트에서 미생물 바이오매스를 평가하기 위한 핵산의 검출 및 정량
US9290802B2 (en) 2012-01-24 2016-03-22 Nalco Company Detection and quantification of nucleic acid to assess microbial biomass in paper defects and machine felts
KR20140131928A (ko) * 2012-01-24 2014-11-14 날코 컴퍼니 종이 결함 및 기계 펠트에서 미생물 바이오매스를 평가하기 위한 핵산의 검출 및 정량
US8613837B2 (en) 2012-01-24 2013-12-24 Nalco Company Detection and quantification of nucleic acid to assess microbial biomass in paper defects and machine felts
CN104471072A (zh) * 2012-07-17 2015-03-25 纳尔科公司 构建工艺样品中微生物的多样性指数和生存力指数的方法
WO2014015044A1 (fr) * 2012-07-17 2014-01-23 Nalco Company Procédé de construction d'indices de diversité et de viabilité de micro-organismes dans des échantillons de traitement
CN104471072B (zh) * 2012-07-17 2016-12-21 纳尔科公司 构建工艺样品中微生物的多样性指数和生存力指数的方法
US9908796B2 (en) 2012-10-23 2018-03-06 Ecolab Usa Inc. Use of oxidizing and non-oxidizing biocides for control of bacteria tolerant to stabilized-oxidant treatment
US10640402B2 (en) 2012-10-23 2020-05-05 Ecolab Usa Inc. Use of oxidizing and non-oxidizing biocides for control of bacteria tolerant to stabilized-oxidant treatment
US9963600B2 (en) 2013-08-19 2018-05-08 Oy Separation Research Ab Hydrophobisation composition, method for hydrophobisation and substrate
WO2015025083A1 (fr) * 2013-08-19 2015-02-26 Oy Separation Research Ab Composition d'hydrophobisation, procédé d'hydrophobisation et substrat
WO2015100123A1 (fr) * 2013-12-23 2015-07-02 Nalco Company Détection et quantification d'acides nucléiques pour évaluer la biomasse microbienne dans des défauts de papier et feutres pour machine à papier
JP2017532977A (ja) * 2014-11-07 2017-11-09 ストラ エンソ オーワイジェイ 微生物の決定のための改善された方法
WO2016071805A1 (fr) * 2014-11-07 2016-05-12 Stora Enso Oyj Procédé amélioré pour la détermination de micro-organismes
US10017806B2 (en) 2014-11-07 2018-07-10 Stora Enso Oyj Method for determination of microorganisms
US11608516B2 (en) 2015-04-15 2023-03-21 Ecolab Usa Inc. Method for determination of diversity and viability thresholds used to assess microorganisms in process samples
EP3476928A1 (fr) * 2017-10-27 2019-05-01 Envall Consulting AB Détection de la croissance bactérienne

Also Published As

Publication number Publication date
AU2003276306A1 (en) 2004-06-07
WO2004042082A9 (fr) 2005-05-26

Similar Documents

Publication Publication Date Title
Schwartz et al. Formation and bacterial composition of young, natural biofilms obtained from public bank-filtered drinking water systems
EP1687481B1 (fr) Procede permettant de controler la presence de micro-organismes generateurs de films biologiques dans l'industrie papetiere
Manz et al. In situ identification of bacteria in drinking water and adjoining biofilms by hybridization with 16S and 23S rRNA-directed fluorescent oligonucleotide probes
WO2004042082A1 (fr) Procede de controle de la presence de micro-organismes nocifs dans l'industrie du papier
Lemos et al. The effect of shear stress on the formation and removal of Bacillus cereus biofilms
Fischer et al. In situ analysis of the bacterial community in the gut of the earthworm Lumbricus terrestris L. by whole-cell hybridization
Si et al. Prevention of multi-species wastewater biofilm formation using vanillin and EPS disruptors through non-microbicidal mechanisms
Bitton et al. Tetrazolium reduction-malachite green method for assessing the viability of filamentous bacteria in activated sludge
WO2008137195A1 (fr) Procédé d'élimination de microbes de surfaces
MacDonald et al. Community analysis of bacterial biofilms in a simulated recirculating cooling-water system by fluorescent in situ hybridization with rRNA-targeted oligonucleotide probes
CA2503648A1 (fr) Inhibition de la formation d'un film biologique par des microbes thermophiles dans des machines a papier ou a carton
Kenzaka et al. Bacterial community composition and activity in urban rivers in Thailand and Malaysia
Lee et al. Membrane biofouling of seawater reverse osmosis initiated by sporogenic Bacillus strain
Di Gioia et al. Aggregation-based cooperation during bacterial aerobic degradation of polyethoxylated nonylphenols
Decamp et al. Abundance, biomass and viability of bacteria in wastewaters: impact of treatment in horizontal subsurface flow constructed wetlands
Forster et al. Rains, drains and active strains: towards online assessment of wastewater bacterial communities
AU2009231362B2 (en) Detection and enumeration of microorganisms
Yu et al. In situ activity recovery of aging biofilm in biological aerated filter: Surfactants treatment and mechanisms study
RU2005103822A (ru) Олигонуклеиды для выявления микроорганизмов
Mattila Biofilms on stainless steels exposed to process waters
Ramothokang et al. ‘Morphological shifts’ in filamentous bacteria isolated from activated sludge processes
Adonadaga et al. In situ identification of filamentous bacteria from activated sludge wastewater treatment plants in Ghana
Wolfaardt et al. The use of DAPI in the quantification of sessile bacteria on submerged surfaces
US20070243532A1 (en) System and Method of Detecting a Microorganism
Riihinen Biofilm management in paper and packaging board manufacturing process Bacterial community analysis by NGS technolog

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): BW GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
COP Corrected version of pamphlet

Free format text: PAGE 3/3, DRAWINGS, ADDED

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP