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WO2000047709A1 - Appareil et procede de conservation de micro-organismes - Google Patents

Appareil et procede de conservation de micro-organismes Download PDF

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Publication number
WO2000047709A1
WO2000047709A1 PCT/GB2000/000378 GB0000378W WO0047709A1 WO 2000047709 A1 WO2000047709 A1 WO 2000047709A1 GB 0000378 W GB0000378 W GB 0000378W WO 0047709 A1 WO0047709 A1 WO 0047709A1
Authority
WO
WIPO (PCT)
Prior art keywords
accordance
storage device
growth medium
microorganism
vessel
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.)
Ceased
Application number
PCT/GB2000/000378
Other languages
English (en)
Inventor
Neil Porter
Frances Mary Giaquinto
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BioDiversity Ltd
Original Assignee
BioDiversity Ltd
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
Application filed by BioDiversity Ltd filed Critical BioDiversity Ltd
Priority to EP00901817A priority Critical patent/EP1165747A1/fr
Priority to JP2000598609A priority patent/JP2002536010A/ja
Priority to AU23112/00A priority patent/AU2311200A/en
Publication of WO2000047709A1 publication Critical patent/WO2000047709A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/58—Reaction vessels connected in series or in parallel
    • 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
    • C12M23/06—Tubular
    • 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
    • C12M23/10—Petri dish
    • 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
    • C12M23/22—Transparent or translucent parts
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis

Definitions

  • the present invention relates to preparing and maintaining cultures of microorganisms, and is particularly applicable to fungi and bacteria.
  • Microorganisms are essential to many important biotechnical processes including the production of foods, fine chemicals such as vitamins and organic acids, pharmaceuticals, enzymes, agrochemicals and biological control agents.
  • microorganisms have yielded drugs which are used for treating infections, disorders of the central nervous system, cardiovascular disease and for suppression of the immune system to prevent rejection following organ transplantation. They hold enormous potential for producing new pharmaceutical compounds .
  • microorganisms As a consequence of the economic and academic importance of microorganisms, microbial genetic resource collections have been established to provide cultures of microorganisms for research.
  • the World Data Center for Microorganisms has been assigned the role of characterising, cataloguing and most importantly preserving microorganisms in a pure, viable and genetically stable condition. Over 500 collections of microorganisms have been registered with the World Data Center.
  • microorganism cultures are stored under conditions that maintain genetic stability. Genetic deterioration of a microorganism during storage can result in a reduction or total loss of its biotechnological properties (known or as yet undiscovered) . This can result in a significant financial loss to a company.
  • a culture can be maintained on a growth substrate by means of repeated sub-culture onto a new substrate as the growth substrate deteriorates.
  • a culture can be maintained on a growth substrate by means of repeated sub-culture onto a new substrate as the growth substrate deteriorates.
  • it is possible to create an environment where metabolism of a culture is severely reduced or halted (Smith, D & Onions, AHS, (1994) "The Preservation and Maintenance of Living Fungi", 2 nd ed. Wallingford, CAB International) .
  • FIG 1 shows a Petri dish 10 which is filled with a layer of agar 12 through which is dispersed a nutrient medium containing sources of carbon, nitrogen, phosphorus, essential vitamins and other elements required for growth.
  • Agar is a natural carbohydrate substance extracted from seaweed.
  • a sample of an organism 14 is inoculated onto the agar at the centre of the Petri dish 10.
  • the Petri dish 10 is then left in a clean environment for a period of 10 days to 2 weeks, or longer if necessary, and maintained at a temperature suitable to promote growth, e.g. 15-25°C. Following that period, the dish will have the appearance as illustrated in Figure 2.
  • a colony of the organism 14 has developed by the growth of filamentous strands along the surface of the agar 12 in all directions from the original sample.
  • a sub-culture sample 16 can be taken from the growing edge, so as to sample the youngest and most viable part of the filaments. That sub-culture sample can then be inoculated at the centre of a further Petri dish of agar for further culturing. The sample 16 can be taken by means of a sterile scalpel. The original Petri dish can then be discarded.
  • Microorganisms and particularly fungi, are inherently genetically variable. For example, as illustrated in Figure 2, whereas the microorganism normally has a green appearance, the zone 18 of the culture identified by chain lines could have genetically segregated so that it has a red appearance. This phenomenon is known as sectoring.
  • This second sample 16' would maintain the full genetic composition of the microorganism, which would reduce the problems resulting from genetic segregation.
  • some sampling error may remain, since a technologist may not be able to identify all genetic modifications by observation, and so important genetic material may be discarded as a result of the sub-culturing method described above.
  • the proportion of materials of different types to be sampled would be a matter for a sampler to identify, which could introduce further errors .
  • microorganism populations are genetically heterogeneous. Consequently, ongoing sub-culture and growth on a synthetic agar medium can act as a selective pressure ensuring that a proportion of the population best suited to those particular conditions of growth become dominant. Desirable properties of microorganisms can be lost as a result. Long term storage of microorganisms by repeated sub-culture is therefore not desirable.
  • cryopreservation at ultra-low temperatures Smith, D (1993) "Tolerance to freezing and thawing", Tolerance of Fungi, Editor - Jennings, DH pp 145-171 published by Marcel Dekker Inc, New York Smith, D (1998) "The use of cryopreservation in the ex-situ conservation of fungi” - Cryoletters 19, 79-90). Little metabolic activity occurs below -70°C but recrystallisation of ice, which can cause cell damage, can occur above -130°C.
  • microorganisms are stored at temperatures below -130°C; in refrigerators (-135°C to -180°C) or in liquid nitrogen vapour at -196°C.
  • Cellular damage due to ice crystal formation can occur if the freezing and thawing rates are not carefully controlled and the use of cryoprotectant chemicals is important to minimise this damage (Smith (1998) previously referred to; Smith D & Thomas VE (1998) "Cryogenic light microscopy and the development of cooling protocols for the cryopreservation of filamentous fungi", World Journal of Microbiology and Biotechnology, 14, 49-57).
  • Genetic selection can also be a problem with this method of storage; only a small amount of culture biomass is taken for cryopreservation and only a proportion of that small amount may be viable when the material is thawed.
  • Cryopreservation is the most expensive in terms of capital equipment required, its running costs and preparation of cultures for storage.
  • the time taken for microorganisms to recover from storage where their cellular metabolic activity has been reduced can take three weeks or longer. It is not possible to do anything with the microorganism before the end of this period. In addition to the inconvenience caused, this time delay can add significantly to the costs of biotechnological processes.
  • a first aspect of the invention has as its object the improvement of sampling techniques to maintain reliability of sub-culturing as a method of maintaining a sample of an organism.
  • agar is somewhat undesirable, in that it is a synthetic growth environment which is in some respects different from the nutrient environment which microorganisms would naturally encounter.
  • agar media are designed to simulate, as closely as possible, the combination of nutrients most amenable to the growth of microorganisms, they remain approximations.
  • the main advantage of agar is that it provides a solid substrate that is not broken down by the microorganism.
  • the invention provides, in a first aspect, a method of sub-culturing which involves maintenance of microbiological material without selection of specific samples thereof.
  • the invention provides, in a second aspect, sub-culturing apparatus for presenting a sub-culture across substantially an entire population of a culture.
  • the invention provides, in a third aspect, a method of generating a metabolite from an organism maintained according to the first aspect of the invention.
  • the invention also provides, in a further aspect, a method of manufacturing a chemical composition from the metabolite generated in accordance with the third aspect of the invention.
  • Figure 1 is a perspective view of a Petri dish in accordance with an example of an existing technique
  • Figure 2 is a plan view of a culture on the Petri dish illustrated in Figure 1;
  • Figure 3 is a perspective view of a receptacle in accordance with a specific embodiment of the present invention.
  • Figure 4 is a longitudinal section of the receptacle illustrated in Figure 3 in an initial condition
  • Figure 5 is a perspective view of an insert of the receptacle illustrated in Figure 3;
  • Figure 6 is a perspective view of an alternative insert to that illustrated in Figure 5;
  • Figure 7 shows a longitudinal section of an end portion of receptacle in accordance with an alternative and specific embodiment of the invention.
  • Figure 8 is a perspective view of a receptacle of a further alternative and specific embodiment of the present invention.
  • Figure 9 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a first stage of use in accordance with a specific exemplary method
  • Figure 10 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a second stage of use in accordance with a specific exemplary method
  • Figure 11 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a third stage of use in accordance with a specific exemplary method
  • Figure 12 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a fourth stage of use in accordance with a specific exemplary method
  • Figure 13 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a fifth stage of use in accordance with a specific exemplary method
  • Figure 14 is a longitudinal sectional view of the receptacle illustrated in Figure 3 in a sixth stage of use in accordance with a specific exemplary method.
  • Figure 15 is a longitudinal sectional view of an arrangement of receptacles as illustrated in Figure 3 for use in accordance with an alternative specific exemplary method.
  • a receptacle 30 has a generally hollow cylindrical body 32 which is open at both ends. As shown in Figure 4, each end of the body 32 has an external screw thread 34, and is closed by a cap 36 having a cooperating internal screw thread 38. The caps 36 may be fitted on the body 32 sufficiently tightly that a seal is formed to prevent ingress of microscopic contaminants into the receptacle 30.
  • the material of the body 32 and the caps 36 is of material which is readily sterilisable for use in biotechnological applications. Moreover, the material is preferably transparent, which allows for observation of the interior of the receptacle 30.
  • a suitable material could be glass, or plastics such as polystyrene, polyethylene, polyamide, polyacrylate. Especially important examples of a suitable material are polycarbonate or polypropylene, which can withstand sterilisation by means of hot water vapour at temperatures up to 121°C.
  • Each cap 36 extends over its respective end of the body 32 to a depth of not less than 25 mm to ensure that the ends of the body remain sterile when one or both of the caps 36 are removed.
  • the thickness of the wall of the cylindrical body 32 is
  • the body may be of any other suitable shape other than that of a cylinder.
  • the internal diameter of the body 32 is 22mm, although this can be varied in alternative embodiments, for example up to 100 mm. Moreover, in the present example, the body 32 is 90 mm in length, but other lengths of body 32 are also envisaged, for instance 50, 150 or 250 mm.
  • An insert 40 as illustrated in Figure 5, comprises a fine mesh 42, supported on a circular collar 44.
  • the insert 40 is placed at one end of the interior of the body 32 (the right hand end as illustrated in Figure 4).
  • the mesh 42 retains the contents of the receptacle 30 in the event of removal of the cap 36 at that end.
  • the collar 44 is of a size suitable for it to form a tight fit within the body 32, to reduce the risk of the insert 40 accidentally falling out of place.
  • the mesh 42 is sufficiently fine as to prevent egress of any growth medium contained in the receptacle therethrough, but not so fine that a filamentous microorganism is impeded from growing therethrough.
  • Figure 6 shows an alternative insert 40' having two crosspieces 42' supported on a collar 44'.
  • the collar 44' is identical with the collar 44 illustrated in Figure 5.
  • the crosspieces 42' extend diametrically and mutually perpendicularly across the collar 44 ' .
  • the crosspieces 42' are operative, in use, to retain any contents of the receptacle 30 to the extent that the cap 36 at that end can be removed and replaced without significant shifting of the medium between the crosspieces 42'. It will be understood that the crosspieces 42' act to impede bulk movement rather than actively preventing it.
  • FIG. 7 of the drawings An alternative example of a receptacle 30' including a push-type fitting between a body 32 ' and a cap 36 ' is illustrated in Figure 7 of the drawings.
  • the second embodiment of the insert 40' as illustrated in Figure 6, has been fitted at the end of the body 32' .
  • the end of the body 32' is tapered on its exterior surface, and a corresponding interior tapered surface is formed on the cap 36' .
  • the cap 36' can then be urged onto the end of the body 32' and, by means of friction and selection of suitable taper angles, the cap 36' can be retained on the body 32', forming a tight seal.
  • Other than this push-fit lid fitting arrangement alternative arrangements are also envisaged, for example a bayonet fitting, and a push and twist fitting.
  • a rectangular membrane 46' is incorporated into the wall of the body 32' .
  • the membrane 46' is of a hydrophobic material, such as polytetrafluoroethylene or polysiloxane, which allows the transfer of gases therethrough, for example oxygen, which is required in many circumstances for the growth of microorganisms.
  • the membrane may be located within one or other of the caps 36'.
  • the material selected for the membrane 46' should be suitable for withstanding sterilisation by means of hot water vapour at temperatures up to 121°C.
  • the membrane extends along the length of the body 32' , up to a distance of 25 mm from each end of the body 32' .
  • a growing medium should contain assimilable sources of carbon, nitrogen and mineral salts.
  • Assimilable sources of carbon, nitrogen and minerals may be provided by simple or complex nutrient sources.
  • complex nutrient sources are used since they reflect more accurately the natural substrates on which the microorganisms grow.
  • the great variety of nutrients present in complex sources may prevent the unwanted selection of genetic variants existing in a microorganism population which can occur by the placement of the population in an unnatural environment.
  • Complex sources of carbon, nitrogen and minerals may be provided by clean (not containing chemical residues such as fungicides or other pesticides) grains, cereals and seeds. Examples of such sources are tabulated below:
  • a medium is formed of a mixture of the above
  • Peptone is a hydrolysed protein which can
  • receptacle 30 is filled with the soaked mixture to a
  • the receptacle 30 is
  • microorganism The microorganism. These conditions include temperatures
  • microorganisms or invertebrate pests such as mites.
  • incubation conditions can include humidity and light regulation.
  • a moderately humid environment will reduce
  • Figure 10 illustrates the expected appearance of a
  • conditions may include humidity and light regulation.
  • Figure 11 shows a receptacle 30 after storage thereof for
  • the microorganism is nearing the end of the receptacle 30
  • the collar 54 is sufficient in one direction to draw the
  • receptacle is discarded and a sterile cap 36 is placed at
  • the present invention is particularly applicable to
  • basidiomycotina basidiomycotina
  • ascomycotina basidiomycotina
  • other sterile mycelia basidiomycotina
  • the mycelium is the source of inoculum. Therefore, by
  • Phlebia Phlebia
  • deflectens is a basidiomycete which is commonly found growing on rotten deciduous wood in British woodland.
  • the sub-culturing apparatus 30 is filled
  • a growth medium 50 consisting of swollen quinoa
  • the quinoa grain is
  • sample 52 is placed at one end of the sub-culturing
  • apparatus 30 is then labelled and stored at 22 °C for four
  • Phlebia deflectens is a basidiomycete that does not
  • sub-culturing receptacle 30 maintains the vigour
  • mycelium is then transferred to a Petri dish containing
  • Phlebia deflectens grows at a moderate rate in the sub-
  • basidiomycetes and ascomycetes grow at a faster rate than
  • the apparatus may be of relatively simple
  • the illustrated embodiment can be stored easily on racks
  • receptacle 30 has an insert 40 as previously described,
  • caps 36 are closed by means of caps 36.
  • a microorganism as shown in Figure 15 can be allowed to
  • a naked flame for example a Bunsen burner flame.
  • a growth stage is typically initiated by the introduction
  • organism is then cultivated by agitation at a desired
  • culture volume can be increased by a factor of ten at
  • the seed stage is used to inoculate production medium in
  • the sub-culturing receptacle 30 described above is advantageous in that it can be used to generate
  • the inoculum may be prepared in a
  • the sub-culturing apparatus can be used as a
  • the microbial biomass mixed with substrate can be any suitable microbial biomass mixed with substrate.
  • the microbial biomass mixed with substrate can be any suitable microbial biomass mixed with substrate.
  • An ideal inoculation level is 3%-
  • a terreus can be maintained and sub-cultured using the
  • the receptacle 30 using the grain quinoa as a growth medium. As described previously, the receptacle
  • the organism can be any organism that has a density of 0.8 g per cm 3 .
  • the organism can be any organism that has a density of 0.8 g per cm 3 .
  • the organism can be any material that has a density of 0.8 g per cm 3 .
  • the organism can be any material that has a density of 0.8 g per cm 3 .
  • the organism can be any material that has a density of 0.8 g per cm 3 .
  • a liquid medium A is prepared in
  • Mevinolin is assayed in both the broth and methanolic
  • the mevinolin can be any organic compound having the inoculum for the production medium.
  • the mevinolin can be any organic compound having the inoculum for the production medium.
  • a small section of growth medium can be
  • biomass can be removed and used in inoculation of further

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  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

La présente invention concerne un appareil de repiquage (30) comportant un récipient pour contenir un milieu de croissance (5) destiné à la culture d'un micro-organisme. Ce micro-organisme est amené à se développer dans un sens prédéterminé, en direction d'une première extrémité du milieu de croissance (50), où un autre appareil (30) peut être placé à proximité. Le micro-organisme peut alors croître dans le milieu de croissance du second appareil. En outre, cette invention concerne un procédé de conservation d'un micro-organisme devant être utilisé dans des processus microbiologiques, et concerne également un procédé de fermentation d'un micro-organisme conservé pour la production de produits biochimiques, comme par exemple des produits pharmaceutiques ou des produits agrochimiques.
PCT/GB2000/000378 1999-02-08 2000-02-08 Appareil et procede de conservation de micro-organismes Ceased WO2000047709A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP00901817A EP1165747A1 (fr) 1999-02-08 2000-02-08 Appareil et procede de conservation de micro-organismes
JP2000598609A JP2002536010A (ja) 1999-02-08 2000-02-08 微生物類保存装置
AU23112/00A AU2311200A (en) 1999-02-08 2000-02-08 Apparatus for preserving microorganisms

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9902757A GB2346379B (en) 1999-02-08 1999-02-08 Method and apparatus for maintaining filamentous microorganisms in a viable andgenetically stable state over a prolonged period
GB9902757.5 1999-02-08

Publications (1)

Publication Number Publication Date
WO2000047709A1 true WO2000047709A1 (fr) 2000-08-17

Family

ID=10847316

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2000/000378 Ceased WO2000047709A1 (fr) 1999-02-08 2000-02-08 Appareil et procede de conservation de micro-organismes

Country Status (5)

Country Link
EP (1) EP1165747A1 (fr)
JP (1) JP2002536010A (fr)
AU (1) AU2311200A (fr)
GB (1) GB2346379B (fr)
WO (1) WO2000047709A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4599877B2 (ja) * 2004-04-13 2010-12-15 東洋製罐株式会社 培養容器および培養方法
DE102010052434A1 (de) * 2010-11-24 2012-05-24 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Verfahren und Vorrichtung zur Überwachung einer kryokonservierten biologischen Probe
CN105002126B (zh) * 2015-08-20 2017-12-29 福建出入境检验检疫局检验检疫技术中心 一种兼性需氧菌的简易批量培养方法
EP4476491A2 (fr) * 2022-02-07 2024-12-18 Sexton Biotechnologies, Inc. Récipient de stockage cryogénique

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR403203A (fr) * 1909-05-21 1909-10-28 Montreux Sa Lab De Dispositif pour cultiver, développer, conserver, emballer et expédier des germes tels que microbes, levures, bactéries, etc., sous une forme non liquide
CH552063A (de) * 1971-04-29 1974-07-31 Bieri Otto Verfahren und vorrichtung zur anzuechtung lyophilisierter anaerober bakterien auf sterilen naehrmedien zu aktiven kulturen.
EP0259116A2 (fr) * 1986-08-28 1988-03-09 Unilever Plc Appareils et méthodes pour cultiver et tester des microorganismes
JPH0330665A (ja) * 1989-06-28 1991-02-08 Mitsubishi Heavy Ind Ltd 自動植え継ぎ装置

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH52063A (fr) * 1911-01-30 1911-10-16 Louis Pfister Raccord tubulaire pour tuyaux de descente ou d'évacuation de l'eau des toits
US2048966A (en) * 1936-04-10 1936-07-28 Brandywine Mushroom Corp Receptacle for mushroom spawn culture
GB1581832A (en) * 1977-03-28 1980-12-31 Univ Strathclyde Cultivation of filamentous fungi
FR2583059B1 (fr) * 1985-06-06 1987-10-09 Orstom Procede de production de spores de champignons filamenteux et dispositif utilise

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR403203A (fr) * 1909-05-21 1909-10-28 Montreux Sa Lab De Dispositif pour cultiver, développer, conserver, emballer et expédier des germes tels que microbes, levures, bactéries, etc., sous une forme non liquide
CH552063A (de) * 1971-04-29 1974-07-31 Bieri Otto Verfahren und vorrichtung zur anzuechtung lyophilisierter anaerober bakterien auf sterilen naehrmedien zu aktiven kulturen.
EP0259116A2 (fr) * 1986-08-28 1988-03-09 Unilever Plc Appareils et méthodes pour cultiver et tester des microorganismes
JPH0330665A (ja) * 1989-06-28 1991-02-08 Mitsubishi Heavy Ind Ltd 自動植え継ぎ装置

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PATENT ABSTRACTS OF JAPAN vol. 015, no. 154 (C - 0825) 18 April 1991 (1991-04-18) *
See also references of EP1165747A1 *

Also Published As

Publication number Publication date
EP1165747A1 (fr) 2002-01-02
GB2346379A (en) 2000-08-09
GB2346379B (en) 2004-02-18
GB9902757D0 (en) 1999-03-31
JP2002536010A (ja) 2002-10-29
AU2311200A (en) 2000-08-29

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