CN105219675A - The isolation cultivation method of a kind of anaerobic bacterium - Google Patents
The isolation cultivation method of a kind of anaerobic bacterium Download PDFInfo
- Publication number
- CN105219675A CN105219675A CN201510681477.5A CN201510681477A CN105219675A CN 105219675 A CN105219675 A CN 105219675A CN 201510681477 A CN201510681477 A CN 201510681477A CN 105219675 A CN105219675 A CN 105219675A
- Authority
- CN
- China
- Prior art keywords
- glue
- sealing
- culture dish
- bacterium
- anaerobic bacterium
- 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.)
- Pending
Links
Abstract
The invention provides the separation of a kind of anaerobic bacterium under common laboratory condition and cultural method, first prepare transparent sealing and cultivate glue and selectivity cultivation glue, again aseptically, glue is cultivated in sealing to be poured in culture dish ware lid, to be cooled solidify after, inoculate anaerobic bacterium to be seeded, be uniformly coated on the surface of sealing cultivation glue and leave standstill, then bringing Selection In property cultivates glue until cover sealing completely to cultivate glue surface, and in central microprotrusion, speed by bottom at the bottom of the culture dish ware after sterilizing down, slowly put into and gently press, venting bubble; Finally, the gap periphery sealing at the bottom of culture dish ware and between culture dish ware lid is cultivated glue and is sealed, and loads and cultivates with in 75% alcohol and the sterilized valve bag of ultraviolet lamp in advance.Method provided by the invention is not only very simple and convenient, and oxygen isolation effect is very good, meets the condition that anaerobic bacterium is required in culturing process completely, and sealing cultivation glue is transparent, is conducive to the growing state observing bacterium colony in culturing process.
Description
Technical field
The invention provides the Simple culture method of a kind of anerobe, adopt the two anti-sandwich assay of culture dish to cultivate anaerobic bacterium, this method can be applicable to the research process such as the separation of most of anaerobic bacterium, diagnosis and counting.
Background technology
Bacterium is that hybrid state exists at occurring in nature, obtain required bacterial strain, must therefrom they be separated.The method of bacteria distribution and purifying is a lot, but ultimate principle is similar, dilutes according to a certain percentage by sample to be separated or concentrate, being inoculated in selectivity and cultivating glue, make the bacterium colony of bacterium with a certain amount of, dispersion state breeding in separation and Culture process as far as possible, and make it grow up to purebred single bacterium colony one by one.
The separation and Culture of bacterium is generally divided into the cultivation of aerophil, the cultivation of anaerobic bacterium, the general cultivation of anerobe, the cultivation of strictly anaerobic bacterium.Be the checkout procedure often run into the qualification of anerobe, separation and Culture and counting in medical science and environmental monitoring, operational requirement is high, required equipment and consumptive material many, isolation identification cost is high.Because part anerobe to be exposed in air very easily dead singularity, pathological material of disease and the particular requirement of detection bodies in transport process, region and common laboratory condition such as to limit at the factor, are unfavorable for isolation identification and the scientific research of anerobe.
At present, the method both at home and abroad for purifying anaerobic bacteria culture is more, but in culturing process, all need utility appliance and a large amount of test consumable.Generally be divided into two classes, the first kind is in anaerobic bacteria culture process, needs encloses container, and adds medicaments (oxygen absorbent) in a reservoir; Second method needs large-scale device or equipment, in culturing process, constantly pass into CO
2with the oxygen in isolated external environment (as CO
2incubator), above method is cultivated anerobe and is needed certain laboratory condition, and in culturing process, resource cost is larger.All these all needs certain anaerobic environment to cultivate this feature according to anaerobic bacterium, and the condition of managing creation one its growth and breeding applicable is carried out.
The weak point of method for cultivation of bacteria of the prior art in culturing bacterium process is, one, on existing market at the bottom of the culture dish ware of microbial culture and ware lid periphery sealing not tight, anerobe is cultivated in earlier stage at the two anti-sandwich assay of culture dish, perfusion sealing need be carried out, in case the penetrating into of oxygen with agaropectin; Its two, after incubation phase bacterium colony choose aspect operation inconvenience, particularly pathogenic bacterium choose aspect, seal gum and selectivity need be separated and cultivate glue and just can choose specific bacterium colony, make troubles to operator.
Summary of the invention
The invention provides and a kind ofly utilize existing culture dish to carry out the isolation cultivation method of anaerobic bacterium by two anti-sandwich assay, to solve the condition that common laboratory does not have separation and Culture anerobe, anerobe is in censorship process or dead, or Mortality causes the technical problem that exists in the separation and Culture, count results of anaerobic bacterium compared with big error.
For this reason, the invention provides the isolation cultivation method of a kind of anaerobic bacterium, comprise the following steps,
S1: the preparation of cultivating glue
The preparation of cultivation glue comprises sealing and cultivates the preparation of glue and preparation two portions of selectivity cultivation glue, and the normal saline that wherein sealing cultivation glue is 0.9% by 16g ~ 20g agar and 1L concentration forms; Selectivity is cultivated glue and is selected different cultivation glue to prepare according to the characteristic of anaerobic bacterium to be seeded.The anaerobic bacterium can cultivated in existing CO2gas incubator all can be used as anaerobic bacterium to be seeded;
S2: the inoculation of anerobe and the sealing of inoculating surfaces
First, sterilizing is carried out to culture dish, aseptically, first cultivate glue with described sealing and pour the culture dish ware lid inner bottom part after sterilizing into, to be cooled solidify after, anaerobic bacterium to be seeded is seeded in sealing cultivate glue surface and be coated with evenly, leaves standstill 5min ~ 10min, till infiltrating in sealing cultivation glue with moisture; Secondly, selectivity is cultivated glue sterilizing and after being cooled to 45 DEG C, glue surface is cultivated in the sealing of pouring the coated bacterial classification of culture dish ware lid into, the thickness that described selectivity cultivation glue is poured into covers described sealing completely and cultivates glue, and in central microprotrusion, speed by bottom at the bottom of the culture dish ware after sterilizing down, slowly put into and gently press, venting bubble; Finally, in the gap periphery at the bottom of culture dish ware and between culture dish ware lid, the sealing cultivation glue slowly pouring 45 DEG C into seals;
S3: the cultivation of anerobe and choosing of bacterium colony
Loaded by product obtained by S2 and cultivate with in alcohol and the sterilized valve bag of ultraviolet lamp successively in advance, disperse to become estranged in air that miscellaneous bacteria is in the growth of seal gum periphery to prevent from cultivating glue, incubation time is 12h ~ 72h, to bacterium colony differential count; When choosing bacterium colony, the product after cultivating is poured in larger culture dish, gently choosing sealing cultivation glue-line with taking the photograph son, the bacterium colony of anaerobic bacterium can be chosen.
Preferably, in S1, described sealing is cultivated the normal saline that glue is 0.9% by 18g agar and 1L concentration and is formed.
Preferably, anaerobic bacterium to be seeded is Intestinal Anaerobic Bacteria.
More preferably, anaerobic bacterium to be seeded is cud clostridium, dislikes oxygen bacterium, methanobacteria, sulphate reducing bacteria, rumen bacteria or cellulose decomposing bacteria.
Preferably, in S2, when sterilizing is carried out to culture dish, by the bottom of culture dish ware down, with culture dish ware lid interior thereof touch, after wrapping under 121 ~ 126 DEG C of conditions sterilizing 0.5h.
Preferably, in S3, first try sterilizing by the wipes of alcohol of 75% to described valve bag is inner, then use the further sterilizing of ultra violet lamp 1h.
More preferably, in S3, product obtained by S2 is cultivated 24h ~ 48h in described valve bag, in case substratum moisture scatter and disappear and in air, miscellaneous bacteria, in the growth of culture dish periphery, also can shorten or the Extending culture time according to colony growth situation.
The two anti-sandwich assay of culture dish provided by the invention is carrying out in a large amount of anaerobic bacterium culturing process, summing up experience, utilize existing culture dish, innovate a kind of novel method carrying out anaerobic bacterium cultivation under routine experimentation room environmental, although it is more similar to the culturing process of general aerophil to use the method to cultivate anerobe, but present method is not only very simple and convenient, and oxygen isolation effect is very good, meets condition required in anaerobic bacterium culturing process completely.
Embodiment
Understand technical scheme of the present invention better to make those skilled in the art can be implemented, below in conjunction with specific embodiment, the invention will be further described.
1. experimental installation:
Method of the present invention is in separation and Culture anaerobic bacterium process, used device is identical with device needed for general aerophil cultural method, culture vessel selects common culture dish, wrap sterilizing for subsequent use, specific operation process by the bottom of culture dish ware down, with culture dish ware lid interior thereof touch, after wrapping under 121 ~ 126 DEG C of conditions sterilizing 0.5h.
2. the preparation of experiment reagent:
It is formulated in the physiological saline of 0.9% for joining 1L concentration by the agar of 16g ~ 20g, is mixed with sparkling and crystal-clear bright sealing and cultivates glue, be convenient to the observation of anaerobic bacterium in culturing process.For anaerobic bacterium to be seeded, according to different anaerobic bacterium kinds, adopt respective ordinary method and require that preparation selectivity cultivates glue.
3. the inoculation of anerobe, rubber seal, the choosing of cultivation and bacterium colony:
First, aseptically, first cultivating glue with described sealing, to pour the culture dish ware lid after sterilizing into inner, to be cooled solidify after, by a certain amount of inoculation, anaerobic bacterium to be seeded is inoculated in culture dish ware lid inside and has been perfused with the surface that glue is cultivated in sealing, coating evenly, leaves standstill 5min ~ 10min, the too short absorption being unfavorable for moisture of time of repose, oversize may part anaerobic bacterium can be dead, therefore with 5min ~ 10min for the best;
Secondly, selectivity is cultivated glue sterilizing, after being cooled to 45 DEG C, pour the seal gum surface of spread bacterial into, described selectivity is cultivated thickness that glue pours into and is covered described sealing completely and cultivate glue, and in central microprotrusion, fast by bottom the culture dish after sterilizing down, slowly put into from side and gently press, venting cultivates the bubble in glue;
Because at the bottom of culture dish ware, diameter is less, and have space between culture dish ware lid, finally, in the gap periphery at the bottom of culture dish ware and between culture dish ware lid, the sealing cultivation glue slowly pouring 45 DEG C into seals;
After aforesaid method inoculation sealing, the gap periphery seal gum of culture dish is exposed to outside, in culturing process, product after inoculation sealing is loaded and cultivates with in 75% alcohol and the sterilized valve bag of ultraviolet lamp in advance, in case the varied bacteria growing in moisture loss and air, incubation time is 12h ~ 72h, and specifically can shorten or the Extending culture time according to colony growth situation, the anerobe colony growth after cultivation is good;
Cultivating glue because of sealing cultivation glue and anaerobic bacterium selectivity is two kinds of different cultivation glue, and pour into for different setting time, when choosing bacterium colony, the product after cultivating is poured in larger culture dish, gently choosing sealing ply cultivation glue with taking the photograph son, layering can choose anaerobic bacterium bacterium colony.
It should be noted that aforesaid operations all aseptically operates.
Embodiment 1
The agar that physiological saline 1L by 0.9% adds 18g forms, and is mixed with sparkling and crystal-clear bright sealing and cultivates glue.
Screening cud clostridium selection type cultivates glue, be mainly used in the separation as Butyrivibrio fibrisolvens and cultivation, be made up of following raw materials according: meat extract 10g, yeast extract paste 1.5g, peptone 10g, water soluble starch 1g, glucose 1g, sodium-acetate 5g, Cys 0.5g, distilled water 1L, after abundant for above-mentioned raw materials mixture mixing, regulate mixture ph between 7.1 ~ 7.2, then carry out sterilizing, after the sterilizing of above-mentioned cultivation glue, every 50ml, adds the Na that 0.5ml concentration is 5%
2s and 0.5ml concentration is the ironic citrate (adding first two reagent all through the ultrafiltration membrance filter miscellaneous bacteria in 0.25 μm of aperture) of 7%.According to aequum after above-mentioned screening cud clostridium mixes with rumen fluid sterilizing volume ratio 1: 1 with selectivity cultivation glue, add 1.8g plain agar by 100ml nutrient solution and make.
Aseptically, first cultivate glue with above-mentioned obtained sealing and pour the inside of the culture dish ware lid after sterilizing into, to be cooled solidify after, by a certain amount of inoculation screening cud clostridium, inoculation method is the normal saline dilution of 0.1ml rumen fluid 10ml, the diluent getting 0.1ml, 0.2ml, 0.3ml tri-kinds amount is again inoculated in respectively and has been perfused with the surface that glue is cultivated in sealing, and coating evenly, leaves standstill 5min; Then after screening cud clostridium being cooled to 45 DEG C with selection type cultivation glue, pour in culture dish ware lid, described screening cud clostridium covers described sealing completely with the thickness that selection type cultivation glue is poured into and cultivates glue, and in central microprotrusion, speed by the bottom of the culture dish ware after sterilizing down, slowly put into from side and gently press, venting selectivity cultivates the bubble on glue surface; And in gap periphery at the bottom of culture dish ware and between culture dish ware lid, the above-mentioned sealing of slowly pouring 45 DEG C into is cultivated glue and is sealed, and available dropper dropwise adds; Finally loaded by the product after inoculation sealing and cultivate with in 75% alcohol and the sterilized valve bag of ultraviolet lamp successively in advance, incubation time is 24h.
What test materials was chosen is Tibetan Yellow Cattle rumen fluid, rumen fluid is obtained by rumen fistula operation, the qualification of bacterial classification is realized by Northernblot hybridization, design of primers is as follows: Butyrivibrio fibrisolvens (Butyrivibrio.fibrisolvens), 5 '-CAGCAGGGAAGATAATGAC-3 ', enumeration is 0.8 ~ 2.11 × 10
2.Cultivated by selective medium, observe colonial morphology, bacterial growth is good.
Embodiment 2
The agar that physiological saline 1L by 0.9% adds 18g forms, and is mixed with sparkling and crystal-clear bright sealing and cultivates glue.
Be separated and dislike oxygen bacterium with cultivating glue, be mainly used in being separated and cultivation as Butyrivibrio fibrisolvens and bacterioide, be made up of following raw materials according: Tryptones 1g, yeast extract paste 0.5g, extractum carnis 0.2g, agar 1.5g, 7.5ml salts solution I, 7.5ml salts solution II, 0.1ml concentration are resazurin solution, the distilled water 78ml of 0.1%.After being dissolved successively by above-mentioned raw materials, then add glucose 2g, 0.07% teichmann's crystals 1ml, finally regulate mixture ph to be 7.After sterilizing, then add 8%Na
2cO
3solution 5ml, 3% cysteine hydrochloride 1ml.
Wherein, above-mentioned salts solution I to be concentration be 0.6% K
2hPO
4solution; Above-mentioned salts solution II is by 0.6%KH
2pO
4, 1.2% (NH
4)
2sO
4, 1.2%NaCl, 0.12%MgSO
47H
2o, 0.12%CaCl
2the mixing solutions of composition.
Aseptically, first cultivate glue with above-mentioned obtained sealing and pour the inside of the culture dish ware lid after sterilizing into, to be cooled solidify after, be separated by a certain amount of inoculation and dislike oxygen bacterium, separation to be seeded disliked oxygen bacterial tubercules gastric juice to be seeded in and be perfused with the surface that glue is cultivated in sealing, coating evenly, leaves standstill 10min; Then after screening bacterium being cooled to 45 DEG C with the sterilizing of selection type cultivation glue, pour in culture dish ware lid, described screening bacterium can cover described sealing completely with the thickness that selection type cultivation glue is poured into and cultivate glue, and add, ensure central microprotrusion, speed by bottom the culture dish after sterilizing down, slowly put into from side and gently press, venting selection type cultivates the bubble on glue surface; And in gap periphery at the bottom of culture dish ware and between culture dish ware lid, the above-mentioned sealing of slowly pouring 45 DEG C into is cultivated glue and is sealed; Finally loaded by the product after inoculation sealing and cultivate with in 75% alcohol and the sterilized valve bag of ultraviolet lamp successively in advance, incubation time is 48h, and the anerobe colony growth after cultivation is good.
Embodiment 3
The agar that physiological saline 1L by 0.9% adds 18g forms, and is mixed with sparkling and crystal-clear bright sealing and cultivates glue.
Be separated rumen bacteria and cultivate glue, be mainly used in common anerobe in initial gross separation cud, as Ruminococcus albus number, yellow coccus number, produce succsinic acid thread bacillus, methane bacillus formicicum etc., be made up of following raw materials according: rumen fluid 40ml, agar 1.5g, glucose 0.025g, cellobiose 0.025g, Zulkovsky starch 0.025g, maltose 0.025g, 7.5ml salts solution I, 7.5ml salts solution II, 0.1ml concentration are resazurin solution, the distilled water 38ml of 0.1%.After above-mentioned raw materials is dissolved successively, carry out sterilizing, after sterilizing, add 8%Na again
2cO
3solution 5ml, cysteine hydrochloride 0.025g, Na
2s0.025g.
Wherein identical with in embodiment 1 of salts solution I and salts solution II.
What test materials was chosen is Tibetan Yellow Cattle rumen fluid, and rumen fluid is obtained by rumen fistula operation, and later separation culturing step is identical with embodiment 2.
The design of primers of table 1 Tibetan Yellow Cattle rumen fluid bacterial classification
Table 2 Tibetan Yellow Cattle rumen fluid bacterial colony count result
In the present embodiment, the qualification of bacterial strain is realized by Northernblot hybridization, the design of primers of Tibetan Yellow Cattle rumen fluid bacterial classification as shown in Table 1, the anerobe colony growth of the cud after the method for embodiment 3 is cultivated is good, and concrete rumen fluid bacterial colony count result is as shown in table 2.
Embodiment 4
The agar that physiological saline 1L by 0.9% adds 18g forms, and is mixed with sparkling and crystal-clear bright sealing and cultivates glue.
Cellulose decomposing bacteria selectivity cultivates glue, be mainly used in the initial gross separation of common fiber element decomposer in cud, as Ruminococcus albus number, yellow coccus number, produce the thread bacillus of succsinic acid, Butyrivibrio fibrisolvens etc., be made up of following raw materials according: cellulose powder 5g, NaNO
31g, Na
2hPO
47H
2o1.18g, KH
2pO
40.9g, MgSO
47H
2o0.5g, KCl0.5g, yeast extract paste 0.5g, hydrolyzed casein (enzymolysis casein) 0.5g, sterilization water 1L, by after the mixing of abundant for above-mentioned raw materials mixture, mixture ph is regulated to be 6.8.
Wherein, above-mentioned cellulose powder preparation needs to carry out pre-treatment before cultivating glue, and concrete pretreatment process is the HCl deepfreeze 12h with 1mol/L, after cleaning with sterilization water.
The rumen fluid of Tibetan Yellow Cattle cellulose-decomposing bacterium obtains, later separation culturing step is identical with the content of embodiment 2, and the anerobe colony growth after cultivation is good.
Simultaneously, same cultivation is carried out to the cellulose-decomposing bacterium of Tibet sheep, do not exist together be rumen fluid is obtained by stomach tube, the qualification of bacterial classification is identical with Tibetan Yellow Cattle, find that the cellulose-decomposing bacterium sum of Tibet sheep is relevant to age, feeding floor interval, drink water situation and fodder grass etc., variation range is 1.4 × 10
5cFU/ml ~ 3.3 × 10
8between CFU/ml.
The above embodiment is only that its protection domain is not limited thereto in order to absolutely prove the preferred embodiment that the present invention lifts.The equivalent alternative or conversion that those skilled in the art do on basis of the present invention, all within protection scope of the present invention, protection scope of the present invention is as the criterion with claims.
Claims (8)
1. an isolation cultivation method for anaerobic bacterium, is characterized in that, comprises the following steps,
S1: the preparation of cultivating glue
The preparation of cultivating glue comprises sealing and cultivates the preparation of glue and preparation two portions of selectivity cultivation glue, the normal saline that wherein sealing cultivation glue is 0.9% by 16g ~ 20g agar and 1L concentration forms, and selectivity is cultivated glue and selected different cultivation glue to prepare according to the characteristic of anaerobic bacterium to be seeded;
S2: the inoculation of anerobe and the sealing of inoculating surfaces
First, sterilizing is carried out to culture dish, aseptically, first cultivate glue with described sealing and pour the culture dish ware lid inner bottom part after sterilizing into, to be cooled solidify after, anaerobic bacterium to be seeded is seeded in sealing cultivate glue surface and coating evenly, leave standstill 5min ~ 10min;
Secondly, selectivity is cultivated glue sterilizing and after being cooled to 45 DEG C, glue surface is cultivated in the sealing of pouring the coated bacterial classification of culture dish ware lid into, the thickness that described selectivity cultivation glue is poured into covers described sealing completely and cultivates glue, and in central microprotrusion, speed by bottom at the bottom of the culture dish ware after sterilizing down, slowly put into and gently press, venting bubble;
Finally, in the gap periphery at the bottom of culture dish ware and between culture dish ware lid, the sealing cultivation glue slowly pouring 45 DEG C into seals;
S3: cultivation and the bacterium colony of anerobe are chosen
Product obtained by S2 is loaded in advance successively with 75% alcohol and the sterilized valve bag of ultraviolet lamp in cultivate, incubation time is 12h ~ 72h, and cultivation terminates to carry out differential count according to colonial morphology and color afterwards;
Choose bacterium colony when carrying out identification of strains, the product after cultivating is poured in larger culture dish, gently choose sealing cultivate glue-line with taking the photograph son, the bacterium colony of the anaerobic bacterium of different shape can be chosen.
2. the isolation cultivation method of anaerobic bacterium according to claim 1, is characterized in that, in S1, described sealing is cultivated the normal saline that glue is 0.9% by 18g agar and 1L concentration and formed.
3. the isolation cultivation method of anaerobic bacterium according to claim 1, is characterized in that, the anaerobic bacterium can cultivated in existing CO2gas incubator all can be used as anaerobic bacterium to be seeded in S1 and carries out enlarged culturing.
4. the isolation cultivation method of anaerobic bacterium according to claim 1, is characterized in that, in S2, anaerobic bacterium to be seeded is Intestinal Anaerobic Bacteria.
5. the isolation cultivation method of anaerobic bacterium according to claim 4, is characterized in that, in S2, anaerobic bacterium to be seeded is cud carboxylic bacterium, dislikes oxygen bacterium, methanobacteria, sulphate reducing bacteria, cud bacterium or cellulose-decomposing bacterium.
6. the isolation cultivation method of anaerobic bacterium according to claim 1, is characterized in that, in S2, when sterilizing is carried out to culture dish, by the bottom of culture dish ware down, with culture dish ware lid interior thereof touch, after wrapping under 121 ~ 126 DEG C of conditions sterilizing 0.5h.
7. the isolation cultivation method of anaerobic bacterium according to claim 1, is characterized in that, in S3, first tries sterilizing by the wipes of alcohol of 75%, then use the further sterilizing of ultra violet lamp 1h stand-by to described valve bag is inner.
8. the isolation cultivation method of anaerobic bacterium according to claim 7, is characterized in that, in S3, product obtained by S2 is cultivated 24h ~ 48h in described valve bag.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510681477.5A CN105219675A (en) | 2015-10-13 | 2015-10-13 | The isolation cultivation method of a kind of anaerobic bacterium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510681477.5A CN105219675A (en) | 2015-10-13 | 2015-10-13 | The isolation cultivation method of a kind of anaerobic bacterium |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105219675A true CN105219675A (en) | 2016-01-06 |
Family
ID=54988947
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510681477.5A Pending CN105219675A (en) | 2015-10-13 | 2015-10-13 | The isolation cultivation method of a kind of anaerobic bacterium |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105219675A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110713899A (en) * | 2019-10-30 | 2020-01-21 | 南通大学 | Culture method for culturing desulfurization vibrio |
| CN111733099A (en) * | 2020-06-18 | 2020-10-02 | 河南金百合生物科技股份有限公司 | Clostridium butyricum culture method for viable bacteria counting and clostridium butyricum counting method |
| CN113817584A (en) * | 2021-08-30 | 2021-12-21 | 北京大学第一医院 | Anaerobic bacteria bedside culture detection system |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1940085A (en) * | 2006-09-28 | 2007-04-04 | 东北林业大学 | Double-layer planar accounting method of bifidobacterium etc. strict anacerobe |
| CN101434924A (en) * | 2008-12-17 | 2009-05-20 | 中南大学 | Method for strictly separating anaerobic methanogenic archaea |
| WO2013037067A1 (en) * | 2011-09-14 | 2013-03-21 | University Of Guelph | Media supplements and methods to culture human gastrointestinal anaerobic microorganisms |
| CN104450592A (en) * | 2014-12-31 | 2015-03-25 | 哈尔滨工业大学 | Method for separating denitrification desulfurizing bacteria based on biodiversity information |
-
2015
- 2015-10-13 CN CN201510681477.5A patent/CN105219675A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1940085A (en) * | 2006-09-28 | 2007-04-04 | 东北林业大学 | Double-layer planar accounting method of bifidobacterium etc. strict anacerobe |
| CN101434924A (en) * | 2008-12-17 | 2009-05-20 | 中南大学 | Method for strictly separating anaerobic methanogenic archaea |
| WO2013037067A1 (en) * | 2011-09-14 | 2013-03-21 | University Of Guelph | Media supplements and methods to culture human gastrointestinal anaerobic microorganisms |
| CN104450592A (en) * | 2014-12-31 | 2015-03-25 | 哈尔滨工业大学 | Method for separating denitrification desulfurizing bacteria based on biodiversity information |
Non-Patent Citations (3)
| Title |
|---|
| 万海清等: "一种分离培养硫酸盐还原菌的改进方法", 《应用与环境生物学报》 * |
| 桂芳: "《微生物学检验实验指导》", 31 August 2009 * |
| 穆军等: "一种分离纯化厌氧细菌的新方法——平皿夹层厌氧法", 《山西大学学报( 自然科学版) 》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110713899A (en) * | 2019-10-30 | 2020-01-21 | 南通大学 | Culture method for culturing desulfurization vibrio |
| CN110713899B (en) * | 2019-10-30 | 2023-04-28 | 南通大学 | Culture method for culturing vibrio desulphurisation |
| CN111733099A (en) * | 2020-06-18 | 2020-10-02 | 河南金百合生物科技股份有限公司 | Clostridium butyricum culture method for viable bacteria counting and clostridium butyricum counting method |
| CN113817584A (en) * | 2021-08-30 | 2021-12-21 | 北京大学第一医院 | Anaerobic bacteria bedside culture detection system |
| CN113817584B (en) * | 2021-08-30 | 2023-12-29 | 北京大学第一医院 | Anaerobic bacteria bedside culture detection system |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105462872B (en) | A kind of compound micro-ecological preparation and preparation method thereof | |
| CN101338283B (en) | Lactobacillus casei and applications thereof in solid-state fermentation | |
| CN108949739A (en) | A kind of complex micro organism fungicide and preparation method thereof for advanced treating high concentration livestock breeding wastewater | |
| CN101401624A (en) | Corn stalk nano-dietary fiber and method of preparing the same | |
| CN101914478B (en) | Bacillus subtilis and application thereof | |
| CN108504602A (en) | A kind of preparation method of selenium-rich lactobacillus | |
| CN106978355A (en) | The coculture of a kind of anaerobic fungi and methane backeria and the application in the feeding native enzyme of milk cow | |
| CN107384811A (en) | A kind of Irpex lacteus and its application | |
| CN105219675A (en) | The isolation cultivation method of a kind of anaerobic bacterium | |
| CN109022313A (en) | One lactobacillus plantarum | |
| CN103555592B (en) | Preparation method of Aspergillus oryzae strain and spore powder | |
| CN104877942B (en) | Series bacillus novel bacterial and its cultural method and application | |
| CN106566788A (en) | Method for biological synthesis of nano-selenium by using silver-resistant Bacillus edaphicus and application thereof | |
| CN111548959B (en) | Klebsiella pneumoniae and application thereof | |
| CN105454422A (en) | A preparing method and applications of a novel Kefir culture | |
| CN101153316A (en) | Method for detecting lactobacillus casei in probiotic bacteria milk product | |
| CN108251334A (en) | The microorganism mixed bacterial and fermentation process of a kind of fermenting lactic acid | |
| CN108624517A (en) | Resistance to sugar saccharomyces cerevisiae of resistance to ethyl alcohol and application thereof | |
| CN109112078B (en) | Brevibacterium strain and application thereof | |
| CN102352320B (en) | Novel myceliophthora thermophila strain and application thereof | |
| CN102701831A (en) | Microbial fertilizer and preparation method thereof | |
| CN102321544B (en) | Filamentous fungus for efficiently degrading lignocellulose in forest litter | |
| CN104877920B (en) | Candida krusei LSA and its application in degradation of ammonia nitrogen | |
| CN104845894B (en) | Pichia farinose NHG and its application in degradation of ammonia nitrogen | |
| Vattamparambil | Anaerobic Microbial Hydrolysis of Agriculture Waste for Biogas Production |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160106 |
|
| RJ01 | Rejection of invention patent application after publication |