CN107058105A - The cultural method of nitrite degradation bacterium - Google Patents
The cultural method of nitrite degradation bacterium Download PDFInfo
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- CN107058105A CN107058105A CN201710248047.3A CN201710248047A CN107058105A CN 107058105 A CN107058105 A CN 107058105A CN 201710248047 A CN201710248047 A CN 201710248047A CN 107058105 A CN107058105 A CN 107058105A
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- degradation bacterium
- nitrite degradation
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- nitrite
- test tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
Abstract
The present invention relates to a kind of cultural method of nitrite degradation bacterium, comprise the following steps：(1) nutrient agar is prepared, (2) activate nitrite degradation bacterium in nutrient broth medium, institute (3) takes the nitrite degradation bacterium solution diluted to distinguish 200 microlitres with liquid-transfering gun.It is an advantage of the invention that：The nitrite degradation bacterium cultivated by the present invention, can effectively remove nitrite, and do not pollute the environment.
Description
Technical field
The present invention relates to a kind of cultural method of nitrite degradation bacterium.
Background technology
It in current sewage, can all contain nitrite, but how clean effective removal to nitrite in sewage,
Still it is in conceptual phase.Pertinent literature is not found through retrieval.
The content of the invention
To overcome the defect of prior art, the present invention provides a kind of cultural method of nitrite degradation bacterium, of the invention
Technical scheme is：
A kind of cultural method of nitrite degradation bacterium, comprises the following steps：
(1) nutrient agar is prepared, specific preparation method is as follows：Weigh 10 parts of peptone, 5 parts of beef extract powder, fine jade
18 parts of cosmetics, 5 parts of sodium chloride is mixed according to proportioning, is heated after mixing in micro-wave oven, the heat time：1-2min, heating
Temperature：70-80 DEG C, untill agar powder melts completely, it is put into high-pressure sterilizing pot and is sterilized, 121 DEG C of sterilising temp, time
30min, is then poured into sterilized desiccation culture ware, each culture dish pours into 15ml in Biohazard Safety Equipment, will be trained well
The culture dish for supporting base is placed in safety cabinet to cooling, cool time：4-5h, chilling temperature：25 DEG C of room temperature, can be put into ice after cooling
Case refrigeration is standby；
(2) by the activation of nitrite degradation bacterium in nutrient broth medium, the preparation side of the nutrient broth medium
Method is as follows：10 parts of peptone is weighed, then 3 parts of beef extract powder, 5 parts of sodium chloride mix, 30 DEG C incubated is put into after mixing
Case culture 2 days, is then carried out in superclean bench to the nitrite degradation bacterium for having activated and having grown in broth bouillon
Flat board is coated with, while nitrite degradation bacterium solution concentration is carried out into gradient dilution, 10-2 powers ,-the 4 of 10 are diluted to respectively
Power ,-the 6 of 10 powers and-the 8 of 10 powers.
(3) take the nitrite degradation bacterium solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into culture dish
Nutrient agar on, it is with spreading rod that nitrite degradation bacterium solution coating is uniform, and with marking pen in culture dish
Bottom is labeled, and in the constant incubator for being then placed in 35 DEG C, is cultivated, after 1-2 days, on observation nutrient agar
The growing state of nitrite degradation bacterium, the nitrite degradation of the different shape grown on culture medium is marked with marking pen respectively
Bacterium, and reference number, then carry out plate streaking to the nitrite degradation bacterium of mark；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, nitrite degradation bacterium on observation flat board
Growth, and dyeing microscopic examination is carried out to the nitrite degradation bacterium that has grown, records its morphological feature, repeat the operation, until
Untill the nitrite degradation bacterium form Economical Purification of micro- Microscopic observation, the nitrite degradation bacterium of purifying is carried out in test tube
Inclined-plane line preservation is carried out in slant medium.
Specific dilution process is in described step (2)：10ml distilled water is taken to be put into test tube with syringe, by distilled water
Sterilized, in Biohazard Safety Equipment, 100 microlitres of strain solution taken with liquid-transfering gun, is injected into the test tube of sterile distilled water,
Now, the strain solution concentration in the test tube is 10-2 powers, and strain solution is diluted to by same method respectively 10 for-4 times
Side, 10-6 powers and-the 8 of 10 powers.
Described step (3) plate streaking method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the top of oese
The nitrite degradation bacterium of mark sequence number is touched, is drawn on nutrient agar, the nitrite degradation bacterium of each sequence number is in training
The areas of base Shang Hua tetra- are supported, oese is placed on alcolhol burner per standardized area and burnt, is repeated.
The preparation method of described step (4) test tube slant culture medium：Weigh 10 parts of peptone, 5 parts of beef extract powder, agar
18 parts of powder, 5 parts of sodium chloride is mixed according to proportioning, is heated after mixing in micro-wave oven, the heat time：1-2min, heating temperature
Degree：70-80 DEG C, untill agar powder melts completely, taken with 10ml syringe in 5ml injecting tubes, test tube is put into 2L's
Beaker is wrapped up, and is then placed in high-pressure sterilizing pot and is sterilized, and is sterilized 121 DEG C, after sterilizing, test tube is taken out by time 30min
Inclined-plane is put, the standard of putting on inclined-plane is the 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane, stood
8h, it is to be condensed after, be put into refrigerator cold-storage.
It is an advantage of the invention that：The nitrite degradation bacterium cultivated by the present invention, can effectively remove nitrite,
And do not pollute the environment.
Embodiment
The invention will now be further described with reference to specific embodiments, advantages of the present invention and feature will be with description and
It is apparent.But these embodiments are only exemplary, do not constitute any limitation to the scope of the present invention.People in the art
Member to the details and form of technical solution of the present invention it should be understood that can enter without departing from the spirit and scope of the invention
Row modifications or substitutions, but these are changed and replacement is each fallen within protection scope of the present invention.
The present invention relates to a kind of cultural method of nitrite degradation bacterium, comprise the following steps：
(1) nutrient agar is prepared, specific preparation method is as follows：Peptone 10g/L, beef extract powder 5g/L are weighed,
Agar powder 18g/L, sodium chloride 5g/L, are mixed according to proportioning, are heated after mixing in micro-wave oven, the heat time：1-2min,
Heating-up temperature：70-80 DEG C, untill agar powder melts completely, it is put into high-pressure sterilizing pot and is sterilized, 121 DEG C of sterilising temp, when
Between 30min, then poured into Biohazard Safety Equipment in sterilized desiccation culture ware, each culture dish pours into 15ml, will be good
The culture dish of culture medium is placed in safety cabinet to cooling, cool time：4-5h, chilling temperature：25 DEG C of room temperature, can be put into after cooling
Refrigerator cold-storage is standby；
(2) by the activation of nitrite degradation bacterium in nutrient broth medium, the preparation side of the nutrient broth medium
Method is as follows：10 parts of peptone is weighed, then 3 parts of beef extract powder, 5 parts of sodium chloride mix, 30 DEG C incubated is put into after mixing
Case culture 2 days, is then carried out in superclean bench to the nitrite degradation bacterium for having activated and having grown in broth bouillon
Flat board is coated with, while nitrite degradation bacterium solution concentration is carried out into gradient dilution, 10-2 powers ,-the 4 of 10 are diluted to respectively
Power ,-the 6 of 10 powers and-the 8 of 10 powers.
(3) take the nitrite degradation bacterium solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into nutrient agar
It is with spreading rod that nitrite degradation bacterium solution coating is uniform on culture medium, and enter rower in the bottom of culture dish with marking pen
In note (mark temperature, date etc.), the constant incubator for being then placed in 35 DEG C, cultivated, after 1-2 days, on observation culture medium
The growing state of nitrite degradation bacterium, the nitrite degradation of the different shape grown on culture medium is marked with marking pen respectively
Bacterium, and reference number, then carry out plate streaking to the nitrite degradation bacterium of mark；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, nitrite degradation bacterium on observation flat board
Growth, and dyeing microscopic examination is carried out to the nitrite degradation bacterium that has grown, records its morphological feature, repeat the operation, until
Untill the nitrite degradation bacterium form Economical Purification of micro- Microscopic observation, the nitrite degradation bacterium of purifying is carried out in test tube
Inclined-plane line preservation is carried out in slant medium.
Specific dilution process is in described step (2)：10ml distilled water is taken to be put into test tube with syringe, by distilled water
Sterilized, in Biohazard Safety Equipment, 100 microlitres of strain solution taken with liquid-transfering gun, is injected into the test tube of sterile distilled water,
Now, the strain solution concentration in the test tube is 10-2 powers, and strain solution is diluted to by same method respectively 10 for-4 times
Side, 10-6 powers and-the 8 of 10 powers.
Described step (3) plate streaking method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the top of oese
The nitrite degradation bacterium of mark sequence number is touched, is drawn on nutrient agar, the nitrite degradation bacterium of each sequence number is in training
The areas of base Shang Hua tetra- are supported, oese is placed on alcolhol burner per standardized area and burnt, is repeated.
The preparation method of described step (4) test tube slant culture medium：Weigh 10 parts of peptone, 5 parts of beef extract powder, agar
18 parts of powder, 5 parts of sodium chloride is mixed according to proportioning, is heated after mixing in micro-wave oven, the heat time：1-2min, heating temperature
Degree：70-80 DEG C, untill agar powder melts completely, taken with 10ml syringe in 5ml injecting tubes, test tube is put into 2L's
Beaker is wrapped up, and is then placed in high-pressure sterilizing pot and is sterilized, and is sterilized 121 DEG C, after sterilizing, test tube is taken out by time 30min
Inclined-plane is put, the standard of putting on inclined-plane is the 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane, stood
8h, it is to be condensed after, be put into refrigerator cold-storage.
Embodiment：The solution with 1L nitrite is taken, wherein, the content of nitrite is 0.138g, accesses the nitrous
After hydrochlorate degradation bacteria 1ml, the content of nitrite is 0.011g/L, and reckoning is learnt, the nitrous acid after purification of this method culture
Salt degradation bacteria reaches more than 90% to the clearance of nitrite.
Claims (4)
1. a kind of cultural method of nitrite degradation bacterium, it is characterised in that comprise the following steps：
(1) nutrient agar is prepared, specific preparation method is as follows：Weigh 10 parts of peptone, 5 parts of beef extract powder, agar powder
18 parts, 5 parts of sodium chloride is mixed according to proportioning, is heated after mixing in micro-wave oven, the heat time：1-2min, heating temperature
Degree：70-80 DEG C, untill agar powder melts completely, it is put into high-pressure sterilizing pot and is sterilized, 121 DEG C of sterilising temp, time
30min, is then poured into sterilized desiccation culture ware, each culture dish pours into 15ml in Biohazard Safety Equipment, will be trained well
The culture dish for supporting base is placed in safety cabinet to cooling, cool time：4-5h, chilling temperature：25 DEG C of room temperature, can be put into ice after cooling
Case refrigeration is standby；
(2) by the activation of nitrite degradation bacterium in nutrient broth medium, the preparation method of the nutrient broth medium is such as
Under：10 parts of peptone is weighed, then 3 parts of beef extract powder, 5 parts of sodium chloride mix, 30 DEG C of constant incubator training is put into after mixing
Support 2 days, flat board then is carried out to the nitrite degradation bacterium for having activated and having grown in broth bouillon in superclean bench
Coating, while nitrite degradation bacterium solution concentration is carried out into gradient dilution, is diluted to 10-2 powers ,-4 times of 10 respectively
Side, 10-6 powers and-the 8 of 10 powers.
(3) take the nitrite degradation bacterium solution diluted to distinguish 200 microlitres with liquid-transfering gun, be injected separately into the battalion in culture dish
Support on agar medium, it is with spreading rod that nitrite degradation bacterium solution coating is uniform, and with marking pen in the bottom of culture dish
It is labeled, in the constant incubator for being then placed in 35 DEG C, is cultivated, after 1-2 days, nitrous on observation nutrient agar
The growing state of hydrochlorate degradation bacteria, the nitrite degradation bacterium of the different shape grown on culture medium is marked with marking pen respectively,
And reference number, plate streaking then is carried out to the nitrite degradation bacterium of mark；
(4) flat board pulled is put into constant incubator, 35 DEG C, cultivated 1-2 days, the life of nitrite degradation bacterium on observation flat board
It is long, and dyeing microscopic examination is carried out to the nitrite degradation bacterium grown, its morphological feature is recorded, the operation is repeated, until micro-
Untill the nitrite degradation bacterium form Economical Purification of Microscopic observation, the nitrite degradation bacterium of purifying is carried out in test tube slant
Inclined-plane line preservation is carried out in culture medium.
2. the cultural method of nitrite degradation bacterium according to claim 1, it is characterised in that in described step (2)
Specifically dilution process is：Take 10ml distilled water to be put into test tube with syringe, distilled water is sterilized, in Biohazard Safety Equipment
It is interior, 100 microlitres of strain solution is taken with liquid-transfering gun, is injected into the test tube of sterile distilled water, now, the bacterial strain in the test tube is molten
Liquid concentration be 10-2 powers, same method, strain solution is diluted to respectively 10-4 powers ,-the 6 of 10 powers and 10
- 8 powers.
3. the cultural method of nitrite degradation bacterium according to claim 1, it is characterised in that described step (3) is put down
Plate scribble method is as follows：In Biohazard Safety Equipment, alcolhol burner is lighted, with the nitrite degradation that mark sequence number is touched at the top of oese
Bacterium, draws on nutrient agar, and the nitrite degradation bacterium of each sequence number will connect in the areas of culture medium Shang Hua tetra- per standardized area
Plant ring and be placed on alcolhol burner and burn, repeat.
4. the cultural method of nitrite degradation bacterium according to claim 1, it is characterised in that described step (4) examination
The preparation method of pipe slant medium：Weigh 10 parts of peptone, 5 parts of beef extract powder, 18 parts of agar powder, 5 parts of sodium chloride, according to
Than being mixed, heated after mixing in micro-wave oven, the heat time：1-2min, heating-up temperature：It is 70-80 DEG C, complete to agar powder
Untill thawing, taken with 10ml syringe in 5ml injecting tubes, the beaker that test tube is put into 2L is wrapped up, and is then placed in height
Pressure autoclave is sterilized, and is sterilized 121 DEG C, time 30min, after sterilizing, and test tube is taken out into pendulum inclined-plane, and the standard of putting on inclined-plane is
The 2/3 of of length no more than whole test tube length of nutrient agar on inclined-plane, stand 8h, it is to be condensed after, be put into refrigerator cold
Tibetan.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710248047.3A CN107058105A (en) | 2017-04-17 | 2017-04-17 | The cultural method of nitrite degradation bacterium |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710248047.3A CN107058105A (en) | 2017-04-17 | 2017-04-17 | The cultural method of nitrite degradation bacterium |
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| CN107058105A true CN107058105A (en) | 2017-08-18 |
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| CN201710248047.3A Pending CN107058105A (en) | 2017-04-17 | 2017-04-17 | The cultural method of nitrite degradation bacterium |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114806971A (en) * | 2022-06-08 | 2022-07-29 | 广州微立旺生物科技有限公司 | Cultivating process based on sulfide bacteria |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673730A (en) * | 2015-03-23 | 2015-06-03 | 大连理工大学 | Brevibacillus laterosporu with function in quickly decomposing nitrite nitrogen and bacteriostasis function and application thereof |
-
2017
- 2017-04-17 CN CN201710248047.3A patent/CN107058105A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673730A (en) * | 2015-03-23 | 2015-06-03 | 大连理工大学 | Brevibacillus laterosporu with function in quickly decomposing nitrite nitrogen and bacteriostasis function and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| PINGMEI YAN ET AL.: "Screening and identification of microorganism degrading nitrite in Chinese sauerkraut", 《ANALYTICAL TECHNOLOGIE》 * |
| 周鲜娇、全丽谦: "一株芽孢杆菌降解亚硝酸盐的特性研究", 《岭南师范学院学报》 * |
| 胡桂学: "《兽医微生物学实验教程》", 31 December 2014, 北京：中国农业大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114806971A (en) * | 2022-06-08 | 2022-07-29 | 广州微立旺生物科技有限公司 | Cultivating process based on sulfide bacteria |
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Application publication date: 20170818 |
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