CN103710287A - Culturing method for ammonia oxidizing bacteria - Google Patents
Culturing method for ammonia oxidizing bacteria Download PDFInfo
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- CN103710287A CN103710287A CN201310745231.0A CN201310745231A CN103710287A CN 103710287 A CN103710287 A CN 103710287A CN 201310745231 A CN201310745231 A CN 201310745231A CN 103710287 A CN103710287 A CN 103710287A
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- oxidizing bacteria
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Abstract
The invention provides a culturing method for ammonia oxidizing bacteria. The culturing method comprises the following steps: inoculating ammonia oxidizing bacteria liquid into a fermentation tank; carrying out fermentation culture for 5-7 days; collecting fermentation liquid, so as to obtain the ammonia oxidizing bacteria, wherein the inoculation amount is 5%; the culturing conditions refer to that a pH (Potential of Hydrogen) value is controlled to be 7.5-8.0 by adding a base liquid, a temperature is 25-30 DEG C, a ventilation amount is 1.5-2.0M<3>/h, a tank pressure is 0.05-0.1MPa, and a rotation speed is 180-200r/min; a culture medium is a nitrobacteria culture medium. The culturing method uses a fermentation tank yeast propagation method, so that infectious microbes are difficult to pollute; various parameters, such as the temperature, the pH value, dissolved oxygen and a stirring rotation speed and the like, can be stably controlled; compared with the traditional opened type yeast propagation method, the culturing method for the ammonia oxidizing bacteria can be used for more economically and effectively preparing an ammonia oxidizing bacteria agent, so that the agent can be rapidly and effectively propagated; the culturing method for the ammonia oxidizing bacteria is suitable for large-scale industrial production and has a wide application prospect in a sewage treatment field.
Description
Technical field
The present invention relates to a kind of method for cultivation of bacteria, the cultural method of especially a kind of ammonia oxidizing bacteria.
Background technology
Ammonia nitrogen is one of topmost pollutent in water body: under meta-alkali environment, ammonia nitrogen can cause toxicity to hydrobiont; Ammonia nitrogen is oxidized to nitric nitrogen and nitrite can cause wawter bloom; The tap water that nitrate and nitrite concentration are high may cause Health hazard to human body.Along with the development of industrial and agricultural production, the pollution of ammonia nitrogen increases the weight of day by day, becomes one of sewage treatment area problem demanding prompt solution, is also one of Environmental indicators of state key monitoring.
The method that ammonia nitrogen is removed in traditional materialization has air stripping method, the selective ion exchange method based on resin or zeolite, chemical precipitation method, medicament neutralisation, emulsion liquid membrane partition method etc., but all exist, nitric efficiency is low, cost is high, be difficult for the defects such as large-scale operation.Therefore, it is to utilize the microorganism in active sludge to complete mostly that current sewage removes ammonia nitrogen technique, and this quasi-microorganism is called as ammonia oxidizing bacteria (Ammonia-oxidizing Bacteria, AOB), is called again nitrite bacteria.
Ammonia oxidizing bacteria relies on ammoxidation to remove ammonia nitrogen, first by ammona monooxygenase, ammonia nitrogen is oxidized to azanol, then is converted into nitrite nitrogen by azanol oxydo-reductase.Subsequently, nitrite nitrogen can be converted into nitrate by nitrite-oxidizing bacteria, is more progressively reduced to nitrogen by denitrifying bacterium, discharges, thereby reach the effect of thorough denitrogenation from water body.Also can be by the direct denitrogenation of mode of short-cut denitrification or Anammox.Therefore, the metabolism of ammonia oxidizing bacteria is the committed step of biological denitrificaion.
Investigator has just found ammonia oxidation phenomenon as far back as 1895.Since the middle of last century, about the physio-biochemical characteristics of ammonia oxidizing bacteria and applied research thereof, be also never interrupted.Yet ammonia oxidizing bacteria microbial inoculum does not have large-scale industrial application at present, and its reason is: it is a kind of chemosynthetic bacteria, poor growth is responsive to the factor of temperature, pH value, dissolved oxygen, heavy metal.In Sewage treatment systems, this class bacterium is difficult to form dominant microflora and impacted by environmental factors, and the ammonia nitrogen removal ability of impair system is the type strain of difficult culturing micro-organisms, carries out very difficulty of purifying list bacterium operation.
Document and the disclosed ammonia oxidizing bacteria enlarged culturing of patent method substantially all utilize open reactive device to carry out at present.CN00808700 discloses the high concentration culturing method of nitrobacteria and denitrifying bacteria in a kind of active sludge, by maintaining alkaline pH value and adding culture promoter, continues 1-2 enriching ammonia oxidation bacterium over the month; CN101240253 discloses the enriching method of High-efficient Nitrobacteria in a kind of active sludge, adopts intermittent activated sludge process, by improving gradually ammonia nitrogen concentration, realizes enrichment; CN200910154684 discloses large-scaled culture method of a kind of high-concentration nitrosobacteria and uses thereof, adopts sequence batch (water inlet in conjunction with the scheme of flocculation sediment, and the longest domestication time is 48 days.Often there is many drawbacks such as length consuming time, easily the relative growth rate of slow, the conditional instability of pollution microbes, reactor start-up, flora is slow in these methods.Therefore, find a kind of rapidly and efficiently value-added cultural method of ammonia oxidizing bacteria that makes, there is very important realistic meaning.
Summary of the invention
Technical problem to be solved by this invention is to provide the cultural method of a kind of ammonia oxidizing bacteria.
For solving the problems of the technologies described above, technical scheme of the present invention is:
The cultural method of a kind of ammonia oxidizing bacteria, ammonia oxidizing bacteria bacterium liquid is inoculated in fermentor tank, fermentation culture is carried out 5-7 days, collect fermented liquid, obtain ammonia oxidizing bacteria, wherein, inoculum size is 5%, culture condition is: adding alkali lye control pH is 7.5-8.0, and temperature is 25-30 ℃, and air flow is 1.5-2.0M
3/ h, tank pressure is 0.05-0.1MPa, rotating speed is 180-200r/min; Substratum is nitrobacteria substratum.
Preferably, the cultural method of above-mentioned ammonia oxidizing bacteria, described nitrobacteria medium component is: ammonium sulfate 2g/L, dipotassium hydrogen phosphate 0.5g/L, sodium-chlor 0.5g/L, sodium bicarbonate 1.5g/L, all the other are tap water.
Preferably, the cultural method of above-mentioned ammonia oxidizing bacteria, described culture condition is: adding alkali lye control pH is 8.0, and temperature is 30 ℃, and air flow is 2.0M
3/ h, tank pressure is 0.1MPa, rotating speed is 200r/min.
Preferably, the cultural method of above-mentioned ammonia oxidizing bacteria, the sodium hydrogen carbonate solution that described alkali lye is 10%.
Preferably, the cultural method of above-mentioned ammonia oxidizing bacteria, is used colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid.
The invention has the beneficial effects as follows:
The cultural method of described ammonia oxidizing bacteria, adopt fermentor tank propagation method, be difficult for pollution microbes, the various parameter stabilities such as temperature, pH value, dissolved oxygen, mixing speed are controlled, compare traditional open pattern that spreads cultivation, can more economically prepare efficiently ammonia oxidizing bacteria microbial inoculum, realize such microbial inoculum and breed quickly and efficiently, and applicable mass industrialized production, has wide practical use at sewage treatment area.
Embodiment
In order to make those skilled in the art better understand technical scheme of the present invention, below in conjunction with the drawings and the specific embodiments, technical scheme of the present invention is described in further detail.
In following embodiment, bacterial classification used is ammonia oxidizing bacteria (Tianjin Kai Ying development in science and technology company limited), and nitrobacteria medium component is: ammonium sulfate 2g/L, and dipotassium hydrogen phosphate 0.5g/L, sodium-chlor 0.5g/L, sodium bicarbonate 1.5g/L, all the other are tap water.
Embodiment 1
Using fermentor tank volume is 30L, adds nitrobacteria substratum 19L, seed liquor 1L.Control parameter is: fermentor tank is automatically added 10% sodium hydrogen carbonate solution and controlled pH=7.5, temperature=25 ℃, and air flow is 1.5M
3/ h, tank pressure is 0.05MPa, rotating speed is 180r/min.Fermentation culture is carried out 5 days, collects fermented liquid, and using colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid is 2.54 * 10
9cFU/ml.
Embodiment 2
Using fermentor tank volume is 30L, adds nitrobacteria substratum 19L, seed liquor 1L.Control parameter is: fermentor tank is automatically added 10% sodium hydrogen carbonate solution and controlled pH=8.0, temperature=30 ℃, and air flow is 2.0M
3/ h, tank pressure is 0.1MPa, rotating speed is 200r/min.Fermentation culture is carried out 7 days, collects fermented liquid, and using colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid is 1.36 * 10
10cFU/ml.
Embodiment 3
Using fermentor tank volume is 200L, adds nitrobacteria substratum 120L, seed liquor 6L.Control parameter is: pH=8.0, and temperature=30 ℃, air flow is 2.0M
3/ h, tank pressure is 0.1MPa, rotating speed is 200r/min.Fermentation culture is carried out 7 days, collects fermented liquid, and using colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid is 1.09 * 10
10cFU/ml.
Embodiment 4
Using fermentor tank volume is 200L, adds nitrobacteria substratum 120L, seed liquor 6L.Control parameter is: pH=8.0, and temperature=27 ℃, air flow is 1.8M
3/ h, tank pressure is 0.08MPa, rotating speed is 200r/min.Fermentation culture is carried out 6 days, collects fermented liquid, and using colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid is 8.75 * 10
9cFU/ml.
Above-mentioned detailed description of the cultural method of this kind of ammonia oxidizing bacteria being carried out with reference to embodiment; illustrative rather than determinate; can list several embodiment according to institute's limited range; therefore in the variation and the modification that do not depart under general plotting of the present invention, within should belonging to protection scope of the present invention.
Claims (5)
1. the cultural method of an ammonia oxidizing bacteria, it is characterized in that: ammonia oxidizing bacteria bacterium liquid is inoculated in fermentor tank, fermentation culture is carried out 5-7 days, collect fermented liquid, obtain ammonia oxidizing bacteria, wherein, inoculum size is 5%, culture condition is: adding alkali lye control pH is 7.5-8.0, and temperature is 25-30 ℃, and air flow is 1.5-2.0M
3/ h, tank pressure is 0.05-0.1MPa, rotating speed is 180-200r/min; Substratum is nitrobacteria substratum.
2. the cultural method of ammonia oxidizing bacteria according to claim 1, is characterized in that: described nitrobacteria medium component is: ammonium sulfate 2g/L, and dipotassium hydrogen phosphate 0.5g/L, sodium-chlor 0.5g/L, sodium bicarbonate 1.5g/L, all the other are tap water.
3. the cultural method of ammonia oxidizing bacteria according to claim 1, is characterized in that: described culture condition is: adding alkali lye control pH is 8.0, and temperature is 30 ℃, and air flow is 2.0M
3/ h, tank pressure is 0.1MPa, rotating speed is 200r/min.
4. according to the cultural method of the ammonia oxidizing bacteria described in claim 1 or 3, it is characterized in that: the sodium hydrogen carbonate solution that described alkali lye is 10%.
5. the cultural method of ammonia oxidizing bacteria according to claim 1, is characterized in that: use colony counting method to detect ammonia oxidizing bacteria quantity in fermented liquid.
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Cited By (6)
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| CN106336017A (en) * | 2016-03-22 | 2017-01-18 | 路域生态工程有限公司 | Method for removing NH4<+>-N in polluted water through high-efficiency activation ammonia oxidizing bacteria |
| CN106635859A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Opening type culture method and applications of Paracoccus FSTB-2 |
| CN107629986A (en) * | 2017-10-31 | 2018-01-26 | 中冶华天工程技术有限公司 | A kind of resistance to low ammonia nitrogen concentration ammonia oxidizing bacteria stirring-type continuously ferments cultural method |
| CN108585190A (en) * | 2018-04-27 | 2018-09-28 | 北京交通大学 | Integral anaerobic ammoxidation handles the device of coal-ethylene glycol sewage |
| CN111349577A (en) * | 2018-12-21 | 2020-06-30 | 中国石油化工股份有限公司 | Culture method of ammonia oxidizing bacteria |
| CN112300969A (en) * | 2020-11-20 | 2021-02-02 | 北京师范大学 | Ammonia oxidizing bacteria culture medium and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106635859A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Opening type culture method and applications of Paracoccus FSTB-2 |
| CN106635859B (en) * | 2015-11-04 | 2020-03-17 | 中国石油化工股份有限公司 | Open culture method and application of paracoccus FSTB-2 |
| CN106336017A (en) * | 2016-03-22 | 2017-01-18 | 路域生态工程有限公司 | Method for removing NH4<+>-N in polluted water through high-efficiency activation ammonia oxidizing bacteria |
| CN107629986A (en) * | 2017-10-31 | 2018-01-26 | 中冶华天工程技术有限公司 | A kind of resistance to low ammonia nitrogen concentration ammonia oxidizing bacteria stirring-type continuously ferments cultural method |
| CN108585190A (en) * | 2018-04-27 | 2018-09-28 | 北京交通大学 | Integral anaerobic ammoxidation handles the device of coal-ethylene glycol sewage |
| CN111349577A (en) * | 2018-12-21 | 2020-06-30 | 中国石油化工股份有限公司 | Culture method of ammonia oxidizing bacteria |
| CN111349577B (en) * | 2018-12-21 | 2022-07-08 | 中国石油化工股份有限公司 | Culture method of ammonia oxidizing bacteria |
| CN112300969A (en) * | 2020-11-20 | 2021-02-02 | 北京师范大学 | Ammonia oxidizing bacteria culture medium and preparation method thereof |
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Address after: 300381 Tianjin, Nankai District Jin Wei South Road, building 76, No. two Patentee after: Tianjin kying technology development Limited by Share Ltd Address before: 300000 Tianjin, Nankai District Jin Wei South Road, building 76, No. two Patentee before: Tianjin Caring Technology Development Co.,Ltd. |