CN101948905A - Method for detecting nitrate reducing bacteria on site in oil field - Google Patents
Method for detecting nitrate reducing bacteria on site in oil field Download PDFInfo
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- CN101948905A CN101948905A CN 201010278200 CN201010278200A CN101948905A CN 101948905 A CN101948905 A CN 101948905A CN 201010278200 CN201010278200 CN 201010278200 CN 201010278200 A CN201010278200 A CN 201010278200A CN 101948905 A CN101948905 A CN 101948905A
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Abstract
The invention relates to a method for detecting nitrate reducing bacteria on site in an oil field, which belongs to the technical field of oil field microbial detection. The method for detecting nitrate reducing bacteria on site in the oil field comprises: 1) making a nitrate reducing bacterium detection bottle: washing a glass bottle, a rubber plug and an aluminum foil cover, rinsing with clear water, washing with distilled water, draining residual water completely, preparing a culture medium according to the requirements of an NRB test, packaging, sterilizing, filling the culture medium in the glass bottle, covering the rubber plug, pressing the aluminum foil cover tightly for sealing, sterilizing the bottle at the temperature of 100 to 140 DEG C with high-temperature steam for 10 to 30 minutes; and 2) and detecting the nitrate reducing bacteria on site the an oil field by using a disappear dilution process: injecting a water sample to be tested into test bottles by a sterile injector gradually to perform dilution culture till there is no bacteria growing in the last test bottle and counting the number of the bacteria in tested water sample. The method has the advantages of simple operation, reliable data, accuracy, quickness, portability, high detection efficiency and the like.
Description
Technical field
The invention belongs to oil field microorganism detection technical field, particularly relate to the detection method of the on-the-spot nitrate reduction bacterium in a kind of oil field.
Background technology
At present, in the every index inspective regulation process of oil field, usually need microbial numbers is detected, can monitoring and evaluating water quality and pollution condition to the The determination in the water sample of oil field.Traditional microorganism detection method is very loaded down with trivial details, time-consuming and be subjected to the envrionment conditions restriction stronger, as the method for plate culture count, need carry out under the aseptic condition of laboratory, and the on-the-spot water sample in oil field is transported to the laboratory, microbe population and type all can be subjected to certain influence.The scene, oil field often has the mensuration that a large amount of water samples need carry out microorganism, and traditional detection method can not satisfy actual needs.
Summary of the invention
The present invention provides the detection method of the on-the-spot nitrate reduction bacterium in a kind of oil field for solving the problem that prior art exists.
The object of the invention provides a kind ofly to be had simple to operately, and data are reliable, accurately fast, is easy to carry, and the detection efficiency height makes things convenient for the detection method of the on-the-spot nitrate reduction bacterium in oil field of characteristics such as the on-the-spot microorganism detection in oil field.
In order to overcome the deficiencies in the prior art, the detection method of the on-the-spot nitrate reduction bacterium in oil field of the present invention adopts following technical scheme:
The detection method of the on-the-spot nitrate reduction bacterium in a kind of oil field is characterized in that: the detection of nitrate reduction bacterium comprises that the detection bottle is made and the on-the-spot detection in oil field;
1) the nitrate reduction bacterium detects the bottle making
(1) cleans
Vial cleans, and rubber plug and aluminium foil lid clean, and use the clear water rinse then, with distilled water vial, rubber plug, aluminium foil lid is cleaned up again, and sky is done residual water;
(2) substratum preparation
At normal temperatures and pressures, substratum is prepared by the requirement of nitrate reduction bacteria test bottles;
(3) encapsulation and sterilization
The substratum of preparation is put in the vial with pipettor, covered rubber plug and aluminium foil lid, with the gland device it is compressed sealing again; To under 100-140 ℃ of condition, carry out high-temperature sterilization 10-30 minute with the high-temperature steam Autoclave through adding a cover the test bottle that seals;
2) the on-the-spot nitrate reduction bacterium in oil field adopts the disappearance dilution method to detect
When the bacterium of the nitrate reduction bacterium in the on-the-spot water sample to be measured in detection oil field is counted, water sample to be measured is injected the test bottle dilution step by step with asepsis injector to be cultivated, to the last till test bottle asepsis growth, calculate the number of bacterium in the water sample to be measured according to the multiple of dilution.
The detection method of the on-the-spot nitrate reduction bacterium in oil field of the present invention can also adopt following technical measures:
The detection method of the on-the-spot nitrate reduction bacterium in described oil field is characterized in: it is that 5-20mL vial, rubber plug and aluminium foil lid are cleaned is to clean 1-3 back rinsing in clear water in washing powder water that vial cleans, and cleans empty dried residual water again with distilled water.
The detection method of the on-the-spot nitrate reduction bacterium in described oil field, be characterized in: the formulation by weight of substratum is peptone 0.5～1g, beef powder 0.1～0.5g, yeast powder 0.1～0.5g, sodium-chlor 0.5～1g, saltpetre 0.05～0.3g, SODIUMNITRATE 0.05～0.3g, ammonium nitrate 0.05～0.2g, mineral substance magnesium 0.0002～0.001g, iron 0.0002～0.0005g, distilled water 100mL.
The detection method of the on-the-spot nitrate reduction bacterium in described oil field is characterized in: before the encapsulation, the pH value of regulating culture medium solution is 7-7.6, and the substratum of preparation is put in the washed glass bottle with the amount of 4-18mL/ bottle with pipettor.
The detection method of the on-the-spot nitrate reduction bacterium in described oil field, be characterized in: water sample to be measured injects test bottle step by step with asepsis injector and dilutes culturing step;
1) prepares several and passed through the clean 1mL injector for medical purpose of sterilization;
2) bundle several test bottles,, compile successively and go up sequence number as row's nitrate reduction bacteria test bottles;
3) get 1mL water sample to be measured with syringe and inject test bottle No. 1, fully concussion; Other gets a syringe and gets 1mL liquid in No. 1 test bottle and inject test bottle No. 2, and fully a last test bottle is arrived in concussion by that analogy;
4) the nitrate reduction bacteria test bottles that will inject the on-the-spot water sample in the oil field indoor cultivation that experimentizes regularly detects reading.
Advantage that the present invention has and positively effect:
The detection method of the on-the-spot nitrate reduction bacterium in oil field, owing to adopted brand-new technology scheme of the present invention, compared with prior art, the detection method of the on-the-spot nitrate reduction bacterium in oil field of the present invention has following characteristics:
(1) detection method of the on-the-spot nitrate reduction bacterium in this oil field, production process is simple, and good reliability is economical and practical.(2) the nitrate reduction bacteria test bottles can be measured the bacterium number of nitrate reduction bacterium rapidly and accurately, uses simple to operately, is easy to carry, and conveniently is used for the detection of oil field microorganism.
The present invention has simple to operate, and data are reliable, accurately fast, is easy to carry, and the detection efficiency height makes things convenient for advantages such as the on-the-spot microorganism detection in oil field.
Embodiment
For further understanding technology contents of the present invention, characteristics and effect, enumerate following example now, and be described in detail as follows:
Embodiment 1
The detection method of scene, a kind of oil field nitrate reduction bacterium comprises and detects the on-the-spot detection of bottle making and oil field, and concrete working method and detection step are:
1. the nitrate reduction bacterium detects the bottle making:
(1) material cleans
The 10mL vial is cleaned twice in washing powder water, rubber plug and aluminium foil cover in washing powder water and to clean once, then with the clear water rinse to there not being foam, with distilled water vial, rubber plug, aluminium foil lid are cleaned up again, empty dried residual water, stand-by.
(2) substratum preparation
At normal temperatures and pressures, press NRB test media preparation explanation preparation substratum.Prescription is:
Somatomedin (peptone 0.5～1g, beef powder 0.1～0.5g, yeast powder 0.1～0.5g), sodium-chlor 0.5～1g, saltpetre 0.05～0.3g, SODIUMNITRATE 0.05～0.3g, ammonium nitrate 0.05～0.2g, trace element (mineral substance magnesium 0.0002～0.001g, iron 0.0002～0.0005g), distilled water 100mL.The pH value of regulator solution makes it reach 7.0～7.6.(3) encapsulation and sterilization
The substratum of preparation is put in the washed glass bottle with the amount of 9mL/ bottle with pipettor, covered rubber plug and aluminium foil lid, with the gland device it is compressed sealing again; To under 121 ℃ of conditions, carry out high-temperature sterilization 20 minutes with the high-temperature steam Autoclave through adding a cover the test bottle that seals.
(4) binding
After the bottle that sealing sterilization is good treated that temperature is reduced to room temperature, n/group bundled with wide adhesive tape.The detection of nitrate reduction bacterium is carried out at the scene, oil field of can taking after finishing.
2. the nitrate reduction bacterium detects the on-the-spot use in bottle oil field:
When the bacterium of adopting the nitrate reduction bacterium number testing bottle to measure the nitrate reduction bacterium in the water sample to be measured is counted, according to disappearance dilution method principle, water sample to be measured is injected the test bottle dilution step by step with asepsis injector to be cultivated, to the last till test bottle asepsis growth, calculate the number of bacterium in the water sample to be measured according to the multiple of dilution.Following method and step are adopted in concrete operations:
(1) prepare several and passed through the clean 1mL injector for medical purpose of sterilization, standby;
(2) get above-mentioned a bundle and tie up the nitrate reduction bacteria test bottles that n test bottle is a row, and compile successively and go up sequence number 1,2,3
(3) get 1mL water sample to be measured with above-mentioned ready syringe and be injected in No. 1 test bottle, fully shake 50 times;
(4) get in addition and take out 1mL liquid in No. 1 test bottle of an asepsis injector after the concussion and be injected in No. 2 test bottles, fully shake 50 times, by that analogy in a last test bottle;
(5) the nitrate reduction bacteria test bottles that will inject on-the-spot water sample brings back to the cultivation that specified temperature is carried out in the laboratory, regularly detects reading.
Claims (5)
1. the detection method of the on-the-spot nitrate reduction bacterium in an oil field is characterized in that: the nitrate reduction bacterium detects and comprises and detect that bottle is made and the on-the-spot detection in oil field;
1) the nitrate reduction bacterium detects the bottle making
(1) cleans
Vial cleans, and rubber plug and aluminium foil lid clean, and use the clear water rinse then, with distilled water vial, rubber plug, aluminium foil lid is cleaned up again, and sky is done residual water;
(2) substratum preparation
At normal temperatures and pressures, substratum is prepared by the requirement of nitrate reduction bacteria test bottles;
(3) encapsulation and sterilization
The substratum of preparation is put in the vial with pipettor, covered rubber plug and aluminium foil lid, with the gland device it is compressed sealing again; To under 100-140 ℃ of condition, carry out high-temperature sterilization 10-30 minute with the high-temperature steam Autoclave through adding a cover the test bottle that seals;
2) the on-the-spot nitrate reduction bacterium in oil field adopts the disappearance dilution method to detect
When the bacterium of the nitrate reduction bacterium in the on-the-spot water sample to be measured in detection oil field is counted, water sample to be measured is injected the test bottle dilution step by step with asepsis injector to be cultivated, to the last till test bottle asepsis growth, calculate the number of bacterium in the water sample to be measured according to the multiple of dilution.
2. according to the detection method of the on-the-spot nitrate reduction bacterium in the described oil field of claim 1, it is characterized in that: vial clean be with 5-20mL vial, rubber plug and aluminium foil lid clean be in washing powder water, clean 1-3 time after rinsing in clear water, clean empty dried residual water again with distilled water.
3. according to the detection method of the on-the-spot nitrate reduction bacterium in the described oil field of claim 1, it is characterized in that: the formulation by weight of substratum is peptone 0.5～1g, beef powder 0.1～0.5g, yeast powder 0.1～0.5g, sodium-chlor 0.5～1g, saltpetre 0.05～0.3g, SODIUMNITRATE 0.05～0.3g, ammonium nitrate 0.05～0.2g, mineral substance magnesium 0.0002～0.001g, iron 0.0002～0.0005g, distilled water 100mL.
4. according to the detection method of the described on-the-spot nitrate reduction bacterium in oil field of claim 1, it is characterized in that: before the encapsulation, the pH value of regulating culture medium solution is 7-7.6, and the substratum of preparation is put in the washed glass bottle with the amount of 4-18mL/ bottle with pipettor.
5. according to the detection method of the described on-the-spot nitrate reduction bacterium in oil field of claim 1, it is characterized in that: water sample to be measured injects test bottle step by step with asepsis injector and dilutes culturing step;
(1) prepares several and passed through the clean 1mL injector for medical purpose of sterilization;
(2) bundle several test bottles,, compile successively and go up sequence number as row's nitrate reduction bacteria test bottles;
(3) get 1mL water sample to be measured with syringe and inject test bottle No. 1, fully concussion; Other gets a syringe and gets 1mL liquid in No. 1 test bottle and inject test bottle No. 2, and fully a last test bottle is arrived in concussion by that analogy;
(4) the nitrate reduction bacteria test bottles that will inject the on-the-spot water sample in the oil field indoor cultivation that experimentizes regularly detects reading.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201010278200 CN101948905A (en) | 2010-09-10 | 2010-09-10 | Method for detecting nitrate reducing bacteria on site in oil field |
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| CN 201010278200 CN101948905A (en) | 2010-09-10 | 2010-09-10 | Method for detecting nitrate reducing bacteria on site in oil field |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421899A (en) * | 2013-08-09 | 2013-12-04 | 中国石油化工股份有限公司 | Specific primer for quickly detecting nitrate reducing bacteria in oil filed and using method of specific primer |
| CN106544292A (en) * | 2016-09-27 | 2017-03-29 | 广东省生态环境技术研究所 | One plant of salt tolerant nitrate reduction bacterium and its application |
| CN106755287A (en) * | 2017-02-07 | 2017-05-31 | 盎亿泰地质微生物技术（北京）有限公司 | A kind of microbial is to content detection method of counting |
| CN106868091A (en) * | 2017-04-07 | 2017-06-20 | 滕州合易食品有限公司 | A kind of improved bacterium viable count method |
| CN110691851A (en) * | 2017-06-02 | 2020-01-14 | 沙特阿拉伯石油公司 | Method for detecting sulfate-reducing bacteria using iron additives as media supplements |
-
2010
- 2010-09-10 CN CN 201010278200 patent/CN101948905A/en active Pending
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| 《全面腐蚀控制》 20100628 胡涛等 渤海某油田微生物NRB和SRB培养及生长条件研究 41-44，48 1-5 第24卷, 第6期 2 * |
| 《应用与环境生物学报》 20070625 谭燕 油井采出液中硝酸盐还原菌的分离培养及对硫酸盐还原的抑制 390-394 1-5 第13卷, 第3期 2 * |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421899A (en) * | 2013-08-09 | 2013-12-04 | 中国石油化工股份有限公司 | Specific primer for quickly detecting nitrate reducing bacteria in oil filed and using method of specific primer |
| CN103421899B (en) * | 2013-08-09 | 2015-01-21 | 中国石油化工股份有限公司 | Specific primer for quickly detecting nitrate reducing bacteria in oil filed and using method of specific primer |
| CN106544292A (en) * | 2016-09-27 | 2017-03-29 | 广东省生态环境技术研究所 | One plant of salt tolerant nitrate reduction bacterium and its application |
| CN106544292B (en) * | 2016-09-27 | 2019-08-23 | 广东省生态环境技术研究所 | One plant of salt tolerant nitrate reduction bacterium and its application |
| CN106755287A (en) * | 2017-02-07 | 2017-05-31 | 盎亿泰地质微生物技术（北京）有限公司 | A kind of microbial is to content detection method of counting |
| CN106755287B (en) * | 2017-02-07 | 2020-05-08 | 盎亿泰地质微生物技术（北京）有限公司 | Method for detecting and counting relative content of microorganisms |
| US11505817B2 (en) | 2017-02-07 | 2022-11-22 | Advanced Energy & Environmental Technologies, Inc. | Method for detecting and counting relative content of microorganism |
| CN106868091A (en) * | 2017-04-07 | 2017-06-20 | 滕州合易食品有限公司 | A kind of improved bacterium viable count method |
| CN110691851A (en) * | 2017-06-02 | 2020-01-14 | 沙特阿拉伯石油公司 | Method for detecting sulfate-reducing bacteria using iron additives as media supplements |
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Application publication date: 20110119 |