CN104046563A - Heterotrophic bacteria test bottle production method - Google Patents
Heterotrophic bacteria test bottle production method Download PDFInfo
- Publication number
- CN104046563A CN104046563A CN201310078446.1A CN201310078446A CN104046563A CN 104046563 A CN104046563 A CN 104046563A CN 201310078446 A CN201310078446 A CN 201310078446A CN 104046563 A CN104046563 A CN 104046563A
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- CN
- China
- Prior art keywords
- test bottle
- bottle
- heterotrophic bacterium
- heterotrophic bacteria
- heterotrophic
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- 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
- C12M23/08—Flask, bottle or test tube
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- 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
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- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
Abstract
The invention relates to a heterotrophic bacteria test bottle production method which belongs to the technical field of application of microbial detection methods, and is characterized in that: at room temperature, respectively taking 0.5% by weight of sodium chloride, 1% by weight of peptone, 0.3% by weight of beef extract and 98.2% by weight of distilled water; respectively washing the sodium chloride, peptone and beef extract with the distilled water, adding into a container, mixing, shaking well, using a 0.5mol / L sodium hydroxide solution to adjust the pH value to 7.4-7.6 to prepare a culture solution required by preparation of a heterotrophic bacteria test bottle; using a liquid packing device to filling every 9.0ml of the prepared culture solution into each penicillin bottle, covering, sealing, sterilizing at high temperature, and cooling to prepare the heterotrophic bacteria test bottle. The heterotrophic bacteria test bottle produced by the method can quickly and accurately determine the bacteria number of heterotrophic bacteria in a water sample, the operation is simple, the working efficiency is greatly improved, and the heterotrophic bacteria test bottle can meet the actual working requirements in water quality detection and water quality control and other aspects of industrial circulating cooling water.
Description
Technical field:
The present invention relates to a kind of production method of heterotrophic bacterium test bottle, belong to using microbe detection method technical field.
Background technology:
In research and production real work, usually need the quantity of microorganism to measure.As the mensuration of the microorganism in the environment such as water body, air, soil is conducive to monitoring and evaluates its pollution level; To the mensuration of microorganism in tap water and in industrial circulating cooling water, can evaluate whether it up to standard; The mensuration of the microbe population in oilfield sewage water filling is contributed to water quality of evaluating sewage water filling etc.Therefore, the mensuration of microbe population has very important directive function and realistic meaning to research and production real work.
The conventional method of microbioassay has the method for plate culture count and disappearance dilution method.The method of plate culture count is can grow the principle design of a bacterium colony according to each microorganism alive, get the bacteria suspension of certain capacity, make a series of doubling dilution, then quantitative diluent being carried out to flat board cultivates, according to the colony number of turning out and extension rate, calculate the viable count in culture; And disappearance dilution method is water sample to be determined to be injected into step by step inoculate dilution in test tube, to the last in a test tube till asepsis growth, then according to microorganism growth situation and extension rate, calculate the number of microorganism in water sample.These two kinds of methods have obvious deficiency, and the method for plate culture count measurement result error is larger, and the operation of disappearance dilution method is very loaded down with trivial details, time-consuming.
The disappearance dilution method of the take heterotrophic bacterium test bottle method that is ultimate principle, has fully absorbed the advantage of disappearance dilution method, and its operation is very simple, growth indication obviously, accurately and reliably, operation does not need professional to grasp to result, thereby has improved widely the measuring method of microorganism.
In industrial circulating cooling water, oilfield sewage water filling, ubiquity heterotrophic bacterium, and heterotrophic bacterium is that a group must be take the general designation of various organism as synthetic self the organic aerobic and facultative anaerobic bacteria of Nutrition and Metabolism.The various organic compound that they can be take in water body etc. carry out growth and breeding as carbon source.Therefore, the existence of heterotrophic bacterium not only can polluted water, but also can cause the obstruction on the heavy corrosion of equipment, pipeline and pipeline, stratum.In order effectively to control the harm of heterotrophic bacterium, need carry out Accurate Determining to heterotrophic bacterium.At present, generally adopt the method for plate culture count to measure the heterotrophic bacterium in water body both at home and abroad, but this method measurement result error is larger, and loaded down with trivial details, time-consuming.
Therefore, there is open defect in traditional detection technique, urgently research and develop a kind of simply, heterotrophic bacterium detects accurately novel method, new tool.Therefore, development has important practical significance for detection of the test bottle of the heterotrophic bacterium in water body.
Summary of the invention:
In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a kind of production method of heterotrophic bacterium test bottle, make produced test bottle can measure rapidly and accurately the bacterium number of the heterotrophic bacterium in water sample, to meet the real work demand of the aspects such as industrial circulating cooling water water quality detection and water quality control.
The present invention realizes above-mentioned purpose by following technical solution.
At normal temperatures, get respectively by weight percentage sodium-chlor 0.5%, peptone 1.0%, extractum carnis 0.3%, distilled water 98.2%; Will sodium-chlor, peptone, extractum carnis dissolve with distilled water successively, join in container and mix, fully shake up, and regulate its pH value to 7.4--7.6 with 0.5mol/L sodium hydroxide solution, are mixed with and produce the required nutrient solution of heterotrophic bacterium test bottle; The molded cillin bottle of 12ml is cleaned up, is dried, with liquid dispenser, prepared nutrient solution is packed in bottle by every bottle of 9.0ml, then add a cover, seal; To with high-temp steam sterilizing pot, under 121 ℃ of conditions, carry out high-temperature sterilization 20 minutes through adding a cover the test bottle of sealing, make heterotrophic bacterium test bottle after cooling.
The present invention has following beneficial effect compared with prior art:
1, the heterotrophic bacterium test bottle that the present invention produces, bacterium number, the growth indication that can measure rapidly and accurately the heterotrophic bacterium in water sample be obvious, simple to operate, can greatly increase work efficiency.
2, nutrient solution is heterotrophic bacterium special culture solution, is only suitable for heterotrophic bacterium growth, and the microorganism of turning out is heterotrophic bacterium, and test job can not be subject to the interference of other microorganism.
3, the bacterium number of the heterotrophic bacterium of the present invention in can Accurate Determining water sample, can be used for the fields such as process water detection, at aspects such as industrial circulating cooling water water quality detection and water quality controls, is with a wide range of applications.
Embodiment:
(take and prepare 1L nutrient solution as example, by that analogy), take respectively by weight percentage sodium-chlor 5.0g, peptone 10.0g, extractum carnis 3.0g at normal temperatures; Will sodium-chlor, peptone, extractum carnis dissolve with distilled water successively, and join in 1L volumetric flask mix, again with distilled water constant volume to 1L scale marks, fully shake up, measure its potential of hydrogen and regulate its pH value to 7.4--7.6 with 0.5mol/L sodium hydroxide solution, being mixed with and producing the required nutrient solution of heterotrophic bacterium test bottle.
The molded cillin bottle of 12ml is cleaned up, is dried, with liquid dispenser, prepared nutrient solution is packed in bottle by every bottle of 9.0ml, with A-13 type medical rubber plug, add a cover, with common antibiotic glass bottle aluminium lid, through special-purpose sealing device, seal, finally will with high-temp steam sterilizing pot, under 121 ℃ of conditions, carry out high-temperature sterilization 20 minutes through adding a cover the test bottle of sealing, make heterotrophic bacterium test bottle after cooling.
When the bacterium that adopts heterotrophic bacterium test bottle to measure the heterotrophic bacterium in water sample to be measured is counted, adopt disappearance dilution method principle, water sample to be measured is injected into dilution in test bottle step by step with asepsis injector to be cultivated, to the last, till a test bottle asepsis growth, can calculate according to the multiple of dilution the number of heterotrophic bacterium in water sample to be measured.
Practical measurement operation can adopt following method and step:
(1) prepare the 1ml injector for medical purpose of number Zhi Xiangtong, with distilled water, clean up rear high-temperature sterilization (under 121 ℃ of high-pressure steam sterilizing pans, 0.105MPa, sterilizing is 15 minutes), cooling standby;
(2) roughly how many according to heterotrophic bacterium in water sample to be measured, line up one group by several above-mentioned made heterotrophic bacterium test bottles, and are numbered with successively sequence number 1,2,3,4
(3) with the asepsis injector after above-mentioned sterilizing, get 1.0mL water sample to be measured and be injected in No. 1 heterotrophic bacterium test bottle, fully shaking 10 seconds;
(4) separately get and in No. 1 heterotrophic bacterium test bottle of an asepsis injector from shaking up, take out 1.0mL liquid and be injected in No. 2 heterotrophic bacterium test bottles, fully shake 10 seconds;
(5) separately get again and in No. 2 heterotrophic bacterium test bottles of an asepsis injector from shaking up, take out 1.0mL liquid and be injected in No. 3 heterotrophic bacterium test bottles, fully shake 10 seconds;
(6) repeat successively aforesaid operations program, until till last bottle;
(7) cultivating in the above-mentioned incubator that adds one group of heterotrophic bacterium test bottle after water sample to be measured to be placed on 30 ℃ or in situ temperature, the variation after 3 days cultivate in observation test bottle, the growing state of judgement heterotrophic bacterium;
(8) judging criterion: the nutrient solution in the heterotrophic bacterium test bottle existing without heterotrophic bacterium is without considerable change, and have the nutrient solution in the heterotrophic bacterium test bottle that heterotrophic bacterium exists to become muddy feculence, therefore, the nutrient solution in heterotrophic bacterium test bottle becomes muddy feculence, indicates heterotrophic bacterium growth;
(9) according to the situation of bacterial indicator growth in heterotrophic bacterium test bottle and the bottle number that has the test bottle of bacteria growing, the 1 heterotrophic bacterium bacterium number that can calculate in water sample to be measured of tabling look-up.
Single bottle of heterotrophic bacterium maximum possible value of table 1
Claims (4)
1. a production method for heterotrophic bacterium test bottle, is characterized in that adding in bottle the nutrient solution of being made by sodium-chlor, peptone, extractum carnis and distilled water, through adding a cover, sealing, high-temp steam sterilizing make.
2. the production method of a kind of heterotrophic bacterium test bottle according to claim 1, is characterized in that at normal temperatures, gets respectively by weight percentage sodium-chlor 0.5%, peptone 1.0%, extractum carnis 0.3%, distilled water 98.2%; Will sodium-chlor, peptone, extractum carnis dissolve with distilled water successively, join in container and mix, fully shake up, and regulate its pH value to 7.4--7.6 with 0.5mol/L sodium hydroxide solution, are mixed with and produce the required nutrient solution of heterotrophic bacterium test bottle.
3. the production method of a kind of heterotrophic bacterium test bottle according to claim 2, is characterized in that the molded cillin bottle of 12ml to clean up, be dried, and with liquid dispenser, prepared nutrient solution is packed in bottle by every bottle of 9.0ml, then adds a cover, seals.
4. the production method of a kind of heterotrophic bacterium test bottle according to claim 3, is characterized in that with high-temp steam sterilizing pot, under 121 ℃ of conditions, carrying out high-temperature sterilization 20 minutes through adding a cover the test bottle of sealing, makes heterotrophic bacterium test bottle after cooling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310078446.1A CN104046563A (en) | 2013-03-13 | 2013-03-13 | Heterotrophic bacteria test bottle production method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310078446.1A CN104046563A (en) | 2013-03-13 | 2013-03-13 | Heterotrophic bacteria test bottle production method |
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| CN104046563A true CN104046563A (en) | 2014-09-17 |
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| CN201310078446.1A Pending CN104046563A (en) | 2013-03-13 | 2013-03-13 | Heterotrophic bacteria test bottle production method |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101029332A (en) * | 2006-11-30 | 2007-09-05 | 白莉 | Bacterium total testing fluid and its testing bottle |
| CN101845389A (en) * | 2009-03-24 | 2010-09-29 | 长江大学 | Method for producing bottles for testing total number of bacteria in drinking water |
-
2013
- 2013-03-13 CN CN201310078446.1A patent/CN104046563A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101029332A (en) * | 2006-11-30 | 2007-09-05 | 白莉 | Bacterium total testing fluid and its testing bottle |
| CN101845389A (en) * | 2009-03-24 | 2010-09-29 | 长江大学 | Method for producing bottles for testing total number of bacteria in drinking water |
Non-Patent Citations (1)
| Title |
|---|
| 易绍金 等: "测试瓶法测定工业循环冷却水中的真菌", 《工业水处理》, vol. 26, no. 7, 31 July 2006 (2006-07-31) * |
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Application publication date: 20140917 |