CN100497654C - Sulfate-reducing bacteria testing fluid and testing bottle - Google Patents
Sulfate-reducing bacteria testing fluid and testing bottle Download PDFInfo
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- CN100497654C CN100497654C CNB2006101283179A CN200610128317A CN100497654C CN 100497654 C CN100497654 C CN 100497654C CN B2006101283179 A CNB2006101283179 A CN B2006101283179A CN 200610128317 A CN200610128317 A CN 200610128317A CN 100497654 C CN100497654 C CN 100497654C
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Abstract
A sulfate reducing bacterium test liquid and its test tube are disclosed. The sulfate reducing bacterium test liquid consists of lactate 2-10wt per mill, chloride 3-10wt per mill, sulfate 1-10 per mill,yeast extract 1-3 per mill,perferrite 0.1-1.0 per mill, vitamins 0.1-0.5 per mill, acetic acid or acetate 1.0-10 per mill and distilled water 955.5-991.8 per mill. Height H1 of test liquid: space height H2=1:0.3-0.8. It's fast, convenient, correct and has long storage time.
Description
One, technical field: the present invention relates to testing method and the Testing apparatus thereof of a kind of bacterium, particularly relate to the test fluid and the test bottle thereof of sulphate reducing bacteria in other materials such as a kind of water quality, mud, settling.
Two, background technology:
Sulphate reducing bacteria is that a class organises can heterotrophic organism, and is extremely wide in distributed in nature.Under anaerobic, sulphate reducing bacteria more is present in water supply, the drainage pipeline in water, mud and the settling, it can utilize the organism that is attached to the metallic surface as carbon source, and utilizes the hydrogen that produces in the bacterial biof iotalm, and sulfate reduction is become hydrogen sulfide.Hydrogen sulfide corrosion is extremely strong, can with metallic matrix generation chemical reaction, cause chemical corrosion.The iron sulfide that reaction generates is deposited on the metallic surface, has changed the surface properties of matrix metal.Simultaneously, at environment intermediate ion (Cl especially
-) synergy under, may cause more serious corrosion.Therefore, people generally believe that sulphate reducing bacteria can bring out corrosion, occupies an leading position in metal bacterial corrosion.
Carried out a series of research at physiology, ecology and biochemical character and the mechanism of corrosion and the safeguard procedures etc. of sulphate reducing bacteria both at home and abroad, but at present, the testing method of sulphate reducing bacteria does not still have national standard method in the water sample, great majority adopt traditional multitube fermentation method, and the key instrument that relates in the multitube fermentation method is: high pressure steam sterilizer, constant incubator, aseptic technique platform, electronic balance, spirit lamp, nichrome wire loop-carrier, culture dish, test tube, suction pipe, beaker, Erlenmeyer flask and sampling bottle etc.
The main operational steps of multitube fermentation method is:
The first step is gathered water sample: draw a certain amount of water sample with the aseptic technique method with the sterilization suction pipe;
The preparation of the second step sterilized water;
The 3rd step vessel, reagent sterilization: will put into autoclave sterilizer after the various vessel cleanings, the sterilization certain hour;
The 4th step culture medium preparation: check and weighing all ingredients, with sterilized water dissolving preparation;
The 5th step stepwise dilution and inoculation water sample;
The 6th goes on foot the cultivation of bacterium;
The 7th step enumeration and report.
Traditional measuring method multitube fermentation method, this method requires very strict to laboratory environment and operator, comprise that expertise from Sampling techniques, laboratory condition, plant and instrument, vessel, reagent quality, operator, operating skill, practical experience are until all observation processes and sequence of control such as quality control.Schedule of operation is loaded down with trivial details, complicated, tediously long, the error that all might cause test result accidentally of any one link.This multitube fermentation method is measured process generally needs 20 days.
Three, summary of the invention:
Purpose of the present invention: overcome the deficiency in the sulfate-reducing bacteria testing method in the prior art, provide a kind of highly selective, fast, have simple and easy sulfate-reducing bacteria testing fluid and a test bottle thereof than long pot life.
Technical scheme of the present invention:
The invention provides a kind of sulfate-reducing bacteria testing fluid, this test fluid is the highly selective liquid nutrient medium, with the weight content is unit, by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1.0～10 ‰ and distilled water 955.5～991.8 ‰ are formed.
Described sulfate-reducing bacteria testing fluid is by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～5 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～0.5 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 5～10 ‰ and distilled water 962～987.8 ‰ are formed.
Described sulfate-reducing bacteria testing fluid is by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 5～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.5～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1～5 ‰ and distilled water 961～987.4 ‰ are formed.
Lactic acid salt is Sodium.alpha.-hydroxypropionate or calcium lactate in the described sulfate-reducing bacteria testing fluid; Muriate is sodium-chlor or Repone K; Vitriol is sal epsom or ferrous ammonium sulphate; Ferrous salt is a ferrous ammonium sulphate; VITAMIN is a vitamins C; Acetate is a sodium acetate.
A kind of sulfate-reducing bacteria testing bottle, this test bottle is equipped with sulfate-reducing bacteria testing fluid, the height H of test fluid in bottle for aseptic, the colourless transparent glass bottle of band plug and aluminium cover in the test bottle
1With the height H of leaving a blank
2Ratio be H
1: H
2=1:0.3～0.8; Described sulfate-reducing bacteria testing bottle, with the weight content is unit, by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1.0～10 ‰ and distilled water 955.5～991.8 ‰ are formed.
Described sulfate-reducing bacteria testing bottle, wherein said test fluid be by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～5 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～0.5 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 5～10 ‰ and distilled water 962～987.8 ‰ are formed.
Described sulfate-reducing bacteria testing bottle, wherein said test fluid be by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 5～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.5～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1～5 ‰ and distilled water 961～987.4 ‰ are formed.
Test bottle of the present invention can be selected 5～15 milliliters of cillin bottles or oral liquid bottle for use.
Working method when using sulfate-reducing bacteria testing fluid of the present invention and test bottle thereof to be used for measuring water quality is: with asepsis injector water sample is drawn 1.0 (or 0.5) milliliter, be injected in the test bottle, be positioned over after shaking up in 35 ℃ of loft drier or the water-bath 14 days, and judged feminine gender or positive reaction according to the variation of liquid in the bottle.The no color of liquid itself in the bottle, if no change is negative, it is positive black precipitate to occur, this is qualitative analysis; According to said method do 2 groups or 3 groups of parallel samples, look into the MPN table, can carry out quantitative test by the stepwise dilution method.Other solid sample such as mud then must be with water dissolution or lixiviate, measures as stated above after being processed into liquid again.
Positive beneficial effect of the present invention:
1. test fluid of the present invention sealing is stored in the vial, has than long storage life, save the multitube fermentation method second and third, four operation steps, asepsis injector need only be disposed and whole operations can be finished, only need 14 days detection time, alleviated labour intensity greatly, shortened the test duration.
2. the present invention then can be reduced to 3～5 days if be used with the hydrogen sulfide production test testing tube detection time.
3. the present invention has exempted gnotobasis condition necessary in the traditional method, makes the biochemical investigation to carry out in general environment, saves a large amount of infrastructure investments.
4. the feature precipitin reaction of reagent is to carry out automatically in aseptic bacterium test bottle in opsonigenous substances and the test fluid, can avoid the pollution and the interference that easily cause in the rapid operating process of traditional method multistep, thereby improves the accurate reliability of detected result greatly; The minimum detectability of present method meets national water quality standard requirement.
5. the present invention can be applicable to the simple and easy quick test of sulphate reducing bacteria in the various water quality, and non-specialised staff's number minute can be learned operation.
6. the present invention is loaded on Bacteria Detection technical productsization, commercialization in the clear-glass bottle with pre-configured mensuration fluid-tight, can be directly used in the mensuration of bacterium, carries, easy to use, measure simple fast, accurately, the shelf time is long.Can be used for the bacterium qualitative and quantitative analysis of multiple materials such as water quality, food, medicine, makeup.
Four, description of drawings:
Fig. 1: the structural representation of sulfate-reducing bacteria testing bottle
Five, embodiment:
The following examples are only in order to further specify the present invention, rather than limit the scope of the invention.
Embodiment one:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 2 ‰, and sodium-chlor 10 ‰, sal epsom 1 ‰, yeast extract 3 ‰, ferrous sulfate 0.5 ‰, vitamins C 0.1 ‰, sodium acetate 5 ‰ and distilled water 978.4 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 3 centimetres, the height H of leaving a blank
2It is 1.5 centimetres.
Embodiment two:
The sulfate-reducing bacteria testing fluid prescription is: by calcium lactate 10 ‰, and Repone K 3 ‰, sal epsom 5 ‰, yeast extract 1 ‰, ferrous ammonium sulphate 0.1 ‰, vitamins C 0.5 ‰, acetate 10 ‰ and distilled water 970.4 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 4 centimetres, the height H of leaving a blank
2It is 1.2 centimetres.
Embodiment three:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 6 ‰, and Repone K 7 ‰, sal epsom 3 ‰, yeast extract 2 ‰, ferrous ammonium sulphate 0.3 ‰, vitamins C 0.3 ‰, acetate 8 ‰ and distilled water 973.4 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 9.0 milliliters every bottle, gets final product through autoclaving.
Sulphate reducing bacteria trial jar implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 9.0 milliliters in the present embodiment, the height H of test fluid
1Be 3.5 centimetres, the height H of leaving a blank
2It is 2.8 centimetres.
Embodiment four:
The sulfate-reducing bacteria testing fluid prescription is: by calcium lactate 4 ‰, and sodium-chlor 5 ‰, sal epsom 7 ‰, yeast extract 3 ‰, ferrous ammonium sulphate 1.0 ‰, vitamins C 0.2 ‰, acetate 5 ‰ and distilled water 974.8 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 9.0 milliliters every bottle, gets final product through autoclaving.
Sulphate reducing bacteria trial jar implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 9.0 milliliters in the present embodiment, the height H of test fluid
1Be 4.5 centimetres, the height H of leaving a blank
2It is 2.7 centimetres.
Embodiment five:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 8 ‰, and Repone K 8 ‰.Sal epsom 10 ‰, yeast extract 1 ‰, ferrous ammonium sulphate 0.5 ‰, vitamins C 0.4 ‰, sodium acetate 1.0 ‰ and distilled water 971.1 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 5 centimetres, the height H of leaving a blank
2It is 2 centimetres.
Embodiment six:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 2 ‰, and sodium-chlor 10 ‰, sal epsom 5 ‰, yeast extract 2 ‰, ferrous sulfate 0.7 ‰, vitamins C 0.3 ‰, sodium acetate 3 ‰ and distilled water 977 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 3 centimetres, the height H of leaving a blank
2It is 1.5 centimetres.
Embodiment seven:
The sulfate-reducing bacteria testing fluid prescription is: by calcium lactate 2 ‰, and Repone K 10 ‰, sal epsom 1 ‰, yeast extract 3 ‰, ferrous ammonium sulphate 0.1 ‰, vitamins C 0.5 ‰, acetate 10 ‰ and distilled water 973.4 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 4 centimetres, the height H of leaving a blank
2It is 1.2 centimetres.
Embodiment eight:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 10 ‰, and Repone K 3 ‰, sal epsom 10 ‰, yeast extract 1 ‰, ferrous ammonium sulphate 1 ‰, vitamins C 0.1 ‰, acetate 1 ‰ and distilled water 973.9 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 9.0 milliliters every bottle, gets final product through autoclaving.
Sulphate reducing bacteria trial jar implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 9.0 milliliters in the present embodiment, the height H of test fluid
1Be 3.5 centimetres, the height H of leaving a blank
2It is 2.8 centimetres.
Embodiment nine:
The sulfate-reducing bacteria testing fluid prescription is: by calcium lactate 5 ‰, and sodium-chlor 8 ‰, sal epsom 3 ‰, yeast extract 2, ferrous ammonium sulphate 0.3 ‰, vitamins C 0.2 ‰, acetate 3 ‰ and distilled water 978.5 ‰ are formed.
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5-7.2, is packaged in aseptic, the water white transparency phial by 9.0 milliliters every bottle, gets final product through autoclaving.
Sulphate reducing bacteria trial jar implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 9.0 milliliters in the present embodiment, the height H of test fluid
1Be 4.5 centimetres, the height H of leaving a blank
2It is 2.7 centimetres.
Embodiment ten:
The sulfate-reducing bacteria testing fluid prescription is: by Sodium.alpha.-hydroxypropionate 7 ‰, and Repone K 5 ‰.Sal epsom 7 ‰, yeast extract 3 ‰, ferrous ammonium sulphate 0.7 ‰, vitamins C 0.3 ‰, sodium acetate 7 ‰ and distilled water 970 ‰ are formed..
Above-mentioned test fluid is prepared the back, and to regulate pH value be 6.5～7.2, is packaged in aseptic, the water white transparency phial by 4.5 milliliters every bottle, gets final product through autoclaving.
Sulfate-reducing bacteria testing bottle implementation method: referring to Fig. 1,1 is aluminium cover among the figure, and 2 is plug, and 3 is the uniform cross section vial, and 4 is test fluid; H
1Be the height of test fluid, H
2Be the height of leaving a blank, test bottle is 4.5 milliliters in the present embodiment, the height H of test fluid
1Be 5 centimetres, the height H of leaving a blank
2It is 2 centimetres.
Claims (7)
1. sulfate-reducing bacteria testing fluid, this test fluid is the highly selective liquid nutrient medium, it is characterized in that: be unit with the weight content, by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1.0～10 ‰ and distilled water 955.5～991.8 ‰ are formed.
2. sulfate-reducing bacteria testing fluid according to claim 1, it is characterized in that: by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～5 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～0.5 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 5～10 ‰ and distilled water 962～987.8 ‰ are formed.
3. sulfate-reducing bacteria testing fluid according to claim 1, it is characterized in that: by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 5～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.5～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1～5 ‰ and distilled water 961～987.4 ‰ are formed.
4. according to each described sulfate-reducing bacteria testing fluid of claim 1～3, it is characterized in that: described lactic acid salt is Sodium.alpha.-hydroxypropionate or calcium lactate; Muriate is sodium-chlor or Repone K; Vitriol is sal epsom or ferrous ammonium sulphate; Ferrous salt is a ferrous ammonium sulphate; VITAMIN is a vitamins C; Acetate is a sodium acetate.
5. sulfate-reducing bacteria testing bottle, this test bottle is characterized in that: sulfate-reducing bacteria testing fluid is housed, the height H of test fluid in bottle in the test bottle for aseptic, the colourless transparent glass bottle of band plug and aluminium cover
1With the height H of leaving a blank
2Ratio be H
1: H
2=1:0.3～0.8; Described sulfate-reducing bacteria testing fluid, with the weight content is unit, by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1.0～10 ‰ and distilled water 955.5～991.8 ‰ are formed.
6. sulfate-reducing bacteria testing bottle according to claim 5, it is characterized in that: described sulfate-reducing bacteria testing fluid is by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 1～5 ‰, yeast extract 1～3 ‰, ferrous salt 0.1～0.5 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 5～10 ‰ and distilled water 962～987.8 ‰ are formed.
7. sulfate-reducing bacteria testing bottle according to claim 5, it is characterized in that: described sulfate-reducing bacteria testing fluid is by lactic acid salt 2～10 ‰, muriate 3～10 ‰, vitriol 5～10 ‰, yeast extract 1～3 ‰, ferrous salt 0.5～1.0 ‰, VITAMIN 0.1～0.5 ‰, acetate or acetate 1～5 ‰ and distilled water 961～987.4 ‰ are formed.
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| CN101368200B (en) * | 2008-07-18 | 2011-09-21 | 哈尔滨师范大学 | Culture medium for sifting motion anaerobic coreduction sulfate and inverse nitrification function bacterial strain and sifting motion method thereof |
| CN102329851B (en) * | 2011-10-08 | 2012-09-26 | 西安长庆化工集团有限公司 | Sulfate reduction bacteria culture medium for oil field sewage treatment system |
| CN103901024B (en) * | 2012-12-27 | 2016-06-08 | 中国石油天然气股份有限公司 | A kind of method evaluating sulfate reducting bacteria inhibition |
| WO2020029112A1 (en) * | 2018-08-08 | 2020-02-13 | 中国石油大学(北京) | Desulfovibrio detection composition, preparation method therefor and use thereof |
| CN110872570B (en) * | 2019-10-22 | 2024-03-19 | 广东第二师范学院 | Solution formula for efficiently producing sulfide by utilizing different soils and method for preparing sulfide |
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Assignee: ZHENGZHOU WATER TESTING TECHNOLOGY CO., LTD. Assignor: Bai Li Contract record no.: 2012410000012 Denomination of invention: Sulfate-reducing bacteria testing fluid and testing bottle Granted publication date: 20090610 License type: Exclusive License Open date: 20070905 Record date: 20120306 |
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