CN101441167A - Method for measuring minimum inhibiting concentration of heavy metal to microorganism - Google Patents
Method for measuring minimum inhibiting concentration of heavy metal to microorganism Download PDFInfo
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- CN101441167A CN101441167A CNA2008101075156A CN200810107515A CN101441167A CN 101441167 A CN101441167 A CN 101441167A CN A2008101075156 A CNA2008101075156 A CN A2008101075156A CN 200810107515 A CN200810107515 A CN 200810107515A CN 101441167 A CN101441167 A CN 101441167A
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Abstract
The invention provides a method for quickly measuring the minimum inhibitory concentration of heavy metals on microbe, comprising: respectively preparing liquid culture medium adding with heavy metal ion gradient concentration which is suitable for growth of the microbe, moving strain in logarithmic phase to sterilized culture medium for culture in sterile conditions, measuring OD value by using UV spectrophotometry, and obtaining the minimum inhibitory concentration which has no increase change in OD values measured several times. The method is favorable to quickly select strains, and is more reliable for measuring the inhibitory of heavy metals on microbe.
Description
Technical field
The invention belongs to the environmental microorganism field, relate to a kind of heavy metal that is used for microorganism minimum inhibition concentration Rapid Determination Method.
Background technology
As screening one step of key that heavy metal is had tolerance and removal effect bacterium, the mensuration requirement of minimum inhibition concentration (MIC) is quick and precision is high.Also there not be now the patent or the invention of special assay method at minimum inhibition concentration.The most approaching is 01103281.2 Rapid Determination Method about the microorganism neurological susceptibility, and it mainly is to judge the neurological susceptibility of microorganism to test substance according to the variation of microbial cell number and aggregated forms.This method measure microorganism to the minimum inhibition concentration of heavy metal because microbial inoculant need be left standstill cultivation on solid medium, so need the long period, generally need two days to week age, so be unfavorable for the needs of rapid screening bacterial classification.Mensuration about minimum inhibition concentration in the document generally is bacterium to be inserted contain in the solid medium of a series of concentration of heavy metal ion, observes its growing state, and MIC is the minimum heavy metal concentration that does not have bacterial growth.This assay method can't demonstrate fully the ability of bacterium tolerance heavy metal, because generally, than solid conditions, the liquid condition is more suitable for bacterial growth.So it is more reliable to the inhibition of bacterium to measure heavy metal under the liquid condition.
Summary of the invention
The object of the present invention is to provide a kind of heavy metal that is used for to microorganism minimum inhibition concentration Rapid Determination Method.This method is beneficial to the needs of rapid screening bacterial classification, and it is more reliable to the inhibition of bacterium to measure heavy metal.
The objective of the invention is to realize in the following manner.
A kind of method of measuring heavy metal to the microorganism minimum inhibition concentration: the fluid nutrient medium that is suitable for growth of microorganism of preparing different heavy metal ion gradient concentrations respectively, again under aseptic condition, the bacterial classification that will be in exponential phase inserts the fluid nutrient medium cultivation of the described different heavy metal ion gradient concentrations of sterilization, and ultraviolet spectrophotometry is measured each OD value respectively; Repeatedly measuring the OD value does not increase the concentration of heavy metal ion that changes and is minimum inhibition concentration.
Under aseptic condition, the bacterial classification that will be in exponential phase inserts in the above-mentioned nutrient culture media of sterilization, places 28 ℃, cultivates in the 150r/min incubator.
Took out bacterium liquid, determined by ultraviolet spectrophotometry OD in every interval 6-8 hour
600Repeatedly measure OD
600Value does not increase the concentration of heavy metal ion that changes and is minimum inhibition concentration.
Beneficial effect of the present invention:
1, the present invention is beneficial to the needs of rapid screening bacterial classification, and being inoculated in behind the fluid nutrient medium only needed to detect in tens hours; With microbial inoculant was left standstill cultivation on solid medium in the past, (generally needing two days to week age) compared with the obvious advantage to need the long period;
2, generally, solid medium is cultivated and can only be judged the neurological susceptibility of microorganism to test substance, i.e. qualitative detection by the variation of microbial cell number and aggregated forms.And the present invention adopts fluid nutrient medium to cultivate in conjunction with ultraviolet spectrophotometry, has reached qualitative and purpose detection by quantitative, and than solid conditions, the liquid condition is more suitable for bacterial growth, so it is more reliable to the inhibition of bacterium to measure heavy metal under the liquid condition.
Embodiment
Following embodiment and embodiment are intended to further specify the present invention, rather than limitation of the invention.
Embodiment 1
Preparation adds the fluid nutrient medium that is suitable for growth of microorganism of heavy metal ion gradient concentration respectively, and under aseptic condition, the bacterial classification that will be in exponential phase inserts in the above-mentioned nutrient culture media of sterilization, places 28 ℃, cultivates in the 150r/min incubator.Take out bacterium liquid in 6-8 hour at interval, under the uv-spectrophotometric instrument, measure OD
600Repeatedly measure OD
600Value does not increase the concentration that changes and is minimum inhibition concentration.
Utilize this measuring method to measure the MIC of ten kinds of bacterium that preserve in the laboratory (separating the endogenetic bacteria from plant black nightshade), the result is as follows:
+: good resistance; +-: has resistance;-: non-resistant.
Measured the minimum inhibition concentration of ten kinds of endogenetic bacterias in chromium, cadmium and zinc ion solution of plant black nightshade, wherein chromium and zinc concentration gradient are set at 2mM, 5mM and 10mM, and cadmium is set at 1mM, 2mM and 5mM.Measure OD three times
600The no marked change in value back be non-resistant, OD
600Value changes and significantly to be good resistance, change but amplitude little be resistance.Nonresistant Cmin is exactly the minimum inhibition concentration of this heavy metal to bacterium.Measuring the used time for three times is 20-24 hour, generally needs two days to a week having saved the time greatly than dull and stereotyped cultural method, so can screen the bacterial classification with environmental applications potentiality more efficiently.
Claims (3)
1, a kind of method of measuring heavy metal to the microorganism minimum inhibition concentration, it is characterized in that, prepare the fluid nutrient medium that is suitable for growth of microorganism of different heavy metal ion gradient concentrations respectively, again under aseptic condition, the bacterial classification that will be in exponential phase inserts the fluid nutrient medium cultivation of the described different heavy metal ion gradient concentrations of sterilization, and ultraviolet spectrophotometry is measured each OD value respectively; Repeatedly measure the concentration that the OD value do not increase the heavy metal ion that changes and be minimum inhibition concentration.
2, a kind of method of measuring heavy metal to the microorganism minimum inhibition concentration according to claim 1, it is characterized in that under aseptic condition, the bacterial classification that will be in exponential phase inserts in the above-mentioned nutrient culture media of sterilization, place 28 ℃, cultivate in the 150r/min incubator.
3, a kind of method of measuring heavy metal to the microorganism minimum inhibition concentration according to claim 1 and 2 is characterized in that, takes out bacterium liquid, determined by ultraviolet spectrophotometry OD at interval in 6-8 hour
600Repeatedly measure OD
600Value does not increase the concentration of heavy metal ion that changes and is minimum inhibition concentration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101075156A CN101441167A (en) | 2008-12-19 | 2008-12-19 | Method for measuring minimum inhibiting concentration of heavy metal to microorganism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101075156A CN101441167A (en) | 2008-12-19 | 2008-12-19 | Method for measuring minimum inhibiting concentration of heavy metal to microorganism |
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| CN101441167A true CN101441167A (en) | 2009-05-27 |
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| CNA2008101075156A Pending CN101441167A (en) | 2008-12-19 | 2008-12-19 | Method for measuring minimum inhibiting concentration of heavy metal to microorganism |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311155A (en) * | 2010-07-05 | 2012-01-11 | 中国科学院城市环境研究所 | Device for inhibiting growth of microbes in water treatment test nutrient liquid |
| CN104237534A (en) * | 2014-09-28 | 2014-12-24 | 淮南师范学院 | Method for rapidly detecting bio-availability of nano-material |
| CN113416647A (en) * | 2021-07-01 | 2021-09-21 | 海南大学 | Method for rapidly screening heavy metal Cd/Cr-resistant microorganisms |
-
2008
- 2008-12-19 CN CNA2008101075156A patent/CN101441167A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311155A (en) * | 2010-07-05 | 2012-01-11 | 中国科学院城市环境研究所 | Device for inhibiting growth of microbes in water treatment test nutrient liquid |
| CN102311155B (en) * | 2010-07-05 | 2014-06-11 | 中国科学院城市环境研究所 | Device for inhibiting growth of microbes in water treatment test nutrient liquid |
| CN104237534A (en) * | 2014-09-28 | 2014-12-24 | 淮南师范学院 | Method for rapidly detecting bio-availability of nano-material |
| CN104237534B (en) * | 2014-09-28 | 2016-09-07 | 淮南师范学院 | A kind of method of quick detection nano material biological effectiveness |
| CN113416647A (en) * | 2021-07-01 | 2021-09-21 | 海南大学 | Method for rapidly screening heavy metal Cd/Cr-resistant microorganisms |
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Open date: 20090527 |