CN102539640A - Method used for testing high throughput acute toxicity of early life stage of fish - Google Patents
Method used for testing high throughput acute toxicity of early life stage of fish Download PDFInfo
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- CN102539640A CN102539640A CN2010106085113A CN201010608511A CN102539640A CN 102539640 A CN102539640 A CN 102539640A CN 2010106085113 A CN2010106085113 A CN 2010106085113A CN 201010608511 A CN201010608511 A CN 201010608511A CN 102539640 A CN102539640 A CN 102539640A
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Abstract
The invention relates to a special method for testing the high throughput acute toxicity of an early life stage of fish, and belongs to the technical field of environment detection and assessment. According to the method, the sensibility of the fish at different life stages on toxic pollutants as well as the influence of conditions such as different quantities of test creatures, different exposed containers and the like on test results are discussed through developing a recombinant dilute solution for toxicity test, thus realizing high throughput on a larval fish acute toxicity test. The method is simple, convenient, economical, rapid and sensitive, has low requirements on experiment operation technical level, and is convenient for popularization and application.
Description
Technical field
The present invention relates to a kind of special high flux acute toxic test method, belong to environment measuring and assessment technique category to the fish early life stage.
Background technology
Along with increasingly sharpening of socioeconomic fast development and industrial and agricultural production activity; Increasing chemicals is produced and is used; It is reported, ten million kind more than of present registered chemical quantity exceedance, the chemical quantity of using in daily life and the production is above 100,000 kinds.These chemicals possibly get into environment or remain in human body, the biosome through number of ways in production and use, ecologic environment and health are caused serious threat.Be the accurately security of evaluating chemical article and environmental sample, the bio-toxicity effect is essential and of paramount importance basic data.
Existing tens of kinds more than of the current bio-toxicity method of testing that generally adopts; International Standards Organization (ISO), the Organization for Economic Cooperation and Development (OECD), Environmental Protection Agency (USEPA), international organizations such as (EU) of European Union and mechanism have formulated standard, standard to the method for part biological toxotest; China has has also issued and implemented the standard method of several bio-toxicity tests in recent years successively; " the water quality material is to fresh-water fishes (zebra fish) determination of acute toxicity method GB/T13267-1991 " is exactly one of them; The method of recommending with International Standards Organization such as ISO has certain comparability, and the toxotest field is widely used at home.But, in the standard method of at present relevant fish acute toxicity test, recommend mostly to use adult fish as biological subject, in practical operation, there are many deficiencies.Such as, because the toxotest of each sample all needs a large amount of fishes of being tried, the nursing relative difficult of adult fish, higher to experimenter's technical requirement; Feeding a large amount of experiments needs lab construction expense, daily servicing expense and the personnel of great number to drop into fish; Test greatlyyer with the adult fish build, it is also corresponding bigger to expose container, be inconvenient to operate, and in addition, the sample volume that needs is also bigger, tests as if the employing continuous-flow type, and required sample volume is bigger, is unfavorable for the test of micro-example; Because differing greatly between the adult fish individuality, often there is certain otherness in measured result, often need carry out a plurality of parallel laboratory tests for reducing individual error, but has increased quantity, sample volume, the experimental expenses of experiment with fish; Traditional adult fish toxotest experiment exposure duration generally was not less than 96 hours, and required time is longer, is unfavorable for the quick test of batch samples; Adult fish is relatively low to the susceptibility of chemical pollutant, and measuring LC50 often needs more a large amount of samples.Therefore, a kind of fast, sensitive, high-throughout fish acute toxicity method of testing of exploitation is very urgent.
Summary of the invention
The objective of the invention is for overcoming deficiencies such as biological subject is individual greatly in traditional adult fish acute toxic test, quantity is few, individual difference is big, the test duration is long, reappearance is low; A kind of easy, economic, quick, high-throughout acute toxic test method is provided, for the acute toxic test of toxic chemical pollutant and environmental sample provides technical guarantee.
The object of the invention is realized in the following manner: to the biological subject fish in the toxotest, develop a kind of special reorganization dilute solution, cultivate newborn prelarva and be used for acute toxic test; Simultaneously, the prelarva through selecting early life stage, adopt microwell plate, carry out acute toxic test, realized the high flux test of fish acute toxicity as the toxic exposure container.
Advantage of the present invention is mainly reflected in the following aspects: a kind of special reorganization dilute solution of cultivating to fish has been developed in (1), is used for fast, stablizes the prelarva of being tried of cultivating early life stage; (2) through relatively, the prelarva having selected to be in early life stage carries out acute toxic test as biological subject, remedied to adopt adult fish to carry out many deficiencies (, poor reproducibility big, sensitivity is low, cost is high, technical requirement is high) of toxotest in the classic method like individual difference; (3) adopt the container of microwell plate, both reduced the volume of experiment testing sample, also realized the high flux of toxotest as toxic exposure; (4) adopt this method both to save experimental period, also reduced experimental cost, experimental result increases than classic method; (5) the present invention is simple to operate, has reduced the requirement to operating personnel's technical merit, also is convenient to popularizing of method.
In sum, the present invention passes through a kind of special reorganization dilute solution to fish cultivation and toxotest of exploitation, has realized that early life stage is tried the quick, stable of fish to be cultivated; Tried the susceptibility of fish through more different life stages to toxic pollutants; Definite employing is in the fish of early life stage as biological subject; Both realize the sensitivity and the stability of acute toxic test, shortened the toxotest time again, also reduced cost; Adopt the exposure container of microwell plate, reduced the volume of testing sample, realized the high flux of test, improved the sensitivity that detects to a certain extent as toxotest.Use this method, guaranteed easy, economic, quick, the high throughput testing of chemical pollutant, environmental sample acute toxicity.
Description of drawings
Fig. 1. prelarva microwell plate method and adult fish classic method acute toxic test result's comparison
The solid post representative is measured zinc ion gained LC50 with the microwell plate high throughput method of prelarva among the figure, and the open column representative is measured zinc ion gained LC50 with the classic method of adult fish.
Embodiment
Embodiment:
The Japanese medaka ovum of gathering through after selecting meticulously, is placed the reorganization dilute solution of preparation, and the constant incubator of putting into 40 ℃ is cultivated, hatching.The prelarva that collection hatches continues to put into dilute solution and cultivated 6 hours, as biological subject.In each hole of 6 orifice plates, hold 5 milliliters of the solution of pollutant to be measured, put into 5 in every hole and tried prelarva, put into 25 ℃ of constant incubators afterwards and expose 12 hours, the prelarva number that record is dead calculates the half fatal rate of pollutant.
Claims (7)
1. the present invention relates to a kind of special high flux acute toxic test method, belong to environment measuring and assessment technique category to the fish early life stage.Reorganization dilute solution through the exploitation toxotest; Discussion is in the susceptibility of the fish of different life stages to toxic pollutants; And the biological subject of varying number, the different influence that exposes condition such as containers to test result, realized the high flux of prelarva acute toxic test.This method is easy, economical, quick, less demanding to the experimental implementation technical merit, easy to utilize.
2. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1 is characterized in that: tried fish and comprise multiple small-sized fingerlings such as zebra fish, Japanese blue or green Medaka, Gobiocypris rarus.
3. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1, its characteristic also is: tried fish and should be the prelarva between birth 0-240 hour.
4. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1; Its characteristic also is: the composition of cultivating the reorganization dilute solution of prelarva; Comprise inorganic component (like sodium chloride, lime chloride, soda mint, magnesium chloride, potassium chloride etc.), organic component (comprising amino acid, glycerine, vitamin, glucose etc.).
5. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1, its characteristic also is: the exposure container of toxotest can be 6 orifice plates, 12 orifice plates, 24 orifice plates, 48 orifice plates, 96 orifice plates etc.
6. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1, its characteristic also is: the Exposure Temperature of acute toxic test can be between 5 ℃-40 ℃.
7. the high flux acute toxic test method of the described a kind of fish early life stage of claim 1, its characteristic also is: the exposure duration of toxotest can be between 6-120 hour.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103116007A (en) * | 2013-01-25 | 2013-05-22 | 南京大学 | Method for screening and identifying wastewater and treating poisonous substances in water |
| CN103667327A (en) * | 2013-12-06 | 2014-03-26 | 中国科学院苏州生物医学工程技术研究所 | Preparation method of transgenetic zebrafish used for detecting environment pollutants |
| CN104062416A (en) * | 2014-07-16 | 2014-09-24 | 南京大学 | Toxicity identification evaluation method for papermaking wastewater |
| CN104374883A (en) * | 2014-11-25 | 2015-02-25 | 安徽省环境科学研究院 | Application of zebra fish in detection of reproduction toxicity of BBP (Butyl Benzyl Phthalate) and method for rapidly detecting reproduction toxicity of BBP |
| CN104833784A (en) * | 2015-05-19 | 2015-08-12 | 山东建筑大学 | Method for determining biotoxicity of water quality based on zebrafish movement velocity change |
| CN104849425A (en) * | 2015-05-19 | 2015-08-19 | 山东建筑大学 | Method for determining water quality biotoxicity |
| CN105301212A (en) * | 2014-07-04 | 2016-02-03 | 中国环境科学研究院 | High-throughput fish ecotoxicity test system |
| CN107907389A (en) * | 2017-09-28 | 2018-04-13 | 同济大学 | The detection method of the bio-toxicity of concrete dynamic modulus cement concrete leachate and preparation method thereof and the leachate |
| CN111896727A (en) * | 2020-07-03 | 2020-11-06 | 浙江省农业科学院 | Gobiocypris rarus embryotoxicity determination method and water body monitoring and early warning method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003083954A (en) * | 2001-09-11 | 2003-03-19 | Yokohama Tlo Co Ltd | Method for testing toxicity in underwater contaminant |
| CN101216471A (en) * | 2008-01-10 | 2008-07-09 | 中国药科大学 | Mode creature method for medicament toxicity research |
| CN101576554A (en) * | 2009-03-06 | 2009-11-11 | 刘征涛 | Dynamic running water type toxicity test device of fishes |
| CN101871927A (en) * | 2010-06-13 | 2010-10-27 | 南京大学 | Online monitoring equipment and method for acute toxicity of water quality |
-
2010
- 2010-12-17 CN CN2010106085113A patent/CN102539640A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003083954A (en) * | 2001-09-11 | 2003-03-19 | Yokohama Tlo Co Ltd | Method for testing toxicity in underwater contaminant |
| CN101216471A (en) * | 2008-01-10 | 2008-07-09 | 中国药科大学 | Mode creature method for medicament toxicity research |
| CN101576554A (en) * | 2009-03-06 | 2009-11-11 | 刘征涛 | Dynamic running water type toxicity test device of fishes |
| CN101871927A (en) * | 2010-06-13 | 2010-10-27 | 南京大学 | Online monitoring equipment and method for acute toxicity of water quality |
Non-Patent Citations (3)
| Title |
|---|
| 梁 峰: "六价铬对黄颡鱼仔鱼和稚鱼的急性毒性效应研究", 《农业环境科学学报》 * |
| 陈璘: "苏州河生态恢复过程中生态毒性解析", 《中国优秀硕士学位论文数据库》 * |
| 陈粉丽: "氯代苯类化合物对斑马鱼胚胎的单一急性毒性", 《环境与职业医学》 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103116007A (en) * | 2013-01-25 | 2013-05-22 | 南京大学 | Method for screening and identifying wastewater and treating poisonous substances in water |
| CN103667327A (en) * | 2013-12-06 | 2014-03-26 | 中国科学院苏州生物医学工程技术研究所 | Preparation method of transgenetic zebrafish used for detecting environment pollutants |
| CN103667327B (en) * | 2013-12-06 | 2015-10-21 | 中国科学院苏州生物医学工程技术研究所 | A kind of preparation method of the transgenic zebrafish for testing environment pollutent |
| CN105301212A (en) * | 2014-07-04 | 2016-02-03 | 中国环境科学研究院 | High-throughput fish ecotoxicity test system |
| CN105301212B (en) * | 2014-07-04 | 2017-09-12 | 中国环境科学研究院 | High flux fish eco-toxicity test system |
| CN104062416A (en) * | 2014-07-16 | 2014-09-24 | 南京大学 | Toxicity identification evaluation method for papermaking wastewater |
| CN104374883A (en) * | 2014-11-25 | 2015-02-25 | 安徽省环境科学研究院 | Application of zebra fish in detection of reproduction toxicity of BBP (Butyl Benzyl Phthalate) and method for rapidly detecting reproduction toxicity of BBP |
| CN104833784A (en) * | 2015-05-19 | 2015-08-12 | 山东建筑大学 | Method for determining biotoxicity of water quality based on zebrafish movement velocity change |
| CN104849425A (en) * | 2015-05-19 | 2015-08-19 | 山东建筑大学 | Method for determining water quality biotoxicity |
| CN107907389A (en) * | 2017-09-28 | 2018-04-13 | 同济大学 | The detection method of the bio-toxicity of concrete dynamic modulus cement concrete leachate and preparation method thereof and the leachate |
| CN107907389B (en) * | 2017-09-28 | 2020-06-26 | 同济大学 | Porous cement concrete leachate, preparation method thereof and detection method of biotoxicity of leachate |
| CN111896727A (en) * | 2020-07-03 | 2020-11-06 | 浙江省农业科学院 | Gobiocypris rarus embryotoxicity determination method and water body monitoring and early warning method |
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Application publication date: 20120704 |