CN105137022A - Zebrafish detoxification and sensitivity evaluation method for conducting on-line water quality monitoring - Google Patents
Zebrafish detoxification and sensitivity evaluation method for conducting on-line water quality monitoring Download PDFInfo
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- CN105137022A CN105137022A CN201510662505.9A CN201510662505A CN105137022A CN 105137022 A CN105137022 A CN 105137022A CN 201510662505 A CN201510662505 A CN 201510662505A CN 105137022 A CN105137022 A CN 105137022A
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Abstract
The invention discloses a zebrafish detoxification and sensitivity evaluation method for conducting on-line water quality monitoring. The method comprises the following specific steps: 1, screening purchased zebrafishes; 2, raising and training the zebrafishes; 3, conducting the sensitivity evaluation; 4, determining the raising and training time; 5, selecting and using the zebrafishes which are raised and trained according to the mentioned method for water quality monitoring. According to the method, the appropriate time for raising and training the purchased zebrafishes is determined through acute toxicity tests and used for water quality monitoring, the sensitivity of tested fishes obtained after the 'detoxification' link is improved, and the accuracy and stability of water quality monitoring results are guaranteed.
Description
Technical field
The present invention relates to a kind of source quality monitoring and warning the raising and train of commercially available zebra fish, detoxification and sensitivity evaluation method thereof, belong to water-carriage system water quality monitoring field, city.
Background technology
From the twenties in last century, biological monitoring technology just starts to apply in environmental quality detecting and assessing.Biological water quality on-line monitoring applying biological mainly comprises fish, water flea, double-deck shell and bacterium.Fish are the biological monitoring species being applied to water environment pollution the earliest, and in recent years along with the quickening of urban development, the pollution of water environment is also day by day serious, and the zebra fish based on International Certification is that sensing of biological monitoring water quality on line technology is at home and abroad widely used.And at present for the water quality biological comprehensive toxicity monitoring and warning ununified standard of fish, because of voluntarily breeding or scientific research institutions' purchase cost higher, zebra fish that monitoring water quality on line adopts is bought from market mostly, and commercially available zebra fish is in order to improve survival rate, shorten culturing time often feeding microbiotic, hormone, the growth-promoting class materials such as feed addictive, the use of these materials may affect the susceptibility of zebra fish to pollutant, affect the Stability and veracity of water quality monitoring result, for this reason the present invention with commercially available zebra fish for object, laboratory is carried out to it and raises and train pollutant Study of Sensitivity before and after detoxification, determine suitable to raise and train the time, the zebra fish that screening susceptibility is higher is using source quality monitoring and warning.
Summary of the invention
Technical matters to be solved by this invention is for the deficiencies in the prior art, provides a kind of water quality monitoring zebra fish detoxification and sensitivity evaluation method, realizes the raising that zebra fish carries out water monitoring data stability and accuracy.
For solving this technical matters, the invention provides a kind of monitoring water quality on line zebra fish detoxification and sensitivity evaluation method, specifically comprising the steps:
1) outsourcing zebra fish screening: be positioned in inspection box by outsourcing zebra fish and observe 24h, in the tap water of water temperature 25 DEG C ± 2 DEG C, continuous aeration, period is feeding not, and in 24h, the mortality ratio of fish is less than 10%, then this batch of fish can enter box for breeding; Otherwise give it up, again buy;
2) zebra fish is raised and train: outsourcing zebra fish being put into water temperature is 25 DEG C ± 2 DEG C, and in the code test water of continuous aeration, turn on light every day 12h, carry out feeding, remove ight soil and swill, carry out raising and train detoxification, record zebra fish raises and train the start time, raises and train the dead fish of period discovery and pulls out in time;
3) sensitivity assessment: using the MLC (median lethal concertration) LC50 value of acute toxicity test 24h, 48h, 72h, 96h as sensitivity assessment index, get and carry out the lower acute toxicity test of local water former water characteristic pollutant series concentration gradient exposure without the zebra fish raising and train and raise and train different time, by GB/T13267-91 water quality-material, the assay method of zebra fish acute toxicity is carried out; Observe the death condition of each experimental group fish of 24h, 48h, 72h, 96h equi-time point, calculate different semilethal rate LC50 value of raising and train zebra fish 24h, 48h, 72h, 96h under specific pollutants of time by statistic software SPSS;
Not raise and train zebra fish for control group, utilize SPSS software significance analysis to check and different raise and train the LC50 value of time fish and the otherness of control group, as p>0.05, think there was no significant difference; During 0.01<p≤0.05, think significant difference; During p≤0.01, otherness is extremely remarkable;
4) time of raising and train is determined: if raise and train LC50 value there was no significant difference compared with control group of time, then continue to raise and train, choose the next one and raise and train the LC50 value of time compared with control group, if still there was no significant difference, repeat said process, until the LC50 value of raising and train time n is compared with control group, significant difference or the time of raising and train reach 30 days and then stop this process; If the time of raising and train reaches 30 days LC50 values there was no significant difference compared with control group, stop raising and train;
If raise and train the LC50 value of time n compared with control group, significant difference, simultaneously compared with the LC50 value of raising and train time n+1, there was no significant difference, then stop raising and train;
If raise and train the LC50 value of time n compared with control group, significant difference, and compared with the LC50 value of raising and train time n+1, still significant difference, then choose and raise and train time n+1 and raise and train time n+2 and carry out difference analysis, until the LC50 value of raising and train time N is compared with control group, significant difference, simultaneously compared with the LC50 value of raising and train time N+1, there was no significant difference, then stop raising and train;
5) choose the zebra fish after raising and train as stated above to monitor for source quality.
Described step 2) zebra fish code test water used in raising and train, containing 0.22mg/LCaCl
2, 0.06mg/LMgSO
4, 0.06mg/LNaHCO
3, 0.006mg/LKCl, pH be 7.6 ± 0.2.
Described step 2) zebra fish raises and train middle zebra fish and raises and train period, and institute's feeding food can be shrimps, laboratory self-control food or commercially available not containing the bait of microbiotic and steroids growth promoting substance, not feeding in 24h and test before test.
Described step 3) in sensitivity evaluation method the selection of specific pollutants according to local water quality characteristic, should choose representative or have the pollutant of the risk of burst to carry out exposure test.
Described step 2) zebra fish raise and train middle zebra fish raise and train period, the mortality ratio of zebra fish fry higher than 10%, then need change a collection of new fish raise and train.
Beneficial effect: the present invention raises and train the time for water quality monitoring by the suitable of acute toxicity test determination outsourcing zebra fish, the test fish after " detoxification " link, susceptibility improves, and ensure that the Stability and veracity of water quality monitoring result.
Embodiment
In order to understand the present invention, further illustrate the present invention with embodiment below, but the present invention is not restricted to this.
On-line monitoring of the present invention zebra fish detoxification and sensitivity evaluation method, specifically comprise the steps:
(1) buy zebra fish a collection of, in inspection box, 24h mortality ratio is less than 10%, enters box for breeding, writing time;
(2) part is used for acute toxicity test research, remainingly puts into laboratory water temperature 25 DEG C ± 2 DEG C, raises and train in the code test water of continuous aeration, every day feeding, remove ight soil and swill, feeding shrimps and laboratory self-control food;
(3) Zn is chosen
2+, Cr
6+, malathion, phenol four kinds of pollutants carry out the acute toxicity test of outsourcing zebra fish, method is carried out fresh-water fishes (zebra fish) acute toxicity regulation according to GB/T13267-91 water quality-material, and pollutant levels setting is with reference to table 1;
Table 1 acute toxicity test concentration gradient sets
(3) the zebra fish refer step (2) adopting laboratory to raise and train 15d, 30d respectively carries out the acute toxicity test of four kinds of pollutants, and before test, 24h stops feeding;
(4) observe the death condition of each experimental group fish of 24h, 48h, 96h equi-time point in acute toxicity test, calculate different semilethal rate LC50 value of raising and train zebra fish 24h, 48h, 96h under specific pollutants of time by mathematical statistics software SPSS;
(5) according to the LC50 of 0d, 15d, 30d, choose suitable water quality monitoring and raise and train the time with zebra fish;
(6) whole 30d phase of raising and train this batch of zebra fish mortality ratio lower than 10%; Otherwise this batch of zebra fish is given it up;
(7) the results are shown in Table 2, under four kinds of pollutants expose, the zebra fish LC50 value of raising and train 15d, 30d is comparatively raised and train has reduction than all in 0 day, illustrate that zebra fish is improved the susceptibility of pollutant, raise and train 0d and 15d by SPSS software analysis, the LC50 value of 30d has significant difference, and the LC50 there was no significant difference of 15d, 30d, show that outsourcing zebra fish need can obtain result more accurately for water quality monitoring after laboratory is raised and train 30 days.
Table 2
The above-mentioned embodiment of the present invention, just illustrates, is not only, and the change within the scope of the present invention all or equivalent scope of the present invention is all surrounded by the present invention.
Claims (5)
1. monitoring water quality on line zebra fish detoxification and a sensitivity evaluation method, is characterized in that, specifically comprise the steps:
1) outsourcing zebra fish screening: be positioned in inspection box by outsourcing zebra fish and observe 24h, in the tap water of water temperature 25 DEG C ± 2 DEG C, continuous aeration, period is feeding not, and in 24h, the mortality ratio of fish is less than 10%, then this batch of fish can enter box for breeding; Otherwise give it up, again buy;
2) zebra fish is raised and train: outsourcing zebra fish being put into water temperature is 25 DEG C ± 2 DEG C, and in the code test water of continuous aeration, turn on light every day 12h, carry out feeding, remove ight soil and swill, carry out raising and train detoxification, record zebra fish raises and train the start time, raises and train the dead fish of period discovery and pulls out in time;
3) sensitivity assessment: using the MLC (median lethal concertration) LC50 value of acute toxicity test 24h, 48h, 72h, 96h as sensitivity assessment index, get and carry out the lower acute toxicity test of local water former water characteristic pollutant series concentration gradient exposure without the zebra fish raising and train and raise and train different time, by GB/T13267-91 water quality-material, the assay method of zebra fish acute toxicity is carried out; Observe the death condition of each experimental group fish of 24h, 48h, 72h, 96h equi-time point, calculate different semilethal rate LC50 value of raising and train zebra fish 24h, 48h, 72h, 96h under specific pollutants of time by statistic software SPSS;
Not raise and train zebra fish for control group, utilize SPSS software significance analysis to check and different raise and train the LC50 value of time fish and the otherness of control group, as p>0.05, think there was no significant difference; During 0.01<p≤0.05, think significant difference; During p≤0.01, otherness is extremely remarkable;
4) time of raising and train is determined: if raise and train LC50 value there was no significant difference compared with control group of time, then continue to raise and train, choose the next one and raise and train the LC50 value of time compared with control group, if still there was no significant difference, repeat said process, until the LC50 value of raising and train time n is compared with control group, significant difference or the time of raising and train reach 30 days and then stop this process; If the time of raising and train reaches 30 days LC50 values there was no significant difference compared with control group, stop raising and train;
If raise and train the LC50 value of time n compared with control group, significant difference, simultaneously compared with the LC50 value of raising and train time n+1, there was no significant difference, then stop raising and train;
If raise and train the LC50 value of time n compared with control group, significant difference, and compared with the LC50 value of raising and train time n+1, still significant difference, then choose and raise and train time n+1 and raise and train time n+2 and carry out difference analysis, until the LC50 value of raising and train time N is compared with control group, significant difference, simultaneously compared with the LC50 value of raising and train time N+1, there was no significant difference, then stop raising and train;
5) choose the zebra fish after raising and train as stated above to monitor for source quality.
2. monitoring water quality on line according to claim 1 zebra fish detoxification and sensitivity evaluation method, is characterized in that: described step 2) zebra fish code test water used in raising and train, containing 0.22mg/LCaCl
2, 0.06mg/LMgSO
4, 0.06mg/LNaHCO
3, 0.006mg/LKCl, pH be 7.6 ± 0.2.
3. monitoring water quality on line according to claim 1 zebra fish detoxification and sensitivity evaluation method, it is characterized in that: described step 2) zebra fish raise and train middle zebra fish raise and train period, institute's feeding food can be shrimps, laboratory self-control food or commercially available not containing the bait of microbiotic and steroids growth promoting substance, not feeding in 24h and test before test.
4. monitoring water quality on line according to claim 1 zebra fish detoxification and sensitivity evaluation method, it is characterized in that: described step 3) in sensitivity evaluation method the selection of specific pollutants according to local water quality characteristic, should choose representative or have the pollutant of the risk of burst to carry out exposure test.
5. the monitoring water quality on line according to any one of claim 1-4 zebra fish detoxification and sensitivity evaluation method, it is characterized in that: described step 2) zebra fish raise and train middle zebra fish raise and train period, the mortality ratio of zebra fish fry higher than 10%, then needs to change a collection of new fish and raises and train.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105891436A (en) * | 2016-06-30 | 2016-08-24 | 福建医科大学 | Biological testing method for comprehensively evaluating water quality of drinking water |
| CN108205053A (en) * | 2018-03-29 | 2018-06-26 | 江苏农牧科技职业学院 | The quick bio monitoring method of Primnesium saltans toxin in water body |
| CN111189999A (en) * | 2019-12-31 | 2020-05-22 | 中国水产科学研究院 | Method for evaluating influence of pollutants on biotoxicity of zebra fish |
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| CN102899383A (en) * | 2012-11-13 | 2013-01-30 | 中国科学院华南植物园 | Application of zebra fish to testing water quality and toxicity and method for applying zebra fish to test water quality and toxicity |
| JP2014178184A (en) * | 2013-03-14 | 2014-09-25 | Hiroshima Univ | Water quality inspection system and fish monitoring system |
| CN103499675A (en) * | 2013-09-30 | 2014-01-08 | 苏州国环环境检测有限公司 | Method for monitoring toxicity of domestic sewage through zebra fishes |
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| CN105891436A (en) * | 2016-06-30 | 2016-08-24 | 福建医科大学 | Biological testing method for comprehensively evaluating water quality of drinking water |
| CN108205053A (en) * | 2018-03-29 | 2018-06-26 | 江苏农牧科技职业学院 | The quick bio monitoring method of Primnesium saltans toxin in water body |
| CN111189999A (en) * | 2019-12-31 | 2020-05-22 | 中国水产科学研究院 | Method for evaluating influence of pollutants on biotoxicity of zebra fish |
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