CN102692478A - Method for detecting toxicity of PFOS (perfluorooctane sulfonate) in water through rotifers - Google Patents
Method for detecting toxicity of PFOS (perfluorooctane sulfonate) in water through rotifers Download PDFInfo
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- CN102692478A CN102692478A CN2012101736051A CN201210173605A CN102692478A CN 102692478 A CN102692478 A CN 102692478A CN 2012101736051 A CN2012101736051 A CN 2012101736051A CN 201210173605 A CN201210173605 A CN 201210173605A CN 102692478 A CN102692478 A CN 102692478A
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Abstract
The invention relates to a method for detecting biotoxicity of PFOS (perfluorooctane sulfonate) in water through rotifers, and belongs to the technical field of environmental detection. Particularly, the technical scheme comprises the steps that rotifers are selected as mode creatures, which are widely spread, quickly propagate, and have short generation time, and survival, propagation, activities and the like are sensitive to the variation of environment physical and chemical factors; the influence of the PFOS on the ecosystem can be evaluated through the change of the population density of rotifers caused by PFOS water solutions of different concentrations; and then the action mechanism of the PFOS can be confirmed through the influence of the PFOS on the life history of rotifers. The method overcomes the difficulties that the period is long and the operation is complicated during the biotoxicity detection for PFOS in water in the prior art, and provides the simple, convenient, sensitive and systemic method for detecting toxicity of PFOS in water; and the method is convenient, quick and low in cost.
Description
Technical field
The present invention relates to a kind of method of utilizing wheel animalcule to detect PFOS toxicity in the water, belong to the environment measuring technical field.
Background technology
Owing to have very unique physicochemical property such as hydrophobic oleophobic property; PFOS (PFOS) is widely used in industry, commerce and the individual consumer's goods; As can be used as hydrophobic oleophobic anti-staining agent (carpet, weaving, upholster, leather, paper products etc.), fire retardant (Aero-Space, fire-fighting), surfactant (fire foam, alkaline cleaner), photoresist (semi-conductor industry), electroplate anti-fog agent, photographic paper electrostatic resistance agent, coating additive etc., and can be used as Insecticides (tech) & Herbicides (tech), lubricant, bonding agent and cosmetics become to grade.At present, in waste water and mud, surface water, underground water, seawater, marine bottom sediment and potable water (tap water), all detected the PFOS of variable concentrations.PFOS remove have antiacid, alkali resistant, the ability of anti-oxidant and anti-reductive agent; Effects such as ability hydrolysis, photodissociation, biodegradation; Can also be through food chain accumulation, its level of accumulating in vivo far above organo-chlorine pesticide with persistence organic pollutants such as bioxin.Mainly concentrate on hydrobionts such as mammal such as mouse and zebra fish, Daphnia magna in recent years about the research of the eco-toxicity that detects PFOS; The PFOS concentration of treatment is far above the concentration that exists of occurring in nature PFOS in the existing method, and the ecology that is difficult to PFOS in the actual response physical environment is disturbed.And experimental period is long, is difficult for observing the influence to the offspring, and experimentation cost is higher.
Wheel animalcule is the one type of animal plankton that is distributed widely in all kinds of water bodys.They are food with algae, bacterium etc., are again simultaneously the good bait of fish seedling and some invertabrate etc.Wheel animalcule breeding is fast, and the assimilatory efficiency of algae food up to 20-80%, is played important pivotal role in the material cycle of aquatic ecosystem and energy Flow.And wheel animalcule and other cladocera and Copepods etc. compete food, influence the composition of community of aquatic organism.The distribution of wheel animalcule extensively, breeding is rapid, generation time is short; Its survival, breeding and behavior etc. are responsive to the variation of environment physical and chemical factor; The standardization etc. that with the wheel animalcule is the biological subject toxicological experiment process of carrying out makes one of its model organism that becomes Aquatic Ecotoxicology research, and is widely used in the monitoring of water environment.1999, in national testing standard (ASTM, 1999) was formally listed Brachionus calyciflorus and brachionus plicatilis as the indicator animal of fresh water and seawater pollution respectively by EPA.At present, what in the research of the Aquatic Ecotoxicology of wheel animalcule, relate generally to is heavy metal ion compound, estrogen, polychlorinated biphenyl, palycyclic aromatic, and pesticide pollutant, and PFOS not appearing in the newspapers to the toxicity of wheel animalcule.
The invention provides and a kind ofly can change the method for judging PFOS toxicity through the life table of wheel animalcule, material is easy to get, lack experimental period, be convenient to observe PFOS to the influence of offspring and whole population, the test consumption is few, cost is low.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing wheel animalcule to detect PFOS (PFOS) toxicity in the water, promptly detect the purpose of PFOS toxicity through the variation that detects wheel animalcule its asexual reproduction strategy, zoogamy strategy and individual size under the situation that PFOS exists; The method with seldom test fluid, in the short period of time, embodied the overall process (like growth, sexal maturity, population growth and reproduction) of each stage of development and the wheel animalcule history of life of wheel animalcule with simple experimentation; In addition, can also estimate the fecundity of first filial generation wheel animalcule.
The technical scheme that the present invention adopted is described below:
A kind of method of utilizing wheel animalcule to detect PFOS (PFOS) toxicity in the water comprises that step is following:
1) cultivation of wheel animalcule
With the wheel animalcule nutrient solution wheel animalcule being carried out " clone " cultivates; With green alga nutrient solution green algate of fresh water that cultivate, that be in the exponential increase phase is bait; Incubation time is more than six months; With the wheel animalcule nutrient solution wheel animalcule is carried out the above preparatory cultivations of two weeks before the experiment, cultivation is carried out under 20 ± 1 ℃, continuous light condition;
2) acute toxicity test
PFOS (PFOS) is made as 100.0,10.0,1.0,0.1 and 5 concentration gradients of 0.01mg/L; Each gradient is provided with 4 repetitions; Other establishes 1 blank, from pre-incubated test tube, draw at random the band amictic egg the female body of wheel animalcule several place double dish to cultivate, collect the wheel animalcule young that hatches behind the 2h; The wheel animalcule young is put into six orifice plates; Add test fluid, test is carried out in 20 ± 1 ℃, the constant incubator of unglazed photograph, adopts probit method to try to achieve its LC50 value respectively;
3) measure the influence of variable concentrations PFOS to wheel animalcule population growth rate
Through the influence of population growth rate experimental evaluation variable concentrations PFOS to wheel animalcule population growth rate; Behind the 96h; To the non-mictic female of wheel animalcule in each test tube, mictic female and not egg count respectively; Return former cultivation vessel then totally and continue to cultivate 24h, afterwards the entrained resting egg of mictic female and the resting egg of test tube bottom deposit are counted;
4) measure the influence of variable concentrations PFOS to wheel animalcule reproduction strategy
Be provided with variable concentrations PFOS, other adds 1 blank (wheel animalcule nutrient solution) and is used for the life table experiment, and the record wheel animalcule produces first piece of time that ovum is required; And it is wide to measure the long body of its body, detects PFOS to its influence of growing; Write down the laying of wheel animalcule, the young number that hatches and parent survival afterwards, and write down the ratio of non-mictic female of the second generation young and mictic female, carry out the mensuration of PFOS in the water (PFOS) toxicity thus.
In the method for the present invention, described wheel animalcule nutrient solution is the MBL nutrient solution.
In the method for the present invention, the PFOS concentration described in the step 4 is 0.01,0.1 and 1.0mg/L.
In the method for the present invention, said wheel animalcule is preferably Brachionus calyciflorus.
Embodiment
To combine embodiment to be elaborated below, more deep understanding arranged with purpose, feature and advantage to the inventive method.Following embodiment specific explanations the present invention, scope of the present invention does not receive the restriction of embodiment.
Embodiment 1
Present embodiment relates to the method for utilizing Brachionus calyciflorus to detect PFOS toxicity in the water, comprises the steps that wherein the PFOS concentration described in the step 4 is 0.01mg/L.
1. source of wheel animalcule and cultivation
At first gather and tried Brachionus calyciflorus; Test indoor wheel animalcule nutrient solution it is carried out " clone " cultivation; With green alga nutrient solution green algate of fresh water (C.pyrenoidosa) that cultivate, that be in the exponential increase phase is bait, and incubation time is more than six months, uses the wheel animalcule nutrient solution prescription (USEPA of improvement before the experiment; 1985) wheel animalcule nutrient solution carries out the above preparatory cultivations of two weeks to wheel animalcule, and the density of throwing something and feeding is 1.0 * 10 every day when cultivating in advance
6CellsmL
-1Green alga, cultivate under 20 ± 1 ℃, continuous light condition (3000lx).
2. acute toxicity test
PFOS (PFOS) is available from Sigma-Aldrich, and (PFOS) is made as 100.0,10.0,1.0,0.1 and 5 concentration gradients of 0.01mg/L, and each gradient is provided with 4 repetitions, and other establishes 1 blank (EPA nutrient culture media).The method of mother liquor dilution is adopted in the preparation of test fluid; Prepare the mother liquor of 1.0g/L earlier with distilled water; Use EPA nutrient culture media (not containing algae food) dilution to obtain the PFOS test fluid of variable concentrations then, from pre-incubated test tube, draw at random the band amictic egg the female body of wheel animalcule several place double dish to cultivate, collect the wheel animalcule young that hatches behind the 2h; Per 10 wheel animalcule young are put into six orifice plates that volume is 10mL; Add the 5mL test fluid, test is carried out in the constant incubator of (20 ± 1) ℃, unglazed photograph, adopts probit method to try to achieve its LC50 value respectively.
3. variable concentrations PFOS is to the influence of wheel animalcule population growth rate
Through the influence of population growth rate experimental evaluation variable concentrations PFOS to wheel animalcule population growth rate, this experiment is carried out in six orifice plates.In each hole, put into 15 of the long wheel animalcule young less than 2h in age, and add each concentration PFOS solution of preparing to 10ml, bait is 1.0 * 10
6The green alga of cells/L, PFOS concentration is set to 5 groups, is respectively 0.125,0.25,0.5,1.0,2.0mg/L and establish 1 control group (nutrient solution does not contain PFOS); All establish 4 repetitions for every group, experiment is carried out in the constant incubator of temperature for (20 ± 1) ℃.In the experimentation, every 12h suspends and is deposited on the algae food of test tube bottom, and every 24h changes and includes 1.0 * 10
6The test fluid of cells/L green alga; Behind the 96h; To the non-mictic female of wheel animalcule in each test tube, mictic female and not egg count respectively; Return former cultivation vessel then totally and continue to cultivate 24h; Afterwards the entrained resting egg of mictic female and the resting egg of test tube bottom deposit are counted, it is not remarkable that population growth rate test findings shows that the PFOS of 0.125mg/L influences difference to wheel animalcule population growth rate, and the PFOS of all the other four kinds of concentration (0.25,0.5,1.0,2.0mg/L) influences significant difference to wheel animalcule population growth rate.
4. variable concentrations PFOS is to the influence of wheel animalcule reproduction strategy
Selecting concentration for use is that the PFOS of 0.01mg/L is used for the life table experiment; The life table experiment is carried out in 24 special hole plastic plates; Put into the long young in an age in every hole, add the test fluid of 1mL, in the 48h of experiment beginning at 2h; Every separated 3h observes once, and the record wheel animalcule produces first piece of time that ovum is required; It is wide to measure the long body of its body during 32h, detects PFOS to its influence of growing; Every afterwards 8h observes once, laying, the young number that hatches and the parent survival of record wheel animalcule, and remove the young to 96 orifice plate, the ratio of record non-mictic female of the second generation young and mictic female; Every interval 24h transfers to the parent of survival in the fresh test fluid; Experimental result shows: the PFOS processing of 0.01mg/L has prolonged the wheel animalcule juvenile stage and has lasted; Reduced the mictic female percent in the population, and then influenced female-male proportion in the population of future generation, egg laying amount has not been had obvious influence.
Embodiment 2
Be the PFOS solution that the PFOS solution of 0.1mg/L replaces 0.01mg/L in the step 4 among the embodiment 1 with concentration, other conditions are with embodiment 1.Experiment shows, the PFOS of 0.1mg/L handles and to have prolonged the wheel animalcule juvenile stage and lasted, shortened the mean lifetime of wheel animalcule, reduced the mictic female percent in the population, and then influences female-male proportion in the population of future generation, and egg laying amount is not had obvious influence.
Embodiment 3
Be the PFOS solution that the PFOS solution of 1.0mg/L replaces 0.01mg/L in the step 4 among the embodiment 1 with concentration, other conditions are with embodiment 1.Experiment shows; 1.0mg/L PFOS handle the growth suppressed wheel animalcule, prolonged the wheel animalcule juvenile stage and lasted, shortened the mean lifetime of wheel animalcule, reduced the mictic female percent in the population; And then influence female-male proportion in the population of future generation, and suppressed the egg laying amount of wheel animalcule greatly.
Embodiment 1,2, and 3 experimental results are as shown in table 1:
Table 1. be exposed to the Brachionus calyciflorus of (mg/L) under the variable concentrations PFOS larval stage time (T), mean lifetime (ML),
The ratio (MF/AF) of average egg laying amount (EQ) and mictic female number/non-mictic female number.
。
Claims (4)
1. method of utilizing wheel animalcule to detect PFOS (PFOS) toxicity in the water comprises that step is following:
1) cultivation of wheel animalcule
With the wheel animalcule nutrient solution wheel animalcule being carried out " clone " cultivates; With green alga nutrient solution green algate of fresh water that cultivate, that be in the exponential increase phase is bait; Incubation time is more than six months; With the wheel animalcule nutrient solution wheel animalcule is carried out the above preparatory cultivations of two weeks before the experiment, cultivation is carried out under 20 ± 1 ℃, continuous light condition;
2) acute toxicity test
PFOS (PFOS) is made as 100.0,10.0,1.0,0.1 and 5 concentration gradients of 0.01mg/L; Each gradient is provided with 4 repetitions; Other establishes 1 blank, from pre-incubated test tube, draw at random the band amictic egg the female body of wheel animalcule several place double dish to cultivate, collect the wheel animalcule young that hatches behind the 2h; The wheel animalcule young is put into six orifice plates; Add test fluid, test is carried out in 20 ± 1 ℃, the constant incubator of unglazed photograph, adopts probit method to try to achieve its LC50 value respectively;
3) measure the influence of variable concentrations PFOS to wheel animalcule population growth rate
Through the influence of population growth rate experimental evaluation variable concentrations PFOS to wheel animalcule population growth rate; Behind the 96h; To the non-mictic female of wheel animalcule in each test tube, mictic female and not egg count respectively; Return former cultivation vessel then totally and continue to cultivate 24h, afterwards the entrained resting egg of mictic female and the resting egg of test tube bottom deposit are counted;
4) measure the influence of variable concentrations PFOS to wheel animalcule reproduction strategy
Be provided with variable concentrations PFOS, other adds 1 blank (wheel animalcule nutrient solution) and is used for the life table experiment, and the record wheel animalcule produces first piece of time that ovum is required; And it is wide to measure the long body of its body, detects PFOS to its influence of growing; Write down the laying of wheel animalcule, the young number that hatches and parent survival afterwards, and write down the ratio of non-mictic female of the second generation young and mictic female, carry out the mensuration of PFOS in the water (PFOS) toxicity thus.
2. method according to claim 1 is characterized in that: described wheel animalcule nutrient solution is the MBL nutrient solution.
3. method according to claim 1 is characterized in that: the PFOS concentration described in the step 4 is 0.01,0.1 and 1.0mg/L.
4. method according to claim 1 is characterized in that: said wheel animalcule is a Brachionus calyciflorus.
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| CN114946720A (en) * | 2022-05-25 | 2022-08-30 | 项尚飞 | Method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus |
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| CN107238687A (en) * | 2017-05-27 | 2017-10-10 | 北京化工大学 | A kind of method of PFOS toxicity in utilization goat's horn crescent moon algae detection water |
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| CN101162224A (en) * | 2007-09-17 | 2008-04-16 | 淮海工学院 | Method for environmental monitoring by fold arm-end rotifer pure line |
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Patent Citations (2)
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| CN1148890A (en) * | 1994-03-19 | 1997-04-30 | 瑞士联邦装备服务集团斯彼茨Ac实验室 | Method and device for determining toxicity and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114946720A (en) * | 2022-05-25 | 2022-08-30 | 项尚飞 | Method for determining acute and chronic toxicity of cephalexin to brachionus calyciflorus |
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