Purpose I would like to test the effects of energy drinks on the heart rate of Daphnia Magna.
Oecd 202 211 Daphnia Magna
3
IBACON GmbH – Arheilger Weg 17 – D-64380 Rossdorf, Germany – all rights protected www.ibacon.com OECD 202: Daphnia sp., Acute Immobilisation Test OECD 211: Daphnia magna Reproduction Test Registration of Plant Protection Products (1107/2009/EC (91/414/EC)), Chemicals (REACH Regulation 1907/2006/EC), Biocides (528/2012/EC), Veterinary Medicinal Products (2001/82/EC) and Medicinal Products for Human Use (2001/83/EC) Indication Water fleas are important grazers feeding on phytoplancton. Therefore, they are part of risk assessments for several kinds of substances like plant protection products, industrial chemicals, biocides and medicinal products for human and veterinary use. Additionally, they are easy to rear and sensitive to water pollution. Test Organisms The water flea Daphnia magna belongs to the Crustacea, subclass Phyllopoda. In the tests, young Daphnia magna from synchronous laboratory culture are used. Fig. 1: Adult Daphnia magna Selection of Study Designs The acute toxicity is tested for most environmental risk assessment schemes in a first step. Depending on the results of the acute study or on further regulatory requirements, the performance of a reproduction study is demanded. Citing an example, plant protection products with a repeated application or a dissipation rate (DT 50 ) greater or equal than 2 days in water require chronic testing. The stability of the test item is one important factor for the selection of the study design. If the test item is not stable under test conditions, it is possible to use a semi-static test design, with renewal of test media in defined time intervals. In special cases e.g. for test items with a very low stability, a flow through test design with a media exchange rate of e.g. five-times per day can be applied (continuous flow of test media through the testing cells with Daphnia, see fig. 2). Fig. 2: OECD 202 / OECD 211: Experimental set up – flow through test design with Daphnia magna Study Details As test medium, reconstituted water (M4) is used. Exposure takes place in a controlled environment room with defined temperature and light regime. Dissolved oxygen, pH and water temperature are measured regularly. The test item concentrations are checked by analytical dose verification. Acute Immobilisation Test (OECD 202) The acute toxicity test determines immobility during an exposure period of 48 hours. Immobilisation and abnormal behaviour is recorded at 24 and 48 hours and compared with control values. The test item is mixed into reconstituted water. 20 Daphnia are divided in four replicates per control and test item concentration. Most usually, a dose- response study with a geometric series of five test item concentrations is examined. This allows the calculation of the EC 50 from immobility data. NOEC (No Observed Effect Concentration) and LOEC (Lowest Observed Effect Concentration) are also determined, if possible. A test with a toxic reference substance is performed twice per year to demonstrate the sensitivity of the test organisms and suitable test conditions.
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Transcript of Oecd 202 211 Daphnia Magna
8/13/2019 Oecd 202 211 Daphnia Magna
http://slidepdf.com/reader/full/oecd-202-211-daphnia-magna 1/2
IBACON GmbH – Arheilger Weg 17 – D-64380 Rossdorf, Germany – all rights protected www.ibacon.com
OECD 202: Daphnia sp., Acute Immobilisation Test
OECD 211: Daphnia magna Reproduction Test
Registration of Plant Protection Products (1107/2009/EC (91/414/EC)), Chemicals (REACH Regulation
1907/2006/EC), Biocides (528/2012/EC), Veterinary Medicinal Products (2001/82/EC) and Medicinal
Products for Human Use (2001/83/EC)
Indication
Water fleas are important grazers feeding on
phytoplancton. Therefore, they are part of risk
assessments for several kinds of substances like
plant protection products, industrial chemicals,
biocides and medicinal products for human and
veterinary use. Additionally, they are easy to rearand sensitive to water pollution.
Test Organisms
The water flea Daphnia magna belongs to the
Crustacea, subclass Phyllopoda. In the tests, young
Daphnia magna from synchronous laboratory
culture are used.
Fig. 1: Adult Daphnia magna
Selection of Study Designs
The acute toxicity is tested for most environmental
risk assessment schemes in a first step. Depending
on the results of the acute study or on further
regulatory requirements, the performance of a
reproduction study is demanded. Citing an example,
plant protection products with a repeated application
or a dissipation rate (DT50) greater or equal than 2
days in water require chronic testing.
The stability of the test item is one important factorfor the selection of the study design. If the test item
is not stable under test conditions, it is possible to
use a semi-static test design, with renewal of test
media in defined time intervals. In special cases e.g.
for test items with a very low stability, a flow
through test design with a media exchange rate of
e.g. five-times per day can be applied (continuous
flow of test media through the testing cells with
Daphnia, see fig. 2).
Fig. 2: OECD 202 / OECD 211: Experimental set up – flow
through test design with Daphnia magna
Study Details
As test medium, reconstituted water (M4) is used.
Exposure takes place in a controlled environment
room with defined temperature and light regime.
Dissolved oxygen, pH and water temperature are
measured regularly. The test item concentrations are
checked by analytical dose verification.
Acute Immobilisation Test (OECD 202)
The acute toxicity test determines immobility during
an exposure period of 48 hours. Immobilisation andabnormal behaviour is recorded at 24 and 48 hours
and compared with control values.
The test item is mixed into reconstituted water. 20
Daphnia are divided in four replicates per control
and test item concentration. Most usually, a dose-
response study with a geometric series of five test
item concentrations is examined. This allows the
calculation of the EC50 from immobility data. NOEC
(No Observed Effect Concentration) and LOEC
(Lowest Observed Effect Concentration) are also
determined, if possible.
A test with a toxic reference substance is performed
twice per year to demonstrate the sensitivity of the
test organisms and suitable test conditions.
8/13/2019 Oecd 202 211 Daphnia Magna
http://slidepdf.com/reader/full/oecd-202-211-daphnia-magna 2/2
IBACON GmbH – Arheilger Weg 17 – D-64380 Rossdorf, Germany – all rights protected www.ibacon.com
Fig. 3: OECD 202: Dose-Response Curve of Immobilisation
of Daphnia magna as observed after 48 hours
Available Guidelines
Council Regulation (EC) No 440/2008, C.2.
Daphnia sp., Acute Immobilisation Test
EPA Ecological Effects Test Guideline OPPTS
850.1010 Aquatic Invertebrate Acute Toxicity
Test, Freshwater Daphnids, April 1996.
OECD Guideline for Testing of Chemicals,
Section 2, No. 202: " Daphnia sp.,
Acute Immobilisation Test", Part I,
adopted. April 13, 2004.
Typical Time Frame (GLP Study)
Range-Finding Test and Protocol .............. 4 weeks
Experimental phase ................................... 4 weeksReporting ................................................... 4 weeks
Total ......................................................... 12 weeks
Reproduction Test (OECD 211)
This prolonged test focuses on sublethal effects like
reproduction ability. During an exposure period of
21 days, the number of offspring as well as
immobility is determined. The test item is mixed
into reconstituted water.
Fig. 4: OECD 211: Reproduction Test with Daphnia magna
In a semi-static test design, the test media are
exchanged regularly to ensure a continuous
exposure with the test item. Appropriate food is
supplied in order to enable survival and
reproduction of the Daphnia.
For special requests it is also possible to perform the
reproduction study in a flow through design asshown in fig. 1 with a continuous exchange of test
media.
Fig.5: OECD 211: Dose-Response Curve of Living Offspring
of Daphnia magna after 21 Day Exposure Period
The test is performed with 10 replicates (static and
semi static design) or 20 replicates (flow-through
design) for each test item concentration and for the
control. Each replicate contains one Daphnia. A
geometric concentration series based on the results
of an acute study or a range-finder is tested. Based
on the test results NOEC and LOEC values for
immobility and reproduction are determined.
The EC50 of the reproduction rate is also calculated.
Typical Time Frame (GLP Study)
Protocol Preparation .................................. 4 weeks
Experimental phase ................................... 8 weeks
Reporting ................................................... 5 weeks
Total ........................................................ 17 weeks
Available Guidelines
OECD Guideline for Testing of Chemicals,
No. 211: " Daphnia magna Reproduction Test", adopted October 02, 2012.
Commission Regulation 440/2008/ECC.20, 2008
EPA Ecological Effects Test Guideline
OPPTS 850.1300 Daphnid Chronic Toxicity
Test, April 1996.
IBACON contact:
Roland KuhlStudy Director
Phone: +49 6154 697 322
Email: [email protected]
Dr. Heiner AlbusManaging Director
Phone: +49 6154 697 347
Email: [email protected]
