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TESTING STRATEGIES Earthworm acute toxicity test (EU Annex V C.8; OECD 207, 1984d; ISO 11268-1, December 1993) The test is designed to assess the effect of chemicals on the survival of the earthworms Eisenia spp. Adult worms are exposed to a range of concentrations of the test substance mixed into the soil. Mortality and effects on biomass are determined after 2 weeks exposure. Where possible, LC50 and EC50 values are determined. Eisenia spp. is considered to be representative of soil fauna and earthworm species. The organism was however selected more for pragmatic reasons (easily cultured in laboratory conditions) than for its sensitivity or it being representative of soil dwelling organisms. Insect larvae acute toxicity test (NF X 31-260 accepted as a new work item ISO/TC/90/SC4/WG2) Oxythyrea funesta is widely distributed in Europe. With a phytophage feeding habit, this organism plays an important role in determining the physical characteristics of soils (structure, texture, aeration,…). Survival of insect larvae (Oxythyrea funesta) exposed to contaminated soils is assessed in a test lasting 10 days. A LC50 is then determined by comparing survival in treated soils with that of the control. Earthworm reproduction test (ISO 11268-2, July 1998, draft OECD, 2000i) The effect of chemicals on the reproduction of adult compost worms (Eisenia fetida or E. andrei) is assessed over a period of 8 weeks. Adult worms are exposed to a range of concentrations of the test substance mixed into the compost. Mortality and growth effects are determined after 4 weeks exposure. The adults are then removed and the number of offspring determined following a further 4 weeks exposure period. The NOEC is determined by comparing the reproductive output of the worms exposed to the test substance to that of the control. Inhibition of reproduction of Collembola (Folsomia candida) (ISO 11267, 1999) Collembola is an important group of arthropods in temperate soils. Several species have been used in toxicity experiments including Folsomia candida for which a standard reproduction test has been developed. A treated artificial soil is used as the exposure medium and a NOEC is determined. Enchytraeidae reproduction test (draft OECD 220, 2000h), ISO/CD 16387, 2001) Enchytraeids are soil dwelling organisms that colonise a wide range of soils. They are easy to handle and breed in laboratory conditions and their generation time is shorter than that of the earthworms. The effect of chemicals on the reproduction of adult enchytraeid worms is assessed over a period of 6 weeks. The principle of the test is the same as for the earthworm reproduction test: adult worms are exposed to a range of concentrations of the test substance mixed into the soil. Mortality and morphological changes are determined after 3 weeks exposure. The adults are then removed and the number of offspring, hatched from the cocoons in the soil is counted after an additional 3 weeks exposure. The NOEC is determined by comparing the reproductive output of the worms exposed to the test substance to that of the control. Effects of pollutants on juvenile land snails (Helix aspersa) (NF X 31255/1, Draft April 2001) Inhibition of growth of the snails is observed through food contamination (1) or soil contamination (2). The French standard protocol will be proposed within ISO as a new work item. 192
TESTING STRATEGIES A soil bioassay for the nematode species Caenorhabditis elegans has been developed with mortality or sublethal endpoints (Løkke and Van Gestel, 1993). Plant assays Inhibition of root growth of higher plants (ISO 11269-1, November 1993) Pre-germinated seeds are planted in control or contaminated soils in laboratory conditions. Growth rates of the roots are determined after an appropriate incubation period (depending on the species: 5 days for Hordeum). Results obtained in contaminated soils are compared to that of the control to determine IC50 or NOEC parameters. Inhibition of emergence and growth of higher plants (ISO 11269-2, December 1995, draft OECD 208 A and B, 2000i) Inhibition of seedling emergence and early growth of higher plants is determined by comparing seedling emergence, biomass and visual detrimental effects on seeds placed on treated soils with seeds placed on control soil. Exposure through soil that has been previously spiked is the general rule. However, foliar application might be more relevant in some cases, depending on the main uses of the substance. In these cases, the effects on plants following deposition of test substance on the leaves and above ground portions of plants could be assessed using other standardised protocols. For example, inhibition of vegetative vigour of higher plants can be determined by comparing biomass and visual detrimental effects in controls plants with those in plants that had been sprayed with the test substance (OECD 208B). 193
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Institute for Health and Consumer P
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LEGAL NOTICE Neither the European C
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OVERVIEW This Technical Guidance Do
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CONTENTS 1 GENERAL INTRODUCTION ...
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4.2.3.4 Use of biodegradation scree
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1 GENERAL INTRODUCTION 1.1 BACKGROU
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GENERAL INTRODUCTION countries in t
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GENERAL INTRODUCTION PNEC is regard
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2 ENVIRONMENTAL EXPOSURE ASSESSMENT
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ENVIRONMENTAL EXPOSURE ASSESSMENT r
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2.2 MEASURED DATA ENVIRONMENTAL EXP
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ENVIRONMENTAL EXPOSURE ASSESSMENT w
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ENVIRONMENTAL EXPOSURE ASSESSMENT o
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ENVIRONMENTAL EXPOSURE ASSESSMENT T
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ENVIRONMENTAL EXPOSURE ASSESSMENT r
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Production ENVIRONMENTAL EXPOSURE A
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Back to processing Product at end o
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ENVIRONMENTAL EXPOSURE ASSESSMENT s
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Explanation of symbols ENVIRONMENTA
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ENVIRONMENTAL EXPOSURE ASSESSMENT s
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ENVIRONMENTAL EXPOSURE ASSESSMENT U
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ENVIRONMENTAL EXPOSURE ASSESSMENT a
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ENVIRONMENTAL EXPOSURE ASSESSMENT F
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2.3.5.1 Adsorption to aerosol parti
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ENVIRONMENTAL EXPOSURE ASSESSMENT I
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2.3.6.1 Hydrolysis ENVIRONMENTAL EX
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2.3.6.3 Photochemical reactions in
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constant of zero. However, if it ca
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ENVIRONMENTAL EXPOSURE ASSESSMENT A
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ENVIRONMENTAL EXPOSURE ASSESSMENT a
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Primary Settler Aeration Tank 2 5 3
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ENVIRONMENTAL EXPOSURE ASSESSMENT I
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ENVIRONMENTAL EXPOSURE ASSESSMENT d
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ENVIRONMENTAL EXPOSURE ASSESSMENT e
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Table 10 Overview of different expo
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ENVIRONMENTAL EXPOSURE ASSESSMENT -
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2.3.8.3 Calculation of PEClocal for
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ENVIRONMENTAL EXPOSURE ASSESSMENT W
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Guidance for calculating PEClocal i
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Concentration (% of initial) 160 14
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Calculation of PEClocalsoil For soi
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ENVIRONMENTAL EXPOSURE ASSESSMENT
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Output ENVIRONMENTAL EXPOSURE ASSES
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3 EFFECTS ASSESSMENT 3.1 INTRODUCTI
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3.2.1.1 Completeness of data New su
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EFFECTS ASSESSMENT • it is clearl
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EFFECTS ASSESSMENT 3.3 EFFECTS ASSE
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Table 16 Assessment factors to deri
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Input data EFFECTS ASSESSMENT The m
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Estimation of the PNEC The PNEC is
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EFFECTS ASSESSMENT biodegradability
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Table 17 Test systems for derivatio
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EFFECTS ASSESSMENT representative o
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In the partitioning method, it is a
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EFFECTS ASSESSMENT As mentioned in
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3.6.2.1 Calculation of PNEC using t
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3.7 EFFECTS ASSESSMENT FOR THE AIR
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EFFECTS ASSESSMENT procedure availa
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EFFECTS ASSESSMENT bioaccumulation
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EFFECTS ASSESSMENT For some chemica
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EFFECTS ASSESSMENT i.e. without enh
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Conversion factors for laboratory a
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EFFECTS ASSESSMENT • relative sen
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EFFECTS ASSESSMENT Earthworms are a
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MARINE RISK ASSESSMENT environment
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MARINE RISK ASSESSMENT of equal rel
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4.2.3.3 Marine biodegradation simul
Page 149 and 150: MARINE RISK ASSESSMENT In addition
Page 151 and 152: Explanation of symbols MARINE RISK
Page 153 and 154: MARINE RISK ASSESSMENT evaluate the
Page 155 and 156: MARINE RISK ASSESSMENT marine water
Page 157 and 158: MARINE RISK ASSESSMENT The addition
Page 159 and 160: MARINE RISK ASSESSMENT Statistical
Page 161 and 162: MARINE RISK ASSESSMENT sediment, fo
Page 163 and 164: Table 27 Assessment factors for der
Page 165 and 166: Table 28 continued Acute and chroni
Page 167 and 168: MARINE RISK ASSESSMENT of different
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Page 175 and 176: MARINE RISK ASSESSMENT The use of t
Page 177 and 178: 4.4.4.2 Assessment of measured BCF
Page 179 and 180: MARINE RISK ASSESSMENT required to
Page 181 and 182: Table 33 Overview of PEC/PNEC ratio
Page 183 and 184: RISK CHARACTERISATION If a refineme
Page 185 and 186: Classified dangerous to the Environ
Page 187 and 188: RISK CHARACTERISATION eaters. This
Page 189 and 190: 6 TESTING STRATEGIES 6.1 INTRODUCTI
Page 191 and 192: TESTING STRATEGIES Performance of a
Page 193 and 194: TESTING STRATEGIES Care must be tak
Page 195 and 196: Algal testing Algae toxicity test (
Page 197 and 198: TESTING STRATEGIES • long-term te
Page 199: Microbial assays TESTING STRATEGIES
Page 203 and 204: REFERENCES Brandes LJ, den Hollande
Page 205 and 206: REFERENCES IPCS (2000). Framework f
Page 207 and 208: REFERENCES OECD (1984f). Activated
Page 209 and 210: REFERENCES Pack S. (1993). A review
Page 211: REFERENCES US EPA (1996). Whole Sed
Page 214 and 215: APPENDIX I volume imported in the E
Page 216 and 217: APPENDIX I MC IV “Wide dispersive
Page 218 and 219: APPENDIX I 210 Stage Main category
Page 220 and 221: APPENDIX I the number of days over
Page 222 and 223: APPENDIX I Stage Main category Ia I
Page 224 and 225: APPENDIX I manufacturing of motor c
Page 226 and 227: APPENDIX I 1. extension of the tabl
Page 228 and 229: APPENDIX I 220 A-tables Estimates f
Page 230 and 231: APPENDIX I PRIVATE USE Not applicab
Page 232 and 233: APPENDIX I IC = 3: CHEMICAL INDUSTR
Page 234 and 235: APPENDIX I IC = 5: PERSONAL /DOMEST
Page 236 and 237: APPENDIX I Table A4.1 continued Com
Page 238 and 239: APPENDIX I IC = 7: LEATHER PROCESSI
Page 240 and 241: APPENDIX I IC = 9: MINERAL OIL AND
Page 242 and 243: APPENDIX I WASTE TREATMENT Table A5
Page 244 and 245: APPENDIX I For the emission factors
Page 246 and 247: APPENDIX I INDUSTRIAL USE Table A3.
Page 248 and 249: APPENDIX I INDUSTRIAL USE Table A3.
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APPENDIX I PRIVATE USE Table A4.5 C
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APPENDIX I PRIVATE USE Table A3.16
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APPENDIX I 246 B-tables Estimates f
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APPENDIX I FORMULATION Table B2.2 f
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APPENDIX I IC = 3: CHEMICAL INDUSTR
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APPENDIX I IC = 5: PERSONAL/DOMESTI
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APPENDIX I IC = 7: LEATHER PROCESSI
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APPENDIX I IC = 9: MINERAL OIL AND
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APPENDIX I IC = 11: POLYMERS INDUST
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APPENDIX I IC =13: TEXTILE PROCESSI
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APPENDIX I IC = 16: ENGINEERING IND
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APPENDIX I Appendix I-a: List of sy
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APPENDIX I No. Use Category No. Fun
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APPENDIX I Appendix I-b: List of sy
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APPENDIX I 290 Organic intermediate
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APPENDIX I Other IC/UC combinations
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APPENDIX II Appendix II Fate of che
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APPENDIX II b) Inherent biodegradab
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APPENDIX II log H % removal -4 -3 -
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APPENDIX III Appendix III Evaluatio
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APPENDIX III Field studies In gener
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APPENDIX IV together the mentioned
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290 Table 1 Selected soil test meth
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292 Table 1 continued Selected soil
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298 Table 1 Selected freshwater sed
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APPENDIX VII Appendix VII Toxicity
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APPENDIX VIII collection), and that
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APPENDIX VIII chemicals (see Sectio
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APPENDIX VIII Multimedia fate model
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APPENDIX VIII Monitoring data Metal
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APPENDIX VIII homeostatic regulatio
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APPENDIX IX based on the proportion
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APPENDIX IX Definition of “blocks
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APPENDIX IX these “blocks” will
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APPENDIX IX and PNECs for the “bl
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APPENDIX XI Appendix XI Environment
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APPENDIX XII Appendix XII Connectio
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APPENDIX XII +40% compared to 1992.
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APPENDIX XIII other substance is be
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APPENDIX XIV Taxonomic group No. of
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