üf IMI european SOCial fund in the m m ministry of education OP Education Czech republic european union youth and sports lor competlHvre» ^na INVESTMENTS IN EDUCATION DEVELOPMENT Dr. Jakub Hofman hofman@recetox.muni.cz EURDFEA.N UNION EUROPEAN REGIONAL DEVELOPMENT f uMĎ INWÍflHe FLUllK OP Research and Development tar Innovation Soil Taxonomy, second edition. • Soil is a natural body comprised of solids (minerals and organic matter), liquid, and gases that occurs on the land surface, occupies space, and is characterized by one or both of the following: horizons, or layers, that are distinguishable from the initial material as a result of additions, losses, transfers, and transformations of energy and matter or the ability to support rooted plants in a natural environment. imporiani pan ot naiure non-renewable source non-replaceable functions of ter. ecosystems base for plant growth nutrient storage - fertility - production start and end of food chains biogeochemical cycles decomposition of organic matter, humification filtration, immobilization and degradation of pollutants water cycling biodiversity treasure WE MUST PROTECT SOIL QUALITY M B fi ■***HA«* **Z*r ""Muco* CO soil = biotic + abiotic = complex bacteria protozoa fungi algae nematoda rotifera annelida arthropodes collembola mollusca Plant root hair Bacterial colonies Mycorrhizal hyphae Actinomycete hyphae and spores Decomposing plant cells biota is important for formation of soil, for soil structure soil fertility organic residues decomposition, release nutrients element cycles air and water regime Clay-organic matter complex Fungal hyphae and spores I 5 -B Sterile soil Soil with bacteria Soil biota Im2 Predators Spiders Harvest men Rovo beetles Mole crickets Grubs Ground bodies Centipedes OOI mm2 Bacteria T mm? Ftotozoa Rotororra • lardigrarte Icm? Mtcrolung, Predacious mites ■ KMnnahxIes htematooesp ]Mites. Collemtmles.Memattitlcs Fungi Pseudosoorpions C«sniipeUe& Spiders Fnadactous miles Mt Ants Earth worms 1dm2 C lit worms Wir t> worms: Tipu lids nehytraeiös Fly larvae Collembotes Pauiopoda Thysanura Sytnpliyla Diplurj Fungi Snails Vvoodltoe Millipedes Insect larvae Roots Litt» 1 ĽITJ FIGURE 4, Schematic survey of the soil fauna community. Squares indicate habitat size and relevant sampling area. Species are arranged according to feeding type. Important species are drawn at scale. (From Eijsack-ers. H. and Van de Bund. C.F., in Interactions Between Herbicides and the Soil, Hance, R.J.. Ed.. Academic Press. London. 1980. 255. With permission.) t|MJj Foiesisdl NirrnalwJa Acaiina ■ • la Enchy» SyinfŕiÉl Oipiopoda LjfiJricxSitrt Ccaeoplem Aianea . Gastropoda PaurOTJOda Other Arthropods FIGURE 3. Mean composition ol tlie soil fauna in a forest, meadow, and arable soil (mean numbers per dm1 for the 0- to 30-cm layer; + means present). (From Eijsackers, H. and Van de Bund, C.F., in Interactions Between Herbicides and the Soil, Hance, R.J., Ed., Academic Press, _London. 1B0, 255. With permission.)_ ACTIVITY BIOMASS 2500 1000 PULMONATA [0.1% NEMAIODA (06 51- ^ANNELIDA 17.7%)/* ZMANNELIUAir.'T.l? PULMOMATA (0.05%) NÉMATOOA ANNELIOA (2.2 T.I ARTHROPOOA (5.5%) ARfHROPpQA 16.2%) Science 11 June 2004: Vol. 304. no. 5677, pp. 1616 - 1618 Soil and Trouble WHEN PEOPLE INTENSIVELY TIU FIELDS and dear cut fdrestv they can damage or de slroy topsail thai took centuries to accumu ■ Late Just how vulnerable toils art depends on underlying conditions. Mismanaged soils in windswept lands can easily turn into desert, for example, and saline soils can become salt-encrusted wastelands This map shows th» main bamers to pro-ductn* farming, along with erosion risk, derived from climatic and soil conditions. Over laid as cross hatching are regions reported to be Wghty or very highly degraded according to a global survey of uA experts published in 1990The hot spots Illustrate e«amptes of the worst soil degradation, from the most com-.T.-ws phyt ,-al 171» /-srpr miwn tnfhcni cat forms, such as that caused by pollution from industrial chemicah and war. An interactive vwsion of this map appears online at www.sciencemag.org/cgi/conteni/ summaiy/304/5677/1614. V»«m IM .1>.IX.»I |.tMMH MM MllfffVMt, PHY5ICAL Of GRADATION CHEMICAL DEGRADATION High and very tugh levels of soil degradation per Global Assessment of Sod ■■■'I Degradston IGLASODI Hisjfrly erodeMe by w*id or water Tew constraints Climate Constraints t i»ti '.ttt fit fid u v H lM«d rr4a Physical Constraints I 1 i*(h«kn(*ji««ilurfin:n. I I Miw r»l ««iwlir* lv I ! UmiiAKiiiiliiaeair _J wnjwStiJ fajaffa Chemical Constraints I_J lawmjifir*i«lfr J ispi ptwarwus ntrt^řm. mti ntprtr wmw J maJt«niMt#>rlifi rytily | lljrt piaUn I I rt«h akjmram I kii .Iv ...I r. J-J Mum.*, W—W u»llu» «m »t^u«^a nirKn tut* Soils h problems • EU Thematic Strategy for Soil Protection (COM/2006/231) defines main threats for soils (sealing, erosion, compaction, salinisation, OM loss, contamination ...) • 3,5 mil. contaminated sites in EU • 0.5 mil. are seriously contaminated and need remediation • Costs related to contaminated sites in EU: 2-17 bil. € (Impact assessment (SEC/2006/620)) Soil protection • we must protect soil quality - policy ^ - legislation - science and education European Commission (2006a): Communication from the commission to the council, the european parliament, the european economic and social committee and the committee of the regions. Thematic strategy for soil protection. COM(2006)231. European Commission (2006b): Proposal for a directive of the European Parliament and of the Council establishing a framework for the protection of soil and amending Directive 2004/35/EC COM (2006) 232. Contamination of soils Soil ecotoxicology FOCUS: THE ENVIRONMENT (SOIL), WHERE RELATIONSHIP BETWEEN ORGANISM AND THE CHEMICAL EXISTS, MUST BE STUDIED TOO !! ^ctrns/,. rf**"**), mm m icology in soil protection FOCUS: Investigate relationships between soil organisms and contaminants ROLE: Scientific basis of soil protection Activities: Provide tools - bioassays for routine praxis: - Chemical and pesticides testing - Testing wastes, sludge, contaminated sites - Soil quality assessment Research of: - Fate and bioavailability - Mixture toxicity - Biodiversity ... M B fi • Very different from aquatic ecotoxicology • Solid matrices are heterogenous • Soil contains all three phases SOLID, LIQUID (pore water) and GAS (soil air) ' • Solid phase especially influences strongly FATE and BEHAVIOUR of chemicals ™ • Depending on soil and chemical properties and depending on TIME, chemical is DISTRIBUTED in soil, chemical SPECIATION occurs • SORPTION is the crucial process and leads to changes of BIOAVAILABILITY - the key factor of soil ecotoxicology • All this changes final TOXICITY and RISKS • All this hampers EXTRAPOLATION possibilities Exposure in soi CHEMICAI entry <4 latrix CHEMICAL in soil spatially distributed; chemical speciation EXPOSURE depends on: Fate of chemical in soil + Organism properties (morphology, physiology, ecology...) ORGANISM Chemical in organisi bolism, eliminate effects 4Mb 10 um 100 nm 10 nm 0.1 nm Clay particle or humic acid 2-0.2 um Organic pollutant Anorganic pollutant 0.1 nm m w m 5- 100^ 90* 80* 75 70 < 60* o E 50- 40* > CO 30 <- mul 20- 10* o 0* [i I V* V >P rt<* \P 4^ ^ ^ 5*^ Soil Fig. 1. Cumulative mortality (mean of three replicates, ± 95*K> CI) of Kiacaia aodrei exposed to 2,jD0O Dig Pb/kg spiked soils for 28 days. From Bradhaag et aLJ2003) M B fi 98 Factors affecting bioavailability Soil properties - Soil composition, organic matter, texture, pH, CEC, moisture, temperature, structure - pores Chemical properties - Chemical structure, Kow, Sw, Koc, pKa, MW, H, pv Organisms properties - physiology (uptake, metabolism, elimination), morphology, ecology Time effects - Aging, sequestration Other chemicals (napr. NAPL) and interactions Degradable/removable fraction Recalcitrant fraction Non-extractable fraction Time 0 Horizon An organic horizon composed primarily 01 recognizable organic malarial m various siages oi decompose - A Horizon The surface horizon. Composed ol various proportions Ol mineral materials aro organic components decomposed beyond recognition ■ E Horizon Zone Ol eluviaiion Mineral horizon resulting Irom intense leaching and chaiac-tenzed by a gray or grayish brown color.- B Horizon Zone ol illuviaiion Hoi ion enriched with minerals. 9 Q day. organic male als. or carbonates leached Irom the ' or E honions.- C Horizon Horizon chracterized by unweaibered minerals thai are the parent material irom which tne soil was lormed - R Horizon Bedrock.- 4Mb B fi ((©)) Why to bother with bioavailability ? • For correct risk assessment: - Soil animals (individuals, communities) - Organisms eating soil (e.g. children) - Plants • Prediction of biodegradation and remediation efficiencies • Legislative framework - Not the total concentrations for limits! • Extrapolation possibilities: - Between different soils - From aquatic to soil tests - From lab experiments to field situation How to measure bioavailability ? ■ Indirect measures Direct (surrogate) measures Chemical *****ohemjoaj measures Solvent extractions 0-? Sol id-phase extractions A . K Correlation Biomimetic Sampling Calibration Devices * K ^ membrane BiOlOgiCSl Toxicity testsT Bioaccumulation ITIGa SU re S Bi o m a rkers C riti cal B ody R esi du es (CBR) LKig. 3. Methods far manuring bmuvajJabilfty. jjyEBSf, Jrf»w""«<«1 rf**"**), ami) i® ® **3>^ Approaches of soil ecotoxicology Bio-indication - (bio)monitoring te-f: *i Ecotoxicity bioassays Response Dose Goal Define and describe relationship between biota conditions and contamination Goal Define safe concentration, describe risks Bioindication in soil ecotoxicology Microbes Doelman, P. and Eijsackers, H.J.P. (2004): Vital Soil - Function, Value and Properties. Elsevier. 358 p. ISBN: 0-444-51772-3. JMI Sttll tCOSVStETTl parameter Microbial indicator Function C-cyding ■ Soil respiration Metabolic quotient tC|OQ>) D«oni position of organic matter Sypl enzyme activity N-cy cling N-minera ligation Nitrification DeJiitrihcatiort N-fixation General activities Itaclenal DMA syntlwsis RNf A measurements Bacterial protein synthesis Community growth physiology Root-activity Mycorrhi/a Biodiversity General biomass Microbial bfomass: direct methods Microbinl biomflss: Indirect methods Microbial quotient Fun^i Fungi-bacteria ratio Protozoa Biodiversity Structural diversity functional diversity Marker lipid? Suppressive™™* to pathogens Bioavailability of Biosensor bacteria contaminants llasmid-cniilaining bacteria Biomarker spedes Incidence and expression of catabolic genes Bioindication in soil ecotoxicology Invertebrates IMI JQQelman, P. and Eijsackers, H.J.P. f(M§ft$)'- Vital Soil" Function, Value and ?A^fcofckrties. Elsevier. 358 p. ISBN: 0-'^«444-51772-3. Indicator system Principle Application Reference Nematode maturity index Nematodes classified on a "colonizer" -"persister" scale Can be applied to all soils; measures general response to stress (metals, acidification, eutrophication) rJongere (1990^ Yeates and Borgers (1999) Predatory mite maturity index Mesost.gmatid mites classified according to on r-K score Mostly limited to forest sails; measures soil properties related to mull/mor humus Ruf (1998) Earthworm life-history Strategies Earthworms classified according lo position in the soil profile and burrowing behav lou r Can be applied to all soils with sufficient number of species; measures aspects of humus lype, pH and cultivation (ploughing) Bouche (1977), Paoletti (1999a) REAL model for earthworms Integrated data base of various aspects related 10 Ihe ecological and agronomical rote of earthworms Very wide application Bouche (1996) Enchytraeid Reaktiprmihl ^ Scores related to responses (o acidity sind humidity assigned to enchytraeids Applicable to situations where effects on soil pH are manifested, for example cement factories Graefe (1993), Beylichetal,(1995) SIVPACS Pollution responses of earthworms, isopods and spiders, comparable lo RIVPACS Data base on species-specific responses not yet operational; at the moment only applied to heavy'metal pollution Spurgeonetal. {1996) Woodlice Iife-#ltr6 Classification of wood lice according to body shape and movement pattern Composition of isopod fauna indicates effects of soil cultivation in agricultural landsca-jes Paoletti and Hassell (1999) Bioindication in soil ecotoxicology Invertebrates ^Qpelman, P. and Eijsackers, H.J.P. //^5&04): Vital Soil - Function, Value and IVRlwberties. Elsevier. 358 p. ISBN: 0-'X^44°4-51772-3. Indicator system Principle Application Reference Macro invertebrate biodiversity Enumeration of species richness of earthworms beetles, isopods, spiders, ants, millipedes, centipedes, etc. Applied in orchards and other agricultural ecosystems lo indicate land use and copper pol lulion PaoLetii and Somaggio (19%), Paoieiti (1999b) Ant functional groups Classification of ants according to groups reflecting susceptibility to stress Wide application; used in evaluation of nature restoration and effects of mining AndecsenOW) Diptera feeding groups Classification of dipteran larvae in five feed log groups Reflects type of organic materials in soil; applicable lo organic soils prov*(1999> Arthropod acidity index Classification of arthropods (Collembola, oclbatids, isopods) according to pFl preference Allows quantitative estimation of soil pi I from invertebrate community structure Van Straalen and VerhMff.1997), Van Straalen (1998) Orlbatid mil? life-history strategies Classification of mites according to reproductive And dispersal strategies Indicates intensity of anthropogenic influence and successional stage of forests and grassland ecosystems Siepel (1994), Siepel (1996) Life-forms of Collembola Classification of Col'erobola according to morphological types reflecting position in the soil profile Indicates profile build-up and ecological processes stratified According to the profile; mostly applicable to forest soils Van Slraalen et al. (1985), Faber (1991) Dominance distribu lion of micro arthropods Lngnnrmal distribution of numbers over species General impression of disturbance; applied to effects of heavy metals and acid rain in forest and grassland soils 1 lagvar (1994) Biological Inde* of Soil Qunlity (BSQ) System of scores assigned to groups of soil micro arthropods Provides indication of biodiversity; wide applicability Parisi (2001), Cardi etaL (2002) ication in soil ecotoxicology Pollutant group Polyeyllc aromatic hydrocarbons* a/aarenes and derivates Persistent organochlorines (t*CBs, rfioxins) Vulnerable animal groups_ ; Remarks Isopods, Collembola Vertebrates (Rodentia and Insectlvora) Little knowledge available. Large inler-species differences in metabolism. Metabolizers expected to be more sensitive than accumulators. Low toxicity to Invertebrates, Effects appear higlwr up in the food-chain Earthworms ore I mpor tnnl in transfer.__ Chlorinated ethyleiws, phenoles and benzenes OiLGTEX Alkyl benwne , sulfonates And other determents Veterinary drugs antibiotics, hormones Copper Zinc Earthworms Earthworms Enchytraeids, nematodes, earthworms Toxicity due to general narcotic effects, probably small inter-species differences. Toxicity partly due to changes in soil structure. Held data scanty. Laboratory data Suggest highest toxicity to pore water-dependent species._ ^__ No data available Earthworms, slugs, snails, přibal Id mires Interactions in decorn poser-micro-organism interactions expected, but not documented. Copper toxicity to earthworms well documented. Cadmium Lead Enchytraelds, nemalodes. earthworms; isopods, soft-bodied sprlngtails_ Oribiitid mites, spiders, somespringtoits, vertebrates (shrews, mole) Toxicity of zijic does not follow the main taxonomic groups of soil invertebrates. Many groups contain sensitive as well as tolerant species^ Herbicides Fungicides Oribalid mites, shrews, mokř Cadmium seems to be most toxic to Invertebrates lhal take up the metal with the food. Due to food-chain accumulation effects appear in predators and vertebrates. Differences between Invertebrate species relatively srnai I, Main I laze rd of lead is hijd wr up In the food chain,_____ Mo group in particular Earthworms, enchytraeids, Isopods j Low toxicity of modern herbicides to animals. Effects are mostly secondary (avoidance of sprayed leaves, loss of food, Increase of Liter cover), Benzimidajcoles, carbamates and organorlns are known for their considerable side-effects on animals Invertebrates Doelman, P. and Eijsackers, H.J.P. (2004): Vital Soil - Function, Value and Properties. Elsevier. 358 p. ISBN: 0-444-51772-3. Pollutant group Vulnerable animal groups Remarks Insecticides Many arthropod groups, in particular beetles, spiders, mesostlgmatid miles and springtails Animals with high surface activity are particularly vulnerable. Large differences between species due to species-speci fic exposures and metabolic capacities. Many secondary effects among detritl vores due to suppression of predators. Acidic precipitation Snails, dipteron larvae, earthworms, some ohbatid miles, some Collembola, some isopods Large differences between species within each group. Earthworms generally avoid add soils. Many Coltembola and mites are acid tolerant, but some are very alkalophillc and suffer from acid precipitation. Radiation Earth worms, oribal td mites Specles-spectftc vulnerability due lo exposure, rather than inherent differences in sensitivity. Permanent soil dwellers and soil ingesiers receive hifth doses. Mote: this table only describes the general Irendsand ignores the many spedes-speciflc sensitivities related to metabolism, micrnhabitnt choice and li fe-cyde soil bioassays in soil protection Sofar, mostly for assessment of hazard of chemicals and pesticides Increase of use for evaluation of hazard of complex mixtures like wastes, sewage sludge, sediments, composts, fertilizers ... Great potential in the future for assessment of soil quality e.g. Before and after the remediation, contaminated sites assessment etc. M B fi ■***HA«* **Z*r ""Muco* CO Why bioassays? Chemical analyses are not able to identify risks properly because: 1) Real exposure is different - bioavailability in particular situation 2) Pollutant mixture - always in real ecosystems 3) Matrix itself has effects or interacts with effects of contaminants 4) Anylytical methods are limited vs. Wide spectrum of possibly toxic chemicals ^ctrns/,. rf**"**), mm m *mZZ*^ N.***^ Why soil bioassays? Eluate tests are not able to predict solid phase exposure WHY? -> real bioavailability effect of matrix involved .^ERS/J- ji'W't*, „J*",R»«»ir. mm @ N.***^ '*Wluhtm Solid material toxicity testi ISO 15799 (2003): Guidance on the ecotoxicological characterization of soils and soil materials ISO 17616 (2008): Guidance on the choice and evaluation of bioassays for ecotoxicological characterization of soils and soil materials Retention function - Biotests with eluates Luminescent Algal inhibition test Umu-test bacteria test Habitat function - Biotests with solids Plant test Earthworm test Collembolan test ^ctrns/,. rf**"**), mm m *mZZ*^ N.***^ EU - test battery IDS' ISO 11268-1 (1997): Soil quality - Effects of pollutants on earthworms {Eisenia fetida). Part 1: Determination of acute toxicity using artificial soil substrate. ISO 11269-2 (2004): Soil quality - Determination of the effects of pollutants on soil flora. Part II: Effects of chemicals on the emergence and growth of higher plants. ISO 16387 (2004): Soil quality - Effects of pollutants on Enchytraeidae -Determination of effects on reproduction and survival. ISO 11267 (1999): Soil quality - Inhibition of reproduction of Collembola {Folsomia Candida) by soil pollutants ISO 11268-2 (1998): Soil Quality - Effects of pollutants on earthworms {Eisenia fetida). Part 2: Determination of effects on reproduction mm ISO 17512-1 (2008): Soil Quality - Avoidance test for evaluating the quality of soils and the toxicity of chemicals. Test with Earthworms {Eisenia fetida/andrei). ^ctrns/,. rf**"**), mm m *mZZ*^ N.***^ Exposure methods • Tested chemical mixed with soil - Artificial soil (OECD, ISO) - Real soil (LUFA 2.2...) • Topic applications, injections, forced feeding ... not so relevant .^ERS/J- a*""""""* Jl**"**. m m @ N.***^ ""^^ What is artificial soil ? Soil component Sphagnum peat (air dried), finely ground and with no visible plant remains Kaolinite clay (air dried), containing not less than 30 % kaolinite Industrial quartz sand (air dried), predominantly fine sand with more than 50 % by mass of particle size 0,05-0,2 mm (amount dependent on calcium carbonate required) Calcium carbonate (CaCO^, pulverised, analytical grade) to obtain an initial pH of 6.0 ± 0.5 Content expressed on % dry mass basis 10 20 70 0.3-1.0 OECD 1984. Guideline for testing chemicals 207. Earthworm acute toxicity test. Is standard medium for many soil bioassays ... Is much more relevant than solution, agar, filter paper ... Should solve problem of high variability of natural soils ... Should resemble natural loamy soil ... Should enable the toxicity extrapolation to natural soils ... hWERS/j. .i^"^ A*«"lK«e, mm m niZZir Nä**^ Soil microbial assays EPA (1996): OPPTS 850.5100 Soil microbial community toxicity test. Ecological effects test guidelines. United States Environmental Agency. EPPO (1994): Decision making scheme for the environmental risk assessment of plant protection products. EPPO Bulletin 24, Chapter 7, Soil Microflora. Lynch, M.R. (1995): Procedures for assessing the environmental fate and ecotoxicity of pesticides. SETAC, Brussels, Belgium. OECD (1999): Proposal for a new guideline 217. Soil microorganisms: Carbon transformation test. OECD guideline for the testing of chemicals. OECD. OECD (1999): Proposal for a new guideline 216. Soil microorganisms: Nitrogen transformation test. OECD guideline for the testing of chemicals. OECD. ISO 14238 (1997): Soil quality - Determination of nitrogen mineralization and nitrification in soil and the influence of chemicals on these processes. International Organization for Standardization. Geneve, Switzerland. 4Mb B fi ((©)) Soil microbial assay according to OECD, ISO .w. Real uncontaminated agricultural soil with indigenous microflora: pHKCI = 7-7.5 Cbi0 = 400 - 700 Mg-Qd-w,1 M Corg = 1.5% BR = 0.5 - 0.7 |jg COz-Ch"1^ sand = 70% 10 g per replic aerobic conditions 60% WHC; 22 C; dark Soil sampling Storage Pre-incubation (7 days) 1 START ■-■ M B fi ■^WA*8- *1«Si«*ř ""tyW* ((©)) Substance application Negative control Positive control 7th day 14th day 21st day 28th day Microbial parameters ts on J/JJCJ obial respiration 100 : S I 10 TT B-resp=370*ppm NOEC=15 ppm £0^=61 ppm -0.49 1 0.01 0.1 1 10 100 Cone Ag (ppm) in soil .^BKSfr .f*>>»U*, „«.«"lR«&, Mr fs§ m Respirometry hWERS/j. .í^"^ „J*"18"*., Ami) I® m niZZíŕ Nematodes pseudocoelomiq cavity gonad muscle nerve cord L hypodermis m verrtraJ nerve cord gonad arm rectum uteru s Caenorhabditis elegans Caenorhabditis elegans ASTM: E2172-01 Standard Guide for Conducting Laboratory Soil Toxicity Tests with the Nematode Caenorhabditis elegans 1.5 ML TEST SOLUTION 2.333 G SOIL 35MM DISH {MIX, EQUILIBRATE AT 20 *C) DAY 1 ADD 10 WORMS DAY 2 imi ((©)) (20-C FOR 24 HR) ■ \ i RINSE WITH LUDOX § a CENTRIFUGE 2500 RPM, 2 MIN. DAY 3 10QMM DISH COUNT WORMS Avoidance test with E. albidus waste Artificial soil Bioaccumul 180 160 140 120 100 ao 60 40 20 0 xperiments with enchytraeids lindane (jig/g) Uptake phase Elimination phase Natural Sou 0 1 2 3 4 5 6 7 B 9 10 11 12 13 14 15 18 17 18 1» 20 TUE (days) B I Earthworms Cocoon formation t 23 days Incubation period * 4 days Eisenia fetida Temperature 25 "C Moisture t 76 % (Venter a Reinecke 1988] Olte/lum development Hatchlings (i 3 hatchlings per cocoon) M 5 i! ^HA**- "Xftuooi*" Earthworm acute toxicity test 14 days 500 g soil + 10 adult Eisenia fetida mortality and weight ^ V.-.'-L: ' ■ 1 - - -- I I J ■ . ■ ■' --->.* '- ¥ v\ r' ■-, .■'/ Glass Petri dish 1 litre glass jar Cow dung Artificial soil 10 adult earthworms f§ P> Earthworm reproduction test • 56 days — • 500 g soil + 10 adult Eisenia fetida • horse manure as food • juveniles extracted using water bath Eisenia fetida reproduction test Soil preparation WHC measurement Water added j Soil weighted to jars 10 adults to 1 jar B fi pi Weighting worms 10 adults from culture Washed E. fetida test - after 28 days E. fetida - 8 weeks After 20 min juveniles appear Water bath, increasing temperature 40°C - 60°C Sieving the soil Collecting and counting juveniles Hand sorting of cocoons Counting Guideline: Species: Substrate: Duration: Parameter: Test vessels: ISO/DIS 17512 (draft) E. andrei LUFASt. 2.2 standard soil 1 - 2 days Behaviour of the worms Dual chamber Risk assessment with earthworms Prüfung der Auswirkungen auf Regenwürmer Labortest mit Kompostwurm Kokons des Kompostwurms einheimische Regenwurmart 1. Akute Toxizität (2 Wochen) Bewertunq: TER = ^- <10 Mortalität, PEC Körpergewicht 2. Einfluss auf die Fortpflanzung (8 Wochen) Bewertung: Anzahl der Jungtiere, Körpergewicht TER = NOEC PEC < 5 3. Auswirkungen im Freiland {1 Jahr) Bewertung: Individuenzahlen, Risiken für Populationen und Lebensgemeinschaften k P iii-iKhLj1 rar M a t ij rbd'jsbalt Folsomia Candida Folsomia Candida Hypoaspis aculeifer Lactuca sativa root growth int protection products risk assessment Verschiedene Konzentrationsstufen im Auf tauftest Wachstumstest Lein Erbse Prüfpflanzen: 6 Pflanzenarten aus unterschiedlichen Familien 1. Stufe: Prüfungen im Gewächshaus • Auflauftest: Auswirkungen auf Keimung und Auflauf • Wachstumstest: Auswirkungen auf den Biomassezuwachs TER < 10 2. Stufe: Weiterführende Versuche Verlängerte Gewächshausversuche Mehr Arten Freilandversuche Microcosms, mesocosms Table 9.1 Classification of Various Semi-Field Tests 1. Model ecosystem segments (= "microcosms") Natural or artificially assembled units; a few centimeters in size — up to approx. I m3 (contents up to a few hundred liters); closed and open systems are both possible. Specialized techniques: Ink-grated techniques: e.g. the plant metabolism box <:f the NATEC (FIGGE, 1992) or small "artificial streams" (CLEMENTS ct al., 1989). e.g. (he Terrestrial Model Ecosystem (TMB) (VAN VORIS et al., 1984; KNACKER et al., 1990, 1991) or the Standardized Aquatic Microcosm (SAM) (TAUB et al., 1986; EPA? 1987). 2. Ecosystem segments in the field (- "mesocosms") Field segments which remain exposed to normal environmental conditions; various sizes ranging between J m3 and several hundred m\ Specialized techniques: Integrated techniques (very rarely in the terrestrial medium): Partial enclosures in lakes or rivers, e.g. plastic bags with algae coenoses (EIDE et al., 1979). Lysimeter (usually about 1 m3 in size), e.g. tests on the mobility of pesticides in natural soil cores (e.g. BBA, 1990a). Semi-field tests (usually tests with beneficial organisms), e.g. effects of pesticides on ground beetles (carabids) in cultivated soil system segments (ABEL & HEIMBACH, 1992). Artificial testing systems, e.g. ''artificial streams" — reconstructions of real streams including sediment (EATON et al., 1985). Natural enclosures, e.g. "Bremerhaven-Caissons" in wadden seas (FARKE et al, 1984). ocosms - TME Microcosms - TME Literature • van Straalen, N.M. and van Gestel, C.A.M. (1993): Soil invertebrates and Microorganisms. In Calow, P. (Ed.): Handbook of ecotoxicology. Oxford : Blackwell scientific publications, p. 251-277. ISBN 0-632-03573-0. Donker, M.H., Eijsackers, H., and Heimbach, F. (1994): Ecotoxicology of soil organisms. CRC Press, Inc., Boca Ranton. ISBN 0-87371-530-6. Tarradellas, J., Bitton, G. and Rossel, D. (1996): Soil Ecotoxicology. Lewis Publishers, Boca Raton, FL. ISBN: 1-56670-134-1. Lokke, H. and van Gestel, C.A.M. (1998): Handbook of soil invertebrate toxicity tests. John Wiley & Sons, Chichester. ISBN 0-471-97103-0. 281 pp. Rombke, J. and Moltmann, J.F. (1996): Applied ecotoxicology. CRC Press LLC, New York. ISBN 0-56670-070-1. • Hoffman, D.J. , Rattner, B.A. (1994): Handbook of Ecotoxicology. Boca Raton, FL, USA : CRC Press. • Suter, G.W. II, Efroymson, R.A., Sample, B.E., and Jones, D.S. (2000): Ecological risk assessment for contaminated sites. Lewis Publishers, Boca Ranton. ISBN 1-56670-525-8. 437p. Doelman, P. and Eijsackers, H.J.P. (2004): Vital Soil - Function, Value and Properties. Elsevier. 358 p. ISBN: 0-444-51772-3. Doran, J.W., Coleman, D.C., Bezdicek, D.F., Stewart, B.A. (1994): Defining soil quality for a sustainable environment. SSSA Special Publication Number 35. Soil Science Society of America, Inc. Pankhurst, C.E., Doube, B.M., Gupta, V.V.S.R. (1997): Biological indicators of soil health. CAB International, Wallingford. ISBN 0851991580. Doran, J.W. and Parkin, T.B. (1996): Quantitative indicators of soil quality: A minimum data set. In Doran, J.W. and Jones, A.J. (Eds.): Methods for assessing soil quality, p. 25-38. SSSA Special Publication Number 49. Soil Science Society of America, Inc., Madison. Doran, J.W. and Jones, A.J. (Eds.): Methods for assessing soil quality. SSSA Special Publication Number 49. Soil Science Society of America, Inc., Madison. Doran, J. W., Parkin, T. B. (1994): Defining and assessing soil quality. In: Defining soil quality for a sustainable environment. SSSA special publication number 35. SSSA, Inc., American Society of Agronomy, Inc. Madison, Wisconsin, USA, 1994, pp. 3 - 21. European Commission (2002): Communication from the Commission to the Council, the European Parliament, the Economic and Social Committee and the Committee of the Regions - Towards a Thematic Strategy for Soil Protection. COM (2002) 179. European Commission (2006a): Communication from the commission to the council, the european parliament, the european economic and social committee and the committee of the regions. Thematic strategy for soil protection. COM(2006)231. European Commission (2006b): Proposal for a directive of the European Parliament and of the Council establishing a framework for the protection of soil and amending Directive 2004/35/EC. COM (2006) 232. European Commission (2006c): Commission staff working document accompanying the communication from the Commission to the Council, the European Parliament, the European Economic and Social Committee and the Committee of the regions. Thematic strategy for soil protection. Impact assessment of the thematic strategy on soil protection. SEC (2006) 260. Literature Van-Camp, L., Bujarrabal, B., Gentile, A-R., Jones, R.J.A., Montanarella, L, Olazabal, C, Selvaradjou, S.K. (2004): Reports of the Technical Working Groups Established under the Thematic Strategy for Soil Protection. Volume V - Monitoring. EUR 21319 EN/5, 872 pp. Office for Official Publications of the European Communities, Luxembourg. EEA (2007): CSI 015 - Progress in management of contaminated sites - Assessment published Aug 2007. European Environmental Agency, www.eea.eu Carlon, C. (2007): Derivation methods of soil screening values in Europe. A review and evaluation of national procedures towards harmonization. European Commission, Joint Research Centre, Ispra, EUR 22805-EN, 306 pp. De Bruijn, J., Crommentuijn, T., van Leeuwen, K., van der Plassche, E., Sijm, D., van der Weiden, M. (1999): Environmental Risk Limits in The Netherlands. RIVM Rapport 601640001. pp. 900. De Vrieš, W. and Bakker, D.J. (1998): Manual for Calculating Critical Loads of Heavy Metals for Terrestrial Ecosystems. Guidelines for Critical Limits, Calculation Methods and Input Data. Report 166. DLO Winand Staring Centre, Wageningen, The Netherlands, 144 pp. hWERS/j. .i^"^ „J*"18"*., mm m