The use of invertebrate soil fauna in monitoring pollutant effects Annette Gomot-de Vauflerl, Nicole Poinsot-Balaguer, Jérôme Cortet, Lucien Gomot, Christine Texier, Daniel Cluzeau

To cite this version:

Annette Gomot-de Vauflerl, Nicole Poinsot-Balaguer, Jérôme Cortet, Lucien Gomot, Christine Texier, et al.. The use of invertebrate soil fauna in monitoring pollutant effects. European Journal of Soil Biology, Elsevier, 1999, 35 (3), pp.115-134. ￿10.1016/S1164-5563(00)00116-3￿. ￿hal-03217681￿

HAL Id: hal-03217681 https://hal.archives-ouvertes.fr/hal-03217681 Submitted on 5 May 2021

HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. The use of invertebrate soil f auna in monitoring poilu tant effects Annette Gomot-De Vauflery, Nicole Poinsot-Balaguer, Jérôme Cortet, Annette Gomot-de Vauflerl, Nicole Poinsot-Balaguer Lucien Gomot, Christine Texier, Daniel Cluzeau

To cite this version:

Annette Gomot-De Vauflery, Nicole Poinsot-Balaguer, Jérôme Cortet, Annette Gomot-de Vauflerl, Nicole Poinsot-Balaguer et al.. The use of invertebrate soil f auna in monitoring poilu tant effects. European Journal of Soil Biology, Elsevier, 1999, 35 (3), pp.115-134. ￿10.1016/S1164-5563(00)00116-3￿. ￿hal-03217681￿

HAL Id: hal-03217681 https://hal.archives-ouvertes.fr/hal-03217681 Submitted on 5 May 2021

HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. The use of invertebrate soil f auna in monitoring poilu tant effects

a Jérôme Cortet *, Annette Gomot-De Vauflerl, Nicole Poinsot-Balaguer\ b C c Lucien Gomot , Christine Texier , Daniel Cluzeau

"Institut méditerranéen d'écologi,e et de paléoécolog-ie, université de Provence, Centre Saint-Jérôme, rase 462, 13397 Marseille cedex 20, France. b Laboratoire de biologi,e et écophysiologie, UFRdes sciences et technique�� place Maréchal-Leclerc, 25030 Besançon cedex, France.

c Laboratoire d 'érologi,e du sol Pl biologi,e des populations, UMR 6553 CNRS: université Rennes-], Station biologi,que, 35380 Paimpont, France. *

Corresponding author: [email protected]

Abstract - A critical review of biological parameters used to indicate pollutant impact on soi! quality was conducted. These param­ eters mention some soi! invertebrates. The value of an indicative organism depends on its life expectancy, life style and specific importance. Nematodes, mites, collembolans, enchytraeids, earthworms, isopods and molluscs are good potential biological indica­ tors. Biological indicators of bioaccumulation and biological indicators of effects (toxicological and ecological) can be distin­ guished. Bioaccumulation studies are difficult to interpret, as wide variations could be found, depending on taxonomie group, habitat, organ studied, soi) type or even pollutant type. Sorne groups, such as Collembola, require in depth bioaccumulation studies. It is suggested to use a pool of macro-concentrators, including at least some earthworm, isopod and gastropod species. Toxicological indicators have been well studied and their lethal and sublethal pollutant effects are well known. However, studies have focused on only a few species, such as the earthworm Eiseniafoetida or the collembolan Folsomia candida. These studies should be extended to other zoological groups, as well as to several species from the same group, to generate a representative test battery. Exposure biom­ arkers and physiological change studies should be emphasised, as they act as very early warning systems of contamination. Data are currently lacking on how soil biological processes malfunction due to pollution. We need to explore the links between pollutant effects on soi! fauna and pollutant effects on soi! functioning. Concerning ecological indicators, studies should develop sampling techniques and parameters, which are specific to ecotoxicological goals. Before-after impact contrai procedures should be carried out, to eliminate the background noise of the study site and only evaluate the influence of pollutants. On the other hand, ecological indices, such as taxonomie diversity or richness, should be used carefully especially concerning chronic pollution. Effects of pollut­ ants on biological cycle studies seem very promising, but need further information on the life history strategies of many species. Furthermore, the pollutant tolerance of rare species should be considered. The differenttypes of biological indicators yield comple­ mentary information on pollutant effects. They ail need standard procedures. In this context, studies should be extended and diversi­ fied, and associate bioaccumulation, toxicological and ecological indicators to provide better information on soi! quality. Ecotoxicology / bioindicators of effects / bioaccumulation / soit ecology / soit quality / soit invertebrates / soit fauna / earthworms / microarthropods / mites / collembolans / isopods / enchytraeids / snails / soit pollution

Résumé - Utilisation de la faune invertébrée du sol comme bio-indicateur des effets des polluants. Une liste critique des para­ mètres biologiques utilisés dans certains travaux pour indiquer l'impact des polluants sur la qualité des sols a été établie. Ces para­ mètres font référence à un ou plusieurs invertébrés du sol. Le rôle d'organisme indicateur dépend de leurs caractéristiques biodémo­ graphiques, de leur mode de vie, de leur taille spécifique. Les nématodes, acariens, collemboles, enchytréides, vers de terre, isopodes. et gastéropodes sont potentiellement des indicateurs biologiques. Indicateurs biologiques de bioaccumulation et indicateurs biolo­ giques d'effets (toxicologiques et écologiques) peuvent être distingués. Les études de bioaccumulation sont difficiles à interpréter, car de fortes variations sont observées. Ces variations dépendent du groupe taxonomique étudié, de l'habitat, de l'organe étudié, du type de sol ou bien encore du type de pollution. Certains groupes, comme les collemboles demandent des études plus poussées à ce . sujet. li est suggéré d'utiliser un pool de macro-concentrateur,. avec au moins des de vers de terres. isopodes et gastéro­ podes. Concernant les indicateurs toxicologiques d'effets. les études les plus nombreuses concernent les effets létaux et sublétaux. Toutefois ces études concernent peu de groupes taxonomiques (essentiellement le vers Eisrnia foetida et k collembnle Folsomia candida). et devraient être étendues à d'autres groupes zoologiques. ainsi qu'à différentes espèces du même groupe. afin de créer une batterie de test, représentatifs. Les travaux concernant les biomarqueurs et les changements phy,iologiques devraient être ampli­ fiés, car ils permettent d'obtenir des ,ystèmes d'alarme très précoces concernant J'impa<.:t des contaminants. Un manque pen,istant de données concerne les conséquences de pollutions sur k fonctionnement des écosystèmes. Il ést en effet important désormais d'explorer les liens entre les effets des polluants ,ur la faunedu sol et les effetsréels de ces mêmes polluants sur le fonctionnemell! des sob. Concernant les indicateurs écologiques. le, travaux entrepris devraient développer des méthodes d'échantillonnage et des paramètres spécifiques au domaine écotoxicologiquc. Ainsi. il est préconi,,{ d'entreprendre des études combinant les approches syn­ chroniques et diachroniques. afin d'éliminer le bruit de fond induit par le site d'étude et évaluer uniquement lïnfluenœ du polluant. En outre. le, indices écologiques. comme la diversité ou lu riche,se taxonomique. doin:nt être utili;,és uvec précautions. particulière­ ment lors des études de pollution chronique. Par ailleurs. le, études conccrnamles effets de polluants sur les cycles biologiques sem­ blent prometteuses, mai, demandent de meilleure, connaissance, concernantles stmtégics de ,ics des différentes espèces étudiées. De plus. la tolérance aux pollutions des rares devrait être fortement prise cn considération. Les différents types d'indicateurs biologiques fournissent des informations complémentaires. Ils demandent tous une standardisatiun des protocoles. Dans cc contexte. les études devraient être développées el diversifiées et associer les indkateur.s de bioaccumulalion avei: les indicateurs biologique, d'effets pour améliorer l'information sur la qualité des sols. Écotoxicologie /bio-indicateurs d'effets/ bioaccumulation / écologie du sol/ qualité du sol/ invertébrés du sol/ faunedu sol/ vers de terre/ microarthropodes /acariens/ collemboles/ isopodes/ enchytraeides / mollus<1ues/ pollution du sol

1. INTRODUCTION ted to environmental management as they examine the effects and consequences of pollutants in the ecosystem. Fauna is an important part of soil environments. It is Several zoological groups can be used: involved in many aspects of organic matter decomposi­ Soi! microfauna (smaller than 0.02 mm). These are tion. partial regulation of microbial activities, nutricnt primarily Protozoa and Nematoda living in the water cycles and crumbly structure. Depending on thcir diet free soi! compartments. They can be used to study soil (saprophagous. phytophagous or prcdation). soi! fauna ecotoxicology. Tests using these organisms are similar are closely linked both to cach other and to micro­ to th ose used in aquatic toxicology [194]. The�e organ­ organisms, plants and soi! [167]. They are a vital key isms have a high hioaccumulation potential 111 J. to understanding the "oil ccosystem. Microcosm tests are currently hcing designed for several Disturbances caused by pollutants in the soi! results Proto;:oa (Cercomonas, Acant/l(lmoeha and Colpoda in bath qualitative and quantitative changes in fauna. ge11era) (57]. Toxicity te�ts in artificial soils using the which affect soi! functioning. Some of soil fauna nematodc Plecrus acw1ti11mus have provided addi­ groups can act as hioindicators1 136]. tional perspectives for studics of soi] contaminants A bioindicator must fulfil the followîng rcquire­ [98]. In situ studîe� using nematode� have yielded ments [83]: several maturity indices which providc data on the soit play an important role in the functioning of the soil functinn. These indices could be uscd to monitor the ecosystcrn; soi! ecology as thcy ret1ect changes in soi! condition 1130. 1311. be widely distributed. common and easy to sample: - Soit mesofauna (from0.02 to 4 mm). Enehytraeids be robust: should not be killed by vcry low levels (oligochaete annelids) are eommon in most soils (pri­ of pollutants; marily in the first 5 cm or soil) and play an important have measurable responses (pollutant concentra­ role in decomposîtîon and humification processes. tion in tissues. characteristk biological disturbances in They are general!y micmphagous and capable of rcus­ growth and fertil ity or resi�tancc J: ing othcr animais (earthworms or microarthropodsJ � have reproducible responses: should produce simi­ excrcments [41. Few ecotoxicological studic� have lar responses to the same lcvels of pollutant exposure been conducted on them. but initi7il reports. particu­ in different site�. larly ex situ studics have been promising [ 121, l50J. ln addition, the selected zoological group!>,should be Springtaib (aptcrygotc insects) and acarids (Arach­ practical for use in both laboratorv and in situ studies. nida) are the rnost common soi! arthropods [25] and Laboratory tests provide a simplified. analytical appro­ play a rolc in soi! functioning. They are vital indi­ ach to obtain information on how an individual rcacts cators for soi] ecotoxicology. and how ils organs or cellular and molecular fonctions Soil macrofauna (larger than 4 mm). Snails and are affected. ln situ ecotox icological studies arc more slugs (terre striai gastropod molluscs) mnve along the complicated than lahoratory tests, but are hettcr adap- soi! surface eating plants and dirL hut part of their bio-

Invertebrate soil fauna as pollution indicators logical cycle (rest, shelter, egg-laying, embryo devel­ which often can lead to biomagnification of pollutants opment, hibernation) takes place in the soil, from during transfers along the food chain. The same pro­ which they absorb nutrients and contaminants. They cess occurs for some organic compounds, particularly have a wide range, are easy to sample, have a high bio­ pesticides. However, no standard procedure exists to accumulation capacity and a phytophagous or saproph­ measure bioaccumulation [ 137]. agous diet which make them very good bioindicators It should be noted that there is no direct relationship [8]. Earthworms (oligochaete annelids) are also good between bioconcentration of a pollutant and intoxica­ indicators, as almost every soil contains at least one tion [ 194]. It is thus necessary to study not only the earthworm species [ 14]. Their range varies depending internai concentration but also the effects of pollutants on depth: epigeous, endogeous and anecic worms can on organisms. be found in temperate zones [116]. In addition, they Concentrations of pollutants are measured using var­ migrate over short distances [82]. They are both resis­ ious chemical extraction techniques followed by gas tant and sensitive. Isopods (crustacean arthropods) are chromatography [ 122] or atomic absorption spectro­ particularly interesting due to their capacity to accu­ photometry analysis [ 173, 196 ]. Pollutant fluxes can mulate metals and their abundance in some soils [82]. also be calculated by kinetics studies (ingestion and They are detritivore or microphytophagous but excretion) of pollutants [94]. strongly linked to humid environments, which could limit their in situ utilisation. Holometabolous insects Although often associated with toxicological tests are often only present in the soil during a particular [163, 182], few bioaccumulation studies have been phase in their development and are generally c:onsid­ made on Collembola [53]. However, Collembola feed ered to be poor accumulators. However, some studies partly on fungal mycelia, which are known to concen­ have been undertaken on the beetle Oxythyrea funesta trate both metals and organic pollutants and radio­ (Cetonidae) for laboratory use [128] and xylophagous nucleotides [84]. In addition, they are often preyed beetles could be used as bioindicators of forest quality upon by other arthropods [58]. [12, 72, 17]. In addition, a laboratory test has been For copper accumulation, Collembola can be ranked standardised using staphylinid adults (Aleochara bilin­ second among detritivorous invertebrate right after iso­ eata) in agriculture [155, 156]. pods [85]. Hunter et al. [90] found that copper concen­ Several works on the use of soil fauna in soi! eco­ tration was 45 times higher in the same species pool at toxicology have already been published [75, 110, 193, a polluted industrial site than that at an unpolluted site. 195, 200] and are published still. However, due to the microarthropods small size, a The goal of the present work is to identify the most large number of organisms must be sampled for test­ significant results obtained for each faunal group in ing, which increases the margin of error. ln addition, terms of soil monitoring, and evaluate how suitable a although interclonal variation in some species (particu­ group might be used as bioindicator, with a particular larly Folsomia candida) does not influence their use­ focus on expected measurable responses. fulness in ecotoxicological tests [33], a wide range of reactions exists between Collembola species. Lupetti We also suggest new requirements and new study et al. [122] found considerable differences between directions, that should be carried out. DDT and PCB concentrations in seven species sam­ For this purpose, we will follow Echaubard [50], pled at the same site. Orchesella cincta was found to who makes a distinction between: be highly vulnerable to DDT concentrations in its tis­ - biological indicators of bioaccumulation; sues. These differences can be explained by the excre­ - biological indicators of effects, including toxico- tion/assimilation ratio and the capacity to eliminate logical and ecological bioindicators. substances from some tissues, which varies between species [83]. Biomarkers reflect either an individual biological reaction or lower level reactions (cellular, molecular, In vivo metal contents in earthworms and isopods genetic) and are thus toxicological indicators of vary depending on species or metal type. Differences effects. However, they cannot always accurately indi­ in concentrations have been observed: cate a genuinely toxic effect, at the individual or the - between species from the same taxa (earthworms: upper levels [103, 202]. [91, 3, 173], isopods: [205, 97, 87]); - between different aged individuals from the same species ( earthworms: [21], isopods: [86] ); 2. BIOINDICATORS OF BIOACCUMULATION - between seasons (earthworms: [92, 2], isopods: [204]). Bioaccumulation occurs when a substance present in Bioaccumulation of metals depends on: the biotope penetrates into an organism where it is - the element (isopods: [88]); stored in various forms. Bioaccumulation of heavy metals by terrestrial organisms has been well studied. - its concentration in the environment [67, 73]; It is a very important phenomenon which provides - the physical and chemical soi! conditions (earth- information on the bioavailability of trace elements worm: [123 ]). .

Table I. Lead concentration in carthworms (mg-kg-1 dry wcight) and concentration factor (ratio of Ph c·

Spccics Tikkurila P()rnaincn Mann-Whitney

Mean± SD Il Median Mean± SD Il Mcdian Ph concentrations in worms Aporrectodea caligino,·" 1 058.0 ± 480.4 10 904.:'i hl .7 ± 42.7 7 'i()_() Lumhric11.1 ruhe/111.1 IJJ.o ± l lb.5 9 129.1 56.2 ± _"\(1_7 5 61 1 ns L castaneu, 75.6 ± 109.4 (, 11.9 88 ..s ± 'il)_/\ 97.2 ns Kruskall-Wallis hetween species Concentration factor /\. caliginosa 0.98 ± 0.38 10 0.92 3.8 ± _l_ 1 3.1 [ .. mhel/u, 0.07 ± 0.05 l) 0.08 J.2 ± 2. l 5 �.� Il L CCL\f{ll1t'llS 005 ± 0.06 6 0.01 :'i -1 ± 5.0 -' ...i J Kruskall-Wallis hctween spccies

Terhivuo et al. l 173 j noted different bioaccumula­ through protein binding, including metallothioneins. tion rates among different earthworm species at a site which can scavenge metals (primarily cadmium) and damaged by lead pollution (tahlc /). store them for long periods [38]. Most studies of terrestrial gastropods (tah/e If) have By evaluating the metal contents in different gastro­ focused on analysing bioaccumulation among some pod specics from different cnvironments. Berger and experimentally contaminated animais or animais sam­ Dallinger 18 j were able to reveal three pollution levels. pled at a polluted site. Gastropods survive in sites In a similar study on cdible snail quality (in fanns and polluted by metals. This resistance stems from their natural environments ). Pihan et al. [ 138] suggcsted a ability to retai n and inactivate toxic metals. either classification according to metal contents (Cu. Zn and through intraccllular compartmentalisation ( confine­ Pb) and studied the relationship bctween metal con­ ment within granules. vesicles. etc. l and excretion. or centrations in snail tissue� and their food supply [71 ].

Tahle Il. Pulmonated land gastropods as contamination indicators.

Rcfcrence Pollutant Cuntamination source Spccics Test duratinn Paramckr..,

[1291 Fe. Ph. Zn ffe!ix /N1mr1lio ..i v,ceks Tcmperaturc effect on Fe. 1 Aug.-Sq,t. 1l/711 Ph.Zn in diffcrelll organ, [29) Cd. Cu. Ph. Cuntaminatcd and uncontaminatcd �ite-; Il. os1wrso Natural ,naib \lctal c,mcentration Zn 1Sep1. 107�1 in nrga11-- [206] Cd. Ph.Zn Roadsidcs Cepoeo hortenv;.\ tJuL-Scpl. i'l7:'i) [371 Cd. Cu. Ph. Contaminatcd fond (lcttuce) H. 11011w1io 3il--J5 d Bioac·c,11nulation. ,·cllular Zn i\1arch !li811 and molecular dis1rihution ui" 1nctals [61 Ph Agar gelose +Ph or +Ph and Ca 1-f. (/lf>l'/"\r/ Bioaccumulation: Ph a..,si,ni­ lation and cxcrdion [761 Ca. Cd. Cu. Ph. Ahandoncd mine Pb/Zn Six ..,1ug -;pccic-; I\atural ...tug, l\ktal ,·onccnlration Zn 1 Sept. i 'l861 in 1i ...... _uc-; 136] diverse Di,crse Castrnpods and land Variahlc Contamination. cellular inwr tchrate, n)mpartmcnt. dctnxification [38] Cel. Zn Agar plates with Cd et Zn H. /J0!1/(lfÙI Subcl'ilular distrihutinn in digestive gland-; [8] Cd. Cu. Ph. Zn Urhan areas A rianto urhustorwn Scasonal ..,amplinf! Meta! concentration hct\vecn ,-,pcciè" in li\:...UL':... (+ influence compari�un of indi\ îdua! "i1.c l I IJ8J Cu.Zn N atural and through food H. u.,per.�o a.\pcrsu i\:atural and raiscd Meta! c·onccntration and li ospcr.,,1 mfv.:ima adult :,,naih in ti-..:-.uc:.... JJ, pomatio l:lioaccurnulation and pPlluliPn r�1L·tnr:... [1 39] Ph Contaminatcd sites and hrceding H. 01pa.\"{/. H. 1101110/iu Natural ami raiscd Ph ,·onccntration in foot adult snails and, Î'-iccra [731 Cd. Ph.Zn Food and suhstratc /)e,ncero.\ rcth·ula!ll111 3 \\ cck:-, Meta[ concentration, in the hoc!\ [711 Cu. Zn Food H. uspersa cHper,\u 10 \Vl'L'k'-. 13 ioaccu mu lati<111 and fi. n.,pcna mnrinw ------�---- . Invertebrate soil fauna as pollution indicators

In addition, gastropods can be used to study metal direct information about environmental contamination. transfers into the ecosystem through the food chain [6]. It thus seems wise to use a pool of macro-concentra­ As they are eaten by many different mammals, they are tors: for example, isopods (e.g. Oniscus asellus) and probably involved in the biomagnification of metals pulmonate gastropods (e.g. Helix aspersa) have pro­ [36, 206]. vided information about the degree of metal contami­ Bioaccumulation of pesticides by terrestrial gastro­ nation (Cu, Cd, Pb, Zn) in sites located near English pods has not yet been well studied, but atrazine and foundries (table Ill, [82]). This author also noted that azinphosmethyl do not appear to be accumulated bioaccumulation varies depending on the organ and is through the food chain [ 157]. particularly high in the digestive gland for Cd, Zn and Pb. Earthworms can have very high and significant Current studies on bioc:oncentration and bioaccumu­ BCFs for organic pollutants, such as chlorophenols lation have provided information about the bioavail­ (particularly, pentachlorophenol), which vary depend­ ability of contaminants found in the surrounding ing on soil type (476 and 956 in two agricultural soils environment and contained in food. To compare results for Lumbricus rube!lus [181]). from different analyses performed on animais sampled in nature or on animais which have been used for con­ Until recently, the indications provided by contami­ tamination experiments, bioconcentration (BCF) or nant concentration analysis in animais sampled in the bioaccumulation (BAF) factors can be calculated using field have not yielded any information about exposure the following ratio: time. To compensate for this, sentine) animals (from standardised cultures) are used either in microcosms, concentration of pollutant in animal mesocosms or on site to evaluate specific exposure (µg·g-1 dry weight) times. Copper accumulation was thus studied in Lum­ bricus rubellus [126] and in Eisenia andrei [171, 172]. coi:icentration of pollutan� in the environment Exposing cultured snails (Helix aspersa) for 30 d at or 111 food (µg-g- dry we1ght) distances from 1 to 200 m away from a busy motorway (Nancy-Metz: 40 000 vehicles per day) demonstrated Concentration factors are often used for aquatic that lead pollution remains high at distances up to 40 m organisms which are bioindicators of accumulation, away and causes lead accumulation in the snail viscera but they are slightly more difficult to use with terres­ ( 16-22 mg-kg-1 dry weight; Pihan and Gomot, unpubl.). trial animais where the environment is more heteroge­ neous. Furthermore, such ratios are only useful for Controlling breeding of some main macro-concen­ non-regulated metals and persistent pesticides. Com­ trator soil fauna species should make it possible to paring metal concentrations (Cd, Pb, Zn) in substrates study pollution over time and define its fluctuations. and food to study metal accumulation in isopods, collembolans, diplopods (]eaf Iitter) and slugs (soil) 1 yielded highly different results (from 14 to 750 mg-kg- 3. BIOINDICATORS OF EFFECTS of Cd and from 490 to 22 000 mg-kg-1 of Zn) when they were experimentally soaked in the same solutions [73]. This experiment illustrates the problems inherent Data based on the biological responses of organisms in between species and between habitat comparisons. to pollutant is used to link pollutant toxicity with eco­ Nevertheless, the overall data indicate that inverte­ logical consequences. Several biological levels exist, brates behave as either macro-concentrators, micro­ from the molecule to the community level, which concentrators or de-concentrators [ 13, 36]. Macro­ affect the degree of toxicological or ecological reac­ concentrator species are the ones that provide the most tions.

Table III. Heavy metal concentrations in the bodies of Helix aspersa and Oniscus asel/us from an uncontaminated site and a contaminated (smelting work) in England (from [82]).

Species Metals Meta! body concentrations (mg-kg· 1 dry weight) lncrease coefficients Uncontaminated site Contaminated site (Kynance Cove Cornwall) (Avonmouth) Helix aspersa Cu 104 ± 28 228 ± 43 2.2 (garden s nai1 J Zn 103 ± 14 418 ± 71 4 Cd 4.88 ± 0.62 49.6 ± 10.7 10.1 Pb 5.79 ± 1.06 121 ± 24 21.8 Oniscus asel/us Cu 115 ± 42 567± 138 4.9 (wood lice) Zn 108 ± 25 488 ± 103 4.5 Cd 5.93 ± 1.01 152 ± 37 2.5 Pb 4.15±0.91 413 ± 150 99.5

Among the biological indicators used (which pro­ NOEC and LOEC are not always considered to be vide data on reaction to pollutant exposure). two main good criteria as they requirc using a large range of categories can be found: concentrations to obtain a value close to reality. LC50 - toxicological indicators (molecules. cells. organ­ and EC50 are more accurate and have a better ecolo­ isms. groups of organisms). A close rclationship exists gical value as they dircctly reflcct fondamental between intoxication and an observed reaction. This changes in stages of an organisms biological processes data helps reveal phenomcna occurring in the upper (growth, reproduction). levcls but has little ccological value. These indicators ln addition. some specics appear to be more sensi­ can act as early warning systems of contamination tive when lethal concentration is tested. while others whose effects are possibly revcrsible 1501: are more sensitive whcn sublethal concentration is - ecological indicators. Possible changes in popula­ testcd. Crommcntuijn et al. f 321 have suggcsted using tion. community or population group structure as a new index to harmonise results. which they called the shown by ccological analysis indicate changes occur­ sublcthal scnsiti,ity index (SSI). It is calculated using ring but provide no information on causes of distur­ the ratio LC 50/EC III or LC,1/NOEC when sublethal bances. concentration cannot hc measurecl. Concerning methodologies. except for mathematical Rccent studies uscd the ILL (incipient lethal lcvel), models. Iwo approaches can be used [ 1471: which is the concentration of toxicant below which - experimental models. with single spccies and mul­ 50 rx- of the exposed organisms can be expected to tispecies tests and bioassays: survive for an indefinitc period f 164 ]. lt normalises the - in situ indicators. based on field studies and biota toxic rcsponse so that the assumption can be made that which live or are introduced in these sites. all comparisons of toxicity within and bctween chcmi­ cals and spccies of organisms arc macle at a steady­ However. the difference bctween these two state with respect to the dose response curve [ 114]. approaches is often arbitrary. For cxample. tïeld obser­ vations need an cxperimental part: and although bio­ lJ ntil now. toxicity tests for tcrrestrial organisms assays are linked to laboratory tests because they are have been bascd on single species to measurc the often standardised. they could be considered to be a impact of toxic products on survival. growth and repro­ particular form of in situ indicator as they use polluted duction. Although three tests have been standardised substrates rather than pure substances. These consider­ (two for eaithworms. one for Collembola). experimen­ ations concur with Forbes and Forbes [63 J. who distin­ tal data arc available ror most soi! falma groups. guish between field indicators and experimental tests, but prcfer the more gencral concept of ecotoxicity 3. 1.1.1. Nematodes tests. where observations or cxperimcntal manipula­ tions are used to evaluatc the fate or cffects of chemical pollutants. without any refercnce to the biological These are the most common soi] invertebrates. Tests organisation levcl. have been performed with Caenorhabditis clegans [42. 141. 187]. but toxic exposure conditions (mainly to Cd) differed too widely to yield correct comparisons of 3.1. Toxicological indicators results. Rccently. experiments have been performed with the 3.1.1. Testing lethal and sublethal effects ncmatode Plcctus acu111i11atus, a bacterivorous species which lives freely in the soi] hut is not associated with The first descriptor used to evaluate the impact of a plant roots or fungal hyphae. Kammenga et al. [98 J pollutant is acute toxicity. Mortality in a population used the juveniles to adults ratio as the test parameter exposed to xenobiotics is a severe reaction. Extreme in an artificial soi! (OECD) under optimal conditions toxicity is expressed as a proportion or a concentration for P. arn111inatus population growth in the soi!. Tests which leads to the death of 50 •Ir, of the tested indivi­ 1 were performed using Cd. Cu. and PCP [98 [. duals. Lethal dose or LD50, cxprcssed in mg·kg- of body weight, depends on the animal. LC50 indicatcs a lethal concentration exprcssed in mg-kg- 1 of substrate. 3.1.1.2. Enchytraeid� It is probably more reasonable to speak of LC 'iofor soi! fauna. Few studies have currently been performed (table IV). Studies on sublethal effects generally calculate EC50 Howcver. thcy have illustrated the complexity involved or the effective concentration for a biological process in ecotoxicological studies in soils and the need for which leads to a 50 '7r inhibition or decrease [ 143]. mcthod standardisation: Rundgren and Augustsson LOEC (lowest observed effect concentration) is the [ 1501 notcd a dccrease in fragmentation rate for Co­ lowest concentration yielding a significant differencc gnettio sphagnetoru111 individuals exposed to increas­ for the studied parametcr compared to the control. ing concentrations of copper in an artificial soi] Occasionally the tenn NOEC (no observed cffect (LUFA). and Sjügrcn et al. 1161] noted an increase in concentration) is used in toxicity tcsting. fragmentation rate in the samc species in a forest soi!.

. Invertebrate soil fauna as pollution indicators

Table IV. Examples of evaluation tests with enchytraeids.

Reference Pollutant Contamination source Species Test duration Parameters Weistheide et al. benomyl Contaminated gelose Enchytraeus crypticus. 30 d Number of hatched (1991), in [194] E. minutus juveniles cocoons

[148] parathion amitrole diuron Substrate E. albidus 56 d LC50 on cocoons

[22] dimethoate pirimicarb Food (oatmeal) E. bigeminus 7 d LC50; growth fenpropirimorph [150] dimethoate Substrate, food (spores) Cognettia 70 d NOEC; LOEC; ECx sphagnetorum (fragmentation rate and number of formed segments) [161] Cu Substrate C. sphagnetorum 15-50 d Fragmentation rate, mode] of dispersal. mini­ mum population size [102] Cu+ Ni Forest soi! C. sphagnetorum 138 d Litter mass loss, soi! (add to other organisms) respiration, change in nitrates [127] dimethoate Artificial soi! (OECD, E. crypticus / mriatus 32 d NOEC on juveniles humic. sandy and clayey)

3.1.1.3. Oligochaetes (earthworms) as they involve several parameters: test duration, fate of pollutant, interaction between pollutant and sub­ This group has undergone considerable lethal and strate [ l O 1]. Maturation of secondary sexual character­ sublethal testing [52, 135]. istics is often combined with earthworm growth rate The species Eisenia fetida andrei from the E. fetida [119, 145, 165, 199]. complex is generally used for lethal testing, as it is genetically relatively uniform [15]. Sorne results are On the other hand, several studies propose to use shown in table V. However, other studies have shown toxicant kinetics and critical body residues (CBR) in relation to toxicity endpoints, which may provide a that E.fetida worms with a long-term history of expo­ sure to cadmium developed resistance to the metal better estimate of toxic dose than soil concentration of [146]. The species Lumbricus terrestris and Allolobo­ contaminants, especially for hydrophobie organic chemicals [60, 61, 114]. phora caliginosa, although more sensitive, are less fre­ quently used as they are harder to raise [39, 56, 79, 80, 127, 168]. 3.1.1.4. Gastropods Sublethal testing is generally performed on cocoon production [24, 119, 132, 145, 185, 188-191. 199]. Experimental studies on metal tox1c1ty have been Juvenile growth studies are more difficult to perform performed on snails and slugs from the stylommato-

Table V. Using the Eiseniafetida group as a toxicological bioindicator of effect.

Reference Pollutant Contamination source Test duration Parameters

[80] Nineteen pollutants fongicides con tact ( fil ter paper). 2 and 14 d LCso and insecticides artificial soi!, artisol [ 1 J [133] Cd, Cu, Ni, Pb, Zn; contact (filter paper), 2 and 14 d LCso ten organic pollutants artificial soi! [145, 199] Dieldrin organic culture medium 90 d Growth, sexual maturity and reproduction

[140] Five herbicides contact (filter paper), brown soi! 2 and 7 d LC50 ; growth (biomass) [184] Cu, pentachlorophenol, artificial soi! 21 and 35 d Growth, reproduction 2,4-dichloroaniline [132] Ten pollutants horse manure + sand 28 and 42 d Growth, reproduction

[185] Cd, Cu, PCP artificial soi! + cow dung 6, 8. 10 and 12 weeks EC50, NOEC. growth, reproduction [109] Parathion, propuxur artificial soil 14, 42 and 56 d Growth, reproduction [186] Cd, Cr, insecticides, fongicides artificial soi! 14, 21 and 35 d LC50; growth, reproduction and herbicides [16] Soi1s and waste artisol 14 d Non-toxic threshold

[60] Pentachlorophenol artificial soi! 28 d LC50; ILL [201] Terbuthylazine, carbofuran food 14, 42 and 56 d Reproduction, respiration (CO2 emissions), excretion (nitrogen)

Table VI. Examples of cxperirncntal studics for dctermination of lcthal and sublethal cffiècts of chemicah in land gastropods pulmonatcs.

Rcference Species Test parametcr Pol lutant Exposure

Duration Fecding te:,,,t, 11511 He/i_r t1sperso Growth; fceding activity; Cd ."\Il d Rat food+ CaCO, +CdCL rcpn>Lluction [ 12.+j Arion arer Growth; f"od consurnption Cu. Hg. Ph. Zn 27 d Natural dièt \\ ith 1.5 ','-( agar+ di!Terent do:,,,age:,, of rnc-tal...., f 1571 ff. (/.\/)('/"\(/ NOAEL. LOAEL; sublethal Twclvc pcstici,ks 12. 1 + d rat fond+ agar + pc:--ticidc.., EC,,,; klha\ity: LC',11 11251 .-1. li/l'i" ()uantitali\ c structure Hg :\(J d Natural dict with 1 � <.; agar+ nf Liigc\tivc tuhuk:-- ditlcrcnt Hg cnnccntrations 1115[ fi. ll.\/Jt'rsa Growth rate; food consumption; CJ. Cu. Ph. Zn 90 d (_jU\cnilcsJ; ,\nificial dict+ dilfrrcnt i"ccumlity 120 d (adults) mctal c·Dnccntrations [69[ Il. mpersu ffl/>er.rn (irowth Cd. Cr. Ph. Zn 21. 28 d hrnd lleli.xcd R· � Cd or contaminatcd soi! !Cd. Cr. Ph. Zn 1 [70[ H. mpe1 '" mper.rn; Growth. \:OEC. EC,,,. EC:,-- Cd 28 d Dier for snails 1Hclixal R'•) + CdCI, H. tnpena mtP.:imu feeding activity ______,. ___ phorous pulrnonate gastropod group (table VI). These According to Krogh and Petersen 1105]. reproduc­ organisrns can survive high levels of heavy rnetals in tion is a better pararneter than mortality and provides their food. However. growth and reproduction are more accuratc information. This parameter integrates inhibited differently depending on the metal and spe­ the possible long-terrn effects of a pollutant by demon­ c1es. strating the possible changes which may occur in the species fuwre reproduction capacities. Schuyterna et al. [ l 57] studied toxicity of pesticides on snails and identified four substance groups accord­ ing to their toxicity through ingestion mixed with food. 3.1.1.6. Isopods In group 1. acephate. atrazine, glyphosate. hexazinone 1 and piclorarnwere not lethal for snails at 5 000 mg-kg • The most frequently used species are Porce!!io The death rate was 13 7c for group 2 ( carbaryl) and scaher [31. 32. 35. 59. 194, 198]. Oniscus ase!lus [31, 75 % for group 3 (fenitrothion and rnethyl parathion). 1941, Tricl1011i.1cï1s pusill11s [1941 and P laevis [1341. These high relative differences in pesticide toxicity According to Crommentuijn et al. [31 ], care must indicate the need for additional studies to evaluate the be taken in studies involving isopods. particularly risks involved in pesticide treatments. P scaher and O. asell11s. Thcse specics can accumulate high concentrations of pollutants and lethal cffects occur only whcn their accumulation capacities are 3.1.1.5. Collernbola fully saturated. Exposurc timc to a pollutant plays a kcy role. The Collembola Folsomia candida is the most fre­ lsopods generally show low sensitivity in sublethal quently used microarthropod both for sublethal and lethal tesh which increases test duration. Drobne [48] esti­ testing. Testing can be performed for organic or metallic mates that growth. reproduction and life cycle studies pollutants. Sorne exarnples are shown in table VII. arc not usually the best for isopods. ln fact, growth F cczndida is currently used as a mode] for comparative rates can vary over a period of several weeks. even for studies of new molecules behaviour. Data obtained on one individual. and females may retain sperm for a this species can also be used for improving testing long lime before reproducing. Furthermore. lite cycles techniques 1106]. are relatively long, often more than 6 to 8 months. Howcver, sublethal tests performed with copper on Although one test using F candida has been standar­ P srnher revealcd neŒative effects on food intakc and 1 dised. most publications on this species indicate widc individual growth at high concentrations ( 1 000mg-kg- ) differences between test results depcnding on strain. while low amounts stimulated juvenile growth rates individual age [33) and soit moisturc content [182]. ln [59]. ln addition. thesc animais arc easy to raise. addition. this parthenogenic species is easily raised. but does not provide an accurate picture of natural environments. This is why other spccies, such as lso­ 3.1.1.7. Arachnids toma viridis are also studied [95]. Tests have also been performed on Orchesella cincw [5. 142. 197]. Onv­ Lethal and sublethal tests were performed on chiurus annatus [7. 177] and/. notabilis [ 177]. Oth.er Platynothrus peltifer for copper and dimethoate [179]. species should also be �tudied to generate a comple­ The rcsults have been encouraging and it may be possi­ mentary test pool. ble to use this acarid. although cmTent study methods

Invertebrate soil fauna as pollution indicators

Table VII. Using Folsomia candida as a toxicological bioindicator of effects.

Reference Pollutant Contamination Duration Variable Parameter [18] DDT Food (yeast) Temperature; Life span. mortality, fecundity, concentration egg viability [176] Seventeen insecticides Artificial soil (sand) 1 d Concentration LDso [170] Paraquat, atrazine Food (yeast) 154 cl Concentration Mortality. moulting frequency, fecunclity. egg viability. incubation

[30] Cel Artificial soi! (OECD) 63 cl Concentration LC5,i: EC50 on growth and egg-laying

[103] Dimethoate Methylated soi! 28 cl Spatial distribution LC50; EC50, NOAEC, LOAEC of food and pollutant on reproduction

[33] Cel, chlorpyrifos, Artificial soi! (OECD) 35 cl Genetie variability LC50; EC50 on growth, juveniles triphenyltin hyclroxicle and population

[32] Cd Food (yeast) 63 d (egg-laying); Specific variability LC50: EC 10, EC50 on egg-laying 392 cl (growth) and growth; SS! [127] Dimethoate Artificial soi! (OECD, 32 cl Soi! type NOEC on juveniles humic, sandy and clayey)

[162] Zn Artificial soi! (LUFA); 42 cl Soi! type LC50; EC50 on growth natural soi! and reproduction

(149] Dimethoate, parathion Artificial soi!, water 14 d Comparison of measure- EC50 on mobility in water ment methocls

[891 Parathion, dimethoate. Water 14 d Di lferences between EC50 on mobility in water carbofuran, oxamyl pesticides

[180] Cd, Zn Artificial soi! 42 d Interaction between Cd LC50; EC50 on growth and Zn and juveniles

[182] Cd Artificial soi! (OECD) 42 d Soi] moisture LC50; EC50, EC10 on biomass .juveniles

[163] Zn Artificial soi! 42 d Tcmperature; LC50; EC50, EC 111 on biomass. concentration juveniles

are time-consuming and difficult(highjuvenile mortal­ To overcome the disadvantages inherent in each type ity, lack of biological synchronisation). of test, several assays should be run using different Work has also been performed on the effect of zoological groups as well as several species from the dimethoate on the predator gamaside Hypoaspis acu­ same group to create a test battery and an evaluation leifer [62, 78, 103, 104). These tests have demon­ grid for toxic product hazards. In addition, the most strated the importance of examining between species sensitive variables for a given species should not be the relationships, particularly predator-prey interactions, only ones examined [99). Severa) studies have already as test results indicatecl that dimethoate essentially been performed using the cadmium model (table VII[): affects the gamaside indirectly due to a clecrease in these test results should be numerically evaluated. number of available prey (i.e. the Collembola Folsomia Dutch scientists have attempted to design an index fimetaria). which integrates toxicological data for several organ­ isms: Van Straalen and Denneman [192) suggested the HC5 index (hazardous concentration for 5 % of the 3.1.1.8. In summary species) to harmonise and correlate toxicological data based on NOEC for different organisms for a given Irrespective of the chosen organism, the tests must pollutant. However, as Hopkin [85) has pointed out, be replicable and standardised according to: this index was designed using a very small number of organisms and suffers froma Jack of toxicological data - species (genetic stability and growth stage); for different organism types. In addition, it fluctuates - breeding techniques and contamination method; widely depending on the species analysed [5). - experiment duration;

- measurement of effects (NOEC, LOEC, EC50, etc.). 3.1.2. Exposure biomarkers and physiological changes Substrate is a vital element, as an organism's reac­ tion is influenced by substrate composition, which can The search for exposure biomarkers which react to interact with pollutants. Every test made with natural, pollutants is currently under way, for both plants and artificial(ISO defined soil) or inert (silica, glass beads) animals. The goal is to define the measurable molecu­ substrate must be double checked by tests using the lar and biochemical changes which occur after expo­ substrate alone as a standard. sure to toxic substances [ 41, 50. 111-1131.

Table VIII. EC,0 values and NOEC for the cffect of cadmium. one of the most toxic among the heavy mctals rolluting the soi!, on the growth. the rerroduction and the survi,al of soil in,ertcbrates.

Refercnccs Srecics Test rararneters 1-:xposurc Rc'-ïult:- {mg·kg ' dry w tl Durati,rn Routes NOEC FC,0 Ncmatodes llJXI Plectus m1rminatus ratio JU, en i le/adu lt .l weeks artitïcial soi! < 10 J2I ± 1.7 10ECDl Annelids Oligochacta [ 1231 Lumhricu.\ nrhe!lu., -;unÎ\•al 6 weeb loamy sand 10 'i(l(J cocoon production 12 weeks 1183] Ei.,eniu Wl(frci survival .l wceks artitïcial soil > 1 ()()() [ 18'il fi 011drei "om,1tic gro\\•th -l2. 'i6. 70. artificial �oil 18--32 3 >- l)(, 8-l d + L'O\\. dung " sexual dc,clopment < 10 � 1 !08 Apterygota - Collcmbola 1 lJ-35 d arti!ïcial soi! 778 -lJ77 [33[ h,!somia condhlu :--urvi\·al grovvth 23 .l'i d .176-807 reproduction 3-l.8 and .<26 159 and> 326 1321 F candido growth 63 d rood 1 687 cgg.., 1 J)7 [321 Orchesc/1" cincw �Jo\vth and cgg.., ô.1 d r,,od > 8'i.5 Cru:-...tacca hopoda 1321 Pon-el/io scaba growth (13 d food 195 Arachnidea Orihatida 1:121 l'iul_rnorhrus 1wl1i/à cg.i;� 6.l d 1·ood 2 LJ .110 Molluscs Gastropoda 1691 lleli.\ osper.-,a a.\/JCr\"a grnwth 28 d food :iO 1-l0 fi. O\'fh'rSCl ll/U.\ill1Cl growth 28 d food 50 120

Current works are examining: ity of measuring DNA adducts by i:'p post-labelling in - induction of proteins in biotransformation and L. terrestris exposed to more or Jess strongly polluted detoxification systems ( cytochrome P450. conjugation soils. Determining DNA adducts was considered to be enzymes. etc.) or more or Jess specific stress proteins a good way of detecting genotoxic substances in soils. (metallothioneins. catalases. etc.): CytologicaL physiological and behavioural modifi­ - specific target molecules (acetylchlolinesterase. cations can also be considered to be valuable hiomark­ DNA. etc.). ers. In a studv of snails whose food was contarninated Tranvik et al. 1178] focused their study on allozyme by Cd for 30 �I. Russel et al. 1151] ohserved hyperplasy polymorphisms in the springtail Orchcse//a bifasciata and necrosis of the epithelium and a disorganisation of and found no significant differences between several the sperrnatozoid flagella in the ovotestis. although the populations subjected to different degrees of metallic ovocytes die! not appear to be affected. ln a study of stress. However. Frati et al. 166] observed differcnt Arion a/er slugs. Marigomez et al. 11251 ohserved sig­ reactions to heavv metals in several loci in tolerant and nificant quantitative modifications in the digestive tube sensitive O. cincta populations. structure and hlood vesse! walls depending on Hg concentration in food and exposure time. Studies on Due to their induction bv metals. metallothioneins isopods have examined the effects or metals on the can be simultaneously respo'nsible for detoxification of digestive gland epithelium and hepatopancreatic reac­ trace elements. such as cadmium. and regulation of tions of Porce!lio srnher149, IOOJ. essential trace elements. They could be used as bio­ markers for environmental pt;llution. Berger et al. 19] A good exarnple of a physiological effect is the neu­ isolated metallothioneins in land snaib (Ariwzta arbus­ rotoxic effect (disturhances in nerve impulse speed) of torum) which had been fcd Cd contaminated food pollutants on earthworms 143--471. Nevertheless. this (lettuce) and sequenced two isofonns (Mta and Mth) in vivo method is rather costly 11()11. which they compared to the structure of other inver­ Behavioural studies are essentially ways of quantita­ tebrates. Other metallothioneins have been identified tively evaluating the effect of pollutants: e.g. measur­ for the Collembola O. cincta 181] and the eanhworm ing the effect on hurrowing hehaviour in earthworms Lwnbricus rubellus [ 169]. exposed to carhamate in microcosms [ 174 ]. On the f Walsh et al. [203] studied early, sensitive markers other hand. Collemhola dispersal can be afected hy for pollution in carthworms. They verified the feasibil- metallic pollution in soils 11601 and some isopods,

Invertebrate soil fauna as pollution indicators such as P. laevis, can identify and avoid leaves contam­ closely studied as they affect species survival. Biomar­ inated with high cadmium sulphate levels [134]. ker studies will help explain how pollutants internet in organisms, both at cellular and subcellular levels, to understand how toxic substances affectgenomes. Stud­ 3.1.3. Malfunction of soil biological processes ies on how soi! processes malfunction should be extended and diversified to involve multi-disciplinary Few studies have compared the interruption of bio­ research teams to examine a wide number of para­ logical process in contaminated soils and soil fauna meters and thus better analyse the different factors biological reactions. These changes are difficult to involved. evaluate in the field as soil biological processes are quite complex and involve many different factors, including soil fauna. However, their activity is essen­ 3.2. Ecological indicators tial for the whole ecosystem and must be maintained for ecosystem survival. Severa! studies have attempted to extrapolate labora­ Laboratory studies have attempted to analyse the tory results to the field. Diekkrüger and Rôske [ 40] different biological processes, particularly changes tried to simulate collembolan population dynamics caused by a pollutant on soil fauna. using Isotoma notabilis in agricultural fields based on data obtained in the laboratory on this species ecophys­ Salminen et al. [154] have demonstrated that the iology. L0kke [120] also tried to link laboratory exper­ herbicide terbuthylazine can provoke changes in iments on the dimethoate mode! with experiments microarthropods introduced into forest soils by effect­ using mesocosms with different collembolan species ing trophic interactions and thus indirectly disturbing (Folsomia candida, F.fimetaria,/. notabilis). soil functioning. However, the same experiments were performed with PCP and did not yield any significant Although the first results have been encouraging, results [153 ]. Komulainen and Mikola [102] observed results obtained in laboratory experiments cannot yet a clear reduction in respiration and nitrificationin soils be extrapolated to the field. Spurgeon [ 166] noted that polluted by heavy metals, although the role of soi! a particular earthworm, Eisenia fetida, could live in mesofauna could not be defined. The effects of differ­ soils containing much higher concentrations of metals ent pesticides on litter-bag microarthropod coloni­ than the LC50 and EC50 values found in the laboratory. sation and maize decomposition in a maize field yield Since it is impossible to control ail the different fac­ complementary information, as some pesticides tors affecting soi! fauna, bioindicators of ecological strongly effect microarthropods but had no visible effectsare used. effect on litter decomposition [28]. Studies to evaluate the in situ impact of different Kula and Rômbke [ 107] have compared methods for types of pollution on animal groups have been per­ testing organic matter breakdown in the field (bait lam­ formed for soil microarthropods and earthworms. ina tests, litterbags and litterboxes, minicontainers, Both synchronie (two similar polluted or unpolluted cotton strip). Different criteria have been used: ecolog­ sites) and diachronie (measurements performed before ical and ecotoxicological (in terms of functional and after pollution) comparisons must be made. In importance fore cosystems, realistic exposure or sensi­ general, these two approaches are combined: this pro­ tivity), practicability (in terms of possibilities of stan­ cedure is well known as before-after impact control dardisation or cost effective) and evaluation (in terms (BACI) design [93]. In fact, different populations often of reproducibility or variation coefficient). Bait lamina reveal natural fluctuations which can be due either to test was recommended to study biological activity standard (climate, photoperiod) or exceptional (severe parameters. The authors pointed out the urgent need cold, drought) abiotic factors or to biotic factors for further studies on the litter decomposition parame­ (between and within species competition, individual ter, especially to improve test reproductiveness. species life cycle). These different factors are like Earthworm activity, which is usually responsible for background noise for a study site and can hide the fragmentation and incorporation of organic malter into effects of a pollutant. They must be quantified as accu­ the soi!, is modified by pollutants [26]. These changes rately as possible. have been measured for Lumbricus terrestris by evalu­ Comparisons are generally based on two types of ating the disappearance of litter or other bait in a criteria: Daniel funnel [ 10]. - those involving population density, i.e. species, population or community abundance or biomass; 3.1.4. Conclusion - those involving population structure, i.e. specific richness or diversity. The lethal effects of severe changes (disappearance Sampling techniques for microarthropods can vary: of certain species) and particularly the sublethal effects boring cores, litter-bags [34] or pitfall traps [74, 96]. on the most sensitive and important phases (growth Results obtained using these techniques are often quite and reproduction) in the animal life cycle should be different.

In fact, different studied species often react differ­ either a trophic or a sexual basis. This is particularly ently to a given pollutant. Frampton et al. [65] demon­ interesting in the field of ecotoxicology. Howevcr. it is strated the different reactions of Collembola specie;. difficult to rank specics whose biological cycle is not subjected to different culture pressures: while Fol.1·0- totally understood. This is wherc lahoratory diYersity mia quadrioculata was shown to benefit from inte­ studies are vital, particularly for studies of a large grated culture systems, the genus LPpidocvrtus reacted numhcr of taxa. better using standard systems. Results analysing the role of mesofauna activity on In addition, apart from the pollutant itself, the pol­ changes in organic matter arc both highly varied and lutant and the site's history can affect the reactions of a irregular, retlecting the cnmplexity of decomposition microarthropod cornrnunity. Sahatini et al. [ 152] found processes which an: only partly dependent on soi! no significant differences hctwcen microarthropod fatma [158. 175]. In addition. no studics using the litter populations in sites treated with atrazine and untreatecl bag method havé been able to signitïcantly C(melatc sites when the study site had been exposcd to the pol­ decompŒition of organic malter and microarthropod lutant for scveral years, as pollution-sensitive specics density. haclprobably vanisheclprior to the study. Currently. there is no standardised method for in situ Diversity indices shoulcl be used carcfully: as studies of rnicroarthropods. Ramade l 143] pointed out for the Shannon index, at Differcnt extraction techniques have been used for sublethal doses, the dominant pollution-sensitive spe­ earthworms: hand sorting [23 ]. formaldchycle [144] cies can decreasc, without disappearing. which para­ and the cthophysical method. cloxicallymay increasc the index. These indices should only be used in extrerncly polluted sites. Most earthwonn studics have only examined lethal toxicity of pollutants l:'i-1-. 55. 117). This is an illustra­ Taxonomie diversity alone is not enough to evaluatc tion of the difficulty of seriously cstimating the rcal a site·s biodiversity. Coenotic diversity, which expres­ toxicity of differcnt pollutants. as a lot of diffcrences ses the distribution of taxa within a species. should occur hctween study protocols. also be consiclered [ 19]. During the Sheffield ecotoxicology workshop, scv­ Cortct [ 271 has demonstrated significant differences eral proposais were made to standardise and co-ordi­ between two eu I turc system;. ( standard and integrated) natc certain rcsearch topics 1:'i 1. 109, 1 181 (tc1h/c IX). for microarthropod biodiversity in Normandy (France) lt should be nnted that field studies primarily use using an index which evaluated taxonomie diversity, r specitïc divcr;.ity, abundance, taxonomie richness and mcthods clcrived f om ccolog:y. Howevcr. it is vital to spatial distribution of taxa [20]. devclop spccific methods for ecotoxicology, whose goals arc differenL Stuclics must be made on diffcrent Normality, in the mathematical scnse of the word, is types of organisms to de�cribc species or groups which not a rcliable criteria cither: highly disturbed commu­ could be used as bioindicators in the field. Thcsc stud­ nities can have a log normal distribution, whosc domi­ ies should lead to the standardisation of rcsearch pro­ nant species or groups are not the samc as those founcl cedures. In the lonŒ run, an evaluation Œrid should be for stressed communities. Hagvar [ 77] i nsists that rare designed which i;,cludes many diffe;ent types of species should also be considered, as they cvolve organisms and would help creatc a biotic index of soi! towards the lower dominance classes if thcv are sensi­ ' quality tu objectively quantify pollution. tive and towards hiŒhcr� clominanœ classes if thev are tolerant. ' Frampton 164] also suggests using an indicative spc­ 4. CONCLUSION cies pool with a knovd1 sensitivity to certain pollutants for field studies. U sing a single species cou Id obscure This paper has reviewecl the main data available inter-specics relationships. whilc consideringf ail the species in a system will not rcveal efects on rare about hov. pollution effects soi! fauna invertebrates species, which can be more affected than abundant used as indicator organisms. Therc is no ideal inclicator spccics. These conclusions shoulcl be treatecl with cau­ l 83): howcvcr, enchytracids. earthworms. Collem bol a, tion, however. as different statistical analvsis tech­ acaricls, isopod-, and pulmonate gastropods have pro­ niques. such as correspondence analysis. no�v make it vided additional data on the three bioindicator cate­ possible to evaluate the influence of hiQhlv� , abundant gorics (bioacrnmulation. toxicological and ccological species compared to rarer specics. consequcnL·cs L The functional aspect must also be considercd, par­ Taxa which could be used as bioindicators, in tcrms ticularly in tcrms of the influence of pollutants on spe­ of rneasurahlc responses. are shown in tah/c X. cies hiological cycle;.. Howevcr. these biological Bioaccurnulation measurements provide data on the cycles must fïrst be characterised and the different taxa hioavailability of pollutants in the environment and must be ranked. Siepel 1159] designed an evaluation make it possible to evaluate species sensitivity to xeno­ grid for rnicroarthropod life cycles which differcnti­ hiotics [31 J. The currcnt Jack of standardiscd measurc­ atcd betwecn twelvc different lite cycle stratcgies on menl proc,:dures for hioaccumulation will hopcfully

Invertebrate soil fauna as pollution indicators

Table IX. Summary of suggestions for field study standard protocols [ 174]. *** Undefined characteristics.

Edwards [51] Lofs fi 18] Kula [108] Site grass, arable land *** grass. arable land, artificial site Soi! description texture, OM, homogeneity texture at least texture, OM, pH, pF History > 5 years without pesticides *** *** Additional parameters temperature, rainfall 2 Density > 100 ind-m2 > 100 ind·m2 > 100 ind-m Representative species at least the six most common species one endogeous + one anecic Lumhricus terrestris; Aporrectodea caliginosa Initial plot size 5 to 10 m2 *** 100 m 2 Number of replicates > 4 per treatment > 4 per trcatment > 4 pcr treatment Pesticide standard pesticide + physical physical and chemical toxicity physical and chemical toxicity and chcmical toxicity parametcrs parameters parameters Dosages used the most important recommended *** *** Treatment dates spring earthworms activity period spring Estimated criteria abundance and biomass abundance and biomass; abundance; biomass with reserves growth stages Sampling methods hand extraction (0.25-1 rn2 ) *30-60 cm; non-destructive extraction; formaldehyde extraction; electric 2 (sample size) formol extraction (0.25-1 m2); mixed *** extraction (> 0.25 m ) 2 extraction (0.25-1 m ) *5-10 cm

Number/duration of sampling > 4/*** ***/ 1 d > 2/*** Experiment duration > 6 months 4-6 months 12 months Sampling date Monthly and > 2 years for remnants 1, 4, 6, 12 months after treatment of pesticides ( 1, 2, 4, 8, 12. 24 months after treatment)

Table X. The use of different taxonomie groups as bioindicators. (-), No available studies; -. use unlikely; +, possible use under certain conditions; ++, interesting for use.

Nematodes Enchytraeids Earthworms Gastropods Collembola Acarida lsopods Bioaccumulation Toxicological Exposure biomakers bioindicators Neurological of effects disturbance Mortality Reproduction Function Ecological Abundance bioindicators Biomass richness Ali groups can be used of effects Diversity

lead to the development of an appropriate methodo­ be continued and expanded to species living in differ­ logy [137]. ent soil layers to design field study methods. Toxicological data from laboratory and field studies Indicators of toxicological or ecological effects pro­ should be used to evaluate environmental hazards vide information which should help develop a series of linked to pollutants. Organism species of ail sizes tests which can be used not only with earthworms [ 16], (microfauna, mesofauna and macrofauna) and diver­ but also with other invertebrate species to localise sity living in the soi] should be used to design micro­ pollutant effects at different levels in the lite cycle cosm and mesocosm models reflecting the diversity (figure 1). and the vital trophic relationships which characterise a living soi!. Single species tests are a step towards envi­ In the near future, the various existing tests and ronmental risk evaluation, but progressively more those soon-to-be standardised should provide a means complicated systems should be designed to analyse of evaluating the environmental quality of different interactions between the environment and the creatures soils. Microcosm and mesocosm experiments should which live there.

___.,-:�-:__�-� - COMMUNITY POPULATION ���- INDl�DUAL ',OR VITAL (C FUNCTION CELL survival integrity growth abundance diversity ! enzyme activity reproduction function distribution dominance mobility age structure · successional \:.·· activity '��>"�-

role in soil

SOIL AND LEACHATE QUALITY

·Figure l. The differcnt ]evcls of polluwnt acti,ity.

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