EcotoxicologicalEcotoxicological teststests..ImpactImpact ofof leachatesleachates fromfrom municipalmunicipal landfills landfills

on underground on underground waterswaters qualityquality..

Agnieszka KuczyAgnieszka Kuczyńńskaska, , Lidia WolskaLidia Wolska,,Jacek NamieJacek Namieśśniknik

GDANSK UNIVERSITY OF TECHNOLOGYGDANSK UNIVERSITY OF TECHNOLOGYCHEMICAL FACULTYCHEMICAL FACULTY

Gdańsk, 05.10.2004 r.Gdańsk, 05.10.2004 r.

BiologicalBiological equilibriumequilibrium andand itsits disturbancesdisturbances atatvariousvarious levelslevels ofof biologicalbiological organizationorganization

Impact of stress-causing factors on the state oforganisms biological equilibrium

no is e

enviro nmental po llutio n

ele ctro magne tic fie ld

phy sico che mica l facto rs (te mpe ra tu re , humidity)

ORGANISM

his to patholog ic al les io ns

g rowth inhibitio n

de ath

b ioch emica l re s po ns es

BiologicalBiological equilibriumequilibrium andand itsits disturbancesdisturbances atatvariousvarious levelslevels ofof biologicalbiological organizationorganization –– contcont..

Hierarchial response of organisms to environmental stressors[Adams S. M., Water, Air, and Soil Pollution, 123, 103 (2000)]

EvaluationEvaluation ofof thethe degreedegree ofof environment environment pollutionpollution

CHEMICAL ANALYSIS• the number of constituents to estimate,

• varied concentration levels,

• fluctuations of pollutant concentrations in time and space,

• complex composition of the matrix,

• complicated, ie. time- and labour-consuming procedures for preparing samples for analysis,

• further pollution of the environment by the chemical reagents used,

• additional costs connected with the necessity to purchase high-purity reagents and the need to use or utilise the surplus,

• problems connected with obtaining appropriate reference materials, necessary for such processes as validation of analytic procedures and calibration of test and measurement devices.

Obtained information isvery detailed, but insufficient for the

assessment of the degreeof environment pollution.

EvaluationEvaluation ofof thethe degreedegree ofof environment environment pollutionpollution –– contcont..

CHEMICAL ANALYSIS TOTAL INDICES BIOASSAYS

Obtained information is very detailed, but

insufficient for assessment of the degree of

environment pollution.

They supply general information, which are not easily connected with the toxicity of the analysed environmental element.

Obtained information isgeneral, but

comprehensive answer is given if the environment impacts infavourably on

living organisms.

BIOTESTBIOTEST (gr. bios – life + lat. testari – to testify)

- an analytical technique relying on observation of a response at a given level (or levels) of biological

organization, as a result of disruption of homeostasis, following changes that take place in a given element of

the environment (appearance of the pollutant).

The purpose of a biotest is to prove the presence of a toxic substance in the environment or to find out how harmful it is by quantitatively evaluating the influence

of this substance on a living organism (based on comparison with a control sample).

BiotestBiotest

phytotests

bacterial tests

based on the application of animal organisms

BiotestBiotest –– contcont..

BacterialBacterial teststests

Microtox®AZUR Environmental (USA)

ToxAlert 100®Merck (Germany)

Colonies of bioluminescence bacterial strains (species: Vibrio fischeri) photographed in the darkness

BacterialBacterial teststests –– contcont..

Scheme of enzyme reaction of luciferin’s oxidation

Scheme of action of bacterial test

ToxkitsToxkits

TheThe productproduct rangerange

test species duration endpoint standard

Tests for freshwater and terrestrial environments

ALGALTOXKIT FTM Selenastrum capricornutum 72h growth inhibition OECD,

ISO

DAPHTOXKIT FTM magna Daphnia magna 24-48h acute toxicity OECD, ISO

DAPHTOXKIT FTM pulex Daphnia pulex 24-48h acute toxicity OECD

CERIODAPHTOXKIT FTM Ceriodaphnia dubia 12h acute toxicity US EPA

THAMNOTOXKIT FTM Thamnocephalus platyurus 24h acute toxicity -

ROTOXKIT FTM 24h acute toxicity ASTM ROTOXKIT FTM short-chronic

Brachionus calyciflorus 48h short-chronic toxicity AFNOR

PROTOXKIT FTM Tetrahymena thermophila 24h chronic toxicity OECD

OSTRACODTOXKIT FTM Heterocypris incongruens 6 dni chronic toxicity -

Tests for estuarine/marine environments

ROTOXKIT MTM Brachionus plicatilis 24-48h acute toxicity ASTM

ARTOXKIT MTM Artemia salina 24-48h acute toxicity -

ToxkitsToxkits –– contcont..

no further need to support a culture all the time

low unit cost of sample analysis

the possibility to test several samples simultaneously

short response time

small volume of the tested sample

small amount of laboratory space occupied by the test package, and finally

the possibility to use the test in the field

TheThe assessmentassessment ofof impactimpact of selected of selected municipal municipal landfillslandfills, , placedplaced inin thethe

PomeranianPomeranian voivodshipvoivodship,, on the on the ecotoxicityecotoxicityof underground watersof underground waters

-- preliminarypreliminary projectproject

((thethe researchresearch performedperformed withinwithin thetheframeworkframework ofof ccontractontract

No.:No.:WFOS/D/366/20/2003)WFOS/D/366/20/2003)

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

Geographic localization of municipal landfills included Geographic localization of municipal landfills included inin tthehe ecotoxicologicalecotoxicological assessment.assessment.

BIERKOWO

LUCIN ŁEBCZ

LINOWIEC

BĄDKI

ROKITKI

RYBSKA KARCZMA

Daphnia – a waterfleaVibrio fischeri

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

BioanalyticalBioanalytical toolstools......organism test standard

acute toxicity

Daphnia magna Daphtoxkit FTM Magna (Crustacean Toxicity Screening Test for Freshwater)

ISO 6341 MicroBio test performed according to producent’s requirements.

Vibrio fischeri ToxAlert 100® (Merck, Germany) PN-EN ISO 11348-2:2002

chronic toxicity

Daphnia magna Assessment of chronic toxicity on the crustacean Daphnia magna

ISO 10706

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

Decree of the Minister of the Environment (dated: 11 February2004) - classification of the state of surface and underground waters,

(Dz.U. Nr 32, poz. 284)

German classification system for sediments (Krebs F., The pT-value as a classification index in aquatic toxicology. GIT Edition

Umweltanalytik-Umweltschutz, 32, 293 (1998))

Categories for management of dredged material Highest dilution

level without effect

Dilution factor

pT-value

Toxicity classes

seven- level

system Colour coding

Three-level assessment system

Original sample 20 0 0 0 1:2 21 1 I I 1:4 22 2 II II

unproblematic

1:8 23 3 III III 1:16 24 4 IV IV

critical

1:32 25 5 V V

≤ (1:64) ≤ 26 ≥ 6 VI VI hazardous

Helcom Recommendation 23/10, 23/11 i 23/12, adopted 6March 2002 having regard to Article 20, Paragraph 1 b) of the

Helsinki Convention

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

Classification of underground waters according to…

Piezometer Decree of the Minister of the Environment

Helcom Krebs

P2 (07.12.2003) V III P2 (23.02.2004) V IV P6 (07.12.2003) V VI P6 (23.02.2004) V VI P7 (07.12.2003) V I P7 (23.02.2004) V I P8 (07.12.2003) V 0 P8 (23.02.2004) V VI P9 (07.12.2003) IV 0 P9 (23.02.2004) IV II

P10 (07.12.2003) IV 0 P10 (23.02.2004) IV II

ClassificationClassification ofof underground underground waterswaters samplessamplesfromfrom thethe landfilllandfill inin LinowiecLinowiec, , accordingaccording to to variousvarious standardsstandards......

EcotoxicityEcotoxicity ofof underground underground waterswaters–– preliminarypreliminary projectproject

anilinechlorobenzene

Chromatogram obtained during the GC-MS analysis of underground water sample collected from a P-6 piezometer

(sampling site: Linowiec, dated: 23.02.2004)

SummarySummary

The results obtained using classical analytical methods cannot be a source of direct data regarding

the toxic effect (both acute and chronic) on living organisms.

However, results obtained using appropriate biotests can be a source of such information.

We should, therefore, consider the possibility of including ecotoxicological tests in examinations of the

quality of underground waters from waste dumps. This, however, requires earlier preparation of a proposal for

classification of underground waters, selection of appropriate ecotoxicological tests and implementation

research.

This presentation was supported by the “Centre of Excellence in Environmental Analysis and

Monitoring” which is a research project supported by the European Commission under the Fifth

Framework Programme and contributing to the implementation of the Key Action “Sustainable Management and Quality of Water” within the

Energy, Environment and Sustainable Development (Contract No.: EVK1-CT-2002-80010).

http://http://wwwwww..pgpg..gdagda..plpl//chemchem/CEEAM//CEEAM/

ThankThank you you for for youryour attentionattention