ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

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A. Kungolos 1 , V. Tsiridis 2 , M. Petala 2 , C. Emmanouil 1 , S. Kipouros 1 , D. Antoniadis 3 , G. Drakopoulos 3 , P. Dimakos 1 and K. Perakis 1 1 Department of Planning and Regional Development, University of Thessaly, 38334 Volos, Greece 2 Department of Civil Engineering, Aristotle University of Thessaloniki, 54006, Thessaloniki, Greece 3 NGO Epidro, Massalias 16, 10680 Athens, Greece Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

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Page 1: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

A. Kungolos1, V. Tsiridis2, M. Petala2, C. Emmanouil1, S. Kipouros1, D. Antoniadis3, G. Drakopoulos3, P. Dimakos1 and

K. Perakis1

1Department of Planning and Regional Development, University of Thessaly, 38334 Volos, Greece

2Department of Civil Engineering, Aristotle University of Thessaloniki, 54006, Thessaloniki, Greece

3NGO Epidro, Massalias 16, 10680 Athens, Greece

Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Page 2: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

How to determine the hazard resulting from pollution?

Chemical approach Chemical analyses of the pollutants

Biomonitoring

Bioassays: effects of chemicals, natural waters, pollutants, wastes,…on aquatic and terrestrial living organisms

Page 3: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Why do we need Monitoring?

Water quality may be affected by: Spills of oil and industrial products from tanks, pipelines

Pesticides from agricultural area, leaching pathogens

Endocrine disrupting chemicals

Neurotoxins, hepatotoxins from algae blooms

Contamination from terrorist attack (toxins, microbes, viruses, radioactive compounds)

Accidents, sabotage etc.

Page 4: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Why do we need Biomonitoring?

It is impossible to analyze all chemical substances, which are brought into nature by human

Scientists suggest that there are more than 100 000 harmful chemicals

Even with the most advanced instruments it is not yet possible to detect them all

With the help of biomonitoring, a much broader view of the possible dangerous effects can be detected

Page 5: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Toxicity testing used in our Lab in the University of Thessaly, Greece

15 min inhibition test of the photobacterium Vibrio fischeri

72 h growth inhibition test of the microalae Pseudokirchneriella subcapitata

24 or 48 h immobilization test of the crustacean Daphnia magna

Page 6: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Normal bioluminescence

Contaminated sample is added!!

Bioluminescence inhibition

Photobacterium Vibrio fischeri (Microtox test)

Photobacterium Vibrio fischeri

Bioluminescence measurement at exposure time 5, 15 and 30 minMicrotox M500 analyzer

Page 7: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Microalgae Pseudokirchneriella subcapitata(Algaltoxkit F)

Immobilized algae

mobilization

Measurement of the optical density (at 670 nm) after 72 h

Page 8: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Crustacean Daphnia magna (Daphtoxkit F magna)

Measurement of the immobilized/dead organisms

Addition of the samples and live organisms in multiwell test plate

Exposure for 24 h at 20 oC

Hatching of the ephippia

Page 9: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Toxicity tests have been used for:

Marine and fresh waters biomonitoring

Toxicity testing of wastewaters and soils

Toxicity testing of fly ash leachates

Toxicity assessments of pure compounds, heavy metals and

pesticides

Wastewater treatment plant applications

Applications of ecotoxicity testing

Page 10: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Correlation between the toxicity of lignite fly ash leachates towards D. magna and the concentration of Cr(VI) detected in the leachates

Correlation coefficient, r= 0.961Level of significance, p<0.01

Page 11: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Investigation of the interactive toxic effects of chemicals on live test organisms

MODE OF INTERACTION

•Additive if there is no significant difference between expected and observed effect

• Synergistic if the observed effect is significantly higher

• Antagonistic if the observed effect is significantly lower

Page 12: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Expected and observed effects

Expectedby a model based on the theory of probabilities

P(E) = P1 + P2 – P1P2/100

where P(E): the theoretically expected additive effect of a binary chemical mixture

Pi : the effect caused by a certain concentration of a

chemical when it acts alone

ObservedThe measured effect of the binary chemical mixture

Page 13: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Combined effect of Cu2+ and Zn2+ on V. fischeri(Synergistic effect)

0

20

40

60

80

100

A B C D E F G

Concentration combinations

Effe

ct,

%

Expected

Observed

Cu2+ Zn2+ (mg/L) (mg/L)A: 0.117 0.432B: 0.117 0.864C: 0.117 1.727D: 0.168 1.727E: 0.252 0.432F: 0.252 0.864G: 0.252 1.727

Page 14: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Comparison between theoretically expected and observed immobilization for the combined effect of metalaxyl-M and copper on D. magna

(antagonistic action)

0

20

40

60

80

100

A B C D E F

Concentration combinations

Imm

obil

izat

ion

, %

ExpectedObserved

Metalaxyl-M Cu (mg/L) (mg/L) A: 10 0.05 B: 10 0.10 C: 30 0.05 D: 30 0.10 E: 60 0.05 F: 60 0.10

Page 15: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

Current project on ecotoxicity testing

Ecotoxicity testing of selected lake waters and treated wastewater effluents

- Lakes Koronia and Karla- Treated wastewater efluents from Tyrnavos region

- Ecotoxicity testing using the photobacterium Vibrio fischeri, the crustacean Daphnia magna and the algae Pseudokirchneriella subcapitata

The research is funded by Ministry of Environment, Energy and Climatic Change/Green Fund programme: Environmental Research-Innovation- National Cooperations 2012”: “Improvement of monitoring water quality with the use of ecotoxicological assays”

Page 16: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

The only way to get an “integrated” answer to the hazard caused by pollutants to the environment, is to expose organisms of the receiving environment to these pollutants

Chemical analyses Bioassays Environmental

Hazard Assessment

+ =

Chemical analysis in many cases deals with a restricted number of compounds due to financial and technical limitations

For a “real hazard” monitoring, phenomena such as bioavailability, synergistic or antagonistic effects on the biota have to be examined

A single toxicity test does not tell whether other important groups of biota are affected by exposure to the same samples. It is thus necessary to use a battery of tests with species representative of the different links in the trophic chains

Page 17: Ecotoxicological assays as a means of monitoring vulnerable aquatic ecosystems

SECOTOX 2013 & CEMEPE Conference(Conference on Environmental Management, Engineering,

Planning and Economics)

Myconos, June 24 – 28, 2013

THANK YOU FOR YOUR ATTENTION!