Aquatic mesocosms for data validation : interest, limitations and recommandations Laurent Lagadic &...

Aquatic mesocosms for data validation :

interest, limitations and recommandations

Laurent Lagadic & Thierry Caquet

Équipe Écotoxicologie et Qualité des Milieux AquatiquesINRA, Rennes

[email protected]

Workshop ‘In situ trialing for ecological and chemical studies

in support of WFD implementation’Pau, France – 14-15 May 2008

Odum (1984) : "bounded and partially enclosed outdoor experimental setups ... falling between laboratory microcosms and the large, complex, real world macrocosms".

Lalli (1990) : "physically confined multitrophic and self-maintaining systems ... whose size is sufficient to enable pertinent sampling and measurements to be made without seriously influencing the structure and dynamics of the system".

Touart (1991) : "an intermediate-sized system ... that can be replicated and manipulated to test both structural and functional parameters".

MesocosmsDefinitions

Studies on populations (including recovery) and between-species interactions Studies on ecological processes

Long-term global scale changesLandscape impacts

BiomonitoringStudies onmechanisms of toxicity(lethal and sublethal effects)

In situ validation and definition ofbiomonitoring strategies in the field

Identification of specific responses

Identification of effect criteriaExperimental validation of tools

OUTDOOR CONDITIONSLABORATORY CONDITIONS

MicrocosmMesocosms

Experimental ponds

Artificial streams

Isolated species

Simplified food-web

Enclosures

Replicability

Complexity - Realism

MesocosmsDefinitions

From Caquet et al. (1989, 2000)

Natural ecosystems

Exposure conditions

Variability - Replicability

Recovery

Effect propagation

Conditions for use of mesocosmsfor data validation

Exposure conditionsInfluence of a tank-mix adjuvant on herbicide (fomesafen)concentration in water

0

50

100

150

200

250

300

350

400

0 48 96

Durée d'exposition (heures)

0

20

40

60

80

100

0 48 96


Con

cent

ratio

n en

fom

esaf

en (

µg/

L)

F1 (4,4 µg/L nominale)




MF1 (4,4 µg/L nominale)




Indoor microcosms Single species tests

No effect of the adjuvant on herbicide water concentration

0

50

100

150

200

250

300

350

400

0 4 8 12 16 20 24

Temps (jours)

Co

nc

en

tra

tio

n e

n f

om

es

afe

n (

µg

/L)

F3 (44,4 µg/L nominale)F4 (222,2 µg/L nominale)MF3 (44,4 µg/L nominale)MF4 (222,2 µg/L nominale)

ApplicationWith adjuvant

Without adjuvant

From Jumel et al. (2002), Lagadic et al. (2007) and Coutellec et al. (2008)

F3C

Cl

O

CONHSO2CH3

NO2

Exposure conditionsInfluence of a tank-mix adjuvant on herbicide (fomesafen)concentration in water

0

50

100

150

200

250

300

350

400

0 48 96


0

20

40

60

80

100

0 48 96


Con

cent

ratio

n en

fom

esaf

en (

µg/

L)









Indoor microcosms Outdoor mesocosmsSingle species tests

Without adjuvant

With adjuvant

No effect of the adjuvant on herbicide water concentration

0

50

100

150

200

250

300

350

400

0 4 8 12 16 20 24

Temps (jours)

Co

nc

en

tra

tio

n e

n f

om

es

afe

n (

µg

/L)

F3 (44,4 µg/L nominale)F4 (222,2 µg/L nominale)MF3 (44,4 µg/L nominale)MF4 (222,2 µg/L nominale)

ApplicationWith adjuvant

Without adjuvant

Obvious effect of the adjuvant

From Jumel et al. (2002), Lagadic et al. (2007) and Coutellec et al. (2008)

F3C

Cl

O

CONHSO2CH3

NO2


From Caquet et al. (2001)

Eff

ect

of

treatm

en

t

CV

Number of replicates

Vari

ati

on

betw

een

mean

s (%

)

-300

-250

-200

-150

-100

-50

0

1 2 3 4 5 6 7 8 9 10 11 12

10%20%

40%

60%

80%

100%

Variability in control mesocosms : CV = (s/x)*100


-175

Number of replicates needed to detect a significant reduction of one given effect criterion according to its variability in control mesocosms(t test; = 0,05, 1 - = 0,80)

HARAP – 1999

Recovery is defined as return of a measured parameter (e.g., the abundance of a population) to the normal range of the controls.

CLASSIC – 2002

“Recovery” means the return of a disturbed system to a status comparable to an undisturbed system.Recovery is possible inherently from within the system (autochthonous) or via recolonization (allochthonous).

Recovery has to be used by experts to determine the "Ecologically Acceptable Concentration" (EAC) for a particular compound.

RecoveryDefinitions

DensityHighLow

Low

High

Recolonisation potential

Adapted from Kedwards (2000)

MolluscsCrustaceans

DipteransColeopteransHeteropterans

EphemeropteransOdonates

RecoveryTheoretical relationship between biological traits and recovery

Stability of age structure HighLowGenetic diversity

HighLowVoltinismUni- Multi-

Dispersal capacity HighLow

Natural stochasticityHigh Low

Natural mortalityHigh Low

RecoveryRole of connectivity between static experimental ecosystems

Open mesocosmstreated (n = 4) & untreated (n = 4)

Covered mesocosmstreated (n = 4) & untreated (n = 4)

Principal Response Curve (PRC) analysis of abundance data

April 22, 2003 : treatment with deltamethrin, and covering

Monitoring of effects for more than one year (April ’03 - June ’04)

Study design

From Hanson et al. (2007)


Zooplankton

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

0.2

0.25

0.3

-50 0 50 100 150

Time after treatment (days)

Cdt

Control OpenTreated OpenTreated Covered

***

*********

* *

* *

*

** Other Rotifers

Cyclopoid Adults

Polyarthra sp.

Gastropus sp.

Daphniidae

Mytilina sp.Asplanchnidae

TestudinellaOstracoda

Other Copepodites

Chydoridae

-2.5

-2

-1.5

-1

-0.5

0

0.5

1

1.5

2

2.5

bK

Open control

Open treated

Covered treated

In both open and covered ponds, recovery occurredafter 40 days

From Hanson et al. (2007)


Benthic macroinvertebrates

Corynoneurinae

Chironominae

Tanypodinae

Baetidae

Physa acutaNematods

Dugesia sp.Lymnaeids

Oligochaetes

EcnomidaeOrthocladiinae

Herpetocypris sp. 1

-5

-4

-3

-2

-1

0

1

2

3

bk

-0.1

0

0.1

0.2

0.3

0.4

0.5

-50 0 50 100 150 200 250 300 350 400

Time after treatment (days)

Cdt

Control Open Treated Open Treated Covered

****

*

*

**

*********

******

***

***

***

***

**

Open control

Open treated

Covered treated

Recovery did not occurin the isolated ponds

Recovery occurred after 60 days in the open mesocosms

From Caquet et al. (2007)

Effect propagationLinking levels of biological organisation

In HCB-exposed snails :

Reduced individual growth

Increased production of eggs

Freshwater snails (Lymnaea) exposed to 3 concentrations of hexachlorobenzene (HCB)in outdoor pondmesocosms

* ** * *

0

5

10

15

20

0 2 4 6 8 10 12Weeks after treatment

Mea

n cu

mul

ated

num

ber

of e

gg-m

asse

s pe

r sn

ail Control

0.5 µg/l

1.25 µg/l

5 µg/l

From Baturo et al. (1995)

Effect propagation

From Caquet (2000) (unpublished)

Chironomini

Notonectidae

Physidae

Dugesiidae Ecnomidae

Asellidae-1,5

-1

-0,5

0

0,5

1

1,5

2

2,5

3

3,5Gastropods

Principal Response Curve (PRC) analysisof the abundance of invertebrate taxa

-0,4

-0,2

0

0,2

0,4

0,6

0,8

1

-5 0 5 10 15 20 25 30Temps écoulé depuis le traitement (Semaines)

Coe

ffic

ient

can

oniq

ue (

Cdt

)

Témoins 0,5 µg/l 1,25 µg/l 5 µg/l

Weeks after treatment

Cdt

Control 0.5 µg/l 1.25 µg/l

Pol

ysac

char

ide

hydr

olys

is e

nzym

e ac

tivity

(µg

gluc

ose

equi

vale

nt/m

in/m

g pr

otei

n)

Glycogen

Corn starch

Rice starch

Hours after treatment

Gly

coge

n co

ncen

trat

ion

(µg/

g fr

esh

wei

ght)

Visceral mass

Mantle

Hours after treatment

From Baturo et al. (1995)

Effect propagationEnhanced glycogen enzymatic breakdownin exposed snails

Effect propagation :Linking levels of biological organisation

Enhanced glycogen

breakdownIncrease in fecundity

Increase in population

density

Effect on CNS (dorsal bodies ?)

Change in neurohormonal control of reproduction

HCB

Explanatory hypothesis

Change in the

structure of gastropod community

From Jumel et al. (2002)

Freshwater snails (Lymnaea) exposed to repeated applicationsof fomesafen (one concentration) in outdoor pondmesocosms

Effect propagation :Linking levels of biological organisation

F3C

Cl

O

CONHSO2CH3

NO2

Effect on reproduction

0

1

2

3

4

0 16 32 48 64

Time (days)

**

*

Ovi

po

sito

ry a

ctiv

ity (

mea

n cu

mul

ated

num

ber

of c

lutc

hes

per

indi

vidu

al)

**

*

Treatments

Time (days)

Fomesafen inhibits reproduction (decreased number of clutches, i.e. fecundity, by individual)

Fomesafen induces :

- enhanced glycogen breakdown

- decrease of sexual steroid levels

Responses of energetic and hormonal biomarkers

0,0

0,2

0,4

0,6

0,8

0 8 16 24 32 40 48 56

Time (days)

Tes

tost

eron

e-lik

e le

vel

(ng/

ovod

iges

tive

glan

d)

0,0

0,2

0,4

0,6

0,8

0 8 16 24 32 40 48 56

Time (days)

Est

radi

ol-li

ke le

vel

(ng/

ovod

iges

tive

glan

d)C

once

ntra

tion

in e

stra

diol

-like

(n

g/ov

otes

tis)

Con

cent

ratio

n in

test

oste

rone

-like

(n

g/ov

otes

tis)

Time (days)

Treatments

0,0

4,0

8,0

12,0

16,0

0 8 12 35 56Time (days)

**G

lyco

gen

conc

entr

atio

n (µ

g/g

fres

h w

eigh

t)

Time (days)

Treatments

From Jumel et al. (2002)

F3C

Cl

O

CONHSO2CH3

NO2

Effect propagation

Conclusions

http://www.rivm.nl./bibliotheek/rapporten/601506009.pdf

Guidance for summarizing and evaluating aquatic micro- and mesocosm studies

de Jong et al., 2008

Many thanks to the people who contributed to the acquisition of the data used in this presentation

Thank you for your attention

Aquatic mesocosms for data validation : interest, limitations and recommandations Laurent Lagadic &...

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