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Consortium "Nîmes University, Catalan Institute for

Water Research and Institute of Groundwater Ecology : a

groundwater and environmental health scientist’s

consortium

ICRA research team:

J. Mas-Pla, M. Petrovic, C. Borrego,

R. Marcé; D. Brusi, A. Menció (UdG)

UNIMES - CHROME team :

Coordinator

Le Gal La Salle C., Roig, B., Meffre P, Benfodda Z,

Verdoux P. IR, L. Sassine (Post doc)

Patner : CHU Nîmes

IGOE team :

Stumpp C. ; Kiecak Aleksandra, Elsner M.,

3

Target the aims of the call

Geologists &

Hydrogeochemist

Environmental

Health

Analytic &

Organist

chemist

Microbiologist &

Geoeocologist

CHROME Team &

Nîmes University

Hospital

Le Gal La Salle C.

Verdoux P. Roig, B.

Meffre P

Benfodda Z Lavigne J.P.

ICRA Team Mas-Pla J.

Brusi D., Menció A. Petrovic M. Borrego C.

Marcé R.

IGOE

Helmholtz

Zentrum

Stumpp C.

Maloszewski P.Elsner M. Maier M.

Promotion of multi-disciplinary work

4

PERSIST – the Aims

Objectives: Gain fundamental information on the behaviour of

pharmaceuticals and resistant microbial communities in a surface

water groundwater continuum

At a field scale

•Occurrence and fate of EOCs in SW

and GW from sources to exploitation

wells

•Persistence and fate of pathogens and

multiresistant microbiomes under the

occurrence of EOCs in SW and GW

•Combined approach with

hydrogeology and natural tracers

At laboratory scale

•Determine pharmaceutical transport

parameters from column and batch

experiments5

Pig Slurry

Two field sites and workpackages

WP2: Empordà Basin

Cattle grazing activities

WP1 : Vistrenque Basin

EOC sources : WWTP/Septic systems

Laboratory experiments

WP3

6

Budget

Projet sur deux ans

Réflexion sur le budget disponible à l’échelle des pays

particiants

7

Choix des Partenaires

Pays

Stake holders

Partenaires

8

Teamwork 2 Pr

1 Doctorant

2 IR

9

Les points

forts

10

Les points

forts

11

Et pour finir !

12

13

Pour la plupart d’entre nous, le danger n’est pas de

viser trop haut et de rater la cible, mais de viser trop

bas et de l’atteindre. (Michel-Ange)

Thank you for your attention

Bezouce Marguerittes Bouillargues

Nîmes Le Cailar Saint Laurent

d’Aigouze

Coordinator : Pr. Corinne Le Gal La Salle (University of Nîmes)

Partner : Pr. Josep Mas Pla (ICRA),

Dr. Christine Stump (IGOE)

Water JPI Pilot Call mid-term report meeting

1st of decembre 2016,

16

17

20 WWTP

50 Groundwater samples

34 pharmaceuticals

searched for

13 Quantified

18

WP1 – Vistrenque Catchment –

Task 1 : Catchment scale survey

0

10

20

30

40

50

60

70

80

90

100

Fre

qu

en

cie

s o

f q

uan

tifi

cati

on

of

EOC

s in

gr

ou

nd

wat

er

sam

ple

s (%

)

1

3

4

5

2

Focus on 5 sites in

the vicinity of streams

and showing

abnormal EOC

concentrations

WP1 – Vistrenque Catchment

Task 3: Transfer from Surface water to groundwater

19

1

10

100

1000

6462

6462

CLO

S H

OP

6129

7

6129

7

6129

7

CHE…

CHE…

CUB

ELLE

2

PER

79…

OLI

VER

OLI

VER

RH

ON

Y

RH

ON

Y 2

SP5

SP5

VIL

MO

RIN

VIL

MO

RIN

Con

cent

rati

on

s (n

g.L-1

)

Carbamazepine

0.1

1

10

100

1000

10000

6462

6462

CLO

S H

OP

6129

7

6129

7

6129

7

CHE…

CHE…

CUB

ELLE

2

PER

79…

OLI

VER

OLI

VER

RH

ON

Y

RH

ON

Y 2

SP5

SP5

VIL

MO

RIN

VIL

MO

RIN

Con

cent

rati

on

s (n

g.L-1

)

Macrolides

0.1

1

10

100

1000

6462

6462

CLO

S H

OP

6129

7

6129

7

6129

7

CHE…

CHE…

CUB

ELLE

2

PER

79…

OLI

VER

OLI

VER

RH

ON

Y

RH

ON

Y 2

SP5

SP5

VIL

MO

RIN

VIL

MO

RIN

Con

cent

rati

on

s (n

g.L-1

)

Sulfamethoxazole

0.1

1

10

100

1000

6462

6462

CLO

S H

OP

6129

7

6129

7

6129

7

CHE…

CHE…

CUB

ELLE

2

PER

79…

OLI

VER

OLI

VER

RH

ON

Y

RH

ON

Y 2

SP5

SP5

VIL

MO

RIN

VIL

MO

RIN

Con

cent

rati

on

s (n

g.L-1

)

Fluoroquinolones

Site 5Site 3 Site 1Site 4

Groundwater Stream water WWTP effluent

Objectives: Combine

with residence time

tracers and

hydrodynamic

modelling

Attenuation rate

from 10 to 1000

Macrolides

FluoroquinolonesSulfametoxazole

Carbamazepine

Focus on site 2

Verify the

hypothesis

Combine with

residence time

tracers and

hydrodynamic

modelling

WP1:Vistrenque Catchment

Task 3: Transfer from surface water

to groundwater

20

0

20

40

60

80

100

120

0

10

20

30

40

50

60

70

juil.-15 oct.-15 janv.-16 mai-16 août-16 nov.-16

Pre

cipitatio

ns (m

m)

Car

bam

aze

pin

e (

ng

/L)

Date

0

20

40

60

80

100

120

0

50

100

150

200

250

300

350

juil.-15 oct.-15 janv.-16 mai-16 août-16 nov.-16

Pre

cipitatio

ns (m

m)

Sulf

ame

tho

xazo

le (

ng

/L)

Date

0

20

40

60

80

100

120

0

50

100

150

200

250

juil.-15 oct.-15 janv.-16 mai-16 août-16 nov.-16

Pre

cipitatio

ns (m

m)

Flu

oro

qu

ino

lon

es

(ng

/L)

Date

Precipitations

ANNE 1

ANNE 2

CANABOU

VISTRE ANNE

0

20

40

60

80

100

120

1

10

100

1000

juil.-15 oct.-15 janv.-16 mai-16 août-16 nov.-16

Pre

cipitatio

ns (m

m)

Mac

rolid

es

(ng

/L)

Date

Sampling along the Vistre river

Primary

source of EOC

Attenuation

processes:

Dilution

Degradation

Sorption

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WP1 – Vistrenque Catchment

Task 2 : Fate of EOC downgradient from a

WWTP

0

10

20

30

40

50

60

70

80

90

100

0 10 20 30 40 50

Car

bam

aze

pin

e (

ng.

L-1)

Distance (km)

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

0 10 20 30 40 50

Mac

rolid

es

(ng.

L-1)

Distance (km)

0

50

100

150

200

250

300

350

400

450

0 10 20 30 40 50

Flu

oro

qu

ino

lon

es

(ng.

L-1)

Distance (km)

0

20

40

60

80

100

120

140

160

180

200

0 10 20 30 40 50

Sufa

met

ho

xazo

le (

ng.

L-1)

Distance (km)

WP1 – Vistrenque Catchment

Task 3: Multidrug resistance bacteria isolated (Vistre)

Pseudomonas fluorescens 56

Pseudomonas putida 26

Pseudomonas chlororaphis 18

Aeromonas hydrophila 8

Pseudomonas oleovorans 5

Parococcus yeei 3

Aeromonas sobria 2

Micrococcus luteus 2

Pseudomonas veronii 2

Bacillus licheniformis 1

Chromobacterium violaceum 1

Paenibacillus spp 1

Pseudomonas mendocina 1

Pseudomonas viridiflava 1

Vibrio mimicus 1

Escherichia coli 5

Enterobacter cloacae 4

Serratia fonticola 1

93%

7% bactéries

environnementales

bactéries humaines

(entérobactéries)

ESBL: Extended Spectrum Beta-lactamases

2 Enterobacter cloacae

1 Escherichia coli 4 Escherichia coli

Derepressed Cephalosporinases

Antibiograms of enterobacteria

Resistance to 1st, 2nd and 3rd

generation cephalosporines

+ penicillins + quinolones

Synergic effect

AMXCEFFOX

CFM AMC TIC

MEC

ETP

CTX TCC CAZ PIP

IPM ATM PPT FEP

AMXCEFFOX

CFM AMC TIC

MEC

ETP

CTX TCC CAZ PIP

IPM ATM PPT FEP

Environmental bacteria

Human bacteria (enterobacteria)

Field characterization, sampling, analytics and environmental issues1

2

Paleogene formations Neogene formations

Sta Magdalena Manol River plainNNW-SSE

0 3 km

0

100

200

300

400m asl

Lower Unit

Upper Unit

Rainfall recharge

EOCs fate and persistence, and resistome characterization

3 Hydrological interpretation and transport processes modelling

Work Package 2 – Empordà Basin – ICRA

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Abstract: 2192 / Boy-Roura et al.

WP2: Spatial variability: most detected antibiotics

Med

iter

ranea

nSea

No clear spatial distribution is observed

Abstract: 2192 / Boy-Roura et al.

WP2: Antibiotics results of the general campaign

47 wells7 river points2 WWTP

ABs detectedGW: 11/53SW: 5/53WWTP: 9/53

Abstract: 2192 / Boy-Roura et al.

WP2: Antibiotics results of 8 monthly campaigns

8 wells2 river points

EOCs detectedGW: 25/53SW: 18/53

High spatial, temporal variability

No conclusive evidence

Message to the managers ?

Abstract: 2192 / Boy-Roura et al.

WP2: GW resistome: quantification of Antibiotic

resistance genes (ARGs)

Relative concentration,

Log (ARGs/16S rRNA)

An

tib

iotic R

esis

tan

ce G

en

es (

AR

Gs)

Fall

Winter

Spring

Sulfonamides

Macrolides

ß-lactams

Fluoroquinolones

Tetracyclines

Antibiotics families

WP 3 - Laboratory migration experiments

Impact of water transit times on the fate of EOCs

HYPOTHESIS

Fate of EOCs strongly depends on water transit time distributions

OBJECTIVES

Determine range of transit times and flow velocities at the pilot field

sites

Study the impact of flow velocity on sorption and degradation rates in

controlled lab experiments

Transfer the lab results to the field sites

for vulnerability assessment

Field-Lab-Field Approach

complementary to WP1 and WP2

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WP3 : Laboratory batch experiments

in abiotic systems: Adsorption

Staticconditions

5 sedimenttypes

10 compounds

Blanks (chemical degradation)

Sorption isotherms

Linear, Freundlich, Langmuir isotherm

Sorption coefficients

E.g. KD Atenolol: E3 > G ≈ VC [µg/L]

S [µ

g/kg]

E3 - Medium sand

V - Silty Sand

G - Technical coarse sand

Example: Atenolol

S = 4.2· C

S = 3.1· C0.92

S = 20· C0.62

WP3

Laboratory column experiments

Dynamic conditions

3 aquifer sediments x 3

flow velocities

Breakthrough curves →retardation factors

R=1

R=1

R=6.2

Bromide,

Uranine,

EOCs

Column

- sediment

from the

field site

Example:

Medium sand (Emporda)

Conclusion from preliminary resultsField observations

In the range of a few to 100 ng/l

Attenuation rate ranging from 1 to 1000

Transfert from surface water from groundwater

High seasonal variability in the Emporda basin

BMR resistance is detected in environmental molecules

Laboratoy experiments

Kd is strongly site dependent (sediments, hydrogeochemistry)

Kd seems not to be velocity dependent

Degradation seems to increase with velocity

Challenges Future work

Confront lab results to field observations

Modelling of transport processes in the field

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• Unsaturated zone observations

• What is the meaning of observed

drug resistance

Abstract: 2192 / Boy-Roura et al.

Collaborations

Research groups:- Laboratory of Virus Contaminants of Water and Food (University of Barcelona), lead by Dr. Rosina Girones within the Water JPI project METAWATER we share our groundwater sampling points for their field work as well as hydrogeological knowledge derived from our field studies.

- Institute of Agrifood Research and Technology (IRTA) – Mas Badia They allow us conduct some field sampling in experimental sites, and provide agriculture knowledge. We inform them about the progress of our project regarding antibiotic occurrence in irrigation water.

- Technological Center BETA – University of Vic collaboration with the analysis of antibiotic content in animal manure samples.

Stakeholders:- Catalan Water Agency (ACA) sharing goals and results of the PERSIST project.

Collaboration, coordination and mobility

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Help in well research and giving access

Facilitate sampling of public supply wells and effluents of wastewater treatment plants.

Interest in results and outcomes of the study

Participation to sampling campaigns

Regular invitations to participate to committee meetings in order to present the

progress of the project.

Give informations about fertilization practices and antibiotic use in livestock farming.

Stakeholder/industry engagement

Persist

Effective collaboration between the PERSIST team partners.

Active mobility of postdoc researchers and IPs

Field-trip visits to the Vistrenque and Empordà field sites

Exchanges about analytical methodoogies

Contribution in columns and batchs experiments

3 papers in peer reviewed journals

2 papers in prep

5 Oral presentations in International (IAH 43rd, San Fransisco) and National meetings (SHF –BRGM).

5 Posters in International (IAH 43rd, San Fransisco) and National meetings (POF III).

3 Public conferences

Dissemination of the results

34

Identified problems or specific risks

Persist

Technical issues in LC/MSMS

Thank you for

your attention

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Funding agencies:

ONEMA – France

MINECO

(Ministerio de Economia

y Competitividad) - Spain

BMBF (Ministry of

Education & Research) -

Germany

!LOGOS

Acknowledgments:

EU JPI-Water Program, Grant JPIW2013-118