SETAC North America 39th Annual Meeting
Fate of Emerging Micropollutants and Mercury in Capbreton Submarine Canyon Sediment in Controlled Experimentations (Biscay Bay, SW France) Alyssa Azaroff(1), Mathilde Monperrus(1), Claire Gassie(2), Emmanuel Tessier(2), Rémy Guyoneaud(2)
4-8 NOVEMBER SACRAMENTO, CA, USA
(1)CNRS/ UNIV PAU & PAYS ADOUR/ E2S UPPA, Institut des Sciences Analytiques et de Physicochimie pour l‘Environnement et les Materiaux – (2) CNRS/ UNIV PAU & PAYS ADOUR/ E2S UPPA, Institut des Sciences Analytiques et de Physicochimie pour l‘Environnement et les Materiaux
INTRODUCTION
MATERIAL AND METHODS EMERGING POLUTTANTS DEGRADATION G14
G10
+
y
G24
MeHg
enriched in 201Hg (96.5%)
Fig. 1. Sampling location of stations studied
SLURRIES INCUBATIONS
Time
• • •
BIOTIC / ABIOTIC ANAEROBIC CONDITIONS at in situ temperature, dark
(6)
Analysis by GC-MS Kinetic times (days) : 0,5, 12, 39, 50, 81, 110
ENRICHMENT IN LIQUID MEDIUM Aerobic / anaerobic conditions
RESEARCH POSTER PRESENTATION DESIGN © 2015
www.PosterPresentations.com
(intensity)
Hg(II)
MeHg
F2
( At sp3 ,a Ra3,/m2 At sp3 ,a ) ( At sp3 ,a Ra3,/m1 At sp1 ,a ) 3/ 2 2 3 ( Ra3,/m1 At s1 At s3 ) N spa ( Ra ,m At s At s ) N spb ( At sp3 ,b Ra3,/m1 At sp1 ,b ) ( At sp3 ,b Ra3,/m2 At sp2 ,b ) 3/2 2 3/1 1 3 ( Ra ,m At s At s3 ) ( Ra ,m At s At s )
Species a
Species b
F1
( Asp3 ,b Rb3,/m1 At sp1 ,b ) ( Asp3 ,b Rb3,/m2 Asp2 ,b ) 3/1 1 3 ( Rb3,/m2 As2 As3 ) N spb ( Rb ,m As At s ) N spa ( Asp3 ,a Rb3,/m2 Asp201,a ) ( At sp3 ,a Rb3,/m1 Asp1 ,a ) 3/2 2 3 ( Rb3,/m1 As1 As3 ) ( Rb ,m As As )
F2 z
F1 s)
z
as
xspMeHg IHg N m N spMeHg
y
y
202 201 199
(m
xspIHg MeHg N m N spIHg
F1 F2
isotope 1 isotope 2 isotope 3
x
(time)
x
Time
Incubation slurries 110 d.
0,5g
HHCB
3
d 1/10 MM20
d 1/10 MM20
1 month
1 month
essay control
essay control
MM20 + µpoll.
MM20 + µpoll. T0 Tf
MM20 + µpoll.
Tf
T0
ODPABA
GC-MS GC-MS DNA DNA
GC-MS GC-MS DNA DNA
% Demethylation
% Methylation
INCUBATIONS
8 6 4 2 0 0 1 2
4
7
11
C
+ µpoll. 1 month + µpoll. 1 month
+ µpoll.
Agar MM20
Agar MM20
Agar MM20
Tf
DNA EXTRACTION OF ISOLATED STRAINS
GC-MS GC-MS DNA DNA
RESULTS 10
B
A
essay control T0
CBZ
ISOLATION ON SOLID MEDIUM Aerobic condition
2
Slurry incubations were spiked with mercury enriched isotopic tracers and with ODPABA and HHCB to determine methylation, demethylation potentials and degradation potentials, respectively.
DEGRADATION TEST
Taxonomic diversity (16s DNA, MiSEQ) were analyzed for initial and final kinetic times to determine microorganisms diversity. Then, microorganisms were isolated by dilution and enrichment in anaerobic and aerobic conditions.
• Methylation is mainly a biotic process
40
(Very low methylation potentials were observed for abiotic control experiments with M around 0.2 %.)
30 20 10 0 1 2
4
7
11
15 Days
• Methylation potentials decreased along the canyon • High demethylation was found under biotic conditions with higher yields in canyon stations (G10, G24) as compared to continental shelf station (G14) (significant yields ranging between 13.9 and 17.0 % were found for the abiotic control.)
INCUBATIONS
Degradation %
Fig. 2 - Yields of 199MeHg (M) and 201Hg(II) (D) formed in biotic condition after two weeks incubation from sediments collected according a distance gradient to the coast (G10, G14 and G24, Fig. 1) 100
Abiotic control
80
Biotic incubation day+ 110
60 40 20 0
G10
G14
G10
G14
G10
G14
ODPABA
G10
60 40 20 0
*** Control
Essay
*
*
Control
Essay
ANAEROBIC
AEROBIC G10
HHCB
ODPABA
CBZ
Degradation %
Degradation %
ENRICHMENT 3
80
CBZ
100 80
G14
60
40 20 0
*below detection limit
*
Net methylation potential were only observed under biotic conditions revealing the involvement of prokaryotes in the fate of Hg species. This was confirmed by taxonomic diversity and sequencing of hgcA(8) gene, a proxy for biotic methylation. Two main families of methylating bacteria have been identified, Desulfobulbaceae and Desulfobacteraceae, suggesting their involvement in the biotic methylation process. Higher methylation potentials were observed for the coastal station whereas demethylation potentials were similar for the three stations suggesting that environmental parameters (e.g. input of continental organic matter) also control methylating microbial activities. Degradation potential was observed under biotic condition for ODPADA and CBZ(9). Aerobic and anaerobic microorganisms of Capbreton canyon sediments might be involved in degradation of emerging micropollutants. Among these, fungi(10) have been isolated from sediment. Analysis of strains isolated on solid medium will permit us to identify which organisms are involved in emerging micropollutants degradation. This work suggests the canyon possess a natural resilience against anthropogenic inputs.
CONCLUSION These results confirm the role of the Capbreton submarine-canyon to trap, to transfer and to transform micropollutants. Degradation potentials reveals the differential resilience of such deep sea sediment for some priority and emerging substances and the key role of microorganisms.
MERCURY TRANSFORMATIONS
50
15
100
Sediment dynamics favor accumulation in terraces and slopes Micropollutants have strong affinity for those sediments
2 ppm
Analysis by double spike ID – ICP-MS(7) Kinetic times (days) : Hg : 0,1, 2, 4, 7, 11, 14
Fig. 3 - HHCB, ODPABA and CBZ degradation potentials (%) in final kinetic time slurry incubation (days 110) and abiotic control (slurry sterilized) from Capbreton canyon sediments (G10, and G14, Fig. 1)
(4,5)
x Time
7 stable isotopes
ISOLATION OF MICROORGANISMS BE ABLE TO DEGRADATE EMERGING SUBSTANCES
1 Mazières, et al, 2014. High-resolution morphobathymetric analysis and evolution of Capbreton submarine canyon head (Southeast Bay of Biscay—French Atlantic Coast) over the last decade using descriptive and numerical modeling. Mar. Geol. 351, 1–12.
IHg
D
HHCB
POLLUTANTS ?
Hg
x
Intensity
M d
QUESTIONS • Methylation / Demethylation yields (M/D) of mercury (Hg)? • Degradation potential of emerging substances? • Isolated microorganisms able to degrade emerging pollutants?
Hg(II)
enriched in 199Hg (91.7%)
Isotpic pattern deconvolution MeHg
CAPBRETON SUBMARINE CANYON
One of the deepest in the world (as big as Grand Canyon, USA) Transfer zone between continent and ocean
(0.2%) (9.9%) 199Hg (16.9%) 200Hg (23.1%) 201Hg (13.2%) 202Hg (29.9%) 204Hg (6.9%)
198Hg
m as s
Priority substances Inorganic mercury (iHg) Organic mercury (MeHg) the most toxic form causing health and environmental issues Emerging substances Synthetic fragrance Galaxolide (HHCB) Synthetic sunscreen Padimate O (ODPABA) Pharmaceutical Carbamazepin (CBZ)
100ppb
CBZ Pharmaceutical
196Hg
++
y
20 ppm
TARGETED MICROPOLLUTANTS
Kinetic transformations of Hg, ODPABA and HHCB were studied independently from three Capbreton Canyon sediment stations according to the distance to the coast (Fig,1).
Enriched isotopic tracers Me201Hg and 199HgII Intensity
ODPABA Synthetic sunscreens
HHCB Synthetic fragrance
DISCUSSION
MERCURY TRANSFORMATIONS
Intensity
Priority substances(1) set by the Water Framework Directive (WFD) are of major interest to evaluate the quality of coastal and marine systems, the final receptors for pollutant emissions. Emerging substances(2,3) not regulated by the WFD, i.e. personal care products and some pharmaceuticals, are of high concern since only scarce information of their occurrence, reactivity and impact are available in the deep sea sediments.
• •
MIRA, UMR 5254, 64600, Anglet, FRANCE – MIRA, UMR 5254, 64000, Pau, FRANCE
* *** ***
Control
Essay
Control
ANAEROBIC
Essay
AEROBIC G14
HHCB
ODPABA
CBZ
Fig. 4 -HHCB, ODPABA and CBZ degradation potentials in the 3rd enrichment in liquid medium in essay (with slurry) and control (without slurry) from Capbreton canyon sediments (G10 and G14, Fig. 1)
EMERGING POLUTTANTS DEGRADATION • HHCB degradation is constant over incubation experiment with similar potential in biotic and abiotic conditions • ODBAPA degradation is higher in biotic condition (98 and 93% in G10 and G14, respectively) compare to abiotic control condition • CBZ degradation is higher in biotic condition (33 and 50 % in G10 and G14, respectively) compare to abiotic control condition • Nevertheless, the degradation potential of CBZ and HHCB suggested those micropollutants are refractory into the Canyon3
REFERENCES (1) Directive 2013/39/EU of the European Parliament and of the Council of 12 August 2013 amending Directives 2000/60/EC and 2008/105/EC as regards priority substances in the field of water policy Text with EEA relevance, 2013. , 226. (2) Hopkins, Z.R., Blaney, L., 2016. An aggregate analysis of personal care products in the environment: Identifying the distribution of environmentally-relevant concentrations. Environ. Int. 92–93, 301–316. https://doi.org/10.1016/j.envint.2016.04.026 (3) Rainieri, S., Barranco, A., Primec, M., Langerholc, T., 2017. Occurrence and toxicity of musks and UV filters in the marine environment. Food Chem. Toxicol., Safety assessment of contaminants of emerging concern in seafood: Contributions of the ECsafeSEAFOOD project 104, 57–68. https://doi.org/10.1016/j.fct.2016.11.012 (4) Cremer, M., Brocheray, S., Gillet, H., Hanquiez, V., 2012. Capbreton canyon: evidence of its formation by differential sedimentation, in: XII International Symposium on Oceanography on the Bay of Biscay, Santander (Spain). (5) Gaudin, M., Mulder, T., Cirac, P., Berné, S., Imbert, P., 2006. Past and present sedimentary activity in the Capbreton Canyon, southern Bay of Biscay. Geo-Mar. Lett. 26, 331. https://doi.org/10.1007/s00367-006-0043-1 (6) Miossec, C., Lanceleur, L., Monperrus, M., 2018. Adaptation and validation of QuEChERS method for the simultaneous analysis of priority and emerging pollutants in sediments by gas chromatography—mass spectrometry. Int. J. Environ. Anal. Chem. 98, 695–708. https://doi.org/10.1080/03067319.2018.1496245 (7) Monperrus, M., Gonzalez, P.R., Amouroux, D., Alonso, J.I.G., Donard, O.F.X., 2008. Evaluating the potential and limitations of double-spiking species-specific isotope dilution analysis for the accurate quantification of mercury species in different environmental matrices. Anal. Bioanal. Chem. 390, 655–666. https://doi.org/10.1007/s00216-007-1598-z (8) Gilmour, C.C., Podar, M., Bullock, A.L., Graham, A.M., Brown, S.D., Somenahally, A.C., Johs, A., Hurt, R.A., Bailey, K.L., Elias, D.A., 2013. Mercury Methylation by Novel Microorganisms from New Environments. Environ. Sci. Technol. 47, 11810–11820. https://doi.org/10.1021/es403075t (9) Barra Caracciolo, A., Topp, E., Grenni, P., 2015. Pharmaceuticals in the environment: Biodegradation and effects on natural microbial communities. A review. J. Pharm. Biomed. Anal., SI: Analytical approaches 106, 25–36. https://doi.org/10.1016/j.jpba.2014.11.040 (10) Vallecillos, L., Sadef, Y., Borrull, F., Pocurull, E., Bester, K., 2017. Degradation of synthetic fragrances by laccasemediated system. J. Hazard. Mater. 334, 233–243. https://doi.org/10.1016/j.jhazmat.2017.04.003
ISOLATION OF MICROORGISMS
CONTACT
• High degradation of HHCB and ODPABA were found in aerobic and anaerobic conditions may mediated by Capbreton sediment microbial communities • No degradation of CBZ with this isolation microbial protocol suggests that micropollutant was not able to be degraded • Results of global diversity (DNA, 16s) and isolation of microbial strains could confirm these hypothesis (data processing) • Microbial and Fungi strains able to degrade those HHCB and ODPABA have been isolated.
Alyssa AZAROFF PhD student UMR IPREM/UPPA
[email protected]
The Micropolit research program "State and evolution of the quality of the South Atlantic coastal environment" is co-financed by the European Union and the Adour Garonne Water Agency. Europe commits to New Aquitaine with the European Regional Development Fund.
