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Pentachlorophenol contamination of Quercus suber forests: A NATO Science for Peace project

SEGH/ISEE 28 th August 2009 University College Dublin. Pentachlorophenol contamination of Quercus suber forests: A NATO Science for Peace project.

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Pentachlorophenol contamination of Quercus suber forests: A NATO Science for Peace project

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  1. SEGH/ISEE 28th August 2009 University College Dublin Pentachlorophenol contamination of Quercussuber forests: A NATO Science for Peace project Iain McLellan1, Andrew Hursthouse1, C Morrison1, C Silva Pereira2,A Hassen3, V Mazzoleni4, M Blahgen5, A Valera2, C Rodrigues2, C Leitão2, H I Ouzari3, A Jaouani3, N Gaâmour3, M D Fumi4, E Novelli4, M Trevisan4 and K Lyamiuoli5 University of the West of Scotland, Paisley, UK Institudo de Biologia Experimental e Tecnológica, Oerias, Portugal Centre de Recherche en Sciences et Technolgies de l’Eau, Solimnen, Tunisia UniversitàCattolica del SacroCuore, Piacenza, Italy University Hassan II, Casablanca, Morocco

  2. Content • Project rationale • Soil microbiota • Chlorophenol presence • Human influence

  3. Project • NATO Science for Peace • “Preventative and remediation strategies for the continuous elimination of ploychlorinated phenols from forest soils and ground waters” • Italy, Morocco, Portugal, Tunisia & United Kingdom • UK • Sampling design / field visits • Chemical analysis / fingerprint • PCP transport • Bioaccumulation

  4. Project Case study evaluation Tunisia Sampling UK Bulk soil simulation Portugal, Tunisia Monitoring & Optimisation Portugal, UK Bioaccumulation UK Dissemination Portugal, Tunisia, UK Project Management Portugal, Tunisia Chemical Analysis UK Microbial Analysis Portugal Bioremediation Portugal

  5. Rationale • Cork • Quercussuber • Portugal, Spain, France, Italy, Tunisia, Morocco, Algeria • Cork taint • Mouldy, earthy aroma • 1 – 5% of bottled wine • Chloroanisoles, chlorophenols • Early contamination • Where do we look? • How do we deal with it? • What can we learn about behaviour?

  6. Pentachlorophenol • Pesticide, herbicide, biocide, preservative • Restricted use • Production ceased within EU • Imports from the United States • Emissions in UK fell 19% 1990 - 2002

  7. Degradation Products

  8. Bioaccumulation & Toxicity • Bioaccumulative in ALL organisms • Germany (1998): urine 1.0ppb • Taiwan (1998): urine 0.08ppb • China (2005): breast milk 2.15ppb • USA (2007): urine 1.27ppb • Repeated human exposure causes headaches, numbness and drowsiness. • Inhibits lytic functions within NK cells • PCP is more toxic than other chlorinated pesticides • Chlorophenol toxicity dependent on: • pH • species • degree of chlorination • position of chlorine atoms • PCP > TeCP > TCP > DCP > MCP • Para- > Meta- > Ortho-

  9. Summary Sediments Depends on soil pH pH Organic Material Clay content Bioavailability Photodegradation - chlorophenols Transformation PCP Soil Fungal methylation converts chlorophenols to chloroanisoles Toxicity Depends on chlorophenol, species and pH. Water

  10. Fungal isolates • Portuguese team • DNA extraction/PCR/Gel electrophoresis • Dr. Rob Samson and JosHoubraken from CBS FUNGAL BIODIVERSITY CENTRE, Netherlands

  11. Summary Sediments Depends on soil pH pH Organic Material Clay content Bioavailability PCP removal Photodegradation - chlorophenols Transformation PCP Soil Fungal methylation converts chlorophenols to chloroanisoles Toxicity Decreases. Depends on chlorophenol, species and pH. Fungal species within soil reduce PCP to less toxic products. Water Bioremediation These species can then be used for in-situ remediation techniques, stopping the transport pathway of PCP.

  12. Chlorophenol determination • Extraction • Soxhlet • Sonication • Analysis • HPLC-UV/Vis • Acetonitrile • 0.1% Phosphoric acid • LC-MS • MeOH with acetic acid • Water with acetic acid • k = (tR – t0)/t0 k2 α • α = k2 / k1 k1 tR2 tR1 t0

  13. Soxhlet extraction

  14. Soxhlet extraction

  15. Soxhlet extraction

  16. Detector 1-210nm PCP & Int Std Calibration 09092008 Retention Time 160 Area 91724 Height 140 1657184 25.172 120 100 80 2563 mAU 1885 3580 9709 2092 16120 60 3357 8961 5739 2287 102630 41429 1765 1193 80315 962 1646 437 1741 239 31414 425659 143 96399 160739 65 444925 141932 194707 43091 14 18764 51795 12.903 13.177 1999 1443 11835 13.783 48805 40 1344 149932 444 14.283 68 20.662 15.703 115 34.973 33.793 27.457 33.297 10.088 10.630 18049 16.497 27.255 21.877 29.417 18.230 30.217 30.002 8.600 2511 20 35.510 5.403 147084 0 3648342 -20 22.727 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 Minutes Sonication extraction • Based on Kähkönenet al, 2008 • 2g soil • 5ml MeOH, sonicated for 15mins • Centrifuged, supernatant removed • Repeat. Supernatants combined • I.S.: 2,4-Dichlorophenol • Method development • 2.5g soil • 2 solvents: MeOH and acetone • Sonication: 15, 30, 45, 60mins • I.S.: 2,4,6-Tribromophenol • Optimised method: • 2.5g soil • 5ml MeOH, sonicatedfor 30mins • Recovery: 86.6% ± 3.6% 2,4,6-TBP PCP

  17. Tunisian samples AînHamraia Belif FejErrih 1 FejErrih 2 RasRajel 1 RasRajel 2

  18. HPLC-UV/Vis

  19. LC-MS • Increased α value • 2007 samples: • AînHamraia • Belif • FejErrih 1 • RasRajel 1 • 2009 sample: • RasRajel 1

  20. Future • Human impact • Other pesticides? • Heavy metals? • Bioaccumulation • Transport

  21. Acknowledgments Society for Environmental Geochemistry & Health UWS NATO SfP 981764 Sylvo-pastoral Institute of Tabarka StazioneSperimentale del Sughero

  22. Any questions?

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