How to influence bioavailability for reducing risks from organic pollutants in bioremediation

Transcription

1 How to influence bioavailability for reducing risks from organic pollutants in bioremediation Jose Julio Ortega-Calvo Instituto de Recursos Naturales y Agrobiologia de Sevilla (CSIC) jjortega@irnase.csic.es

2 How to influence bioavailability to reduce risks associated to bioremediation of organic pollutants OUTLINE - BIOAVAILABILITY vs. BIODEGRADABILITY - ENHANCED PHASE EXCHANGE: BIOSURFACTANTS - DESORPTION & BIODEGRADATION METHODS - EFFECTS OF PLANTS:EXUDATES - COMBINED EFFECTS PLANTS & BIOSURFACTANTS - CONCLUSIONS

3 ES&T (2015) 49, SETAC Europe setac.org sesss10.setac.org

4 Bringing different worlds together Ortega-Calvo et al. ES&T, ,

5

6 Ortega-Calvo et al. ES&T, ,

7 Why PAHs?

8 Because ARE UBIQUITOUS & PERSISTENT, RECOGNIZED AS PRIORITY POLLUTANTS, ARE THE REASON FOR REMEDIATION ARE BIODEGRADABLE, SIGNIFICANT POTENTIAL FOR BIOREMEDIATION ARE HYDROPHOBIC, WITH SPECIFIC LIMITATIONS TO BIODEGRADATION DUE TO LOW BIOAVAILABILITY.

9 BIODEGRADABLE BIODEGRADED -TOXICITY AT POLLUTED SITE -LOW LEVELS OF NUTRIENT & e- ACCEPTORS -LOW TOTAL CONCENTRATION OF SUBSTRATE -LOW BIOAVAILABILITY: SORPTION, NAPLs

10

11 ATTACHMENT ES&T 2011, 45: ES&T 2017, 51: ENHANCED PHASE EXCHANGE BIODEGRADATION ES&T 2011, 45: ES&T 2014, 48: ES&T 2008, 42: ES&T 2015, 49: ES&T 2016, 50: ES&T 2018, 52: MOBILIZATION Ortega-Calvo et al., Soil Biological Communities and Ecosystem Resilience, Sustainability in Plant and Crop Protection, M. Lukac et al. (Eds.), DOI: / _19.

12 Effect of Pseudomonas aeruginosa 19SJ biosurfactants on surface tension: critical micelle concentration 80 Surface tension (mn/m) CMC RE (µg/ml)

13 Effect of biosurfactants from Pseudomonas aeruginosa 19SJ on partitioning of pyrene from a NAPL (HMN) Biosurf. Solids Part. rate C eq C eqsolids (µg/ml) (mg/ml) (ng/ml/h) (ng/ml) (ng/mg) 0 0 0, , , , , M. Garcia-Junco et al., Environ. Sci. Technol. 2003, 37,

14 Microbial biosurfactants influence on bioavailability of sorbed pyrene: role of desorption kinetics E. Congiu & J.J. Ortega-Calvo, Environ. Sci. Technol., 2014, 45:

15 Environmental sample DESORPTION BIODEGRADATION TENAX SAMPLE SUSPENSION SAMPLE SUSPENSION NaOH TRAP - TENAX EXTRACTION - HPLC ANALYSIS - MC EXTRACTION - HPLC ANALYSIS (native PAH) - 14 CO 2 MEASUREMENTS ( 14 C-PAH)

16 Desorption kinetics using Tenax extraction Sediment or soil suspension Liquid medium Tenax Biocide (formaldehide) S t / S o = F rap exp (-K rap t) + F slow exp (-K slow t) S t and S o are the sorbed amounts at time t (h) and at the start of the experiment, respectively. F rap and F slow are the rapidly and slowly desorbing fractions. K rap and K slow (h -1 ) are the rate constants of rapid and slow desorption. M. Bueno-Montes et al., Environ. Sci. Technol. 2011, 45:

17 DESORPTION OF PAHs WITH TENAX IN GREENHOUSE SOIL 1. COMPLETE DESORPTION KINETICS OF PAHs 2. DESORPTION OF PAHs EXTRACTING F rap as T 20 (ISO 16751) T 20 = 97.3 % F rap Benzo(a)pyrene T 20 = 99.9 % F rap Phenanthrene S t / S 0 = F rap * exp (-K rap * t) + F slow * exp (-K slow * t) The study of the complete desorption kinetics is very important in determining the magnitude of the different desorbing fractions present in this soil With this test we can assess quickly bioavailability through F rap in a high number of samples during the greenhouse experiment

18 BIOACCESIBILITY ASSAY A. 14 CO 2 production B. HPLC analysis Teflon-lined stoppers Biometer flask Syringe sampling Teflon-lined stopper 250 ml Erlenmeyer flask Main body 14 CO 2 Lateral body 40 g Sample 40 g sample + radiolabelled PAH Alkali trap

19 Bioaccessibility of phenanthrene in creosote-polluted soil Phen 60 Phenanthrene (mg kg -1 ) CO 2 12 C-Phen % 14 C mineralized Time (days) J.L. Niqui-Arroyo et al., IUPAC Ser. Biophys. Proc. Env. Syst. Vol 3 (2011)

20 River sediment (Stokholm, industrial pollution) Forest soil (Cádiz, background pollution) R. Posada-Baquero et al., Environ. Geochem. Health 2008, 30,

21 EFFECT OF SURFACTANTS ON BIODEGRADATION OF PAHs SOILREM SOIL- AUTOCTHONOUS POP. Phen Pyr Native PAH conc. (mg/kg) Initial F slow Control +Brij35 Phen Pyr Benzo(a)pyr M. Bueno-Montes et al., Environ. Sci. Technol. 2011, 45:

22 EFECT OF PLANTS ON BIOAVAILABILITY OF PAHs

23 Breakthrough curves of Pseudomonas putida G7 transported through sand columns: exudates from Helianthus annuus vs. other DOM sources High TOC (100 mg/l) Low TOC (10 mg/l) C. Jimenez-Sanchez et al. Environ. Sci. Technol., 2015, 49:

24 Motility pattern mediated by taxis affects bacterial dispersal and biodegradation rate in porous media C. Jimenez-Sanchez et al. 2018, 52:

25

26 M. C. Tejeda-Agredano et al., Soil Biol. Biochem. 2013, 57:

27 GREENHOUSE BIOAVAILABILITY EXPERIMENT 1. BIODEGRADATION 1.A 1.B MEASURE THE TOTAL PAHs CONCENTRATION WITH SOXHLET EXTRACTION AND HPLC ANALYSIS TO STUDY THE EFFECT OF PLANTING ( SUNFLOWERS) AND BIOSURFACTANTS ADDITION ( RHAMNOLIPIDS AT 7 mg/g) SEPARATE BIODEGRADATION EXPERIMENT UNDER LABORATORY CONDITIONS USING AN EXCESS OF NUTRIENTS, RADIORESPIROMETRY DETERMINATIONS WITH 14C-PYRENE AND ANALYSIS OF RESIDUAL CONCENTRATIONS OF NATIVE PAHs 2. BIOAVAILABILITY Chemical method :Desorption extraction. Kinetics and single-point extraction at 20 h (ISO 16751) SAMPLE SUSPENSION NaOH TRAP

28 RESULTS IN DIFFERENT ASSAYS A B NO SIGNIFICANT EFFECTS A SIGNIFICANT EFFECT AFTER THE ADDITION OF THAMNOLIPIS C D D20 = T 20 /C total A SIGNIFICANT INCREASE IN THE RAPIDLY DESORBABLE FRACTION WAS OBSERVED AFTER RHAMNOLIPIDS ADDITION FOR MINERALIZABLE AND COMETABOLIZABLE PAHs

29 Study of biodegradation experiment under laboratory conditions BIODEGRADATION EXPERIMENT USING AN EXCESS OF NUTRIENTS, RADIORESPIROMETRY, SHAKING, DETERMINATIONS WITH 14C-PYRENE AND ANALYSIS OF RESIDUAL CONCENTRATIONS OF NATIVE PAHs With this experiment we can say that the concentration was lower under laboratory conditions THE EFFECT OF BIOSURFACTANT WAS OBSERVED ONLY IN THE PRESENCE OF PLANTS DUE TO ROOT COMPONENTS OR EXUDATES COULD PROMOTE THE BIOSURFACT ACTION PAH Soil Soil + rhamnolipid Planted soil Planted soil + rhamnolipid Pyrene D 20 a Min. rate (µg/kg/h) b 1.5 ± 0.1a d 1.7 ± 0.2ab 3.9 ± 0.9bc 5.3 ± 0.01c Min. extent (%) b 40 ± 6 38 ± 8 48 ± 6 40 ± 7 C 0 (mg/kg) b 4.0 ± 0.2A e 6.3 ± 0.3A 5.0 ± 1.0A 13.8 ± 4.4 A C f (mg/kg) b 1.0 ± 0.1Aa 0.5 ± 0.1Bab 0.6 ± 0.2Bab 0.4 ± 0.01Bb C 210d (mg/kg) c 2.8 ± 0.8A 2.4 ± 0.2AB 3.7 ± 0.4A 2.7 ± 0.2AB Benzo(a)pyrene D 20 a C 0 (mg/kg) b 1.9 ± 0.5A 1.6 ± 0.01A 1.8 ± 0.15A 2.5 ± 0.5A C f (mg/kg) b 0.8 ± 0.1Aa 0.6 ± 0.04Bab 0.7 ± 0.05Aab 0.5 ± 0.01Bb C 210d (mg/kg) c 1.6 ± 0.2A 1.8 ± 0.01A 1.5 ± 0.5A 1.6 ± 0.04AB

30 RESULTS IN DIFFERENT ASSAYS LIFE15 ENV/IT/ PARTNER OUR ROLE: BIOAVAILABILITY ASSESSMENT THE OBJECTIVE OF THIS PROJECT IS TO OPTIMIZE A BIOREMEDIATION METHOD WHERE THE TRANSFORMATION MADE BY CONSORTIA OF FUNGI AND BACTERIA IS FINALIZED BY THE FINAL STEP OF REVEGETATION IN A SOIL HIGHLY CONTAMINATED BY PAHS, BTEX AND ALKANES

31 BIOAVAILABE CONCENRATION FOR MINERALIZABLE AND COMETABOLIZABLE PAHs IN LIFE PROJECT A No significant effect was observed in cometabolizable PAHs B C

32 CONCLUSIONS We used different bioremediation approaches, operating on the bioavailability of PAHs, as a proof-of-concept for the use of desorption extraction methods to estimate bioavailability. Tenax extraction during 20 hours has resulted a reliable and robust method to determine bioavailability in a wide set of operational conditions ranging from a different time scale to dissimilar treatments (planting, biosurfactant application, etc.).

33 Acknowledgements Magdalena Grifoll (Universidad de Barcelona) Joop Harmsen (Wageningen Environmental Research (Alterra) Projects from Spanish Ministry of Economy, Industry and Competitiveness (CGL R and CGL R), Andalusian Government (RNM 2337) and European Commission (LIFE15 ENV/IT/000396)

34 Thank you for your attention