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Innovative System for the Biochemical Restoration and Monitoring of Degraded Soils

BENEFICIARIES:. Innovative System for the Biochemical Restoration and Monitoring of Degraded Soils. BIOREM LIFE11 ENV/IT/000113 . Expected start date : 01/ 01 /2013. Expected end date: 31/03/2015. Project objectives.

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Innovative System for the Biochemical Restoration and Monitoring of Degraded Soils

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  1. BENEFICIARIES: Innovative System for the Biochemical Restoration and Monitoring of Degraded Soils BIOREM LIFE11 ENV/IT/000113 Expected start date: 01/01/2013 Expectedend date: 31/03/2015

  2. Project objectives The main objective of the ‘BIOREM’ project is to demonstrate an innovative, integrated methodology for the restorationand biochemical monitoring of degraded soils. In terms of soil remediation, the project will demonstrate that the BIOREM strategy can successfully restore degraded soils, enhancing their physical-chemical properties and biochemical activity, and increasing fertility. In terms of soil monitoring, the project promises faster and dynamic monitoring of soil conditions.

  3. Actions

  4. B. Implementationactions Identification of the demonstrationplots (360 m2each) Italy: 1) Imola (BO): overexploitedclayeysoil: Imola 2 ) Overexploitedsandy and weaklydevelopedsoil:Tebano 3) Matera: soilcharacterizedby high erosion and low water input:Fusetto 4) Matera: overexploitedsoilwithpoorvegetable cover: Fontana 5) Matera: poorlydevelopedsemi-aridsoil; Albicocco Spain: 6) Santomera: soilwith 50% vegetation cover: SantomeraCanas 7) Santomera: soilwith 25% vegetation cover: SantomeraEntrada 8) Abanilla: xericsoilwithscarcecontent in organicmatter and vegetation (5-10%): Abaran 9) Los Cuadros, Cieza: stressedsoil: Boqueron 10) Cartagena: sandy and stonysoilwith a low and fragile vegetable cover: Cartagena

  5. B. Implementationactions Action carried out until now: • Soil characterizations • Sampling and characterization of soils immediately after the application of the remediation strategies (T0)

  6. Innovative diagnosticmethodology Soilmonitoring Physical-chemicalparameters (e.g. stabilityofaggregates, total organiccarbon, etc.) theychangeveryslowly Biologicalparameters: Microbial community structure (genomicanalysis) Proteinexpressedbymicroorganisms (metaproteomicanalysis) Biochemicalparameters: enzymeactivities (b-glucosidaseactivity) Plantmonitoring Superoxidedismutaseenzymeexpression and activity (biomarkerof stress conditionsforplants) Each soil sample consists of three subsamples collected at 0-20 and 20-40 cm, mixed, homogenised, sieved (2 mm) and stored dried at room temperature until chemical analysis, and stored at -80°C until biological analysis. Very sensitive

  7. BIOINDICATORS AT ECOSYSTEM LEVEL Biochemical and microbiological soil characteristics have been widely proposed as indicators of soil quality • Microbialbiomassquantityandbiodiversity • Microbialbiomassactivity: 1) generalparameters (ATP, CO2, Dehydrogenase) • 2) specificparameters (hydroliticenzymes) SoilEnzymeactivities decomposition of the organic substances mineralization and release of nutrients fertility and productivity of soil response of the soil to ecological and functional recovery practices after natural or human stresses monitor the progress of environmental clean up and restoration They have an effect on:

  8. Enzymeactivities b-Glucosidase enzymes catalyze the final limiting step of cellulose degradation Carbon cycle Phosphatase enzymes catalyze the hydrolysis of various organic phosphate esters Phosphorus cycle Dheydrogenase enzymes catalyze the oxidation of organic compounds with the removal of two hydrogen atoms that are transferred to the molecule of NAD+ R-H2 + NAD+ NADH + R + H+ Indicator of global microbial metabolism R-O-PO32- + H2O R-O-H + HO-PO32-

  9. 4) Dead intact cell 1) Non-proliferating cells (V Lysis Dead Lysis 2) Living cell Enzyme-substrate complex (i) 5) Dead disintegrated cell Secretion Growth/Division Release 6) Free extracellular enzymes Clay minerals 3) Living cells Humicacids Adsorbed enzymes 7) Clay-enzyme complex Entrapped enzymes 8) Humus-enzyme complex Location of Enzymes in Soils -Intracellular enzymes -Extracellular enzymes Proteins/enzymes that bind to clay minerals and/or humic substances They are more stableto some environmental stresses such as extreme temperature, digestion by proteases, etc., than free enzymesand can persistforextendedperiods, therebyproviding a long-termperspectives in indicating the hystoryof the sample and not just a snapshotof the timeofsampling

  10. Stable Organic Matter -Humus >1000 years old humus is resistant to microbial attack Function of Humus • Promotes infiltration of air and water • Holds water and nutrients • Helps establish and maintain a strong soil physical structure • Provides nutrients (N and P) • Buffers effects of contaminats • Entraps enzymes

  11. Humic-enzymecomplexes Due totheir long persistence in soil, theyreact and complywith the surroundingphysicalenvironmenttakingalsoitscharacteristics Humo-enzymecomplexescouldrepresent a reserveofbiochemicalenergy and nutrientscapabletoreactivate the ecosystemfunctionalityeven in heavystressedsituations, thusrepresenting the necessaryconditionsforsoilresilience Sincetheyhavebeenfound in a greatvarietyofnaturalsoils or biologically and physicallydegradedsoils, itmaybesupposedthattheyrepresent a sortofbiologicalbarrierbeforefinal and irreversiblesoildegradation

  12. Soil SoilextractPPi 0.1M, pH 7.0 Ultrafiltration Fraction >10^4 out Fraction <10^4 IEF Extracellular b-glucosidaseActivity Pyrophosphate Extractable Carbon (PEC) ActiveHumic Carbon Humic-b-glucosidase Activity Howtostudyhumic-enzymecomplexes Total enzymeactivities

  13. 1 2 Band 2 1 0 8 Enzymeactivities in the IEF bands Buffer and substrate Gel extrusion and Band cutting Shaking 17h, 37°C Band removal Humicb-glucosidaseactivity: b-glucosidaseactivity in the stablehumiccomplex (bands 1 and 2) after IEF Supernatant enzyme test Specific Humic b-glucosidase activity: Enzyme activity of the stable humic complex (bands 1 and 2) with respect to the focalised carbon content in the same bands Activehumiccarbon: carbonamountcalculatedfrom the IEF peakareasfocused in the pH range4.5-4.2 (bands 1 and 2) after IEF

  14. B.1Demonstrationof the innovative soilmonitoringmethodthrough a characterizationof the initialsoilconditions Soilcharacterization The effect of sampling depth (0-20 and 20-40 cm) is generally evident, since a decreasing gradient of microbial metabolism, extracellular activity and C content with depth is showed. Spanish sites showed a higher soil quality and functionality with respect to Italian sites.

  15. B. Implementationactions B.2 Demonstrationof the additioneffectofexogenousorganicmatterto 10 differenttypesofsoil B.3 Demonstrationof BIOREM revegetationstrategies Demostrationplots (360 m2 per site) • In each plot the following treatments (in triplicate) have been applied: • Organic matter (compost) (3 plots) (A, in yellow)(12 kg m-2May-June) • Organic matter (compost) + vegetation (Lentiscus and Pinus) (3 plots) (B, in green) • Vegetation(Lentiscus and Pinus) (3 plots) (C, in blue) (June-September) • Control soil (3 plots) (D, in orange) Duringall the phasesof the actions, photos and videos havebeentaken

  16. CompostAmek(12 kg m-2)

  17. Compost ALSIA (12 kg m-2)

  18. CompostOrganicos (12 kg m-2)

  19. B. Implementationactions B.3 Demonstrationof BIOREM revegetationstrategies Responsible: PANTANELLO The areas for plantation have been mechanically prepared with a subsoiler. The controlled revegetation strategy has been carried out from June to September 2013. The plants Pinohalepensis(P)and Pistacialentiscus (L) were alternatively planted by PANTANELLO (ALSIA), AMEK and ORGANICOS at a distance of about 1 m in 20 sub-plots

  20. Analyses on soil samples collected immediately after the application of the remediation strategies (T0) Italiansites Albicocco Fontana Fusetto Spanishsites Abaran Boqueron Cartagena Santomera entrada Santomera cañas Total OrganicCarbon HumicCarbon (Pyrophosphate pH7.0) Dehydrogenaseenzymeactivity Extracellularb-glucosidaseactivity Humicboundb-blucosidaseactivity Total OrganicCarbon HumicCarbon (Pyrophosphate pH7.0) Dehydrogenaseenzymeactivity Extracellularb-glucosidaseactivity Italian site Tebano Italian site Imola Total OrganicCarbon Dehydrogenaseenzymeactivity

  21. Total OrganicCarbon Italy Spain South North

  22. Microbialactivity(Dehydrogenase) Italy Spain North South

  23. Humiccarbon (PPi pH7.0) Italy Spain North South

  24. Extracellularb-glucosidaseactivity Italy Spain North South

  25. Humicboundb-glucosidaseactivity (IEF bands pH4.2-4.5) Italy Spain South

  26. B. Implementationactions B.4 Demonstrationof the innovative biochemicalmonitoringmethodology Soilsamplesfromall the 40 sub-plotswillbeanalyzedeverysixmonths: -immediatelyafter the applicationof the remediationstrategies (3-5 months) 30/06/2013 -6 monthslater(10 months) 31/12/2013 by the end of the January -12 monthslater(16 months) 30/06/2014 -18 monthslater(22 months) 31/12/2014 Duringthesemonths, ALSIA, AMEK and ORGANICOS willperform the ordinary management and empiricalmonitoringof the involvedsoils. Analysisof 960 composite soilsamples (10 sites x 4 treatments x 2 depth x 4 samplingtimes x threereplicates) Responsible: CNR-ISE

  27. Todo… -All the workers in BIOREM project need (by a manager or the director) a statement certifying  that the employee is engaged in the BIOREM project for "X" (estimate, reported in the proposal) number of hours. • -In all the cost sheets (consumable, travel, etc..) it is not necessary fill in the columns H and I (or J and K) , but only the columns K and L (or M and N), being our national currency "€". • -The invoices must preferably report the BIOREM • references (written by the  supplier)  and not a "stamp“ • Make and send us photo of the equipment bought with • BIOREM funding where is well visible the "LIFE" logo • (pasted on the equipment) • -Remind to report in the time sheets also the hours spent in • other "LIFE projects“ • -CEBAS and ALSIA: document about the not recovery of VAT • -ORGANICOS: personnel

  28. Activities in the next 6 months • Technical part: • -Soil sampling after six months fromthe application of the remediation strategies (T1) (by the end of January 2014) -Complete the T0 soil sample analysis -Analyze the T1 soil samples • Deliverables • -Mid -term project report (31/03/2014): • Answer to questions by EU • Documents until “the end of February????” • -Project DVD (30/04/2014) • -Environmental report from sampling campaigns (31/05/2014) • Administrative part and others: • -New financial statement (file excell) • -Preparation of “BIOREM protocol” • - 3rd Coordination meeting in “........?.........” in September 2014

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