1. LIFE12 ENV/IT/000352 BIONAD “Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle” Start: 01/01/2014 End: 30/06/2016 07/08/2015 18-month meeting INESCOP premises

2. LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle The University of Florence Department of Chemistry “Ugo Schiff” Organic Chemistry

3. The University of Florence Laboratory No 8 Prof. Roberto Bianchini Room 161 LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

4. B6 : naturalised dyes in leather dyeing at pilot scale level (M13-M21) C2 : depuration of dyeing effluents using bacteria at laboratory level (M3-M9) C3: monitoring of quality improvement for leather dyed with nat. dyes (M13-M21) C4 : depuration of dyeing effluents at pilot scale level (M19-M30) C6: monitoring of quality assessment for leather dyed with nat. dyes (M13-M18) ACTIONS OF THE EITHGTEEN MONTHS LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

5. Determination of COD and BOD Recycling of bacterial biomass Action of bacterium strain E. coli on dye effluents Considerations on the mechanism of action of E. coli Assessment of dye reduction (visible analysis) Potential reuse of dyeing effluent after depuration Laboratory scale ; period: 3 - 9 months Depuration of dyeing effluents using E. coli UNIFI Action C.2 Extension of action requested in the amendment Use of fungi cultures for the degradation of dyes Selection of high performing bacteria LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

6. Recaps Action C.2 Shaker E. coli: DH5  Centrifuge Shaking at 37  C Separation of the biomass Nutrient only After E. coli growth Mixed dyes Selection of best performing E. coli. LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

7. Action C.2 Depuration of dyeing effluents using white rot fungi Laboratory scale; period 3-9 months UNIFI ● The fungus used was Funalia trogii ● Eukaryote type microorganism ● More complex metabolism compared to bacteria ● Wide pool of enzymes, responsible for degradation ● Greater adaptability to the environment compared to bacteria ● Wide range of substrates to target LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

8. Action C.2 F. trogii growing in MEAF. trogii liquid growth in BRM Shaking at room temperature (10 d)F. trogii after liquid growth LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

9. Action C.2 Blending the fungusF. trogii in CZAPEKAutoclave sterilizing dyes LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

10. Action C.2 Adding the fungus to dyes F. trogii with dye solutions LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle

11. Ongoing results Action C.2 LIFE12 ENV/IT/000352 “BIONAD“ Naturalised dyes replacing commercial colorants for environmentally friendly leather dyeing and water recycle DR202DB27 Mix 4 dyes