1. Innovative integrated methodology for the use of decontaminated river sediments in plant nursing and roadbuilding Activity of partner UniFi

2. Activities done:  Dissemination – done  Sediment monitoring (BioTox) – done  Microbial diversity analysis (in progress)

3. Dissemination: SICA Winter School – Piacenza, Feb., 2014 Workshop REMIDA – ARPA Umbria, Terni, Oct., 2013 Proceedings of the REMIDA workshop Mar 2014 Presentation at the UniFi institution – Mar., 2014

4. Sediment toxicity monitoring: BioTox test The sediments toxicity analysis is assayed with the BioTox test (Aboatox Oy, Turku, Finland) according to the ISO standard method (ISO 11348e3, 1998).  Bio-indicator: Vibrio fischeri, a luminescent marine bacterium. _ Sensitivity to contaminants  N sample bacteria _ Luminescence  amount of contaminants  Advantages: _ Standardization _ Many kind of substances (soils and sediments; liquids or solids) _ Economical and rapid _ Correlation with the effects on higher organisms (Qureshi et Al., 1998)

5. BioTox test: analysis procedure Suspension of 2 g of sieved (<2 mm) sediments in 8 ml of 2% NaCl Shake for 5min by hand and settling for 30min Adjustment of pH and conductivity Reconstituition freeze-dried V. fischeri cells Addition of 300 ml of the bacterial suspension to 300 ml of samples (solid/supernatant) Measure of bioluminescence Calculation of the inhibition of bioluminescence index (INH%) after 15 or 30 minutes < 20% INH% > 20%

6. BioTox test: preliminary results Untreated sediments dredged in November, 2013: pore water (toxicity of lecheates) and sediment slurry Legend 1: Incile 2: Sandy; low contamination 3: Sandy-clay; middle contamination 4: Clay-silty; middle contamination Sediment slurry 1234 Mean INH% 51.6620.8800 Pore water 1234 Mean INH% 0000 Toxic!

7. BioTox test: preliminary results Untreated sediments dredged in November, 2013: dilituition series on 1 and 2 samples Legend 1: Incile 2: Sandy; low contamination 1 1:11:21:41:81:161:321:641:128 2921172022700 2 1:11:21:41:81:161:321:641:128 00191711100

8. BioTox test: preliminary results Treated sediments sampled in November 28, 2013: torbid samples None samples showed toxicity! Legend 1: Paspalum vaginatum + Eisenia foetida 2: Tamarix gallica + Paspalum vaginatum + Eisenia foetida 3: Spartium junceum + Paspalum vaginatum + Eisenia foetida 4: Nerium oleander + Paspalum vaginatum + Eisenia foetida 5: Phragmites australis + Paspalum vaginatum + Eisenia foetida 6: No plants used as control area 123456 Mean INH% 000000

9. BioTox test: preliminary results Treated sediments sampled in January 28, 2014: pore water (toxicity of lecheates) and sediment slurry The last sampling confirmed the non toxicity of the treated sediments! Torbid samples 1 (0-30cm)2 (0-30cm)3 (0-30cm)4 (0-30cm)5 (0-30cm)6 (0-30cm) 000000 1 (30-60cm)2 (30-60cm) 3 (30-60cm)4 (30-60cm)5 (30-60cm)6 (30-60cm) 001.907.7500 Pore water 1 (0-30cm)2 (0-30cm)3 (0-30cm)4 (0-30cm)5 (0-30cm)6 (0-30cm) 000000 1 (30-60cm)2 (30-60cm) 3 (30-60cm)4 (30-60cm)5 (30-60cm)6 (30-60cm) 000 0 00

10. Conclusions The solid phase of untreated sediments (samples 1 and 2) is toxic The pore water is not toxic, indicating that contaminants are strongly retained by the solid phases Phytoremediated sediments are non toxic either the solid phase and the lecheates There is no difference between 0-30 cm (high plant root colonization) and 30-60 cm (low plant root colonization) Microbial activity seems the main factor in de-toxification of treated sediments

11. Microbial diversity analysis … in progress Two aims: 1) Understanding the progress of microbial diversity 2) Amend phytoremediated sediments with plant growth promoting rhizobacteria (PGPR)

12. Additional activities Test the performance of phytoremediated sediments as a suitable substrate in real plant nursery companies

13. Thanks for your attention!