Andreja%20palatinus microlitter%20monitoring

Photo: Eco Vitae, Boštjan Mljač

Overview of the proposed DeFishGear Monitoring and Assessment Strategy of micro litter Andreja Palatinus, IWRS dr. Manca Kovač Viršek, IWRS Dr. Elen Kaberi, HCMR Dr. Cristina Zeri, HCMR Dr. Catharina Tsangaris, HCMR Giluio Pojana, Ca‘Foscari Dr. Marilyine Pflieger, UNG WP4 Working Group Meeting on Marine Litter 12th May 2014, Athens Greece

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Main goal of WP5 • To choose common approach to assess microplastic pollution on sea surface (accumulation zones, fishery areas, river outflows), beach sediment and ingested by biota (commercially available fish); • Analyse chemical composition of found microlitter; • Analyse POPs on pellets and larger pieces; • Start research activities on nanoplastic (<0,3mm) analysis.

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Partnership Institute for Water of the Republic of Slovenia The National Institute of Chemistry, Ljubljana, Slovenia ARPA Emilio-Romagna, Italy Caâ&#x20AC;&#x2DC;Foscari University of Venice, Italy University of Nova Gorica, Slovenia Institute for Oceanography and Fisheries, Croatia Hydro-engineering Institute of the Faculty of Civil Engineering, Sarajevo Bosnia&Herzegovina

Agricultural University of Tirana, Albania

Hellenic Centre for Marine Research,Greece Euro-Mediterranean Center or Climate Change, Italy The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

WP5 Timeline 5.1 - Defining an assessment and monitoring strategy for microplastics in the Adriatic 5.2 - Pilot survey I based on modeling results (sea surface, beach sediment) 5.3 - Data collection, evaluation, processing, validation of modeling 5.4 - Pilot survey II in fishing areas (sea surface, biota) 5.5 - Data analysis and reporting The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Sampling (planned) â&#x20AC;˘ 2014 seasurface + riverine outflow + beach sediment sampling locations will be determined by the calculation of the accumulation zones of pollution by CMCC (except Bosnia & Herzegovina and Slovenia)

â&#x20AC;˘ 2015 seasurface + biota sampling locations will be in fishing areas (except Bosnia & Herzegovina and Slovenia)

Foto: CMCC The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Seasurface sampling • IWRS & ARPA experience • Manta net (Eriksen) • Out of the wake zone using a spinnaker boom or „A“ frame • 30 min, speed 2.5 - 3 knots • Only in calm weather (1-2 beaufort) • June-October and if possible November-March Precaution to minimise contamination & Metadata recording as in TG ML Guidelines The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Foto: Eriksen

Country

Year 2014

Year 2015 (fishery area)

Type of sample

Number of sampling days

Number of samples

Number of sampling days

Number of samples

Sea surface

2 days

10

2 days

10

River outflow surface

1 day

2

Sea surface

3 days

15

3 days

15

River outflow surface

1 day

2

Sea surface

1 day

5

1 day

5

River outflow surface

1 day

2

Greece

Sea surface

2 days

10

2 days

10

Slovenia

Sea surface

1 day (2 days) 5 (10)

1 day (2 days)

5 (10)

Bosnia Herzegovina

Sea surface

?

?

?

Croatia

Italy

Albania

?

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Seasurface sample analysis • Microplastics separation • Microplastics analysis (chemical) Sample separation method: • Sample dilution followed by dissecting stereomicroscope

Photo: NIC

– Stereomicroscope (80x zoom) – Particles categorized by the size, type, shape and color (MSFD Technical Subgroup on Marine Litter, 2013)

• Results expression – particles/m3 and particles/m2 and also weight/m3 and weight/m2 The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Chemical caracterization

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Beach sediment sampling • HCMR experience

• sandy beaches on the top of the shore (strand line) • Upper 5cm of the surface Photo:HCMR sediment • At least 5 replicates, separated by at least 5m • Sampling separately the two size categories: 1-5mm and 20μm-1mm • before May or after September (October, November – depending of the weather) The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Country

Year 2014 Type of sample

Number of sampling Number of days samples

Croatia

Beach sediment

Italy

Beach sediment

1 beach (before and 10 after summer) 1 beach (before and 10 after summer)

Albania

Beach sediment

-

-

Greece

Beach sediment

30

Slovenia

Beach sediment

3 beaches (before and after summer) 1 beach (before and after summer)

Bosnia Herzegovina

Beach sediment

?

?

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

10

Seabed sample analysis • Microplastics separation • Microplastics analysis (chemical) Photo: HCMR

• Density separation:

– A known volume (normally 50 ml) of sediment is added to a separating funnel using a metal spoon and 200 ml of saturated NaCl added. A stopper is added and the mixture agitated by hand for 2 minutes, then allowed to settle for 2 minutes. The supernatant is then transferred to suction filtration via a Buckner funnel and passed through a 10μm retention filter paper. Filter papers are removed and stored in sealed petri dishes prior to examination under the microscope. The NaCl separation procedure is repeated three times with each sediment sample to ensure a high recovery of buoyant debris.

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Biota WP4 • HCMR & IOF Split experience • Commercially available fish (benthic and pelagic) species (additionally mussels & Caretta caretta)

– Species, which fulfill the requirements of an indicators species such as sand eel, sprat, herring, and sardines, should be considered

• A sample size of at least 50 specimen per species • Potential use of already ongoing fisheries analysis The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Photo: HCMR

• Timing: spring 2015 • IT - mussels; • ALB, CRO, GR, SI - commercially available fishes

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Biota sample analysis Microplastics separation • Gut examined with a dissecting microscope • Digestive tract slit open using scissors and examined immediately • Depending on the size of the organism the entire gut or samples of gut wall can be examined • Fragments of unusual appearance removed with forceps, placed on clean filter papers in petri dishes and sealed • Results: Nr of items/g of intestine, size (100s µm), color, shape Categorisation and identification of particles • Fragments categorized according to size, shape, color etc • Identification of polymere type via spectroscopy (FT-IR, RAMAN or NIR Raman) The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

POPs analysis • International Pellet Watch studies –IPW(Endo et al., 2005; Ogata et al., 2009; Hirai et al, 2011) • sampling procedure for sediment (OSPAR Commission) • Focus mostly on PE pellets (100-200 pellets (at least 50) per location/beach) • Later floating litter WP4

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

POPs analysis Main steps of the procedure: • Extraction of POPs in organic solvent by Soxhlet and/or ASE (Accelerated Solvent Extractor); • Purification by SPE (Solid Phase Extraction) on florisil based sorbent; • Concentration step: Evaporation and redissolution in appropriate solvent; • Analysis by GC-MS (20 PCBs), GC-ECD (DDTs, lindane and its isomers, endosulfan).

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Nanoplastics (0.03-0.3mm) • Ca‘ Foscari UNIVE experience and work • Experimental part of the project Aim: • Development of specific protocols for the determination of nanoplastics by modification/adaptation of methodologies and protocols already available (proposed/applied) for microplastics, as those recently recommended by European Commission Technical Subgroup on Marine Litter The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Nanoplastics • data to be obtained and reported for a correct assessment of nanoplastic pollution and identification of impacts on marine biota • metrics (number, mass) • size distribution • shape (sphere, fiber, platelet, etc.)

• chemical composition (polymer composition)

• impurities (organic, inorganic)

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Nanoplastics • Critical steps to be evaluated: – sampling protocols (size and matrix dependant) – extraction procedures (size and matrix dependant) – purification techniques (i.e. separation from natural particles) – instrumental identification

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

• techniques to be evaluated according to: – applicability to the specific size range – statistical significance – simplicity (wide applicability in different labs) – matrix (sediment, water, biota)

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Thank you very much for your attention Andreja Palatinus - Andreja.palatinus@izvrs.si dr. Manca Kovač Viršek – manca.virsek@izvrs.si Dr. Elen Kaberi - ekaberi@hcmr.gr Dr. Christina Zeri - chris@hcmr.gr Dr. Catharina Tsangaris - ctsangar@ath.hcmr.gr Dr. Giulio Pojana- jp@unive.ti Dr. Maryline Pflieger - marilyne.pflieger@ung.si

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Sample collection Rinsing the net

with the filtered sea water or tap water

End volume of sample

max. 250ml

Type of jar used (material)

glass

Fixation on the boat

70 % alcohol

Slikca vzorca z ladje

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

â&#x20AC;˘ Microplastics in beaches can be sampled at the same time as macro debris on beaches, or in parallel with any other routine intertidal monitoring (e.g. for chemical contaminants, biota) â&#x20AC;˘ Reporting units: items/ml of sediment (LMP), items/g of sediment (SMP), relative abundance of main colors and shapes

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)

Sample collection

SMP sampling*

LMP sampling

Collect from the top 5cm Collect from the top 5cm of sand using a metal of sand using a metal spoon. spoon.

Procedure End volume of sample

max. 250ml

Type of jar used (material)

Metal/glass/paper

Fixation

-

Max. 5 l Pass the sample through a 1 mm metal sieve.

Metal/glass/paper -

*The SMP should be collected prior to the LMP samples **Recommendation: extend the protocol for meso-debris (525mm) using a 5mm sieve to separate debris from beach sediment. Preferably the sieves could be stacked together.

The Project is co-founded by the European Union, Instrument for Pre-Accession Assistance (IPA)