andreja%20palatinus microlitter%20monitoring

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

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


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.


Partnership

Institute for Water of the Republic of Slovenia

The National Institute of Chemistry, Ljubljana, Slovenia

ARPA Emilio-Romagna, Italy

Ca‘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


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


Sampling (planned)

• 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)

• 2015 seasurface + biota

sampling locations will be in

fishing areas (except Bosnia &

Herzegovina and Slovenia)

Foto: CMCC


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

Foto: Eriksen

Precaution to minimise contamination & Metadata recording as in TG ML Guidelines


Country Year 2014 Year 2015 (fishery area)

Type of sample Number of

sampling

days

Number of

samples

Number of

sampling

days

Number of

samples

Croatia Sea surface 2 days 10 2 days 10

River outflow

surface

1 day 2

Italy Sea surface 3 days 15 3 days 15

River outflow

surface

1 day 2

Albania 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 ? ? ? ?


Seasurface sample analysis

• Microplastics separation

• Microplastics analysis (chemical)

Sample separation method:

• Sample dilution followed by dissecting stereomicroscope

– 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

Photo: NIC


Chemical caracterization


Beach sediment sampling

• HCMR experience

• sandy beaches on the top

of the shore (strand line)

• Upper 5cm of the surface

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)

Photo:HCMR


Country Year 2014

Type of sample Number of sampling

days

Number of

samples

Croatia Beach sediment 1 beach (before and

after summer)

10

Italy Beach sediment 1 beach (before and

after summer)

10

Albania Beach sediment - -

Greece Beach sediment 3 beaches (before

and after summer)

30

Slovenia Beach sediment 1 beach (before and

after summer)

10

Bosnia -

Herzegovina

Beach sediment ? ?


Seabed sample analysis

• Microplastics separation • Microplastics analysis (chemical)

• 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.

Photo: HCMR


Biota

• 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

WP4

Photo: HCMR


• Timing: spring 2015

• IT - mussels;

• ALB, CRO, GR, SI - commercially available fishes


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)


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


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).


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


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)


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


• techniques to be evaluated according to:

– applicability to the specific size range

– statistical significance

– simplicity (wide applicability in different labs)

– matrix (sediment, water, biota)


Thank you very much for your attention

Andreja Palatinus - [email protected] dr. Manca Kovač Viršek – [email protected]

Dr. Elen Kaberi - [email protected] Dr. Christina Zeri - [email protected]

Dr. Catharina Tsangaris - [email protected] Dr. Giulio Pojana- [email protected]

Dr. Maryline Pflieger - [email protected]

mailto:[email protected]










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


• 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)

• Reporting units: items/ml of sediment (LMP),

items/g of sediment (SMP), relative abundance of main colors and shapes


*The SMP should be collected prior to the LMP samples **Recommendation: extend the protocol for meso-debris (5-25mm) using a 5mm sieve to separate debris from beach sediment. Preferably the sieves could be stacked together.

Sample collection SMP sampling* LMP sampling

Procedure Collect from the top 5cm

of sand using a metal spoon.

Collect from the top 5cm of sand using a metal

spoon.

End volume of sample max. 250ml Max. 5 l

Pass the sample through a 1 mm metal sieve.

Type of jar used (material)

Metal/glass/paper Metal/glass/paper

Fixation - -

andreja%20palatinus microlitter%20monitoring

Documents

italy sea surface

european union

greece sea surface

albania sea surface

river outflow surface

fishing areas sea surface

sea surface accumulation

modeling results sea