Cristina%20fossi

Are macro and micro‐ Are macro and micro‐plastics impacting marine organisms in the Pelagos Sanctuary? in the Pelagos in the

M. Cristina Fossi ‐ University of Siena ‐ Fossi@unisi.it

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Can Plastic Affect l i ff

Mediterranean Biodiversity?

Mediterranean Biodiversity?

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Marine Litter: a Global Challenge The main legal and institutional frameworks affecting the Mediterranean on this topic are: (1) Local Agendas 21;

(2) national legislation on waste management and environmental protection; (3) the Barcelona Convention and its Protocols; (4) the Mediterranean Strategy for Sustainable Development (MSSD); (5) MEDPOL of UNEP; (6) the EU Environmental Strategy for the Mediterranean and Horizon 2020; (7) the EU Marine Strategy Directive; (8) the EU Thematic strategy on the Prevention and Recycling of Waste; (9) the IMO MARPOL 73/78 Convention – Annex V; (10) the GPA and the Regional Seas Programme of UNEP; (11) the Basel Convention

GAP

There is a general lack of available d t i ildlif ff t d b data on marine wildlife affected by marine litter in the Mediterranean.

UNEP/MAP Barcelona Convention RAP on Marine litter in Mediterranean i li i di (Istanbul 2013)

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Gap Information are required about macro‐plastic and micro plastic inputs, Mediterranean distribution micro‐plastic inputs Mediterranean distribution (Pelagos Sanctuary) and the potential effects on marine organisms. marine organisms.

? BEST INDICATOR SPECIES ?

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Do microplastics threat Do microplastics Pelagos l Sanctuary??

The Pelagos g Agreement establishing g g the Sanctuaryy for marine mammals in the Mediterranean Sea, is an international governmental agreement between France, Italy and the Principality of Monaco entered into force in 2002 to insure a favorable entered into force in 2002, to insure a favorable conservation status of marine mammals by protecting them and their habitat from direct and indirect negative impacts of human activities, in compliance li with ith a management plan. t l With about 87,500 sq. km, most of which lie in high Seas, the Pelagos Sanctuary is registered as a Specially p y Protected Area of Mediterranean Importance (SPAMI).

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Aim of the Project Aim of the Project Do microplastics threat p The Pelagos The Pelagos Sanctuary?

Are baleen whales Are baleen whales exposed to microplastic threat? threat?

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Marine organisms as sentinel species: micro‐ species: micro‐plastic Case studies: Mediterranean fin whale and basking shark Case studies: Mediterranean fin whale and basking shark Aim: exploring the toxicological effects of micro‐plastics in large filter feeders species Further implication: indicators of micro plastics in the pelagic in the pelagic Further implication: indicators of micro‐plastics environment in the implementation of the EU Marine Strategy Framework Directive (Descriptor 10)

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Balaenoptera physalus 8

300 liters of water daily

70,000 liters of water with each mouthful

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Microplastics and contaminants

Adsorption of POPs on on microplastics surface Plastic additives released in the environment

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Experimental work The work is implemented through four steps: Step 1 collection/count of microplastics collection/count of microplastics in Pelagos in Pelagos Step 1‐ Sanctuary (Mediterranean Sea); Step 2‐ detection of phthalates in superficial neustonic/planktonic samples; Step 3 ‐ detection of phthalates in stranded Mediterranean fin whale; Mediterranean fin whale; Step 4 ‐ detection of phthalates and biomarkers responses (CYP1A1, CYP2B, lipid peroxidation) in skin biopsies of fin whales collected in the Pelagos biopsies of fin whales collected in the Pelagos Sanctuary (n=18) and Sea of Cortez (n=7).

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Do Microplastics threat p the Pelagos the Pelagos Sanctuary?

Yes!

LIGURIA Microplastics (Items/m3)

Microplastic particles in superficial neustonic/planktonic samples (items/m3) collected in the Pelagos Sanctuary (Ligurian Sea and Sardinian Sea) and mean DEPH and MEPH concentrations (ng/g).

MPM14 MPM15

0-0.1

MPM13

0 11-1 0.11 1

MPM12 MPM11

MPM16 MPM8

1.01-5

MPM7 MPM9

5.01-10

Ligurian Sea MPM5 MPM6

MPM3

MPM10 MPM4 MPM18 MPM23

MPM24

Sardinian Sea MPM25

MPM17

Phthalates concentration in superficial neustonic/planktonic samples

MPM26 MPM19 MPM20

MPM21

SARDINIA

AREA

DEHP (ng/g)

MEHP (ng/g)

n

mean

sd s.d.

n

mean

sd s.d.

Ligurian Sea

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18.38

44.39

14

61.93

124.26

Sardinian Sea

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23.42

32.46

9

40.30

41.55

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Microplastics in Ligurian Sea Microplastics in Ligurian Microplastics in

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Microplastics in Sardinian Sea

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Microplastics and MEPH Microplastics and MEPH –– DEPH i Pelagos in Pelagos in P l S t Sanctuary

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Phthalates as plastic tracer?

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Are baleen whales exposed to microplastics to microplastics l threat? h ?

MEHP concentration in stranded fin whales SPECIES

TISSUE

Mean MEHP (ng/g)

Balaenoptera physalus

Blubber

57.97 57 97

Yes! DEHP concentrations (ng/g) in blubber samples of five stranded fin whales collected along the Italian coasts during the period July 2007 – June 2011 in five different locations.

San Rossore PI (Male) g/g MEPH 53.98 ng/g Orbetello GR (Male) MEPH 51.84 ng/g

Castelsardo SS (nd) MEPH 1.00 ng/g Amalfi SA (Female) lf ( l ) MEPH 99.93 ng/g Palinuro SA (nd) MEPH 83.12 ng/g

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Presence and effects of contaminants in fin whale skin biopsies t i t i fi h l ki bi i

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Contaminants in fin whale skin biopsies k b 90

MEHP

30000

80

OCs

25000

70 20000

ngg/g d.w.

ng/g d.w. n

60 50

15000

40

10000

30 20

5000 10 0

0

Pelagos Sanctuary

Sea of Cortez

Pelagos Sanctuary

Sea of Cortez

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No ormalized ffold expresssion

Endocrine interference iin fin whale skin biopsies fi 12 h l ki bi i 10 8

**

Estrogen Receptor α

Males F Females l

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(** = p< 0.05) 0.05)

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**

2 0 Pelagos Sanctuary Undesirable Biological Effect

Gulf of California

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Differences between areas e e ces bet ee a eas Cluster dendrogram: phthalates, Ocs and biomarkers responses in skin bi i f fi h l ll di h P l S dS f biopsies of fin whales collected in the Pelagos Sanctuary and Sea of Cortez

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Messico

80000 600 000

Sardegna g

2

1

40000 20000 2

Height

100000 0

120000 140000

Cluster Dendrogram

Liguria g

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Do fin whales like microplastics ?

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Are other Are other large filter feeders large filter feeders exposed to p microplastics threat?

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BASKING SHARK SHARK

Approx. 12600 items/day!

A 12600 i /d !

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MEHP concentration in basking sharks SPECIES

TISSUE

M Mean MEHP ( / ) MEHP (ng/g)

Basking sharks

muscle

12.97

Phthalate as plastic tracer hh l l i 26

Marine organisms as sentinel species: macro‐ species: macro‐plastic Case study: the Mediterranean Loggerhead turtle (Caretta caretta) Aim: exploring the toxicological effects of macro‐plastics Further implication: indicators of macro‐plastics p p in the marine environment in the implementation of the Descriptor 10 of MSFD

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found 483 pieces of marine litter with a i litt ith total mass of 62.37g

145 plastic items in the stomach

22 loggerhead turtles out of 31 animals had ingested marine ingested marine debris (71%)

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MSFD objective: to achieve the Good Environmental status for the marine environment for 2020 Caretta caretta has been proposed as a target species in the Mediterranean sea in the MSFD (Descriptor 10) to evaluate indicator IV of GES: trends in the amount and d composition iti off litter litt ingested i t d by b marine i animals i l (e.g ( stomach t h analysis) l i)

2020 Stomach content in sea turtles in 2013 sea turtles in 2013

Plastic items = ?

Stomach content in sea turtles in 20XX

Time scale

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Effects of litter in free in free‐‐ranging turtles? turtles?

Comet assay (Frenzilli et al., 1999) ENA assay (Pacheco & Santos 1997) Diffusion assay (Singh et al., 1988) Polycyclic aromatic hydrocarbons (Marsili et al., 1997) Organochlorines (Marsili & Focardi, 1996)

WHOLE BLOOD EXCRETA Porphyryns (Grandchamp et al., 1980) Plastic fragments

PLASMA LPO, Lipid peroxidation (Bird & Draper, 1984) γGT, ALT, AST (commercial kits, Polymed) VTG, Vitellogenin (Goksoyr et al., 1991) EST E EST, Estradiol (Abraham, 1969) di l (Ab h 1969) BChE, Butyrylcholinesterase (Ellman et al., 1961) AChE, Acetylcholinesterase (Ellman et al., 1961)

CARAPACE Trace elements (Hg, Pb, Cd) (Stoeppler & Backhaus 1978) (Stoeppler & Backhaus, 1978)

SKIN BIOPSY

Protein expression of CYP1A (Fossi et al., 2008, modified) Biopsy slices treated with PAHs and PBDEs (Fossi et al., 2009)

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Take Home Message

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The workshop Th kh recommended d d that th t baleen b l whales h l and d other large filter feeders should be considered in national and international marine debris strategies (e.g. (e g Descriptor 10 (marine litter) in the EU Marine Strategy Framework Directive) as critical indicators of the presence and impact of microplastics in the marine environment.

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Take Home Message

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MSFD: Descriptor 10 Marine Litter Marine Litter

The potential use of these species in the implementation of the Descriptor 10 (marine litter) in the EU Marine Strategy Framework Directive (MSFD), as presence and impact p of micro‐litter in the p pelagic g sentinel of the p environment

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Marselle

Venice

Leghorn Barcelona Naples

Dubrovnik

Algiers

Antalya

Gibraltar La Valletta

Tunis

Heraklion Tripoli

Larnaca

Athens

Beirut Tel Aviv

Alexandria

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Acknowledgements g This project was partially supported by the Italian Ministry for Environment and Territory

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Thank you for your attention!

Thank you for your attention!

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