INDICIT tools

GA n°11.0661/2016/748064/SUB/ENV.C2

INDICIT tools

Tools Versions September 2018

GA n°11.0661/2016/748064/SUB/ENV.C2

1

2

Video-tutorial

Special focus on

entanglement3

Protocol for collecting data

on litter ingestion

GA n°11.0661/2016/748064/SUB/ENV.C2

1 VIDEO-TUTORIAL

Video

https://www.youtube.com/watch?v=fNnh_pAKIxU

GA n°11.0661/2016/748064/SUB/ENV.C2

2PROTOCOL FOR COLLECTING DATA

ON LITTER INGESTION

https://indicit-europa.eu/protocols/

Deliverable 2.6

https://indicit-europa.eu/protocols/

GA n°11.0661/2016/748064/SUB/ENV.C2

Protocol for collecting data on litter ingestion

Origin:• Protocol from 2013 MSFD guidance on monitoring marine litter in

European seas (Fulmar, Turtles)

• Improvements from experience, results, collaborators’ comments

Objectives:• Collecting, classifying and quantifying the debris ingested by sea turtles

• Assessing body condition

• Evaluating the possible causes of death and impacts caused by debris

• Particular focus on Entanglement

Focus• Example on Caretta caretta, applicable to all species (except Dermochelys

coriacea for some manipulations → biometric measures)

• Detail Plastic categories (+ other types of debris)

In brief

Comments to make the protocol evolve are welcome!!

GA n°11.0661/2016/748064/SUB/ENV.C2

Sea turtles are protected species• Only authorized people can handle live and dead animals or parts of them.

→ Contact responsible Authorities.• CITES permit asked for sending/receiving specimen or samples


Special remarks

Sanitary precautions• Be careful to minimize risks for manipulator as for alive

animal (zoonosis)• Mark-off place of intervention to keep away and protect

bystanders• Wear protective suit with glasses, gloves (a cut-resistant

pair of gloves below 2 pairs of gloves) and rubber boots• Carefully separate and disinfect or thrown materials.

2 types of manipulations• Necropsy of dead turtle: Collect debris from the entire digestive tract

(B1, p.11)• Faeces analysis on alive turtle: Collect debris from excretions

(C1, p.13)• General information common to the 2 protocols

GA n°11.0661/2016/748064/SUB/ENV.C2


Common datasheets

GA n°11.0661/2016/748064/SUB/ENV.C2

Associated Standard Database

2 tabs: Necropsy / Faeces

1 line per individual

→ Data banking will be managed by other procedures (ex. Clean Atlantic)

Data shared in private area


Common database

GA n°11.0661/2016/748064/SUB/ENV.C2

~ 5H / individual for protocol necropsyPreferable to be 2 manipulators


Other tools

GA n°11.0661/2016/748064/SUB/ENV.C2


THE PROTOCOL STEP BY STEP

GA n°11.0661/2016/748064/SUB/ENV.C2

• “Basic” parameter: Fundamental for using the indicator and assessing distance to GES

• “Optional” parameters: Acquire knowledge on the factors influencing litter ingestion in sea turtles


Basic and optional parameters

Black / Red

Black / Grey

GA n°11.0661/2016/748064/SUB/ENV.C2

A1. ON SITE:

-> Describe discovery placeLocation, coordinates

-> Give contact coordinatesName, Tel…

-> Identify the individualSpecies, Tag, (standard) Identification code → all samples contents

-> Take picturesUse a scale; identify the picture

Image fro

m IN

DIC

IT video

-tuto

rial


A. GENERAL INFORMATION

GA n°11.0661/2016/748064/SUB/ENV.C2

Circumstances:

• Stranding

•Bycatch/fisheries

• Found at sea

•Dead at the Recovery Centre


A2 to A3. DESCRIBE ANIMAL BODY CONDITION

→ By-catch engine

→ Stranding cause

• Bycatch• Entanglement• Anthtopogenic trauma•Natural predation• Petrol / oils•Natural disease•Others

From Alfaro-Shigueto et al., 2011

• Longline• Trawl•Drifting net•Other

GA n°11.0661/2016/748064/SUB/ENV.C2

Status: From 1 to 5

• Status 1 → “Alive turtle” Protocol → Faeces (C1), except if dead after the recovery (e.g., at the rescue center). In this case, report to tab “Necropsy”

• Status 2 to 4 → Necropsy

• Status 5 reported but not considered in statistics



GA n°11.0661/2016/748064/SUB/ENV.C2

Impacts of litter

• Probable cause of death or stranding

• Main injuries + Affected body part

• Entanglement characterization (described latter)

• Health status

• Fat reserves (in Other descriptive parameters)


From Thomson et al., 2009

Overall evaluation from:- External examination- Internal examination


GA n°11.0661/2016/748064/SUB/ENV.C2


Biometric measures• Curved (CCL) and straight (SCL) carapace lengths (0.1 cm)

• Weight (grams)

Min Standard MAX


With biometric measure, test in litter ingestion according to stage and the area where turtles may ingest plastics

GA n°11.0661/2016/748064/SUB/ENV.C2

DIGESTIVE TRAC(NECROPSIES)

STRANDING CAUSE

FAECES ANALYSIS

DEAD TURTLES

ALIVE TURTLES

IND1: Debris ingestion (>1mm)

IND2: Entanglement on

marine debris

IND3: micro-debris ingestion (1-5mm)

SAMPLING DEBRIS

Fig

ure

13.

Rem

oval

of

the p

lastr

on

. (a

) C

utt

ing

th

rou

gh

so

ft t

issu

e a

t th

e

pla

str

on

-cara

pace j

un

cti

on

; (b

) E

levati

on

of

the f

ron

t ed

ge u

sin

g a

ho

ok,

an

d e

xcis

ion

of

skele

tal

mu

scle

tis

su

e;

(c)

Ven

tral

vie

w o

f a c

arc

ass w

ith

th

e p

lastr

on

rem

oved

.

F

ore

lim

bs

Exam

ine

the

fore

limbs

to

assess

muscle

m

ass

and

identify

any

fore

ign

bodie

s.

Make a

cir

cum

fere

ntial

incis

ion

thro

ugh t

he s

kin

aro

und e

ach f

ore

limb

(Fig

ure

14a).

E

levate

th

e c

audal

aspect

of

the s

capula

(c

ora

coid

pro

cess)

and e

xcis

e t

he c

onnective t

issue o

f th

e d

ors

al

surf

ace o

f th

e p

ecto

ralis

majo

r m

uscle

gro

up f

or

each f

ore

limb (

Fig

ure

14b).

In

a h

ealthy tu

rtle

, pecto

ralis

m

ajo

r m

uscle

s s

hould

be r

ed t

o p

ink,

firm

and a

mple

. A

s a

nim

als

beco

me

em

acia

ted,

muscle

s a

trophy a

nd b

ecom

e p

ale

. R

ota

te t

he

flip

pe

r m

ed

ially

.

Severa

l com

ple

te

rota

tions

should

re

move

the

flip

per

by

tearing

the

connective t

issues (

Fig

ure

14c).

A

ll underlyin

g s

tructu

res s

hould

be left inta

ct

(Fig

ure

14d).

P

reserv

e t

he f

ore

limbs i

f skele

tal chro

nolo

gy t

ests

or

toxic

olo

gy

scre

enin

g is t

o b

e c

onducte

d.

F

or

skele

tal

chro

nolo

gy,

str

ip t

he f

ore

limb o

f m

uscle

and c

onnective t

issue t

o

expose

the

hum

eru

s.

C

ross-s

ectional

sam

ple

s

of

the

mid

-shaft

hum

eru

s

should

be c

olle

cte

d f

or

measure

ment.

For

toxic

olo

gy scre

enin

g,

rem

ove a

50 x

50 x

25 m

m (

if p

ossib

le)

pie

ce o

f re

pre

senta

tive s

kele

tal

muscle

. A

com

mo

n s

ite f

or

muscle

harv

esting i

s t

he

fore

limb

s.

In

ad

ditio

n,

co

llect

a r

ep

resenta

tive s

am

ple

of

ca

rdia

c m

uscle

by

excis

ing

a

suitable

siz

ed

full

thic

kness

pie

ce

of

the

ventr

icle

.

Consult

30

GA n°11.0661/2016/748064/SUB/ENV.C2


• Various possible sampling → from the cleaner to the dirty (e.g., cranial

cavity first)

• Preferable to be 2 manipulators to turn/maintain the animal body

OesophagusStomach

Intestines

Clamp and separate each section

B) SAMPLING FROM DEAD INDIVIDUALS:(DIGESTIVE TRACT ANALYSIS)

GA n°11.0661/2016/748064/SUB/ENV.C2


• Various possible sampling → from the cleaner to the dirty (e.g., cranial

cavity first)

• Preferable to be 2 manipulators to turn/maintain the animal body

Stomach + Intestines (here 2 parts)

1 mm sieve → Analyse or frieze

(cutaneous papillae)

Oesophagus

B) SAMPLING FROM DEAD INDIVIDUALS:(DIGESTIVE TRACT ANALYSIS)

GA n°11.0661/2016/748064/SUB/ENV.C2


(1mm mesh)

→ Analyse or frieze

Direct observation Filtering water tank

C) SAMPLING FROM ALIVE INDIVIDUALS:(FAECES ANALYSIS)

GA n°11.0661/2016/748064/SUB/ENV.C2


D) DEBRIS ANALYSIS AND CLASSIFICATION:LITTER CLASSIFICATION:

→ Adapted from MSFD classification

GA n°11.0661/2016/748064/SUB/ENV.C2


Occurrence (1/0)

By category in each digestive section / faeces• Dry mass (0.01 gram)• Number of fragments• Number of types of items

For plastic only• Total dry mass• Total number of fragments• Total volume of items• Color per category• Number per size category

D) DEBRIS ANALYSIS AND CLASSIFICATION:LITTER ANALYSIS:

GA n°11.0661/2016/748064/SUB/ENV.C2


→ specific analysis for micro (1-5 mm)(optional protocol)

D) DEBRIS ANALYSIS AND CLASSIFICATION:LITTER ANALYSIS:

Debris > 1mm

micro Macro

2 sieve train

MACRO

DEBRIS(>5mm)

5mm

SIEVE (rigid)

DIET

SAMPLE-20ºC

• IDENTIFICATION

• CLASSIFICATION

• COUNT

• WEIGH

Visual extraction

1mm

SIEVE (flexible)

MICRO

DEBRIS(1-5mm)

DIGESTION(KOH, H2O2, etc.)

GA n°11.0661/2016/748064/SUB/ENV.C2


• Training sessions

• Specific material or overall kit

• Share your observations in column “Notes”

• Other ideas• Forum (share pictures, ask questions…)

Forward the needs and share experience

GA n°11.0661/2016/748064/SUB/ENV.C2


Probably high impact on marine biota

Developing a new Litter impact indicator:

• See online Feasibility study→ Need standardize typology→ Passive (litter) Active (fishing)→ Requires specific protocol and database

Sea turtle = a good indicator taxa• Available competent network• Recurrent observation of entanglement• Recognized by experts as fundamental and feasible

3 SPECIAL FOCUS ON ENTANGLEMENT

GA n°11.0661/2016/748064/SUB/ENV.C2


Probable cause of death/stranding

Main injuries & affected body part

Litter causing entanglement

(multiple choice)• Fishing nets• Monofilament• Ropes• Plastic bags• Raffia• Other

GA n°11.0661/2016/748064/SUB/ENV.C2

Thank you

INDICIT tools

Documents

Transcript of INDICIT tools