indicit tools
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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
GA n°11.0661/2016/748064/SUB/ENV.C2
2PROTOCOL FOR COLLECTING DATA
ON LITTER INGESTION
https://indicit-europa.eu/protocols/
Deliverable 2.6
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
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
Common database
GA n°11.0661/2016/748064/SUB/ENV.C2
~ 5H / individual for protocol necropsyPreferable to be 2 manipulators
Protocol for collecting data on litter ingestion
Other tools
GA n°11.0661/2016/748064/SUB/ENV.C2
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
A. GENERAL INFORMATION
GA n°11.0661/2016/748064/SUB/ENV.C2
Circumstances:
• Stranding
•Bycatch/fisheries
• Found at sea
•Dead at the Recovery Centre
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
A2 to A3. DESCRIBE ANIMAL BODY CONDITION
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)
Protocol for collecting data on litter ingestion
From Thomson et al., 2009
Overall evaluation from:- External examination- Internal examination
A2 to A3. DESCRIBE ANIMAL BODY CONDITION
GA n°11.0661/2016/748064/SUB/ENV.C2
Protocol for collecting data on litter ingestion
Biometric measures• Curved (CCL) and straight (SCL) carapace lengths (0.1 cm)
• Weight (grams)
Min Standard MAX
A2 to A3. DESCRIBE ANIMAL BODY CONDITION
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
Protocol for collecting data on litter ingestion
• 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
Protocol for collecting data on litter ingestion
• 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
Protocol for collecting data on litter ingestion
(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
Protocol for collecting data on litter ingestion
D) DEBRIS ANALYSIS AND CLASSIFICATION:LITTER CLASSIFICATION:
→ Adapted from MSFD classification
GA n°11.0661/2016/748064/SUB/ENV.C2
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
→ 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
Protocol for collecting data on litter ingestion
• 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
Protocol for collecting data on litter ingestion
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
Protocol for collecting data on litter ingestion
Probable cause of death/stranding
Main injuries & affected body part
Litter causing entanglement
(multiple choice)• Fishing nets• Monofilament• Ropes• Plastic bags• Raffia• Other