Cephalopods have been defined as “advanced invertebrates” due to the complexity of their nervous system and to their sophisticated behavioural repertoire. However, until recently, the protection and welfare of this class of invertebrates has been mostly disregarded by EU regulations on the use of laboratory animals. The inclusion of “live cephalopods” in the Directive 2010/63/EU has been prompted by new scientific knowledge on the “sentience” of animals used for experimental or other scientific purposes, a fundamental criterion to which animal species are included or not under the protective umbrella of the Directive. In this scenario, the imminent publication of the “Guidelines for the Care and Welfare of Cephalopods in Research” as an initiative by the CephRes-FELASA-Boyd Group is a sign of ethical progress in the consideration of animals in research, and is likely to have a significant impact on both scientific and practical aspects of research with these animals

Alleva, Enrico

Berry, Alessandra

Carere, Claudio

Vitale, Augusto

Annali dell Istituto Superiore di Sanità

Annali dell'Istituto Superiore di Sanità

C
o
m
m
e
n
t
a
r
y
267
Ann Ist Super Sanità 2015 | Vol. 51, No. 4: 267-269 
DOI: 10.4415/ANN_15_04_04
Key words
•  cephalopods
•  animal welfare 
•  sentience 
•  Directive 2010/63/EU 
•  guidelines 
Commentary
EU guidelines for the care and welfare  
of an “exceptional invertebrate class” 
in scientific research
Alessandra Berry1, Augusto Vitale1, Claudio Carere2,3 and Enrico Alleva1
1Dipartimento di Biologia Cellulare e Neuroscienze, Istituto Superiore di Sanità, Rome, Italy 
2Centro Ittiogenico Sperimentale Marino (CISMAR), Dipartimento di Scienze Ecologiche e Biologiche,  
Università della Tuscia, Tarquinia (Vt), Italy 
3Laboratory of Experimental and Comparative Ethology, University Paris 13, Sorbonne Paris Cité, France
Abstract
Cephalopods have been defined as “advanced invertebrates” due to the complexity of 
their nervous system and to their sophisticated behavioural repertoire. However, un-
til recently, the protection and welfare of this class of invertebrates has been mostly 
disregarded by EU regulations on the use of laboratory animals. The inclusion of “live 
cephalopods” in the Directive 2010/63/EU has been prompted by new scientific knowl-
edge on the “sentience” of animals used for experimental or other scientific purposes, a 
fundamental criterion to which animal species are included or not under the protective 
umbrella of the Directive. In this scenario, the imminent publication of the Guidelines 
for the care and welfare of cephalopods in research as an initiative by the CephRes-
FELASA-Boyd Group is a sign of ethical progress in the consideration of animals in 
research, and is likely to have a significant impact on both scientific and practical aspects 
of research conducted with these animals.
“Animals have an intrinsic value which must be respected. 
There are also the ethical concerns of the general public as 
regards the use of animals in procedures. Therefore, animals 
should always be treated as sentient creatures ….” [1]
INTRODUCTION
Europe has a long history of legislation on the use of 
laboratory animals that – in some countries – dates back 
to the 19th century (e.g. Denmark in 1891; Germany in 
1883; and UK, one of the first to regulate, in 1876). 
In 1986 the EU Directive 86/609/EEC (Council of the 
European Communities, 1986) and the European Con-
vention 123 (Council of Europe, 1986) were published 
with the main aim of harmonising local regulations on 
animals used for experimental and other scientific pur-
poses. However, both the aforementioned Directive 
and the Convention were meant to protect only live 
vertebrates. 
In the following period of twenty-four years (1986-
2010) the scientific community has achieved significant 
progress in experimental techniques, and these have 
been paralleled by changes in the ethical approaches 
to animal experimentation, leading to new insights into 
animal welfare. Moreover, the acquisition of new scien-
tific knowledge on the capacity of animals to experience 
and express pain, suffering, distress and lasting harm 
has resulted in the need to improve the welfare of ani-
mals used in scientific procedures by raising the mini-
mum standards protecting those animals still needed 
to be used for scientific purposes. The result of these 
new contexts has been the publication in 2010 of the 
European Directive 2010/63 on the protection of ani-
mals utilised in scientific procedures. Among the most 
important changes brought about by the new Directive 
it should be mentioned the welfare and protection of 
foetal forms of mammals (as from the last third of their 
normal development) and independently feeding larval 
forms; such protection also extends to genetically modi-
fied animals by defining their creation and maintenance 
as a procedure (see Article 3). 
Noteworthy, and differently from the EU Direc-
tive 86/609/EEC and Convention 123, the Directive 
2010/63/EU brings under its scope the welfare and 
protection of a whole class of invertebrates such as 
“live cephalopods” (both adults and juveniles). This is 
a major change in the legislation on the use of animals 
Address for correspondence: Alessandra Berry, Dipartimento di Biologia Cellulare e Neuroscienze, Istituto Superiore di Sanità, Viale Regina Elena 
299, 00161 Rome, Italy. E-mail: alessandra.berry@iss.it. 
Alessandra Berry, Augusto Vitale, Claudio Carere and Enrico Alleva
c
o
m
m
e
n
t
a
r
y
268
in scientific procedures [2] that takes into account 
advances not only in research techniques but also the 
improved understanding and assessment of animal wel-
fare, particularly in relation to invertebrates [3-5]. In 
this scenario, the recent publication of the Guidelines 
for the care and welfare of cephalopods in research [6] 
is a sign of ethical progress in considering animals in 
research, and is likely to have a significant impact on 
both scientific and practical aspects of research with 
these animals.
As reviewed by Smith and co-workers [2], in 1993 
the UK pioneered the concern for the use of inverte-
brates in research by including the welfare of Octopus 
vulgaris under the scope of the Animals (Scientific Pro-
cedures) Act - ASPA -1986. Likewise, scientific proce-
dures involving cephalopods and decapod crustaceans 
are regulated in Switzerland while Norway regulates 
the scientific use of squids, octopuses, decapod crus-
taceans and honeybees. The Australian Government 
National Health and Medical Research Council’s Code 
of Practice regulates the use of “cephalopods such as 
octopus and squid”; the New Zealand Animal Welfare 
Act includes “octopus, squid, crab, lobster and crayfish” 
under its scopes while the Canadian Council on Animal 
Care in science includes “cephalopods and some other 
higher invertebrates [that] have nervous systems as well 
developed as some vertebrates” insofar as they may ex-
perience from little to severe pain, stress, discomfort 
or other suffering (for further details see [2] and refer-
ences therein).
WHY CEPHALOPODS SHOULD BE 
PROTECTED AND HOW TO BASE SUCH  
A CHOICE: THE ISSUE OF SENTIENCE
Why to protect a class of invertebrates? The concern 
for animal welfare in the Directive 2010/63/EU is fo-
cused on the ability of animals to experience and ex-
press pain, suffering, distress and lasting harm. This 
concept calls into question the complexity of the anato-
my and function of an animal’s nervous system that are 
in turn related to the “psychophysical” wellbeing and 
eventually to the sentience of the individuals. 
Sentience can be roughly defined as the individual 
capacity to feel, perceive and interpret a situation or a 
certain context. In a more refined definition, sentient 
beings are characterised by the capacity to assess the 
actions of other organisms in relation to themselves and 
third parties, to remember some of their own actions 
and the related consequences, to evaluate risks, to pos-
sess some feelings and a certain degree of awareness 
[7]. 
As pointed out by Donald M. Broom “It is of sci-
entific and practical interest to consider the levels of 
cognitive ability in animals, which animals are sentient, 
which animals have feelings such as pain and which 
animals should be protected. (…) These abilities can be 
taken into account when evaluating welfare. There is 
evidence from (…) cephalopods and decapod crusta-
ceans of substantial perceptual ability, pain and adre-
nal systems, emotional responses, long- and short-term 
memory, complex cognition, individual differences, de-
ception, tool use, and social learning. The case for pro-
tecting these animals would appear to be substantial” 
[7]. Keeping in mind these considerations, EU State 
Members have certainly made a major step forward by 
deciding to include the welfare of live cephalopods un-
der the scope of the Directive 2010/63/EU. Finally, it is 
worth emphasising that the very concept of “sentience” 
is the fundamental criteria to which animal species are 
included or not under the protective umbrella of the 
Directive.
THE NEED FOR GUIDELINES TO PROTECT 
THE WELFARE OF CEPHALOPODS USED  
IN RESEARCH
The use of cephalopods in scientific research, particu-
larly in the field of neuroscience, has a centenarian his-
tory. The reason for this success has its origin in some 
main features characterising this “exceptional class of 
invertebrates” [8]. First of all a complex and centralised 
nervous system resulting in a phenotypic plasticity and 
in a sophisticated behavioural repertoire that has been 
suggested to be evolved from their unique “embodi-
ment” [8-10]. Moreover, it should be mentioned that 
their tractability for studies dealing with cellular and 
molecular mechanisms of learning and memory (e.g. 
long-term potentiation), likewise anatomical features 
facilitating physiological studies (e.g. squid giant axon 
and synapse) make cephalopods very suitable for scien-
tific purposes.
As mentioned before, the Directive 2010/63 now in-
cludes cephalopods under the umbrella of animal spe-
cies protected in Europe under the new legal text. As 
for other species, the Directive involves the justification 
and authorisation of projects dealing with cephalopods 
from local competent authorities and the revision of 
such projects includes a harm-benefit assessment and 
adherence to the 3Rs Principle [11].
To support project evaluation and compliance with 
the new EU law, specific guidelines for the care and 
welfare of cephalopods in research have then been 
developed following an international consensus based 
on an initiative by CephRes, FELASA and the Boyd 
Group [6]. The aim of the guidelines is to provide infor-
mation as practical and effective support for research-
ers, animal care committees, veterinarians and techni-
cians as well as animal care staff involved in research 
with these species.
Topics covered by the guidelines include (but are not 
limited to): implications of the Directive for cepha-
lopod research; the application of the 3Rs Principle; 
project authorisation process; harm‐benefit assessment 
and severity classification. Detailed aspects of require-
ments, care and welfare for the main laboratory spe-
cies currently used are provided on: supply, capture 
and transport; housing conditions (e.g. water quality 
control, lighting requirements, vibration/noise sensi-
tivity); cephalopod care (including tank main features, 
feeding, environmental and social enrichment); assess-
ment of health and welfare; prevention of disease and 
treatment; general anaesthesia and analgesia and meth-
ods of humane killing. In addition, specific sections 
dealing with risks for operators, education and training 
requirements for carers, researchers and veterinarians 
Guidelines for the welfare of cephalopods in research 
C
o
m
m
e
n
t
a
r
y
269
are reported. These guidelines will have an undoubted 
positive impact on the scientific environment by “guid-
ing” investigators in the process of project authorisa-
tion possibly by also providing new inspiration for re-
search on cephalopods. Personnel directly in charge of 
monitoring cephalopods’ psychophysical health such as 
veterinarians, animal care staff and technicians will also 
greatly benefit from the guidelines, since they currently 
rely only upon few aquatic medicine courses covering 
invertebrates and specific review papers on this topic. 
Furthermore, as detailed in the review by Fiorito and 
co-workers, the guidelines have a number of implica-
tions for different groups including not only working 
categories directly involved in the animal experimen-
tation but also funders, journal editors and reviewers 
[12]. As an example, funders could evaluate grants in-
volving the regulated use of cephalopods to make sure 
that the proposed studies comply with the Directive 
and any national Codes of Practice related to care and 
welfare. Likewise, editors and reviewers of scientific 
journals will need to ensure that papers submitted for 
publication, when necessary, make reference to compli-
ance with the Directive (in the philosophy of the AR-
RIVE guidelines).
The Guidelines for the care and welfare of cephalo-
pods in research is a multifaceted and comprehensive 
document that will set an example for future similar 
initiatives.
Accepted on 8 October 2015.
REFERENCES
1. Council of Europe. Directive 2010/63/EU of the Europe-
an Parliament and of the Council of 22 September 2010 
on the protection of animals used for scientific purposes. 
Official Journal of the European Union L 276, 20 October 
2010. Available from: http://ec.europa.eu/environment/
chemicals/lab_animals/legislation_en.htm.
2. Smith J, Andrews PLR, Hawkins P, Louhimies S, Ponte 
G, Dickel L. Cephalopod research and EU Directive 
2010/63/EU: Requirements, impacts and ethical review. 
J Exp Mar Biol Ecol 2013;447:31-45. DOI: 10.1016/j.
jembe.2013.02.009
3. Horvath K, Angeletti D, Nascetti G, Carere C. Inverte-
brate welfare: an overlooked issue. Ann Ist Super Sanità 
2013;49:9-17. DOI: 10.4415/ANN_13_01_04
4. Mather JA, Anderson RC. Ethics and Invertebrates: a 
cephalopod perspective. Dis Aquat Organ 2007;75:119-
29. DOI:10.3354/dao075119
5. Moltschaniwskyj N, Hall K, Marian J, Nishiguchi M, 
Sakai M, Shulman D, Sinclair B, Sinn D, Staudinger M, 
Van Gelderen R, Villanueva R, Warnke K. Ethical and 
welfare considerations when using cephalopods as experi-
mental animals. Rev Fish Biol Fisher 2007;17:455-76. DOI 
10.1007/s11160-007-9056-8
6. Fiorito G, Affuso A, Basil J, Cole A, De Girolamo P, 
D’Angelo L, Dickel L, Gestal C, Grasso F, Kuba M, 
Mark F, Melillo D, Osorio D, Perkins K, Ponte G, Sha-
shar N, Smith D, Smith J, Andrews PLR. Guidelines for 
the care and welfare of cephalopods in research. A con-
sensus based on an initiative by CephRes, FELASA and 
the Boyd Group. Lab Anim 2015;49(2 Suppl.):1-90.
7. Broom DM. Cognitive ability and sentience: which 
aquatic animals should be protected? Dis Aquat Organ 
2007;75:99-108. DOI:10.3354/dao075099
8. Zullo L, Hochner B. A new perspective on the orga-
nization of an invertebrate brain. Commun Integr Biol 
2011;4:426-9. DOI: 10.4161/cib.4.1.13804
9. Kröger B, Vinther J, Fuchs D. Cephalopod origin and 
evolution: A congruent picture emerging from fossils, 
development and molecules: Extant cephalopods are 
younger than previously realised and were under major 
selection to become agile, shell-less predators. Bioassays 
2011;33:602-13. DOI: 10.1002/bies.201100001
10. Albertin CB, Simakov O, Mitros T, Yan Wang Z, Pun-
gor JR, Edsinger-Gonzales E, Brenner S, Ragsdale CV, 
Rokshar DS. The octopus genome and the evolution of 
cephalopod neural and morphological novelties. Nature 
2015;524:220-4. DOI:10.1038/nature14668
11. Russell WMS, Burch RL. The principles of humane experi-
mental technique. Wheathampstead, (England): University 
Federation for Animal Welfare; 1959 (Reprinted in 1992).
12. Fiorito G, Affuso A, Anderson DB, Basil J, Bonnaud L, 
Botta G, Cole A, D’Angelo L, De Girolamo P, Dennison 
N, Dickel L, Di Cosmo A, Di Cristo C, Gestal C, Fonse-
ca R, Grasso F, Kristiansen T, Kuba M, Maffucci F, Man-
ciocco A, Mark FC, Melillo D, Osorio D, Palumbo A, 
Perkins K, Ponte G, Raspa M, Shashar N, Smith J. Smith 
D, Sykes A, Villanueva R, Tublitz N, Zullo L, Andrews P. 
Cephalopods in neuroscience: regulations, research and 
the 3Rs. Invert Neurosci 2014;14:13-36. DOI: 10.1007/
s10158-013-0165-x


EU guidelines for the care and welfare of an “exceptional invertebrate class” in scientific research

Cephalopods have been defined as "advanced invertebrates" due to the complexity of their nervous system and to their sophisticated behavioural repertoire. However, until recently, the protection and welfare of this class of invertebrates has been mostly disregarded by EU regulations on the use of laboratory animals. The inclusion of "live cephalopods" in the Directive 2010/63/EU has been prompted by new scientific knowledge on the "sentience" of animals used for experimental or other scientific purposes, a fundamental criterion to which animal species are included or not under the protective umbrella of the Directive. In this scenario, the imminent publication of the Guidelines for the care and welfare of cephalopods in research as an initiative by the CephRes-FELASA-Boyd Group is a sign of ethical progress in the consideration of animals in research, and is likely to have a significant impact on both scientific and practical aspects of research conducted with these animals

Alessandra Berry

Augusto Vitale

Claudio Carere

Enrico Alleva

Directory of Open Access Journals

EU guidelines for the care and welfare of an "exceptional invertebrate class" in scientific research

EU guidelines for the care and welfare of an “exceptional invertebrate class” in scientific research

Abstract

Similar works

Full text

Available Versions

Annali dell'Istituto Superiore di Sanità

Directory of Open Access Journals