Operating protocol for Animal Welfare Body of SZN (OBA)

The Animal Welfare Body (OBA) of Stazione Zoologica Anton Dohrn (Resolution of the Administration Council of the SZN n. 8 of 30/1/2020) operates in application of Art. 25 of Legislative Decree n. 26 of 4 March 2014 "Implementation of Directive 2010/63/EU on the protection of animals used for scientific purposes" (Italian GU, 14 March 2014).

The SZN OBA operates according to the principles and in compliance with Legislative Decree 26/2014, Directive 2010/63/EU and the "A working document on Animal Welfare Bodies and National Committees to fulfil the requirements under the Directive”.

The SZN OBA has the following tasks:

i. To manage the procedures related to Legislative Decree 26/2014 facilitating the SZN researchers and collaborators - engaged in research activities with animals – improving the understanding of the areas and ensuring the correct application of the regulation;

ii. To protect and promote the welfare of animals used for scientific, exhibition or educational purposes at SZN facilities;

iii. To monitor the projects and activities concerning animal experimentation, verifying their ethical and scientific correctness and compliance with the regulations in force;

iv. To formulate Ethical clearance for projects or research activities that involve the use of aquatic organisms and that do not fall within the purposes of Legislative Decree 26/2014;

v. To inform and promote training and guidance for researchers and collaborators involved in research activities with animals.

In particular the SZN OBA:

• advises the staff dealing with animals in the application of the "3Rs" principle (replacement, reduction and refinement)
• encourages the professional updating of the staff involved in the use of animals;
• formulates  an opinion on research projects using animals for scientific purposes, on their possible modifications and revisions and - if the case - forwards them to the Ministry of Health (National Competent Authority) following their development and outcome;
• it provides advice on the reintegration programs of animals, where this is compatible with the study;
• intervenes on the adoption and monitoring of procedures excluded from the scope of Legislative Decree 26/2014.

We hereby remind that the lack of compliance with Italian law (DLgs 26/2014) will result in civil, administrative and penal sanctions. 

How does it work

SZN researchers, technologists and collaborators, may submit a Project Proposal for the use of organisms for scientific and/or educational research purposes at any time by sending the proposal to the email address This email address is being protected from spambots. You need JavaScript enabled to view it..

Proposals must be submitted by filling in forms that indicate all the information required. The forms are available at the link.

The SZN's AWB considers the following proposal modalities:

a. Request for Authorization pursuant to Art. 31 of Legislative Decree 26/2014 and for Authorization pursuant to Art. 33 of Legislative Decree 26/2014;
b. Request for Notification of the killing of animals for the sole purpose of using their organs or tissues;
c. Protocol request for project OUTSIDE the application of Legislative Decree 26/2014 (e.g., for studies in nature, behavioral observations, other purposes).

After preliminary examination of the individual proposals a. and b., OBA will send to the PI further paperwork to be filled (dichiarazione sostitutiva di certificazione e allegato IX).

In order to facilitate the collection of proposals, the OBA operates according to a calendar of meetings distributed throughout the year and proposals must be submitted by the 20th of each month, to be then examined by the SZN OBA at the first useful meeting.

The OBA expresses an opinion on each proposal received and attributes its status (i.e. "approved", "suspended pending review" or “rejected").

Things to consider for a research project using animals included in Legislative Decree 26/2014:

The initiation of a research and/or study in the SZN facilities involving the use of animals belonging to the list of species included in Legislative Decree 26/2014 (vertebrates that for SZN are represented by fish, aquatic reptiles, birds and mammals and Cephalopods), including national, European and international project activities, prior to the startup of the proposal it is subject to the verification by OBA of the following aspects:

i. The need for research and the likely contribution in terms of long-term positive effects on the health of humans, other animals or the environment (e.g., social relevance of the research).

ii. The necessity of the use of animals compared to alternative methods, evaluating the application of replacement, even partial, of the animals and/or of the foreseen procedures. Where the use of animals is essential, this must be justified (replacement).

iii. The respect of the principle of the maximum possible reduction of the number of animals used, and/or with the adoption of those methodologies that allow the maximization of the results obtained through the use of the smallest number of organisms, maintaining the forecast of reliable and robust results (reduction).

iv. The need to refine procedures in order to maximize the protection of the welfare of the animals used. In particular, in case the procedures applied may cause pain, suffering, distress or prolonged harm in species with the ability to experience and express them (i.e. sentience), the proposed experimental protocol will be evaluated taking into account the maximum consideration the degree and duration of suffering, the sample size (ie. number of individuals), and the planned and foreseeable measures to reduce the probability of harm to the animals (refinement).

For particular projects involving animals caught and and consequently killed or recovered already dead, the approval cannot disregard the use of animals from fishing activities authorized according to the health regulations in force, as well as the careful and precise evaluation of the purpose of the research.

Live specimens belonging to protected species on which diagnostic and therapeutic procedures are provided for recovery, may be the subject of research projects to be submitted for OBA approval by the SZN, provided that the activities of recovery of specimens are not affected by the research project and the manner of maintenance and handling, are in compliance with the regulations in force (e.g., permits: Local Health Authority, "Guidelines for the recovery of sea turtles" and CITES).

All proposals for research projects involving the use of animals for scientific purposes must obtain authorization - where applicable – from the NCA or the positive opinion of the SZN OBA as far as relevant and the case, before starting the research. The researcher will then have to wait for a response from the OBA and/or the Competent Authority before starting the activities; the negative opinion of the AWB on a specific project means that it cannot be carried out in the SZN facilities.

SZN's OBA list of evaluations (last update, 31 October 2020)

Ethical Clearance :

1. Acidification effects on herbivorous fish (Sarpa salpa)
2. Neural genes on cephalopods and deep-sea fish
3. Migration of Thunnus thynnus and Xiphias gladius
4. Distribution of Tetrapturus belone in Mediterranean Sea
5. Octopus vulgaris genome
6. Natural antioxidant agents effects in Danio rerio
7. Role of endogenous ouabain in various species
8. Tursiops truncatus calves management
9. Brain sizes in cephalopods
10. An exploratory study on Tool use in Octopus vulgaris
11. Gene expression and Learning and memory in Octopus vulgaris

Proposals submitted to the NCA:

Proposals not accepted: none

1. Neurogenesis in numerous species of aquatic vertebrates

Proposals under evaluation: none

Members of the Animal Welfare Body

Dott.ssa Claudia Gili - Responsabile del Benessere e della Cura degli Animali utlizzati negli esperimenti

Dott. Andrea Affuso - Medico Veterinario Designato

Dott. Graziano Fiorito - Componente comunità scientifica interna alla SZN

Prof. Bruno Cozzi - Universtià di Padova, esperto esterno

Prof.ssa Barbara De Mori - Università di Padova, esperto esterno

Reference: Anna Palumbo

Laboratory equipped for studies of the biochemistry of marine organisms including adaptive responses and immune system.

It includes an area for analysis of proteins and nucleic acids, and classic and basic biological laboratory equipments.

Systems for the experimentation

• UV Spectrophotometer UV, recording spectra of UV/visible and enzyme kinetics.
• electrophoresis on polyacrylamide and agarose gel.
• Centrifuges.
• Chromatography.

Analysis

• Measure of the response of marine organisms (adult and embryonic and larval stages) to environmental factors.
• Morphological characterization and evaluation of biochemical parameters (doses of enzyme activities, evaluation of the cellular redox state).
• Gene Expression, proteins and post-translational modifications induced by stress.
• Purification and biological characterization of proteins and metabolites from marine organisms.

Supervisor: Dr. Giovanna Romano

Scientific activities

The Laboratory of Chemical Ecology offers instrumentation and support to conduct studies of Chemical Ecology on the marine plankton. The research activities carried out in the laboratory are focused in particular on the study of the effects that secondary metabolites produced by diatoms induce in planktonic organisms and in marine model organisms such as sea urchins. The laboratory is also equipped for the extraction and the first fractionation of bioactive compounds and for the study of their biological activity.

Experimental systems

• Rotating wheel for copepod rearing and experimentation.
• Thermostated chamber for microalgae cultivation and incubation of copepods in controlled conditions.
• Basic facility for microalgae cultivation.

Analysis

• Sorting, isolation and identification of live phytoplankton and zooplankton species from natural samples.
• Determination of ingestion and reproduction rates of marine organisms.
• Biological assays to test diatoms and their metabolites using copepods and sea urchins.
• Analysis of assay results by light microscopy.
• Culturing of microalgae in different media.
• Preparation of algal pellet by centrifugation.
• Extraction and first fractionation of secondary metabolites from algal pellets.
• Biochemical and colorimentric assay to study the metabolism of oxylipins in diatoms.

Equipments

• Chemical hood (total expulsion).
• Stereo-microscopes.
• Upright and inverted microscopes.
• Fluorescence microscope equipped with a PC and video camera.
• Benchtop refrigerated centrifuges.
• Rotavapor.
• Spectrophotometer.
• Sonicator.
• Ph meter.
• Oxygraph.
• Recirculating water bath.

Coordinator: Christophe Brunet

Activity

The laboratory of Biogeochemistry and Physiology of Plankton is a multidisciplinary laboratory that uses and develops different approaches dedicated to the study of biogeochemistry and physiology of marine phytoplankton. The laboratory covers research activities related to biological oceanography, biogeochemistry and ecophysiology of marine phytoplankton. Mostly, the laboratory BioFilP implements activities and techniques for the study of the responses of marine microalgae to environmental forcing, with the aim to understand the pelagic ecosystem functioning and the potential use of microalgae in "Blue Growth".

Experimental systems

• Room and cultivation system for microalgae, with development of different types of illumination systems suitable for microalgal growth.
• Pelagic mesocosms for in situ experiments.

Analysis

• Photosynthetic rate and regulation of photosynthesis in microalgae.
• Pigments of photosynthetic organisms.
• Bio-optics of water masses and microalgal cultures.
• Antioxidant activities in microalgae.
• Concentrations of macronutrients in the water masses and in microalgal cultures.
• Spectral light in the water masses.

Equipment

• HPLC analysis for pigments.
• PAMs: Dual-pam and PSI.
• Autoanalyzer for nutrient concentration analysis.
• Spectrophotometer.
• Spectroradiometer in situ probe.
• Bio-optical in situ probe (AC-9).
• Laminar flow hood.
• Room and cultivation system for microalgae.