5th RESAMA Days

The study of acoustic communication in fish has developed over the last decades. We now know that there are a multitude of means of sound production in fish using organs initially intended for other functions (swimming bladder, pharyngeal teeth, fin bones, etc.). Specializations have also developed to improve hearing sensitivity and the range of perceived frequencies. In cichlids, particularly in Maylandia zebra, recent studies have highlighted the importance of sounds in territory defense, partner choice and specific recognition. The detailed analysis of the signals made it possible to highlight differences in sound production between day and night relating to the number of sounds and the number of sound pulses per sound.

Marking animal populations is an essential tool in pet stores: it allows optimized management of the breeding plan as well as individual genetic, health and behavioral monitoring of animals. Since 2018, regulations have required the marking of wildlife individuals with RFID PIT Tags. At CRB Xenopes, after years of effective practice of this type of subcutaneous marking, the animals began to lose their chips. We present here the various test protocols which were carried out to find the cause of this rejection. It was discovered that parylene, an anti-migration coating coating the chips, was the triggering agent: it is a polymer that induces the formation of connective tissue at the tag/surrounding tissue interface, rendering the chip immobile at the insertion site. This leads to increased internal friction forces leading to lesions at the chip exit points. Flea loss occurs in both males and females but with a latency period in the latter. With this observation, we do not recommend the use of RFID tags with parylene coating for subcutaneous markings in xenopus animals.

Telemetry and biologging are preferred monitoring tools for studying the ecology of species movements. Constantly evolving technologically, they make it possible to remotely measure their behavior as well as the environment in which they evolve and the way in which this environment affects their condition. The presentation will illustrate the different technologies and methodologies used in aquatic environments and the main types of associated marking, with a particular focus on the refinement measures applied from capture to release.

Although included since 2013 in the legislation relating to animal experimentation, cephalopods do not yet have a specific reference in Annexes III and IV.

Although well-being is now defined in many laboratory species, a clear reference to “pain” and an intensity scale are only available for a few aquatic species.

The preliminary work on juvenile cuttlefish (Sepia officinalis) presented during the RESAMA 2022 days is part of an approach to constructing a tool for assessing the well-being of this experimental species.

The Nice Cancer & Aging Research Institute (IRCAN) studies the molecular mechanisms underlying the aging process as well as the diseases linked to this process (e.g., cancer) in order to advance our fundamental understanding of this multifactorial process as well as to develop innovative solutions for healthy aging. With the vision that modern biology must develop unconventional research models that enable major discoveries, IRCAN has set up the ANTIAGE platform. This is the first marine invertebrate platform within a Faculty of Medicine in France, which allows academic and private users to study the impressive resistance to environmental stress, extreme regeneration and longevity of cnidarians.

Stress management in fish is now a major societal concern, both in scientific farms and in those set up for aquaculture purposes. Measuring stress levels involves different techniques, one of which focuses on cortisol, the major stress hormone in fish. Here, we review the different cortisol measurement and dosage techniques, from the most invasive to the least invasive.

The study of zebrafish lines mutant for collagen VI, a major component of the extracellular matrix in muscle, requires carrying out forced swimming experiments in order to evaluate the swimming and metabolic performances of mutant and wild individuals. In these behavioral experiments, taking into account animal well-being is central, both to ensure the comfort of the animal being tested and to ensure the validity of the results obtained.

Presentation of the aquatic animal facility of the Interdisciplinary Center for Biology Research (CIRB) housed at the Collège de France (Paris), which accommodates zebrafish (Danio rerio). Short presentation of the members of the platform, the user teams and their scientific projects, then recent events which allowed the introduction of a new species within the animal facility: the medaka (Oryzias latipes). Presentation of the new breeding cabinets installed in the conventional zone.

It will simply be a matter of presenting the medaka and xenopus breeding structure of the Watchfrog laboratory animal facility.

An essential tool of the ENES laboratory, the experimental ethology platform of the ENES/CRNL team at the University of St Etienne is a structure making it possible to immediately make available to research teams animal models that can be maintained in captivity in dedicated spaces. Versatile, this installation is equipped with 200m² of animal room and 150m² of experimentation area. Capable of evolving according to research projects, we are able to maintain birds, fish, reptiles and mammals in conditions consistent with the rules for using animals in scientific research.

The XenUB platform is one of the nine technological platforms of the Bordeaux Institute of Oncology (https://www.bricbordeaux.com/technologies-et-cellule-de-transfert/). The platform has space and equipment for experimentation on the oocyte and embryo of the Xenopus amphibian. Its aim is to provide internal research teams at the University as well as external teams with the possibility of using the xenopus model for their research project. This model has contributed in the past to a large part to our knowledge in Developmental Biology. It is now more widely used to quickly model a large number of human pathologies such as cancer, congenital heart disease, craniofacial malformations or even kidney diseases and ciliopathies. Coupled with the use of chemical screening, this model constitutes a formidable tool for the pharmacological development of active substances.

We are developing several projects including the modeling of rare diseases such as Xeroderma pigmentosum or oculocutaneous albinism. We have shown the interest of the model for testing anti-cancer molecules on neural development (Capdevielle et al. (2020) Neuro-Oncology 22 (4): 550-562) or the function of inhibitory genes of the WNT signaling pathway (Mosca et al. (2021) Liver Cancer 11 (2): 126-140).

When carried out as part of an economic activity, the transport of live vertebrate animals is regulated for various reasons, in particular that of animal protection.

Regulation (EC) No. 1/2005 relating to the protection of animals during transport and related operations schematically provides for 3 levels of requirements which combine, from transport of less than 65 km (obligations of results, supplemented by obligations of means), through transport limited to 8 hours (subject to prior Type 1 authorization from 65 km, and qualification conditions for personnel preparing the animals and driving the vehicles), to culminate in transport that can exceed 8 hours (subject to Type 2 authorization, adding to the previous requirements the approval of vehicles, their traceability and emergency plans).

Non-tuberculous mycobacteria (NTM) are naturally resistant to most antibiotics and are very difficult to eradicate. We use zebrafish larvae to study the pathogenesis of several NTMs. Our practices in handling larvae and exploiting experimental data have been optimized, allowing us to evaluate the effectiveness of new therapeutic approaches to NTM infections.

The first recommendations on fish health monitoring and protection measures have just been published by the joint FELASA - AALAS working group. This presentation details the notion of epidemiological unit (EU), the concepts of SMOP and SLOM, risk analysis, examples of animal facilities using scenarios as well as a focus on the 2 key documents: Health Monitoring Report (HMR) and the description of the animal facility (EU).

The Smooth Xenopus is a southern African amphibian widely used in research. As a result of its wide export, repeated introductions have taken place on several continents which have led to the creation of invasive populations. The species is considered the third most invasive amphibian in the world due to its characteristics that generate impacts on ecosystems. The life program aimed to study the methods of controlling the French population. The main results are presented. Emphasis is placed on the need to consider biosecurity procedures for research facilities as well as monitoring the evolution of the European regulation on invasive alien species.

One of the 3 main wild animal models studied in the Biogéosciences laboratory animal facility is the Gammaridae model. The development of this breeding and the tools allowing its study had to be done with two major constraints:

- limited financial resources implying the search for inexpensive solutions for their design

- very little information available at the beginning of 2000 on the maintenance of this amphipod in captivity, implying local development.

The study of wild animals at Biogéosciences does not stop only at these three species, others will be quickly addressed during this presentation, in order to give a global vision of the research carried out there.

In a changing human society, animals have difficulty finding their place: food for 300,000 years, companionship for 15,000 years (dog and wolf), breeding for 8,500 years, object of study since the acclimatization societies (1854), attraction in zoological parks, subject of naturalist passions, attraction for breeding or use of animal products. Today, it no longer has its place in zoos, in circuses, on farms, on plates and unfortunately in the “natural” environment either. At the dawn of the 6th great mass extinction, wildlife species have never benefited from as much protection status as they currently do. The use of animals through these different aspects is governed by a whole set of international, European and national regulatory texts. This presentation aims to clarify the confusion that may still exist in people's minds and establish the framework in which each of us must fit in during our professional and/or leisure activities. Herpetofauna (Calotriton of the Pyrenees) will accompany the presentation of precise and concrete examples. We will address the basic regulatory triptych to know perfectly, protected species, breeding and use of animals for scientific purposes.

The Experimental and Predictive Ecology Research Center, Ecotron IleDeFrance, is a service unit dedicated to experimentation in terrestrial and aquatic ecology, both with animal and plant models. Through this presentation, we will give you a brief overview of our unit and our work in animal experimentation at the aquatic level.

The INRAE ​​experimentation station at Saint Seurin sur L’Isle is attached to UR 1454 EABX. Research is carried out there on migratory amphihaline fish and aquatic plants. Minimally invasive methodologies are used for the capture and transportation of wild fish. Sturgeon stocks are housed for the restoration of the endemic species Acipenser sturio. Their conservation in a closed circuit is becoming systematized, with constraints associated with their long life cycle.

The Anses “Virology, Immunology and Eco-toxicology of Fish” (VIMEP) unit is located at the Brest-Iroise Technopôle in Plouzané (Finistère). In addition to its cellular virology, molecular biology and immunology laboratories, it has approved experimental facilities allowing it to carry out contamination under controlled conditions with biological or physico-chemical agents on freshwater or seawater fish. The unit develops research activities relating to two main strategic axes, fish viruses and eco-toxicology, through a certain number of projects at regional, national and international level.

A CNRS unit was established in 1948 in the small village of Moulis in Ariège. First created as an underground laboratory, the unit then has a natural cave set up for research, which makes it possible, among other things, to accommodate cave-dwelling species in breeding to obtain their entire reproduction cycle and to increase the number of individuals for experiments. In the 2000s, the unit developed on the basis of a project focused on experimental ecology, in particular through the construction of terrestrial and aquatic continental freshwater mesocosms, which were backed by breeding facilities for invertebrates, fish, amphibians and small reptiles which make it possible to carry out measurements in controlled climatic conditions and to supply organisms for experiments within these mesocosms and in the natural environment. These facilities are accessible to researchers from the unit but also to the international scientific community.

Two series of experiments were carried out in a controlled environment to experimentally validate and refine the protocol for implantation of PIT tags on Planer lampreys collected from the natural environment. The prospects for applying this protocol to improve the management of wild populations with regard to ecological continuity will be discussed through an example of a study carried out in the Vosges du Nord Regional Natural Park.

The Thonon-les-Bains ground experimental structure is included in the OLA structure which brings together 4 types of services. The experimental ground structure allows multiple experiments with a strong impact on salmonids. This strong footprint has led us to develop grid tests taking into account animal welfare for animals kept in breeding (routine) and animals in experiments.

In recent years, the stickleback model has been highlighted for its ability to be used in the laboratory as well as in biomonitoring of aquatic environments. Several uses of the three-spined stickleback are discussed in this presentation from the point of view of the refinement of the techniques but also of the procedures used depending on the environment or the objectives sought.

The LAMNA project led by APECS (Association for the Study and Conservation of Selacians) in collaboration with UMR LEMAR (UBO/CNRS/IRD/Ifremer). This project focuses on the porbeagle shark (Lamna nasus, Bonnaterre 1788), a species classified as vulnerable by the IUCN and subject to a moratorium in the European Union. The objective of acquiring knowledge on the use of the Trégor coastal zone by the porbeagle species involves the deployment of tags and the taking of biological samples from females 130 to 250 cm long caught at sea.

The development of the technique of taking DNA by swab from the cave-dwelling Astyanax mexicanus was carried out in the laboratory on a very small number of breeding individuals. This minimally invasive sampling technique is compared to fin sampling for molecular biology aspects, considered more invasive. Although the quantity of DNA is lower, these samples allow the extraction of genomic DNA and 18 mitochondrial molecular markers. The aim is to reduce the impact on wild populations studied in the natural environment for which it is difficult to verify the safety of fin sampling a posteriori. This made it possible to set up capture/recapture experiments intended to refine the size estimates of these natural populations, probably very reduced by refining the practice of DNA sampling.