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1. The tropical coral Pocillopora acuta displays an unusual chromatin structure and shows histone H3 clipping plasticity upon bleaching [version 2; peer review: 2 approved]

Author

Patrick Masanet, Christoph Grunau, Pascal Romans, Jeremie Vidal-Dupiol, Céline Cosseau, Guillaume Mitta, Kelly Brener Raffalli, and David Roquis

Subjects
Pocillopora acuta, Pocillopora damicornis, invertebrate epigenetics, chromatin structure, Histone H3 clipping, eng, Medicine, Science
Abstract

Background: Pocillopora acuta is a hermatypic coral with strong ecological importance. Anthropogenic disturbances and global warming are major threats that can induce coral bleaching, the disruption of the mutualistic symbiosis between the coral host and its endosymbiotic algae. Previous works have shown that somaclonal colonies display different levels of survival depending on the environmental conditions they previously faced. Epigenetic mechanisms are good candidates to explain this phenomenon. However, almost no work had been published on the P. acuta epigenome, especially on histone modifications. In this study, we aim at providing the first insight into chromatin structure of this species. Methods: We aligned the amino acid sequence of P. acuta core histones with histone sequences from various phyla. We developed a centri-filtration on sucrose gradient to separate chromatin from the host and the symbiont. The presence of histone H3 protein and specific histone modifications were then detected by western blot performed on histone extraction done from bleached and healthy corals. Finally, micrococcal nuclease (MNase) digestions were undertaken to study nucleosomal organization. Results: The centri-filtration enabled coral chromatin isolation with less than 2% of contamination by endosymbiont material. Histone sequences alignments with other species show that P. acuta displays on average ~90% of sequence similarities with mice and ~96% with other corals. H3 detection by western blot showed that H3 is clipped in healthy corals while it appeared to be intact in bleached corals. MNase treatment failed to provide the usual mononucleosomal digestion, a feature shared with some cnidarian, but not all; suggesting an unusual chromatin structure. Conclusions: These results provide a first insight into the chromatin, nucleosome and histone structure of P. acuta. The unusual patterns highlighted in this study and partly shared with other cnidarian will need to be further studied to better understand its role in corals.

Published
2022
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4. Species and population genomic differentiation in Pocillopora corals (Cnidaria, Hexacorallia)

Author

Didier Aurelle, Marine Pratlong, Nicolas Oury, Anne Haguenauer, Pauline Gélin, Hélène Magalon, Mehdi Adjeroud, Pascal Romans, Jeremie Vidal-Dupiol, Michel Claereboudt, Camille Noûs, Lauric Reynes, Eve Toulza, François Bonhomme, Guillaume Mitta, Pierre Pontarotti, Institut méditerranéen d'océanologie (MIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Ecologie Marine et BIOdiversité (EMBIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Institut méditerranéen de biodiversité et d'écologie marine et continentale (IMBE), Avignon Université (AU)-Aix Marseille Université (AMU)-Institut de recherche pour le développement [IRD] : UMR237-Centre National de la Recherche Scientifique (CNRS), Institut Pythéas (OSU PYTHEAS), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Aix Marseille Université (AMU), Université de La Réunion (UR), Ecologie marine tropicale dans les Océans Pacifique et Indien (ENTROPIE [Réunion]), Institut de Recherche pour le Développement (IRD)-Université de La Réunion (UR)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Universités (COMUE), Interactions Hôtes-Pathogènes-Environnements (IHPE), Université de Perpignan Via Domitia (UPVD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Sultan Qaboos University (SQU), Laboratoire Cogitamus, Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), ANR-11-LABX-0061,OTMed,Objectif Terre : Bassin Méditerranéen(2011), ANR-11-IDEX-0001,Amidex,INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE(2011), ANR-12-ADAP-0016,ADACNI,Processus adaptatifs chez les cnidaires: étude intégrative de la réponse au stress thermique et au changement climatique, des gènes aux populations(2012), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), and ANR-10-LABX-0041,TULIP,Towards a Unified theory of biotic Interactions: the roLe of environmental(2010)

Subjects
Genetic Structures, species delineation, Sequence Analysis, DNA, Plant Science, General Medicine, RAD sequencing, Anthozoa, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Species Specificity, Insect Science, Genetics, genetic structure, Animals, Animal Science and Zoology, Metagenomics, Pocillopora, clonal reproduction, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, coral
Abstract

International audience; Correctly delimiting species and populations is a prerequisite for studies of connectivity, adaptation and conservation. Genomic data are particularly useful to test for species differentiation for organisms with few informative morphological characters or low discrimination of cytoplasmic markers, as in Scleractinians. Here we applied Restriction site Associated DNA sequencing (RADsequencing) to the study of species differentiation and genetic structure in populations of Pocillopora spp. from Oman and French Polynesia, with the objectives to test species hypotheses, and to study the genetic structure among sampling sites within species. We focused here on coral colonies morphologically similar to P. acuta (damicornis type β). We tested the impact of different filtering strategies on the stability of the results. The main genetic differentiation was observed between samples from Oman and French Polynesia. These samples corresponded to different previously defined primary species hypotheses (PSH), i.e. PSHs 12 and 13 in Oman, and PSH 5 in French Polynesia. In Oman, we did not observe any clear differentiation between the two putative species PSH 12 and 13, nor between sampling sites. In French Polynesia, where a single species hypothesis was studied, there was no differentiation between sites. Our analyses allowed the identification of clonal lineages in Oman and French Polynesia. The impact of clonality on genetic diversity is discussed in light of individual-based simulations.

Published
2022
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5. Low vulnerability of the Mediterranean antipatharian Antipathella subpinnata (Ellis & Solander, 1786) to ocean warming

Author

Godefroid, Mathilde, Zeimes, Tom, Bramanti, Lorenzo, Pascal, Romans, Bo, Marzia, Toma, Marguerita, Danis, Bruno, Dubois, Philippe, Guillaumot, Charlène, Godefroid, Mathilde, Zeimes, Tom, Bramanti, Lorenzo, Pascal, Romans, Bo, Marzia, Toma, Marguerita, Danis, Bruno, Dubois, Philippe, and Guillaumot, Charlène

Abstract

Antipatharians (black corals) are major components of mesophotic ecosystems in the Mediterranean Sea. The arborescent species Antipathella subpinnata has received particular attention as it is the most abundant and forms dense forests harbouring high levels of biodiversity. This species is currently categorized as “Near Threatened” in the IUCN Red List, due to increasing fishing pressure and bottom-trawling activities. Yet, the effects of ocean warming have never been investigated for this species, nor for any other antipatharians from temperate regions. Our study aimed at evaluating the effects of increasing seawater temperatures on A. subpinnata, by combining predictive distribution modelling with a physiological tolerance experiment. During the latter, we exposed A. subpinnata for 15 days to different temperature conditions spanning the current seasonal range to forecasted temperatures for 2100, while measuring biological endpoints such as oxygen consumption rates and different signs of stress (tissue necrosis, total antioxidant capacity). Unexpectedly, no stress was found at organism nor cellular level (wide thermal breadth) suggesting low susceptibility of this species to mid-term temperature increase. If the response to the 15-days heat stress is representative of the response to longer-term warming, ocean warming is unlikely to affect A. subpinnata. The species distribution model predicted the presence of A. subpinnata at depths that correspond to temperatures colder than its maximum thermal tolerance (as determined by the physiology experiment). This suggests that the presence of A. subpinnata at shallower depths is not limited by physiological constraints but by other ecological factors including interspecific competition., Godefroid M. and Zeimes T. contributed equally., SCOPUS: ar.j, info:eu-repo/semantics/published

Published
2022

6. Population genomics of Pocillopora corals: insights from RAD-sequencing

Author

François Bonhomme, Marine Pratlong, Eve Toulza, Pauline Gélin, Didier Aurelle, Hélène Magalon, Guillaume Mitta, Michel R. Claereboudt, Pierre Pontarotti, Anne Haguenauer, Nicolas Oury, Mehdi Adjeroud, Lauric Reynes, Pascal Romans, Camille Noûs, Jeremie Vidal-Dupiol, Institut méditerranéen d'océanologie (MIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Institut méditerranéen de biodiversité et d'écologie marine et continentale (IMBE), Avignon Université (AU)-Aix Marseille Université (AMU)-Institut de recherche pour le développement [IRD] : UMR237-Centre National de la Recherche Scientifique (CNRS), Institut de Mathématiques de Marseille (I2M), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), Ecologie marine tropicale des océans Pacifique et Indien (ENTROPIE [Nouvelle-Calédonie]), Ifremer - Nouvelle-Calédonie, Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut de Recherche pour le Développement (IRD [Nouvelle-Calédonie])-Université de la Nouvelle-Calédonie (UNC), Université de La Réunion (UR), Centre de recherches insulaires et observatoire de l'environnement (CRIOBE), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU), Interactions Hôtes-Pathogènes-Environnements (IHPE), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Perpignan Via Domitia (UPVD), Sultan Qaboos University (SQU), Laboratoire Cogitamus, Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche pour le développement [IRD] : UR226, Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), ANR-11-LABX-0061,OTMed,Objectif Terre : Bassin Méditerranéen(2011), ANR-11-IDEX-0001,Amidex,INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE(2011), ANR-12-ADAP-0016,ADACNI,Processus adaptatifs chez les cnidaires: étude intégrative de la réponse au stress thermique et au changement climatique, des gènes aux populations(2012), ANR-10-LABX-0046,EGID,EGID Diabetes Pole(2010), ANR-10-EQPX-0007,LIGAN PM,Plate forme Lilloise de séquençage du génome humain pour une médecine personnalisée(2010), ANR-10-LABX-0041,TULIP,Towards a Unified theory of biotic Interactions: the roLe of environmental(2010), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Institut de Recherche pour le Développement (IRD [Nouvelle-Calédonie])-Ifremer - Nouvelle-Calédonie, Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de la Nouvelle-Calédonie (UNC), Université de Perpignan Via Domitia (UPVD)-École Pratique des Hautes Études (EPHE), Université de Perpignan Via Domitia (UPVD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, Mitochondrial DNA, Lineage (evolution), [SDV]Life Sciences [q-bio], Population, Biology, RAD sequencing, 010603 evolutionary biology, 01 natural sciences, Population genomics, 03 medical and health sciences, genetic structure, 14. Life underwater, Pocillopora, education, coral, 030304 developmental biology, 0303 health sciences, education.field_of_study, Genetic diversity, species limits, biology.organism_classification, Genetic marker, Evolutionary biology, Genetic structure
Abstract

Scleractinian corals are of great ecological interest as ecosystem engineer species. Accordingly, there is a wealth of studies on their adaptive abilities facing climate change. Such studies should rely on precise species and population delimitation. Nevertheless species delimitation in corals can be hindered by the lack of adequate genetic markers, by hybridization, and by morphological plasticity. Here we applied RAD sequencing to the study of species delimitation and genetic structure in populations of Pocillopora spp. from Oman and French Polynesia with the objectives to test primary species hypotheses based on mitochondrial DNA sequencing, and to study the genetic structure among sampling sites inside species. Regarding the varying levels of missing data observed among samples we tested different filtering strategy. The main genetic differentiation was observed between samples from Oman and French Polynesia, which also corresponded to different mitochondrial lineages and species hypotheses. In Oman, we did not observe any clear differentiation according to the main mitochondrial lineages considered here, nor between sampling sites. In French Polynesia where a single mitochondrial lineage was studied, we did not evidence any differentiation according to sampling sites. These results provide an additional example of the importance of using independent nuclear markers for the study of species delimitation. Our analyses also allowed the identification of clonal lineages in our samples, and to take them into account in our interpretations. We used simulations to study the impact of clonal reproduction on the distribution of statistics of genetic diversity and genetic structure among loci.

Published
2021

7. Sea anemone and clownfish microbiota diversity and variation during the initial steps of symbiosis

Author

Raphaël Lami, Pascal Romans, Natacha Roux, Patrick Masanet, David Lecchini, Kévin Magré, Pauline Salis, Vincent Laudet, Biologie intégrative des organismes marins (BIOM), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre de recherches insulaires et observatoire de l'environnement (CRIOBE), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biodiversité et Biotechnologies Microbiennes (LBBM), PIERRE FABRE-EDF (EDF)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Excellence CORAIL (LabEX CORAIL), Université des Antilles (UA)-Institut d'écologie et environnement-Université de la Nouvelle-Calédonie (UNC)-Université de la Polynésie Française (UPF)-Université de La Réunion (UR)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École des hautes études en sciences sociales (EHESS)-Université des Antilles et de la Guyane (UAG)-Institut de Recherche pour le Développement (IRD), Université de Perpignan Via Domitia (UPVD)-École Pratique des Hautes Études (EPHE), Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de La Réunion (UR)-Université de la Polynésie Française (UPF)-Université de la Nouvelle-Calédonie (UNC)-Institut d'écologie et environnement-Université des Antilles (UA), and Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École pratique des hautes études (EPHE)

Subjects
0301 basic medicine, 030106 microbiology, Biodiversity, Zoology, lcsh:Medicine, Sea anemone, Article, 03 medical and health sciences, Symbiosis, RNA, Ribosomal, 16S, Animals, 14. Life underwater, Microbiome, lcsh:Science, [SDU.STU.OC]Sciences of the Universe [physics]/Earth Sciences/Oceanography, Multidisciplinary, biology, Host (biology), Microbiota, lcsh:R, biology.organism_classification, Mucus, Biological Evolution, Haliangiaceae, Perciformes, 030104 developmental biology, Sea Anemones, Heteractis magnifica, lcsh:Q, [SDV.EE.IEO]Life Sciences [q-bio]/Ecology, environment/Symbiosis
Abstract

Clownfishes and sea anemones form an intriguing long-term association, but the mechanism underlying this symbiosis is not well understood. Since clownfishes seem to cover themselves with sea anemone mucus, we investigated the microbiomes of the two partners to search for possible shifts in their compositions. We used a 16S rRNA gene sequencing strategy to study the dynamics of the microbiota during the association between the clownfish Amphiprion ocellaris and its host Heteractis magnifica under laboratory conditions. The experiment conducted in aquaria revealed that both clownfish and sea anemone mucus had specific signatures compared to artificial sea water. The microbiomes of both species were highly dynamic during the initiation of the symbiosis and for up to seven days after contact. Three families of bacteria (Haliangiaceae, Pseudoalteromonadacae, Saprospiracae) were shared between the two organisms after symbiosis. Once the symbiosis had been formed, the clownfishes and sea anemone then shared some communities of their mucus microbiota. This study paves the way for further investigations to determine if similar microbial signatures exist in natural environments, whether such microbial sharing can be beneficial for both organisms, and whether the microbiota is implicated in the mechanisms that protect the clownfish from sea anemone stinging.

Published
2019
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8. The tropical coral

Author

David, Roquis, Céline, Cosseau, Kelly, Brener Raffalli, Pascal, Romans, Patrick, Masanet, Guillaume, Mitta, Christoph, Grunau, and Jeremie, Vidal-Dupiol

Published
2021

9. A star is born again: Methods for larval rearing of an emerging model organism, the False clownfish Amphiprion ocellaris

Author

David Lecchini, Vincent Laudet, Laurence Besseau, Nancy Trouillard, Natacha Roux, Rémi Pillot, Pauline Salis, Pascal Romans, Valentin Logeux, Kévin Magré, Biologie intégrative des organismes marins (BIOM), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Centre de recherches insulaires et observatoire de l'environnement (CRIOBE), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Excellence CORAIL (LabEX CORAIL), Université des Antilles (UA)-Institut d'écologie et environnement-Université de la Nouvelle-Calédonie (UNC)-Université de la Polynésie Française (UPF)-Université de La Réunion (UR)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École des hautes études en sciences sociales (EHESS)-Université des Antilles et de la Guyane (UAG)-Institut de Recherche pour le Développement (IRD), Okinawa Institute of Science and Technology Graduate University, Academia Sinica, Université de Perpignan Via Domitia (UPVD)-École Pratique des Hautes Études (EPHE), Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de La Réunion (UR)-Université de la Polynésie Française (UPF)-Université de la Nouvelle-Calédonie (UNC)-Institut d'écologie et environnement-Université des Antilles (UA), ANR-19-CE14-0010,SENSO,Le dialogue environnement-hormones dans le contrôle du cycle de vie du poisson clown Amphiprion ocellaris.(2019), and ANR-19-CE34-0006,Manini,Effet des stress anthropogéniques sur le recrutement larvaire des poissons coralliens(2019)

Subjects
0106 biological sciences, 0301 basic medicine, rearing methods, Coral reef fish, Ecology (disciplines), ved/biology.organism_classification_rank.species, Zoology, Captivity, anemonefishes, Biology, 010603 evolutionary biology, 01 natural sciences, Predation, 03 medical and health sciences, Laboratory Animal Science, [SDV.BA.ZV]Life Sciences [q-bio]/Animal biology/Vertebrate Zoology, Genetics, Animals, Resource Article, 14. Life underwater, Animal Husbandry, Model organism, Ecology, Evolution, Behavior and Systematics, geography, Larva, coral reef fish, geography.geographical_feature_category, ved/biology, fungi, Fishes, Coral reef, eco‐evo‐devoexperimental model, 030104 developmental biology, Amphiprion ocellaris, Molecular Medicine, Animal Science and Zoology, Developmental biology, Developmental Biology
Abstract

As interest increases in ecological, evolutionary, and developmental biology (Eco‐Evo‐Devo), wild species are increasingly used as experimental models. However, we are still lacking a suitable model for marine fish species, as well as coral reef fishes that can be reared at laboratory scales. Extensive knowledge of the life cycle of anemonefishes, and the peculiarities of their biology, make them relevant marine fish models for developmental biology, ecology, and evolutionary sciences. Here, we present standard methods to maintain breeding pairs of the anemonefish Amphiprion ocellaris in captivity, obtain regular good quality spawning, and protocols to ensure larval survival throughout rearing. We provide a detailed description of the anemonefish husbandry system and life prey culturing protocols. Finally, a “low‐volume” rearing protocol useful for the pharmacological treatment of larvae is presented. Such methods are important as strict requirements for large volumes in rearing tanks often inhibit continuous treatments with expensive or rare compounds., To properly use anemonefish as a laboratory model species, it is required to have breeding pairs, husbandry with broodstock room, larval rearing tanks, and low‐volume rearing tanks for experimental purposes, and finally living prey cultures (nauplii of artemia, rotifers, and algae), Research Highlights This paper describes how to set up a rearing system for anemone fishes at the laboratory scale as this species is becoming a relevant marine fish model to tackle Eco‐Evo‐Devo questions. We detail two rearing methods, one consisting of classical rearing conditions and the other one consisting of low‐volume conditions (500 ml).

Published
2021
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10. The tropical coral Pocillopora acuta displays an unusual chromatin structure and shows histone H3 clipping plasticity upon bleaching

Author

Céline Cosseau, Pascal Romans, Guillaume Mitta, Kelly Brener Raffalli, Patrick Masanet, David Roquis, Jeremie Vidal-Dupiol, Christoph Grunau, Agroscope, Interactions Hôtes-Pathogènes-Environnements (IHPE), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Perpignan Via Domitia (UPVD), Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), and Aquarium de Canet-en-Roussillon

Subjects
Coral bleaching, viruses, Pocillopora acuta, Medicine (miscellaneous), Hermatypic coral, Pocillopora damicornis, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Histone H3, 0302 clinical medicine, Histone H3 clipping, Nucleosome, 14. Life underwater, 030304 developmental biology, chromatin structure, 0303 health sciences, biology, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], fungi, virus diseases, Epigenome, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, digestive system diseases, Chromatin, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Histone, Evolutionary biology, biology.protein, invertebrate epigenetics, 030217 neurology & neurosurgery
Abstract

Background: Pocillopora acuta is a hermatypic coral with strong ecological importance. Anthropogenic disturbances and global warming are major threats that can induce coral bleaching, the disruption of the mutualistic symbiosis between the coral host and its endosymbiotic algae. Previous works have shown that somaclonal colonies display different levels of survival depending on the environmental conditions they previously faced. Epigenetic mechanisms are good candidates to explain this phenomenon. However, almost no work had been published on the P. acuta epigenome, especially on histone modifications. In this study, we aim at providing the first insight into chromatin structure of this species. Methods: We aligned the amino acid sequence of P. acuta core histones with histone sequences from various phyla. We developed a centri-filtration on sucrose gradient to separate chromatin from the host and the symbiont. The presence of histone H3 protein and specific histone modifications were then detected by western blot performed on histone extraction done from bleached and healthy corals. Finally, micrococcal nuclease (MNase) digestions were undertaken to study nucleosomal organization. Results: The centri-filtration enabled coral chromatin isolation with less than 2% of contamination by endosymbiont material. Histone sequences alignments with other species show that P. acuta displays on average ~90% of sequence similarities with mice and ~96% with other corals. H3 detection by western blot showed that H3 is clipped in healthy corals while it appeared to be intact in bleached corals. MNase treatment failed to provide the usual mononucleosomal digestion, a feature shared with some cnidarian, but not all; suggesting an unusual chromatin structure. Conclusions: These results provide a first insight into the chromatin, nucleosome and histone structure of P. acuta. The unusual patterns highlighted in this study and partly shared with other cnidarian will need to be further studied to better understand its role in corals.

Published
2021
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11. A star is born again: Methods for larval rearing of an emerging model organism, the False clownfish Amphiprion ocellaris

Author

Natacha, Roux, Valentin, Logeux, Nancy, Trouillard, Rémi, Pillot, Kévin, Magré, Pauline, Salis, David, Lecchini, Laurence, Besseau, Vincent, Laudet, Pascal, Romans, Natacha, Roux, Valentin, Logeux, Nancy, Trouillard, Rémi, Pillot, Kévin, Magré, Pauline, Salis, David, Lecchini, Laurence, Besseau, Vincent, Laudet, and Pascal, Romans

Abstract

As interest increases in ecological, evolutionary, and developmental biology (Eco-Evo-Devo), wild species are increasingly used as experimental models. However, we are still lacking a suitable model for marine fish species, as well as coral reef fishes that can be reared at laboratory scales. Extensive knowledge of the life cycle of anemonefishes, and the peculiarities of their biology, make them relevant marine fish models for developmental biology, ecology, and evolutionary sciences. Here, we present standard methods to maintain breeding pairs of the anemonefish Amphiprion ocellaris in captivity, obtain regular good quality spawning, and protocols to ensure larval survival throughout rearing. We provide a detailed description of the anemonefish husbandry system and life prey culturing protocols. Finally, a “low-volume” rearing protocol useful for the pharmacological treatment of larvae is presented. Such methods are important as strict requirements for large volumes in rearing tanks often inhibit continuous treatments with expensive or rare compounds., source:https://onlinelibrary.wiley.com/doi/10.1002/jez.b.23028

Published
2021

12. Unveiling asexual reproductive traits in black corals: polyp bail-out in Antipathella subpinnata

Author

Rémi Pillot, Lorenzo Bramanti, Martina Coppari, Pascal Romans, Lara Fumarola, Marzia Bo, Giorgio Bavestrello, Laboratoire d'Ecogéochimie des environnements benthiques (LECOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Observatoire océanologique de Banyuls (OOB), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, biology, Asexual reproduction, 010604 marine biology & hydrobiology, [SDV.BA]Life Sciences [q-bio]/Animal biology, Antipatharians, Zoology, Aquatic Science, biology.organism_classification, Antipatharians, Asexual reproduction, Bailout, Aquaria, 010603 evolutionary biology, 01 natural sciences, Black coral, Bailout, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Antipathella subpinnata, 14. Life underwater, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, Aquaria
Abstract

Cnidarians are known to undergo reverse development as a survival mechanism against adverse environmental conditions. Polyp bail-out consists in the polyps’ detachment from the mother colony due to stressful conditions, followed by a complete tissue and cells rearrangement and in some cases in a regression into a simple, ciliated form. Here we describe a massive polyp bail-out event occurred in the mesophotic black coral Antipathella subpinnata in reared conditions. This is the first report of a bail-out event in this species providing new insights into the life cycle and ecology of black corals.

Published
2020
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13. Metabolomics Reveal That Octocrylene Accumulates in Pocillopora damicornis Tissues as Fatty Acid Conjugates and Triggers Coral Cell Mitochondrial Dysfunction

Author

Alice M. S. Rodrigues, Didier Stien, Rémi Pillot, Fanny Clergeaud, Philippe Lebaron, Sonja K. Fagervold, Karine Lebaron, Pascal Romans, Laboratoire de Biodiversité et Biotechnologies Microbiennes (LBBM), PIERRE FABRE-EDF (EDF)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Observatoire océanologique de Banyuls (OOB), and Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Coral, Cell, Pocillopora damicornis, [SDV.BID]Life Sciences [q-bio]/Biodiversity, 010402 general chemistry, 01 natural sciences, Analytical Chemistry, chemistry.chemical_compound, Metabolomics, Tandem Mass Spectrometry, medicine, Animals, Chromatography, High Pressure Liquid, chemistry.chemical_classification, [SDU.OCEAN]Sciences of the Universe [physics]/Ocean, Atmosphere, biology, Fatty acid metabolism, Molecular Structure, 010401 analytical chemistry, Fatty Acids, Fatty acid, biology.organism_classification, Anthozoa, 0104 chemical sciences, Mitochondria, Octocrylene, medicine.anatomical_structure, chemistry, Biochemistry, Acrylates, Toxicity
Abstract

International audience; Octocrylene (OC) is an ingredient used in many sunscreens and cosmetics worldwide. Our group evaluated the toxicity of OC in corals. Adult Pocillopora damicornis coral was treated with OC at concentrations of 5, 50, 300, and 1000 μg/L. Most polyps were closed at concentrations of 300 μg/L and higher. Further, metabolomic profiling provided crucial information regarding OC accumulation in coral tissues and OC toxicity. First, we demonstrated that OC was transformed into fatty acid conjugates via oxidation of the ethylhexyl chain, yielding very lipophilic OC analogues that accumulate in coral tissues. Second, the differential analysis of coral profiles revealed higher levels of 15 acylcarnitines, suggesting abnormal fatty acid metabolism related to mitochondrial dysfunction. The formation of OC analogues suggests that OC concentrations measured in the environment, and organisms may have been largely underestimated. Overall, these results call for an in-depth evaluation of OC toxicity and the reevaluation of the actual OC accumulation rate in the ocean’s food chain, including OC–fatty acid conjugates.

Published
2019
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14. Staging and normal table of postembryonic development of the clownfish ( Amphiprion ocellaris )

Author

Pascal Romans, Valentin Logeux, Bruno Frederich, Natacha Roux, Vincent Laudet, Pauline Salis, David Lecchini, Olivier Soulat, Anne Lambert, Biologie intégrative des organismes marins (BIOM), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre de recherches insulaires et observatoire de l'environnement (CRIOBE), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE)-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique Fonctionnelle de Lyon (IGFL), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Université de Liège, Laboratoire d'Excellence CORAIL (LabEX CORAIL), Université des Antilles (UA)-Institut d'écologie et environnement-Université de la Nouvelle Calédonie (UNC)-Université de la Polynésie Française (UPF)-Université de La Réunion (UR)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-École pratique des hautes études (EPHE)-École des hautes études en sciences sociales (EHESS)-Université des Antilles et de la Guyane (UAG)-Institut de Recherche pour le Développement (IRD), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Université des Antilles (UA)-Institut d'écologie et environnement-Université de la Nouvelle-Calédonie (UNC)-Université de la Polynésie Française (UPF)-Université de La Réunion (UR)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École des hautes études en sciences sociales (EHESS)-Université des Antilles et de la Guyane (UAG)-Institut de Recherche pour le Développement (IRD), Université de Perpignan Via Domitia (UPVD)-École Pratique des Hautes Études (EPHE), École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de La Réunion (UR)-Université de la Polynésie Française (UPF)-Université de la Nouvelle-Calédonie (UNC)-Institut d'écologie et environnement-Université des Antilles (UA), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), and Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École pratique des hautes études (EPHE)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de La Réunion (UR)-Université de la Polynésie Française (UPF)-Université de la Nouvelle Calédonie (UNC)-Institut d'écologie et environnement-Université des Antilles (UA)

Subjects
0301 basic medicine, Coral reef fish, Ontogeny, Zoology, Pomacentridae, developmental stages, Global Warming, 03 medical and health sciences, 0302 clinical medicine, [SDV.BA.ZV]Life Sciences [q-bio]/Animal biology/Vertebrate Zoology, Animals, Juvenile, pigmentation, 14. Life underwater, Damselfish, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Research Articles, geography, coral reef fish, geography.geographical_feature_category, Ecology, biology, Hatching, fungi, Fishes, Coral reef, biology.organism_classification, Perciformes, 030104 developmental biology, postembryonic, ontogeny, Models, Animal, Evolutionary developmental biology, 030217 neurology & neurosurgery, Research Article, Developmental Biology
Abstract

Background The clownfish Amphiprion ocellaris is one of the rare coral reef fish species that can be reared in aquaria. With relatively short embryonic and larval development, it could be used as a model species to study the impact of global changes such as temperature rise or anthropogenic threats (eg, pollution) on the postembryonic development at molecular and endocrinological levels. Establishing a developmental table allows us to standardize sampling for the scientific community willing to conduct experiments on this species on different areas: ecology, evolution, and developmental biology. Results Here, we describe the postembryonic developmental stages for the clownfish A. ocellaris from hatching to juvenile stages (30 days posthatching). We quantitatively followed the postembryonic growth and described qualitative traits: head, paired and unpaired fins, notochord flexion, and pigmentation changes. The occurrence of these changes over time allowed us to define seven stages, for which we provide precise descriptions. Conclusions Our work gives an easy system to determine A. ocellaris postembryonic stages allowing, thus, to develop this species as a model species for coral reef fishes. In light of global warming, the access to the full postembryonic development stages of coral reef fish is important to determine stressors that can affect such processes., Key Findings Seven developmental stages have been identified to describe the larval development of the clownfish Amphiprion ocellaris.Clownfish larvae undergo two distinct developmental growth phases that correspond to growth and metamorphosis.A dichotomous key determination has been created to assist users in identifying the various developmental stages.

Published
2019
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15. Gene expression plasticity and frontloading promote thermotolerance in Pocillopora corals

Author

Oliver Rey, François Bonhomme, Guillaume Mitta, Anne Haguenauer, K. Brener-Raffalli, Marine Pratlong, Pauline Gélin, Pierre Pontarotti, Didier Aurelle, Mehdi Adjeroud, L. Feuillassier, Eve Toulza, Michel R. Claereboudt, Hélène Magalon, Jeremie Vidal-Dupiol, Pascal Romans, Rémi Pillot, Interactions Hôtes-Pathogènes-Environnements (IHPE), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Perpignan Via Domitia (UPVD), Institut de Recherche pour le Développement (IRD), Laboratoire d'Excellence CORAIL (LabEX CORAIL), Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de La Réunion (UR)-Université de la Polynésie Française (UPF)-Université de la Nouvelle-Calédonie (UNC)-Institut d'écologie et environnement-Université des Antilles (UA), Ecologie marine tropicale des océans Pacifique et Indien (ENTROPIE [Perpignan]), Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche pour le développement [IRD] : UR226, Institut méditerranéen de biodiversité et d'écologie marine et continentale (IMBE), Avignon Université (AU)-Aix Marseille Université (AMU)-Institut de recherche pour le développement [IRD] : UMR237-Centre National de la Recherche Scientifique (CNRS), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Sultan Qaboos University (SQU), Ecologie marine tropicale dans les Océans Pacifique et Indien (ENTROPIE [Réunion]), Centre National de la Recherche Scientifique (CNRS)-Université de La Réunion (UR)-Institut de Recherche pour le Développement (IRD), Centre National de la Recherche Scientifique (CNRS), Institut méditerranéen d'océanologie (MIO), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Université de Perpignan Via Domitia (UPVD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut de Recherche pour le Développement (IRD)-Université des Antilles et de la Guyane (UAG)-École des hautes études en sciences sociales (EHESS)-École Pratique des Hautes Études (EPHE), Centre de recherches insulaires et observatoire de l'environnement (CRIOBE), Université de Perpignan Via Domitia (UPVD)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut de Recherche pour le Développement (IRD)-Université de La Réunion (UR)-Centre National de la Recherche Scientifique (CNRS), Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), and ANR-12-ADAP-0016,ADACNI,Processus adaptatifs chez les cnidaires: étude intégrative de la réponse au stress thermique et au changement climatique, des gènes aux populations(2012)

Subjects
0106 biological sciences, Genetics, Regulation of gene expression, 0303 health sciences, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], Biology, biology.organism_classification, 010603 evolutionary biology, 01 natural sciences, Transcriptome, 03 medical and health sciences, 13. Climate action, Heat shock protein, DNA methylation, Gene expression, 14. Life underwater, Epigenetics, Pocillopora, Gene, 030304 developmental biology
Abstract

Ecosystems worldwide are suffering from climate change. Coral reef ecosystems are globally threatened by increasing sea surface temperatures. However, gene expression plasticity provides the potential for organisms to respond rapidly and effectively to environmental changes, and would be favored in variable environments. In this study, we investigated the thermal stress response in Pocillopora coral colonies from two contrasting environments by exposing them to heat stress. We compared the physiological state, bacterial and Symbionaceae communities (using 16S and ITS2 metabarcoding), and gene expression levels (using RNA-Seq) between control conditions and heat stress (the temperature just below the first signs of compromised health). Colonies from both thermal regimes remained apparently normal and presented open and colored polyps during heat stress, with no change in bacterial and Symbionaceae community composition. In contrast, they differed in their transcriptomic responses. The colonies from Oman displayed a more plastic transcriptome, but some genes had a higher basal expression level (frontloading) compared to the less thermotolerant colonies from New Caledonia. In terms of biological functions, we observed an increase in the expression of stress response genes (including induction of tumor necrosis factor receptors, heat shock proteins, and detoxification of reactive oxygen species), together with a decrease in the expression of genes involved in morpho-anatomical functions. Gene regulation (transcription factors, mobile elements, histone modifications and DNA methylation) appeared to be overrepresented in the Oman colonies, indicating possible epigenetic regulation. These results show that transcriptomic plasticity and frontloading can be co-occurring processes in corals confronted to highly variable thermal regimes.

Published
2018
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16. Effects of intracoelomic tagging procedure on white seabream (Diplodus sargus) behavior and survival

Author

Barbara Koeck, Anaïs Gudefin, Julien Loubet, Pascal Romans, and Philippe Lenfant

Subjects
High variability, Captivity, Zoology, Anatomy, Diplodus, Aquatic Science, Biology, biology.organism_classification, White (mutation), Ethogram, medicine.anatomical_structure, medicine, Body cavity, Surgical incision, Ecology, Evolution, Behavior and Systematics, Sargus
Abstract

The effects of intracoelomic tagging procedure (i.e. implantation by surgical incision into the abdominal cavity or laparotomy) on wild white seabream (Diplodus sargus) were evaluated by the comparison of the behavior of tagged and untagged control fish. Activity budgets were made to examine their behaviors, using an instantaneous scan sampling method. This method consists in recording the occurrence of specific behaviors at a regular time interval. An ethogram was previously established, summarizing the different possible behaviors for white seabream in captivity conditions. The occurrence of the isolated behaviors from the ethogram between tagged and untagged groups and the temporal evolution of these behaviors were tested. The activity budgets of individual fish showed no significant differences, but high variability in behavioral patterns. No differences were detected for the occurrence of the different behaviors between the tagged and control groups, except for socializing behavior. Tagged fish were more often grouped than control fish. One fish lost its tag after 17 days, due to suture shedding (i.e. the loss of surgical monofilament used to close a wound by joining the edges). After 18 days, suture inspection showed that the incision regions had not healed, but no other fish had lost sutures. No mortality was recorded during the experiment, regardless of fish group. After 48 days, the tagged fish presented neither visceral damage nor inflammation. Incision regions presented no inflammation and had totally healed with only one out of the two or three suture points we made persisting for each tagged fish. In conclusion, neither tag presence in the body cavity nor the tagging procedure cause any adverse effects on the behavior and survival of wild seabream in captivity conditions. Therefore, we consider that the monitoring of tagged white seabreams is representative of untagged fish and reflect their natural behavior. Additionally, as white seabream recover quickly after surgery and present no adverse behavioral effects due to intracoelomic tag implantation, we recommend maintaining fish for as short a time as possible in captivity conditions to avoid extra stress, and to release them as soon as they recover from the tagging procedure.

Published
2013
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17. Effects of post-settlement mortality on size and parasite load in juvenile Diplodus vulgaris and D. sargus in the Mediterranean

Author

Serge Planes, Suzanne C. Mills, David Lecchini, and Pascal Romans

Subjects
Ecology, biology, Mortality processes, Zoology, Growth, Diplodus, Aquatic Science, Oceanography, biology.organism_classification, Parasite load, Life history theory, Abundance (ecology), Parasite hosting, Juvenile, Diplodus vulgaris, Parasites, Life history, Selection, Ecology, Evolution, Behavior and Systematics, Sargus
Abstract

This study explored how mortality processes influence life history traits of recently set- tled juvenile fishes (Diplodus vulgaris and D. sargus) in the Mediterranean. Mortality was estimated by measuring the decrease in abundance of a cohort from settlement to 4 mo post-settlement. Growth and parasite load were simultaneously measured over time to determine those phenotypes more sus- ceptible to mortality. Diplodus spp. juvenile mortality was between 71 and 81% over the 4 mo period. A decrease in the coefficient of variation in size over time suggests that mortality may influence growth of both Diplodus species. A decrease in parasite abundance of both Diplodus species 60 d after settlement suggests that fish with higher parasite loads may have been more susceptible to mor- tality. Overall, mortality did not act randomly in D. vulgaris and D. sargus cohorts, as the variance in both size and parasite abundance of recently settled juveniles was not comparable to that of the adult population; only larger, parasite-free individuals survived.

Published
2009
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18. Towards a vitrification-based cryopreservation protocol for the coral Pocillopora damicornis L.: Tolerance of tissue balls to 4.5 M cryoprotectant solutions

Author

Dominique Barthélémy, Patrick Masanet, Lionel Feuillassier, Florent Engelmann, and Pascal Romans

Subjects
Glycerol, medicine.medical_specialty, Ethylene Glycol, Molar concentration, Cryoprotectant, Pocillopora damicornis, General Biochemistry, Genetics and Molecular Biology, Cryopreservation, chemistry.chemical_compound, Cryoprotective Agents, medicine, Animals, heterocyclic compounds, Vitrification, Dimethyl Sulfoxide, biology, Coral Reefs, Methanol, General Medicine, biology.organism_classification, Anthozoa, Surgery, chemistry, cardiovascular system, General Agricultural and Biological Sciences, Ethylene glycol, Nuclear chemistry
Abstract

In this study, we tested the tolerance of tissue balls (TBs, 100–400 μm in diameter) from the coral Pocillopora damicornis produced using mechanical excision to exposure to cryoprotectant (CPA) solutions. TBs were treated for 20 min at room temperature with individual, binary, ternary or quaternary CPA solutions with a total molarity from 2.0 to 5.0 M. Four CPAs were used: ethylene glycol (EG), dimethylsulfoxide (Me 2 SO), methanol (Met) and glycerol (Gly). In some experiments, the molarity of the CPA solutions was increased and decreased in a stepwise manner. The tolerance of TBs following CPA treatment was evaluated using two parameters. The Tissue Ball Regression (expressed in μm/h) measured the diameter regression of TBs over time. The % Undamaged TBs quantified the proportion of TBs, which remained intact over time after the CPA treatment. TBs tolerated exposure to binary solutions with a total molarity of 4.0 M containing 2.0 M EG + 2.0 M Met and 2.0 M EG + 2.0 M Gly. TBs displayed tolerance to ternary solutions with a total molarity up to 3.0 M, containing each CPA at 1.0 M. Quaternary solutions with a total molarity of 4.0 M containing each CPA at 1.0 M were not tolerated by TBs. When the molarity of the CPA solutions was increased and decreased in a stepwise manner, TBs withstood exposure to a CPA solution with a total molarity of 4.5 M, containing 1.5 M EG + 1.5 M Gly + 1.5 M Me 2 SO. This study confirmed the interest of using TBs to test CPA solutions, with the objective of developing a vitrification-based cryopreservation protocol.

Published
2015

19. Survival of tissue balls from the coral Pocillopora damicornis L. exposed to cryoprotectant solutions

Author

Patrick Masanet, Florent Engelmann, Lucie Martinez, Dominique Barthélémy, Pascal Romans, Isabelle Engelmann-Sylvestre, and Lionel Feuillassier

Subjects
Glycerol, Ethylene Glycol, Cryoprotectant, Coral, Pocillopora damicornis, Biology, General Biochemistry, Genetics and Molecular Biology, Cryopreservation, chemistry.chemical_compound, Cryoprotective Agents, Animals, Tissue ball, Dimethyl Sulfoxide, Chromatography, Dimethyl sulfoxide, Methanol, General Medicine, Anatomy, biology.organism_classification, Anthozoa, Propylene Glycol, chemistry, Toxicity, General Agricultural and Biological Sciences, Ethylene glycol
Abstract

In this study, the tolerance of tissue balls (TBs, 100–300 μm in diameter) from the coral Pocillopora damicornis produced using mechanical excision to exposure to cryoprotectant (CPA) solutions was tested. TBs were treated for 20 min at room temperature with solutions of ethylene glycol (EG), methanol (Met), glycerol (Gly) or dimethyl sulfoxide (Me2SO) at concentrations between 1.0 and 4.5 M. Two parameters were used to evaluate the survival of TBs following CPA treatment. The Undamaged Duration of Tissue Balls (expressed in h) corresponded to the time period during which the membrane surface of TBs remained smooth and their motility was preserved. Tissue Ball Regression (expressed in μm/h) corresponded to the size reduction of TBs over time. TBs tolerated exposure to all CPAs tested at the three lower concentrations employed (1.0 M, 1.5 M and 2.0 M). No survival was achieved following exposure to a 4.5 M CPA solution. At concentrations of 3.0 and 4.0 M, higher Undamaged Duration of Tissue Balls and lower Tissue Ball Regression were obtained following treatment with EG compared to the other three CPAs. Our experiments show that TBs constitute a good experimental material to evaluate CPA toxicity on corals using large numbers of samples. Performing preliminary experiments with TBs may allow reducing the number of tests carried out with less easily available coral forms such as planulae, thereby preserving larval stocks.

Published
2014

20. Tolerance of apexes of coral Pocillopora damicornis L. to cryoprotectant solutions

Author

Patrick Masanet, Dominique Barthélémy, Isabelle Engelmann-Sylvestre, Lionel Feuillassier, Pascal Romans, and Florent Engelmann

Subjects
Glycerol, Ethylene Glycol, Sucrose, Molar concentration, Cryoprotectant, Pocillopora damicornis, General Biochemistry, Genetics and Molecular Biology, chemistry.chemical_compound, Cryoprotective Agents, Osmotic Pressure, Botany, Animals, Dimethyl Sulfoxide, solutions, Cryopreservation, biology, Dimethyl sulfoxide, Methanol, Osmolar Concentration, Trehalose, General Medicine, biology.organism_classification, Anthozoa, Adaptation, Physiological, Propylene Glycol, chemistry, cardiovascular system, Coral, Apex, General Agricultural and Biological Sciences, Ethylene glycol, Nuclear chemistry
Abstract

In this study, we investigated the tolerance of Pocillopora damicornis apexes to treatments with solutions containing penetrating and non-penetrating cryoprotective agents (CPAs). CPAs were employed individually or in binary, tertiary or quaternary solutions. In some experiments apexes were treated successively with two CPA solutions with increasing total concentration. P. damicornis apexes withstood exposure for up to 30 min to solutions containing 0.6-0.8 M sucrose (Suc) or trehalose (Tre). When apexes were treated with binary cryoprotectant solutions containing Suc and ethylene glycol (EG), methanol (Meth), dimethyl sulfoxide (Me2SO) or glycerol (Gly), the CPAs employed in combination with Suc could be ranked in the following order of decreasing tolerance: EG > Meth > Me2SO > Gly. P. damicornis apexes tolerated exposure to complex CPA solutions containing Suc, Me2SO, EG and/or Meth with a total molarity of 2.45 M. In experiments where two successive CPA solutions were employed, apexes withstood treatment with the second, more concentrated solution at 0 degrees C for up to 10 min. These preliminary results pave the way to the development of a cryopreservation protocol for P. damicornis apexes.

Published
2014

21. Temporal changes in the growth of two Mediterranean cold-water coral species, in situ and in aquaria

Author

Nadine Le Bris, Simon Pareige, Pascal Romans, Elwyn de la Vega, Franck Lartaud, Marc de Rafélis, Erwan Peru, Lionel Feuillassier, Marjorie Bideau, Karine Nedoncelle, Laboratoire d'Ecogéochimie des environnements benthiques (LECOB), Observatoire océanologique de Banyuls (OOB), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Biominéralisations et environnements sédimentaires (BES), Institut des Sciences de la Terre de Paris (iSTeP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Canyon, Mediterranean climate, geography, geography.geographical_feature_category, biology, Coral, media_common.quotation_subject, [SDU.STU]Sciences of the Universe [physics]/Earth Sciences, Oceanography, biology.organism_classification, Lophelia, Mediterranean sea, Ecosystem, 14. Life underwater, Reproduction, ComputingMilieux_MISCELLANEOUS, media_common, Madrepora oculata
Abstract

In situ growth patterns of two-species of reef-building cold-water corals were investigated for the first time at different temporal scales, based on the redeployment of coral nubbins in their natural environment. Lophelia pertusa and Madrepora oculata were collected in November 2010 and May 2011 from the Lacaze-Duthiers canyon in the northwestern Mediterranean Sea (520 m depth). Three in situ growth experiments were performed from November 2010 to May 2011, May to September 2011 and November 2010 to September 2011. For comparison, aquaria experiments over comparable lengths of time were conducted with coral colonies collected in November 2010. In the canyon, new polyps from M. oculata and L. pertusa displayed similar growth rates in summer (5.8±3.8 and 7.3±1.7 mm yr−1 respectively), but M. oculata growed significantly slower during winter/spring than L. pertusa (4.1±1.8 and 8.4±2.7 mm yr−1 respectively). Budding rates however (the rate of new polyp addition per mother polyp per year) were similar between both species in winter/spring (45±40% for M. oculata and 48±72% for L. pertusa), but were significantly lower in summer for M. oculata (14±19%) compared to L. pertusa (58±94%). This seasonal difference in the growth between L. pertusa and M. oculata might reflect differences in species-specific physiology (such as reproduction) or feeding strategy, or a higher sensitivity of M. oculata to the variability of food supply in the Lacaze-Duthiers canyon resulting from periodic cascading events. The comparison of in situ and aquaria growth experiments showed no significant differences for budding and new polyp growth rates, which supports the validity of aquaria experiments for these kinds of investigations. However the budding rates observed were consistently lower in coral maintained in aquaria than those in in situ conditions, a finding which is to be considered when extrapolating laboratory based investigation results to the naturally occurring coral ecosystem.

Published
2014
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22. A new approach for assessing cold-water coral growth in situ using fluorescent calcein staining

Author

Nadine Le Bris, Pascal Romans, Frédéric Alcala, Marc de Rafélis, Franck Lartaud, Erwan Peru, Marjorie Bideau, Lionel Feuillassier, Simon Pareige, Laboratoire d'Ecogéochimie des environnements benthiques (LECOB), Observatoire océanologique de Banyuls (OOB), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Biominéralisations et environnements sédimentaires (BES), Institut des Sciences de la Terre de Paris (iSTeP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), and Compagnie Maritime d'Expertise (COMEX)

Subjects
0106 biological sciences, In situ, geography, geography.geographical_feature_category, 010504 meteorology & atmospheric sciences, biology, Ecology, 010604 marine biology & hydrobiology, Scleractinia, [SDU.STU]Sciences of the Universe [physics]/Earth Sciences, Coral reef, Aquatic Science, biology.organism_classification, 01 natural sciences, Calcein, chemistry.chemical_compound, Animal science, Mediterranean sea, Lophelia, chemistry, 14. Life underwater, Reef, ComputingMilieux_MISCELLANEOUS, 0105 earth and related environmental sciences, Madrepora oculata
Abstract

Research on the biology and ecology of cold-water corals (CWCs) is still in its infancy. The growth patterns of CWCs in their natural environments are poorly known. Growth rate investigations on these deep-sea reef builder species are needed to predict recovery times following damage to their ecosystems. This study investigates a new approach for analysing CWC growth rate, suitable for in situ application. Lophelia pertusa and Madrepora oculata (Scleractinian) were collected from the Lacaze-Duthiers canyon in the northwestern Mediterranean Sea (520 m depth), marked and then either redeployed in situ for 6 months, or maintained in aquaria for growth rate comparison at a constant temperature of 13 °C, corresponding to their habitat conditions. Two different types of staining (calcein and manganese) and three different exposure times (30, 60 and 240 min) were tested. The results show that calcein offers rapid incorporation and easy detection, making it particularly suitable for skeletal growth rate investigations compared with other chemical staining. In situ linear polyp growth rates of 7.5 ± 1.2 mm y-1 and 3.5 ± 2.1 mm y-1 were measured in new polyps of L. pertusa and M. oculata , respectively. Those values were significantly higher in young polyps than in older ones, where they decreased to 1.3 ± 1.5 mm y-1 and 1.2 ± 1.2 mm y-1 . Beyond the study of coral reef growth processes, this approach offers a methodological basis for habitat quality assessment which could be used in the management of deep-sea marine protected areas (MPA).

Published
2013
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