Rearing of research model organisms

The unit takes care of short and long-term maintenance of Mediterranean model species, mainly sampled in the Gulf of Naples and used for the purposes of Scientific Research (various echinoderms, molluscs, tunicates, cnidarians, sponges), under controlled conditions. It also cures the maintenance in closed loop systems of non-autochthonous echinoderm species.

Breeding

The unit is developing effective systems for the breeding of the two most commonly used model species (Ciona intestinalis and Paracentrotus lividus) and various research projects are conducted aimed at the breeding of various species of Mediterranean invertebrates.

Laboratory services for research

Collection and supply of organs / tissues / gametes and preparation of batch cultures of embryos at different stages of development and larvae. The technicians of the unit actively cooperate with the SZN researchers to meet their experimental needs to their full potential. A mass production service and supply of phytoplankton crops is also available for the purposes of the scientific research.

New model species (“candidate models”)

Identification of new species of model animals (invertebrates and vertebrates) is ongoing.
The aim of these activities is the identification and the creation of a "bank" of phyto- and zoo-planktonic and benthic organisms to be used as models, as food for other organisms, etc.

Mesocosms and Facilities for Research in the Marine Environment

At the Mare Unit, technicians are frequently involved in the development of experimental set-ups specifically constructed according to the needs expressed by various research groups, in response to required formalities. For this purpose, automatic systems have been developed for trophic studies on molluscs and echinoderms, set-up were designed to simulate the effect of turbulence on the stratification of marine sediments, microcosms were realized for the management of echinoderms, molluscs, cephalochordates and sponges under controlled conditions. The set-ups are realized according to own original projects and they remain an intellectual property of the Unit, while the services rendered allow the SZN researchers to count on a continuous and due cooperation in support of their research. These activities will be improved in the next future, with the installation of new facilities that will include, among others, programmable automatic systems for the breeding of model organisms (SZN Patent).

Area di ricerca: Life Sciences – Immunity and Infection: Microbiology, Veterinary Medicine
Principal Investigator: Sandra Hochscheid
PhD Student: Antonino Pace
External Supervisor: Ludovico Dipeneto, Dipartimento di Medicina Veterinaria e Produzioni Animali, Università degli Studi di Napoli Federico II

Surveillance and research on infective diseases in wildlife populations may identify changes in ecosystem balance and emerging threats to human and animal welfare. Health assessment in coastal organisms can be used to indirectly monitor marine ecosystem status, investigate the effects of human activities on animal health, and identify risks to humans utilizing the same habitat for food, work, or recreation.
Fishes are susceptible to a wide variety of bacterial pathogens. Many of these bacteria are considered to be saprophytic in nature, becoming pathogenic when fishes are physiologically unbalanced, nutritionally deficient, or there are other stressors (e.g. poor water quality, overstocking). The systemic investigation is critical to the identification of potential pathogens and to a better understanding of their effects on host species.

What we do
We perform the microbiological and parasitological survey on marine organisms of the Gulf of Naples and the surrounding area in order to determine the health status and obtain information on the quality of the environment of origin

Partners
Dipartimento di Medicina Veterinaria e Produzioni Animali, Università degli Studi di Napoli Federico II - Istituto Zooprofilattico Sperimentale del Mezzogiorno

Staff
Sandra Hochscheid
Andrea Affuso
Antonino Pace

2017

Maibam C, Mutalipassi M and Zupo V (submitted) Gonopore sexing technique allows determination of sex ratios in the marine shrimp Hippolyte inermis Leach. (Integrative and Comparative Ecology).

Mutalipassi, M., Di Natale, M., Mazzella, V., & Zupo, V. (2017). Automated culture of aquatic model organisms: Shrimp larvae husbandry for the needs of research and aquaculture. Animal, 1-9. doi:10.1017/S1751731117000908.

Ruocco N, Caramiello D, Glaviano F, Zupo V, Fontana A, Costantini M (submitted) Experimental evaluation of the feeding rate, growth and fertility of the sea urchins Paracentrotus lividus. Invertebrate Reproduction and Development.

Ruocco N,  Costantini S, Zupo V, Romano G, Ianora A, Fontana A, Costantini M (2017) High-quality RNA extraction from the sea urchin Paracentrotus lividus embryos.  PLoS ONE 12(2): e0172171. doi:10.1371/journal.pone.0172171

Zupo V., Glaviano F, Caramiello D, Mutalipassi M (in press) Effect of five benthic diatoms on the survival and development of Paracentrotus lividus post-larvae in the laboratory. Aquaculture.

Zupo V, Glaviano F, Paolucci M, Ruocco N, Polese G, Di Cosmo A , Costantini M, Mutalipassi M (submitted) Roe enhancement of Paracentrotus lividus: nutritional effects of fresh and formulated diets. Aquaculture Nutrition.

Zupo V, Mutalipassi M, Fink P. and Di Natale M. (2017) Effect of Ocean Acidification on the Communications among Invertebrates Mediated by Plant-Produced Volatile Organic Compounds. Glob J Ecol 1(1): 012-018.

Zupo V, Mutalipassi M, Glaviano F, Buono AC, Cannavacciuolo A & Fink P. (Submitted) Induction of larval settlement in a decapod crustacean: effect of seagrass leaves and an epiphyte diatom on the metamorphosis of the shrimp Hippolyte inermis Leach. Marine Biology.

2016

Migliaccio, O., Castellano, I., Di Cioccio, D., Tedeschi, G., Negri, A., Cirino,P., Romano, G., Zingone, A. and Palumbo, A. (2016). Subtle reproductive impairment through nitric oxide-mediated mechanisms in sea urchins from an area affected by harmful algal blooms. Scientific Reports 6. Article number: 26086.

Motta, C.M., Cerciello, R., De Bonis, S., Mazzella, V., Cirino, P., Panzuto, R., Ciaravolo, M., Simoniello, P., Toscanesi, M., Trifuoggi, M. and Avallone, B. (2016). Potential toxicity of improperly discarded exhausted photovoltaic cells. Environmental Pollution 216, 786-792.

2015

Crocetta, F., Marino, R., Cirino, P., Macina, A., Staiano, L., Esposito, R., Pezzotti, M. R., Racioppi, C., Toscano, F., De Felice, E., Locascio, A., Ristoratore, F., Spagnuolo, A., Zanetti, L., Branno, M. and Sordino, P. (2015). Mutation studies in ascidians: A review. Genesis, 53: 160–169. doi: 10.1002/dvg.22837.

Manzo S., Cirino P., Schiavo S. , Oliviero M. , Paglialonga A., Ciaravolo M. (2015). I ricci di mare nella ricerca ecotossicologica, possibili strategie per la disponibilità continua di gameti: Risultati preliminari. Atti in: Giornate di Studio, 6° Edizione: Ricerca ed Applicazione di Metodologie Ecotossicologiche in Ambienti Acquatici e Matrici Contaminate. ISPRA – Atti 2015. ISBN 978-88-448-0711-5

2014

Caputi L., Crocetta F, Toscano F, Sordino P & Cirino P. (2014). Long-term demographic and reproductive trends in Ciona intestinalis sp. A. Marine Ecology AEP, DOI: 10.1111/MAEC.12125

Ciaravolo M., Di Cosmo A, Polese G, Cirino P. (2014). The Marketing of the Paracentrotus lividus: Gonadal Growth vs Maturation. In: Aquaculture Europe 14 Abstracts: 257. San Sebastian (Spagna), 14/10/2014.

Della Torre C., Bergami E, Salvati A, Faleri C, Cirino P, Dawson KA, Corsi I. (2014). Accumulation and Embryotoxicity of Polystyrene Nanoparticles at Early Stage of Development of Sea Urchin Embryos Paracentrotus lividus. Environmental Science & Technology, 48(20): 12302- 11.

Sorrenti G., Bagnoli A, Miraglia V, Crocetta F, Vitiello V, Ristoratore F, Cirino P, Sansone G, Sordino P. (2014). Investigating sperm cryopreservation in a model tunicate, Ciona intestinalis sp. A Cryobiology 68 (1) : 43-9.

2013

Cirino P., Ciaravolo M., de Girolamo P., Paglialonga A., Rosica A.M. (2013). The “Captive Gonadal Cycle” of the Sea Urchin Paracentrotus lividus. In: Aquaculture Europe 13 Abstracts. Trondheim (Norway), 09-12/08/2013.

Cirino P, Ciaravolo M, Paglialonga A. (2013). "Restorative" Handling of Under-Sized Paracentrotus lividus. In: Aquaculture Europe 13 Abstracts. Trondheim (Norway), 09-12/08/2013

Cirino P. (2013). Dissection of the endostyle in Ciona intestinalis. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

2012

Cirino P., Toscano A. (2012). Spawning of gametes in Paracentrotus lividus. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Cirino P., Francone M., Ristoratore F., Zanetti L. (2012). Preparation of Ascidian hemolymph for the culture of Ciona cells. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Cirino P. (2012). Induced spawning of gametes in Ciona intestinalis. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Cirino P., Toscano A. (2012). Fertilization in Paracentrotus lividus. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Cirino P., Ciaravolo M., Paglialonga A. (2012). Sea urchins in a long-term maintenance project: results and perspectives. In: Aquaculture Europe 2012 (AQUA 2012) Abstracts : 216 . Prague (Czech Republic) 1-5th September.

2011

Cirino P., Ciaravolo M, Paglialonga A, Toscano A. (2011). A long-term laboratory maintenance system for promoting out-of-season maturation in the sea urchin Paracentrotus lividus (Lamarck, 1816). In: Aquaculture Europe 2011 (AE2011: Mediterranean Aquaculture 2020) Abstracts : 195. Rodhes, Greece, October 18-21 2011. Best Poster Award

Sardo A, Rossi R, Soprano V, Ciminiello P, Fattorusso E, Cirino P, Zingone A. (2011). The impact of Ostreopsis ovata on Mytilus galloprovincialis and Paracentrotus lividus. In: ICOD Ecology abstracts. Villefranche sur mer, France, 6-8/4/2011

2010

Cirino P, Brown E. (2010). Protocol for fertilization tests in Ciona intestinalis. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

L. Manfra, E. De Nicola, C. Maggi, E. Zambianchi, D. Caramiello, A. Toscano, D. Cianelli and A. M. Cicero (2010). Exposures of rotifers, crustaceans and sea urchins to produced formation waters and seawaters in the Mediterranean Sea. Journal of the Marine Biological Association of the United Kingdom, pag. 1-7; doi:10.1017/S0025315410001037.

Lasbleiz R, Leveque L, Schires G, Cirino P. (2010). Materials and packaging techniques for the shipment of Ciona intestinalis. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Cirino P, Lasbleiz R, Leveque L, Schires G. (2010). Health and maturation status of Ciona intestinalis. In: ASSEMBLE virtual tool box protocols www.assemblemarine.org.

Sardo A, Rossi R, Soprano V, Ciminiello P, Fattorusso E, Cirino P, Zingone A. (2010). Feeding Mytilus galloprovincialis and Paracentrotus lividus with Ostreopsis ovata: toxicity and pathological aspects. In: 14th International Conference on Harmful Algae Abstracts. Hersonissos-Crete, Greece, 1-5 November 2010

Cirino P. (2010). Optimization of conditioning / re-acclimatization after transport of Ciona intestinalis. WS 2 Assemble: Remote access & Culture techniques Workshop (OOB): October, 2010 Observatoire Océanologique in Banyuls, FR. 

Toscano A., Cirino P. (2010). Allevamento di stadi larvali di riccio di mare. Metodologie di studio del Plancton marino. Manuali e Linee guida ISPRA/SIBM Vol. 56 (cap.50): 601-608.

Reference: Luigia Santella

Laboratory equipped to study the cellular and molecular mechanisms of cell cycle and fertilization of marine organisms. It includes a live cell imaging facility, areas for microinjection, cellular and molecular biology, live-image analysis, in addition to basic laboratory equipments.

Systems for experimentation

The laboratory utilizes two species of starfish (Astropecten aranciacus and Asterina pectinifera) and sea urchin (Paracentrotus lividus) as model systems for scientific research in collaboration with Meda and MaRe units (RiMAR Section, SZN) for sampling and animal keeping facility.

 Analysis

• Study of the mechanisms responsible for the generation, propagation, and decay of intracellular calcium signals in eggs of marine animals during meiotic cell cycle and fertilization, aiming to the identification of the role of intracellular calcium reservoire (e.g. endoplasmic reticulum, nucleus, mitochondria, acid vesicles, etc) and ion release/resorption mechanism.
• Study of the causal relations between the external environment (seawater salinity, pH, temperature, environmental contaminants, etc.) and the morphological and functional changes of oocytes and eggs and their effects on fertilization and early stages of embryonic development

• Analysis of calcium signaling, cytoskeleton ultrastructure (in collaboration with AMOBIO unit, RiMAR Section), polyspermy, and progression of the division of the zygote.

• Analysis of the molecular mechanisms that regulate meiotic maturation and activation of the eggs; effects of microinjected proteins blocking specific functions, antibodies and antisense RNA of target genes identified by transcriptome of seastar and sea urchins.

Equipments

• 2 Set up (CCD cameras) multichannel (brightfield and fluorescence) for live cell fluorescence imaging
• 1 Set up (Confocal Spinning Disk) multichannel (brightfield and fluorescence) for live fluorescence imaging
• Live cell imaging during microinjection UV photoactivation of caged compounds
• Upright fluorescent microscope for microinjection into cytoplasm and nucleus of starfish oocytes and sea urchin eggs
• Stereomicroscopes for gametes quality evaluation and their manipulation (enucleation etc)
• Incubator to keep gametes under controlled temperature