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First published online February 15, 2006
Journal of Experimental Biology 209, 801-809 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02068

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Giant liposomes as delivery system for ecophysiological studies in copepods

Isabella Buttino^1,*, Giuseppe De Rosa², Ylenia Carotenuto¹, Adrianna Ianora¹, Angelo Fontana³, Fabiana Quaglia², Maria Immacolata La Rotonda² and Antonio Miralto¹

¹ Stazione Zoologica `Anton Dohrn' Villa Comunale, 80121 Napoli, Italy
² Dipartimento di Chimica Farmaceutica e Tossicologica, Università degli Studi di Napoli Federico II, Via D. Montesano, 49, 80131 Napoli, Italy
³ Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche (ICB-CNR), Via Campi Flegrei, 34, 80078 Pozzuoli, Italy

^* Author for correspondence (e-mail: buttino{at}szn.it)

Accepted 29 December 2005

Giant liposomes are proposed as a potential delivery system in marine copepods, the dominant constituent of the zooplankton. Liposomes were prepared in the same size range as the food ingested by copepods (mean diameter of about 7 µm). The encapsulation of a hydrophilic and high molecular mass fluorescent compound, fluorescein isothiocyanate-dextran (FitcDx), within the liposomes provided a means of verifying copepod ingestion when viewed with the confocal laser-scanning microscope. Females of the calanoid copepod Temora stylifera were fed with FitcDx-encapsulated liposomes alone or mixed with the dinoflagellate alga Prorocentrum minimum. Control copepods were incubated with the P. minimum diet alone. Egg production rates, percentage egg-hatching success and number of faecal pellets produced were evaluated after 24 h and 48 h of feeding. Epifluorescence of copepod gut and faecal pellets indicated that the liposomes were actively ingested by T. stylifera in both experimental food conditions, with or without the dinoflagellate diet. Ingestion rates calculated using ³H-labelled liposomes indicated that females ingested more liposomes when P. minimum was added to the solution (16% vs 7.6% of uptake). When liposomes were supplied together with the algal diet, egg production rate, egg-hatching success and faecal pellet production were as high as those observed for the control diet. By contrary, egg production and hatching success were very low with a diet of liposomes alone and faecal pellet production was similar to that recorded in starved females. This results suggest that liposomes alone did not add any nutritive value to the diet, making them a good candidate as inert carriers to study the nutrient requirements or biological activity of different compounds. In particular, such liposomes are proposed as carriers for diatom-derived polyunsaturated aldehydes, which are known to impair copepod embryo viability. Other potential applications of liposomes as a delivery system of drugs and nutrients in copepod mass cultivation, or as carriers of pollutants to study copepod physiology in ecotoxicological experiments, are also discussed.

Key words: Temora stylifera, Prorocentrum minimum, giant liposome, diatom, unsaturated aldehyde, copepod egg viability, delivery system, confocal microscopy, feeding experiment

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				I. Buttino, G. De Rosa, Y. Carotenuto, M. Mazzella, A. Ianora, F. Esposito, V. Vitiello, F. Quaglia, M. I. La Rotonda, and A. Miralto Aldehyde-encapsulating liposomes impair marine grazer survivorship J. Exp. Biol., May 1, 2008; 211(9): 1426 - 1433. [Abstract] [Full Text] [PDF]