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First published online April 18, 2008
Journal of Experimental Biology 211, 1426-1433 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.015859
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Aldehyde-encapsulating liposomes impair marine grazer survivorship

Isabella Buttino1,*, Giuseppe De Rosa2, Ylenia Carotenuto1, Marialuisa Mazzella2, Adrianna Ianora1, Francesco Esposito1, Valentina Vitiello1, 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


Figure 1
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  		Fig. 1. 2-trans,4-trans decadienal loaded into liposomes, expressed as µg mg–1 (means ± s.d., N=3) of the total lipids during storage at 4°C for 15 days.

 

Figure 2
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  		Fig. 2. Temora stylifera daily faecal pellet production (A) and percentage female survival (B) for couples fed liposomes at a lipid concentration of either 7.5 µg ml–1 lipids (Lipo A) or at 4.0 µg ml–1 lipids (Lipo B) supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1 and compared with a control diet of P. minimum alone (Pro) at the same cell concentration. Values are means ± s.e.m. (N=10).

 

Figure 3
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  		Fig. 3. Temora stylifera couples fed blank liposomes at a lipid concentration of 2 µg ml–1 (Lipo C) and liposomes encapsulating decadienal (DD) at 2.9 ng ml–1 (LipoDD) supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1, and the control diet P. minimum alone (Pro) at the same cell concentration. (A) Daily egg production rates per female; (B) percentage egg hatching success; (C) faecal pellet production rates per couple; (D) percentage female survival. Values are means ± s.e.m. (N=10).

 

Figure 4
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  		Fig. 4. Calanus helgolandicus females fed blank liposomes at a lipid concentration of 6.3 µg ml–1 (Lipo Cal) and liposomes encapsulating decadienal at 3.6 ng ml–1 (LipoDD Cal) supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1, and the control diet of P. minimum alone (Pro) at the same cell concentration. (A) Daily egg production rates per female; (B) percentage egg hatching success; (C) faecal pellet production rates per female; (D) percentage female survival. Values are means ± s.e.m. (N=10).

 

Figure 5
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  		Fig. 5. Calanus helgolandicus embryos produced by females fed for 8 days with liposomes encapsulating decadienal at a concentration of 3.6 ng ml–1 (LipoDD Cal; A) and blank liposomes at a lipid concentration of 6.3 µg ml–1 lipid (Lipo Cal; B) and supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1. Embryos were stained with the vital fluorescent probe Annexin V-FITC. Green fluorescence indicates that nuclei were apoptotic (asterisks). Magnification: x200.

 

Figure 6
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  		Fig. 6. Calanus helgolandicus females fed for 9 days with liposomes encapsulating decadienal at a concentration of 3.6 ng ml–1 (LipoDD Cal; A,B) and blank liposomes at a lipid concentration of 6.3 µg ml–1 lipid (Lipo Cal; C) and supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1. TUNEL was used to detect apoptosis. Green fluorescence indicates that tissues were apoptotic. Arrow in C indicates the same region of the gonad as in B but no green fluorescence is present. Magnification: (A,C) x100; (B) x250.

 

Figure 7
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  		Fig. 7. Temora stylifera females fed for 6 days with liposomes encapsulating decadienal at a concentration of 2.9 ng ml–1 (LipoDD; A,B) and blank liposomes at a lipid concentration of 2.0 µg ml–1 lipid (Lipo C; C) and supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1. TUNEL was used to detect apoptosis. The green fluorescence in the gonad (arrow in B) indicates that tissues were apoptotic. In C, the arrow indicates the same region of the gonad as in B but no green fluorescence is present. Magnification: (A,C) x100; B x250).

 

Figure 8
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  		Fig. 8. Temora stylifera females fed for 10 days with liposomes encapsulating decadienal at a concentration of 2.9 ng ml–1 (LipoDD; A,B) and supplied with Prorocentrum minimum at a concentration of 8000 cells ml–1. TUNEL was used to detect apoptosis. Green fluorescence indicates that tissues were apoptotic. Magnification: (A) x100; (B) x250.

 

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