A simple method for mass-production of liposomes, in particular large liposomes, suitable for delivery of free amino acids to filter feeding zooplankton.

In marine fish larviculture the live feed organisms are often enriched in order to enhance their nutritional value. One of the challenges is to enhance the phospholipids (PL) content, and another is to enhance the content of specific water soluble nutrients, like free amino acids (FAA). There are a few studies where this has been achieved by the use of liposomes. The aim of this study was to develop a simple method for mass-production of liposomes within a size range of 1-5 mum a size range suitable to feed live food organisms. Furthermore, the liposomes should have a high FAA concentration and be stable under conditions typical for short-time enrichment of live feed organisms. The method used in the present study is based on a combination of a reverse-phase evaporation method for preparing liposomes and re-hydration of freeze-dried, empty liposomes. The liposomal membrane was made of soy phosphatydilcholine and was loaded with a highly concentrated free amino acids solution. Most of the liposomes produced were 2-8 mum in diameter and the FAA encapsulation efficiency was 42.6%. Two experiments simulating 2 hr of live food enrichment were used to evaluate the liposomes. The results showed the liposome did not disintegrate or aggregate when suspended in seawater and that only 9% of the FAA content of the liposomes was lost after 2 hr suspension. The developed method was easy and reliable, producing tens of grams of liposomes per batch.

Hyperthermia induced atrial natriuretic peptide expression and deviant heart development in Atlantic salmon Salmo salar embryos.

Heart abnormalities are increasingly recognized as a problem in salmon aquaculture. Fish in early life-stages are particularly susceptible to teratogens, including elevated water temperature. Recently, heat-induced mRNA expression of the cardiac hormone atrial natriuretic peptide (ANP), which is known to be involved in modulation of cardiac growth and regulation of cardiac homeostasis, was demonstrated in Atlantic salmon (Salmo salar) embryos by RAP-PCR. The relation between heat sensitive ANP expression and heart abnormalities was explored in two experiments. In an experiment with short-term exposure, salmon eggs were heat shocked at 16 degrees C at eight different embryonic stages from gastrulation till completion of somitogenesis. The RT-PCR results showed that the ANP mRNA expression was down-regulated at the onset of heart formation at the gastrula stage, while the transcription became heat inducible from the fusioning of the heart tube around the 15th-20th somite stage and onwards. This was confirmed by whole-mount in situ hybridization, which also showed that ANP is exclusively expressed in the heart of Atlantic salmon embryos. In a second long-term experiment, salmon embryos were incubated at either 10 degrees C (high temperature) or 8 degrees C (controls) from fertilization till first feeding, and subsequently reared within normal conditions to an average size of 52 g. The long-term hyperthermic embryos showed up-regulated ANP transcription at the approximately 9th and approximately 20th somite stage and at the completion of somitogenesis. The cardiosomatic index [CSI; (ventricle weight/body weight) *100] demonstrated a significant decrease in the relative heart weight of fish incubated at 10 degrees C during the embryogenesis compared with controls. In these fish, aplasia of septum transversum was observed in 2 of 25 fish, resulting in abnormally shaped hearts situated partly within the abdominal cavity. Altogether, our results demonstrate that hyperthermia both induce deviant development of heart and associated structures and up-regulation of ANP transcription during embryogenesis. A possible role of ANP in development of heart malformations is thus suggested.