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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Oscars, Astronotus ocellatus, have a dietary requirement for vitamin C
D M Fracalossi1, M E Allen, D K Nichols
1Department of Zoological Research, National Zoological Park, Smithsonian Institution, Washington, DC 20008, USA.
Abstract:
We found that vitamin C is an essential nutrient for an Amazonian ornamental fish, the oscar (Astronotus ocellatus). This was demonstrated by the absence of L-gulonolactone oxidase activity, the enzyme responsible for the biosynthesis of vitamin C, in liver or kidney of oscars and by a feeding trial in which oscars without vitamin C dietary supplementation developed clinical deficiency signs. Fish weighing 29.2 +/- 1.9 g were divided into four groups, and each group was fed a casein-based semipurified diet containing 0, 25, 75 or 200 mg ascorbic acid equivalent (AA)/kg diet for 26 wk. Vitamin C was supplemented in the diets as L-ascorbyl-2-polyphosphate, a mixture of phosphate esters of ascorbate, which is more stable to oxidation than AA. At the end of 26 wk, fish fed no AA had significantly lower weight gain than fish fed the AA-supplemented diets (P < 0.05). Oscars without dietary AA supplementation gained only 37% of their initial weight, compared with 112, 102 and 91% gained by fish fed 25, 75 and 200 mg AA/kg diet, respectively. After 25 wk without dietary supplementation of AA, fish began to develop clinical deficiency signs, including deformed opercula and jaws, hemorrhage in the eyes and fins, and lordosis. Histology indicated that fish without AA supplementation had deformed gill filament support cartilage and atrophied muscle fibers. Collagen content of the vertebral column was significantly lower in fish devoid of dietary AA (P < 0.05). Liver AA concentration varied in proportion to dietary concentration of AA. The minimum dietary AA concentration tested in this study, 25 mg AA/kg diet, was sufficient to prevent growth reduction and AA deficiency signs in oscars.
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