. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 272 MORAN AND MANAHAN slope of the regression line of oxygen consumed per hour against number of larvae in each chamber. The error of each estimate was calculated as the standard error around the slope of the regression line. Biochemical anal\sis The conversion and interpretation of mass-to-energy equivalents in marine invertebrates has long been studied, and remains difficult ( Paine. 1971: Gnaiger and Bitter- lich, 1984; Jaeekle and Manahan, 1989a; Moreno et 2001). Previously in our laboratory, ash-free
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 272 MORAN AND MANAHAN slope of the regression line of oxygen consumed per hour against number of larvae in each chamber. The error of each estimate was calculated as the standard error around the slope of the regression line. Biochemical anal\sis The conversion and interpretation of mass-to-energy equivalents in marine invertebrates has long been studied, and remains difficult ( Paine. 1971: Gnaiger and Bitter- lich, 1984; Jaeekle and Manahan, 1989a; Moreno et 2001). Previously in our laboratory, ash-free dry weight was used successfully as an index of total organic content in larvae of the red abalone, H. rufesceiis (Jaeckle and Mana- han. 1989a: Shilling et ill., 1996); techniques and instru- mentation were identical to those used in the present study. For both species of abalone studied here, we attempted to measure ash-free dry weight but found that the samples never reached a stable dry weight, even after 3 months of drying at 60 °C. As a result, we used direct measurements of carbohydrate, lipid. and protein to determine the general patterns of energy utilization throughout development. Carbohydrates. Known numbers of animals (500-1000, depending on developmental stage) were aliquoted into microcentrifuge tubes and centrifuged; seawater was aspirated, and the samples were frozen at —80 °C for later analysis. Total carbohydrates were quantified using the methods of Holland and Gabbott (1971). In brief, samples of eggs (day 0) and metamorphically competent larvae (day 8) were extracted with cold 5<7r trichloroacetic acid. The supernatant was hydrolyzed for 2 h at 95 °C in \M HC1, then carbohydrates were spectrophotometrically quantified with a ferricyanate reduction reaction using glucose as a standard. Protein. Samples of known numbers of eggs or larvae were processed as above, and protein was analyzed with the methods of Bradford (1976) as modified by Jaeck
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