Chitin is thought to be abundant in marine environments, but examination of the role of chitin in nutrient cycling has been hampered by the lack of an adequate assay for measuring concentrations at ambient levels. We developed a simple assay using the lectin, wheat germ agglutinin (WGA), which has an affinity for the N-acetylglucosamine (NAG) residues found in chitin. The specificity of the assay was confirmed by enzymatic hydrolysis with chitinase and competitive inhibition with chltotriose. The lectin bound specifically to chitin even when samples contained high concentrations of cellulose, clay, and bacteria. Concentrations of suspended chitin were 4 to 21 pg 1-' in Delaware Bay, USA, and 4 to 10 pg 1-' in the subarctic Pacific. The assay was also applied to sediment trap samples collected in the subarctic Pacific. We found that the chitin flux accounted for less than 1 % of carbon and nitrogen fluxes above 500 m. Using fluorescently-labelled WGA and epifluorescence microscopy, we were able to differentiate detritus, zooplankton fecal pellets and possibly fungi from nonchltinous particles.
The physiological basis of the carotenoid-I4C-labeling method for the determination of growth rates ( p , d-') of specific groups of microalgae was established in the laboratory and the method was tested in the subarctic Paclfic and in Chesapeake Bay (USA). '%labelin9 patterns of carotenoids in a variety of algal species grown in batch cultures were descnbed successfully with a s~mple precursor-pigment model whose free parameters, the specific rate of carotenoid synthesis (p,,,,, d-') and the precursor and the pigment turnover rates (d-l), were determined by least squares analysis. A U xanthophylls except peridinin had turnover rates that did not differ significantly from zero; the turnover rate of peridinin was 0.7 p. Precursor turnover rates varied from about 5 p for fucoxanthin to 36 p for lutein. We propose to use the precursor-pigment model to calculate p, , , from the amount of I4C incorporated into carotenoids and values of carotenoid precursor turnover rates, which are assumed to be known a priori. A well-constrained estimate of the fucoxanthin precursor turnover rate is presented here. It was shown for laboratory cultures that the carotenoid-labeling method is capable of measuring specific rates of carotenoid synthesis and that these rates equal rates of cell growth only when growth is balanced. We demonstrated that pigment synthesis and carbon flxation can be unbalanced in natural phytoplankton populations due to the effects of light perturbations and growth under a natural photocycle. We recommend that labeling experiments last 24 h to average rates of synthesis over the die1 photoperiod since rates of carotenoid synthesis and cell growth can be unbalanced at any time during the photoperiod. The field experiments also demonstrated that the carotenoid-labeling method is a powerful tool to study the physiological ecology of natural populations of phytoplankton.
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