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Abstract. RNA/DNA ratios in individual herring (Clupea harengus) larvae (collected from Kiel Bay, Baltic Sea, in 1989) were measured and proved suitable for determining nutritional status. Significant differences between fed and starving larvae appeared after 3 to 4 d of food deprivation in larvae older than 10 d after hatching. The RNA/ DNA ratio showed an increase with age or length of the larvae and was less pronounced in starving larvae compared to fed larvae. The individual variability of RNA/ DNA ratios in relation to larval length of fed larvae and of larvae deprived of food for intervals of 6 to 9 d is presented. Based on the length dependency and the individual variability found within the RNA/DNA ratios, a laboratory calibration is given to determine whether a larva caught in the field has been starving or not. An example for a field application is shown.

We investigated whether nutrient limitations of primary producers act upward through food webs only in terms of density effects or if there is a second pathway for nutrient limitation signals channelled upward to higher trophic levels. We used tritrophic food chains to assess the effects of nutrient-limited phytoplankters (the cryptophyte Rhodomonas salina) on herbivorous zooplankters (the calanoid copepod Acartia tonsa) and finally zooplanktivores (larval herring Clupea harengus) living on the herbivores. The primary producers' food quality had a significant effect on fish condition. Our experimental phosphorus-limited food chain resulted in larval fish with a significantly poorer condition than their counterparts reared under nitrogen-limited or nutrient-sufficient conditions. Our results show that mineral nutrient requirements of consumers have to be satisfied first before fatty acids can promote further growth. This challenges the match/mismatch hypothesis, which links larval fish survival probability solely to prey availability, and could imply that reduced nutrient releases into the environment may affect fish stocks even more severely than previously believed.

The RNA/DNA ratio is a useful indicator of the nutritional condition of fish larvae. The presented analytical procedure is an improvement of Clemmesen's (Meeresforschung 32: 134-143, 1988) methodology which involves purification of fish larvae tissue homogenates and subsequent fluorescence-photometric measurements using specific nucleic acid dyes. The modifications concern the homogenization and nucleic acid extraction procedures. A 'shaking mill' was compared to a potter Elvehjem microhomogenizer and a reduction in the washing and purification steps was achieved. Treatment of samples with ribonuclease A and subsequent fluorescence measurement using ethidium bromide was given preference compared to the DNA-bisbenzimidazole determinations due to problems arising from high self-fluorescence of the samples and the influence of 'quenching' substances disturbing the DNA-bisbenzmidazole determinations. Different RNase concentrations and their influences on RNA and DNA were checked. Recovery rates of standard RNA and DNA 'spikes' were determined. Fish larvae samples were analysed with the previous and the improved modified procedure and a correction factor to compare results measured with the 2 procedures was calculated. With the presented method the physiological condition of individual larvae and the amount of variabhty can be determined.

The ratio of tissue RNA to DNA (R/D) is a widely used index of recent growth and nutritional condition in larval and juvenile fish. To date, however, no standard technique for measuring nucleic acids has been adopted. Because methodological details can affect the estimate of R/D, researchers using different analytical protocols have been unable to compare ratios directly. Here, we report on the results of an international interlaboratory calibration of 4 spectrofluorometric protocols to quantify nucleic acids. Replicate sets of 5 tissue samples and 2 standards (common standards) were supplied to each of 5 researchers for analysis with their own methods and standards. Two approaches were evaluated for mitigating the observed differences in values: 1) the use of common nucleic acid standards and 2) standardizing to a common slope ratio (slope of DNA standard curve/slope of RNA standard curve or m DNA /m RNA ). Adopting common standards slightly reduced the variability among protocols but did not overcome the problem. When tissue R/Ds were standardized based on a common m DNA /m RNA slope ratio, the variance attributed to analytical protocol decreased dramatically from 57.1% to 3.4%. We recommend that the ratio of the slopes of the standard curves be provided to facilitate intercomparability of R/D results among laboratories using different spectrofluorometric methods for the analysis of nucleic acids in fish.