Microbial RNA and DNA synthesis in marine sediments

A technique for measuring rates of RNA and DNA synthesis in sedimentary microbial communities has been adapted from methods developed for marine and freshwater microplankton research. The procedure measures the uptake, incorporation and turnover of exogenous [2, ³H]-adenine by benthic microbial populations. With minor modification, it is applicable to a wide range of sediment types. Measurement of nucleic acid synthesis rates are reported from selected benthic marine environments, including coral reef sediments (Kaneohe Bay, Oahu, Hawaii), intertidal beach sands (Oahu and southern California) and California borderland basin sediment (San Pedro Basin), and comparisons are made to selected water-column microbial communities. Biomass-specific rates of nucleic acid synthesis in sediment microbial communities were comparable to those observed in water-column assemblages (i.e., 0.02 to 2.0 pmol deoxyadenine incorporated into DNA [ng ATP]^-1 h^-1 and 0.2 to 8.9 pmol adenine incorporated into RNA [ng ATP]^-1 h^-1). DNA synthesis rates were used to calculate carbon production estimates ranging from 2 g C cm^-3 h^-1 in San Pedro Basin sediment (880 m water depth) to 807 g C cm^-3 h^-1 in coral reef sediment from the Kaneohe Bay. Microbial community specific growth rate, (d^-1), estimated from DNA synthesis rates in surface sediments ranged from 0.1 in San Pedro Basin to 4.2 in Scripps Beach (La Jolla, California) intertidal sand.     

Partitioning radiolabeled thymidine uptake by bacteria and meiofauna using metabolic blocks and poisons in benthic feeding studies

Techniques exist which allow for the measurement of in situ grazing rates of meiobenthos on sedimentary bacteria and microalgae. Radiolabeled substrates are incorporated into microbes which serve as food for meiofauna and which themselves also become labeled during feeding. However, during in situ grazing experiments, meiofauna may become radiolabeled by a variety of non-feeding processes. Proper controls to correct for these extraneous routes of labeling have been developed in the present study. The use of [methyl-³H] thymidine (³HTdR) in studies of meiofaunal grazing on bacteria has two unique advantages: (1) it is incorporated only into prokaryotic macromolecules, and (2) bacterial incorporation of ³HTdR may be selectively blocked by several inhibitors which are non-toxic to meiofaunal grazers. Coupled with formalin-killed control treatments, the use of these inhibitors makes it possible to accurately determine the partitioning of radiolabel into meiofauna during grazing into adsorptive, absorptive and grazing components. A saturated solution of nalidixic acid and 5-deoxythymidine was found to be most effective in inhibiting water-column bacterial uptake and incorporation of ³HTdR, and had no toxic effects on meiofauna. The inhibitor was found to immediately block bacterial incorporation of ³HTdR and was as effective at 20% saturation as at 100%. The presence of sediment reduced the effectiveness of this inhibitor by 50%. Solutions of the inhibitor with excess undissolved material, however, completely blocked sediment bacterial uptake of ³HTdR. Employing these techniques during in situ grazing experiments showed that up to 83% of total meiofaunal uptake of ³H-label may be attributable to non-grazing processes. Experiments conducted in slurried sediments yielded grazing rates which were the same as those from intact cores. Furthermore, meiofaunal grazing rates on multiple food sources (e.g. bacteria and diatoms) may be determined synoptically by adding isotopically-distinct substrates (e.g. ³HTdR and H¹⁴CO₃) to the same experimental incubation.University of Texas Marine Science Contribution No. 698.     

Reproduction of the estuarine mysid Taphromysis bowmani (Crustacea: Malacostraca) in fresh water

We have monitored the incorporation of ³H-glycine into, and the excretion of, soluble tissue and extrapallial fluid proteins in the hardshell clam Mercenaria mercenaria in an attempt to follow some of the metabolic events that occur antecedent to shell deposition. After incubating at 20°C for 48 h, clams were killed and the distribution of incorporated and unincorporated tritium in seawater, mantle fluid, hemocoelic-tissue fluid, extrapallial fluid and tissue was determined. Most of the incorporated tritium was in the insoluble tissue proteins. Much more incorporated tritium was found in the hemocoelic-tissue fluid fraction than in the extrapallial fluid. We assume that most of the radioactivity we followed was due to free or incorporated radioactive glycine. The ratio of ³H-protein to ³H-glycine was greater in the extrapallial fluid than in the hemocoelic-tissue fluid, suggesting either protein secretion into or glycine removal from the extrapallium. We also observed that both ³H-protein and ³H-glycine concentrations were higher in the mantle fluid than in the external sea water, although the ratios of ³H-glycine to ³H-protein in these two fluids were not different.     

The use of RNA:DNA ratios to predict growth limitation of juvenile summer flounder (Paralichthys dentatus) from Delaware and North Carolina estuaries

Nutritional indices were used to develop biochemical correlates of feeding and growth rates for juvenile summer flounder, Paralichthys dentatus (Linnaeus), from North Carolina (NC) and Delaware (DE). Six parameters (Fulton's condition K=10⁴xweight/(length³), wet weight/dry weight, [protein], [RNA], [DNA], and RNA:DNA) were related to feeding and growth rates of fish from previously reported 10 to 14-d experiments at temperatures ranging from 2 to 20 °C with varying feeding levels (0 to 100% and libitum). RNA:DNA ratios were the best predictors of growth rates, but inclusion of a temperature term improved the relationship between RNA:DNA ratios and growth rate for Delaware fish. Feeding rates were poorly correlated with all parameters. RNA:DNA ratios of fish in the laboratory changed significantly within 1 d of starvation and refeeding at 16 °C. RNA:DNA of juvenile summer flounder collected from one site in Indian River Bay, DE and two sites in the Newport River Estuary, NC, between January and June 1992 were used to estimate in situ growth rates following settlement. Predicted growth rates in both estuaries were close to maximum (suggesting ad libitum feeding) until early May. Growth rates of juveniles from Delaware were <0% d^-1 from December through early March, and were higher (0.6 to 3% d^-1) from April through early June. However, growth rates of DE juveniles during May were <50% of maxinum. North Carolina juveniles had growth rates of 2 to 5% d^-1 from February through early April. Juveniles from one of the Newport River sites (a marsh habitat) were also severely growth limited (<20% of maximum) after April. Prolonged periods of sub-optimal growth may be important to survival and recruitment of juvenile summer flounder in northern mid-Atlantic estuaries. A model is presented which illustrates the potential impact that small changes in temperature and growth limitation can have on recruitment success in both delaware and North Carolina estuaries.     

Diel variation of bacterial productivity in seagrass (Zostera capricorni) beds measured by rate of thymidine incorporation into DNA

Diurnal variations occurred in bacterial growth rates in the sediment and water column associated with seagrass (mainly Zostera capricorni Aschers) beds in Moreton Bay, Queensland, Australia. Studies were carried out in March and June 1981. Cell production rates increased by 5- to 10-fold during the morning and decreased during the afternoon. No nocturnal variation was observed. Daily bacterial cell production rate in the aerobic zones of the seagrass bed was estimated to be 43 mg C m^-2. A minimum of 100 mg C m^-2 d^-1 would be required to support the bacterial production. This represents about 10% of net primary production. The incorporation of tritiated thymidine into DNA was used to measure bacterial growth. The validity of the method is discussed.     

Microbial populations in surface films and subsurface waters: Amino acid metabolism and growth

Sea surface microlayer (film) and subsurface microbial populations (biomass and activities) were studied in the Damariscotta (Maine) estuary in May and September, 1987. Dissolved free and combined amino acids (DFAA, DCAA), bacterial numbers, microbial ATP, bacterial and microbial DNA synthesis (via³H-thymidine and³H-adenine), and amino acid (³H-glutamic acid) metabolism were measured. DFAA and DCAA were typically enriched in the surface microlayers relative to surface waters, although utilization of glutamic acid was usually more rapid in subsurface waters, as was incorporation of thymidine. Bacteria represented 12 to 40% of the microbial biomass as determined by ATP, except during microalgal blooms in the microlayer. Bacteria were generally not enriched in the surface films, although ATP usually was enriched. Rain input appeared to deplete population densities but stimulated population activities. Two stations which contained similar microbial populations (as estimated by bacterial counts, chlorophylla and ATP) showed very different microbial activities, apparently due to the effects of a substantial rainstorm on one of the stations. The bacterially-dominated processes utilizing thymidine and glutamic acid were enhanced approx five-fold after the rain. Autotrophic carbon production increased approx two-fold, while total microbal community DNA synthesis (as estimated by adenine incorporation into DNA) increased nearly tenfold. The observations of this study indicate that surface microlayers in the temperate waters off the coast of Maine contain highly active heterotrophic and autotrophic populations. The microbial community responds rapidly to changes in nutrient and dissolved organic matter concentrations resulting from both seasonal and temporal effects, including rain and runoff.     

Vertical profiles of bacterial abundance,productivity and growth rates in coastal sediments of the central Great Barrier Reef lagoon

Vertical patterns of bacterial densities, productivity and specific growth rates in coastal muds, quartz sands and muddy sands of the central Great Barrier Reef lagoon were examined in summer (February) and autumn (May) 1988. Variations in these parameters with station location, sediment depth and season were complex, exhibiting significant main and interaction effects in most instances. Some trends were apparent despite the large and complex variations. Bacterial densities did not vary seasonally, ranging from 2.9 to 38.1×10⁹ cells g^-1 dry wt, averaged over sediment depth (0 to 20 cm) and seasons. Trend analysis revealed that densities decreased with increasing sediment depth. Bacterial production (tritiated thymidine incorporation into DNA) was high, ranging from 0.4 to 5.7 gCm^-2 d^-1 (integrated over 10 cm depth), as were specific growth rates (grand mean, =0.25 d^-1; range=0.004 to 1.3 d^-1). Both were generally higher in summer than in autumn. Vertical profiles of productivity and specific growth rates revealed actively growing bacterial assemblages down to 20 cm depth. Factors which may account for these very abundant and productive communities are: (1) subsurface accumulations of detritus exported from adjacent mangrove forests, and (2) physical disturbance from tidal scouring and severe climate (e.g. cyclones, wet-season floods). Disturbance events occur frequently enough to inhibit the development of highly sulphidic conditions, but stimulate production of bacterial types (aerobes, fermenters) capable of incorporating labelled thymidine into their DNA.     

Species-specific grazing rates of heterotrophic dinoflagellates in oceanic waters,measured with a dual-label radioisotope technique

A dual-isotope method was developed to measure grazing rates and food preferences of individual species of heterotrophic dinoflagellates from natural populations, collected from the Slope, Gulf Stream, and Sargasso Sea and from a transect from Iceland to New England, in 1983. The isotope method measures the grazing rates of microzooplankton which cannot be separated in natural populations on the basis of size. Tritiated-thymidine and ¹⁴C-bicarbonate were used to label natural heterotrophic and autotrophic food, respectively. Nine oceanic dinoflagellate species in the genera Protoperidinium, Podolampas, and Diplopsalis fed on both heterotrophic and autotrophic food particles with clearance rates of 0.4 to 8.0 l cell^-1 h^-1, based on ³H incorporation, and 0.0 to 28.3 l cell^-1 h^-1, based on ¹⁴C incorporation. Two dinoflagellate species, Protoperidinium ovatum and Podolampas palmipes, fed only on ³H-labelled food particles. Several species of dinoflagellates fed on bacteria (<1 m) which had been prelabelled with ³H-thymidine. The clearance rates of heterotrophic dinoflagellates and ciliates were similar and within the range of tintinnid ciliate clearance rates reported in the literature. As heterotrophic dinoflagellates and ciliates can have comparable abundances in oceanic waters, we conclude that heterotrophic dinoflagellates may have an equally important impact as microheterotrophic grazers of phytoplankton and bacteria in oceanic waters.Partially supported by a grant from the National Science Foundation, OCE-81-17744     

Testicular DNA synthesis inhibition—an in vivo system for the detection of mutagenic and carcinogenic chemicals

By binding covalently to DNA chemical mutagens and carcinogens inhibit replication, which can be measured as a decrease in thymidine incorporation into DNA. This DNA synthesis inhibition (DSI) has been determined in testicular cells of mice for a large number of compounds and has been found to correlate very well with their known mutagenic and carcinogenic properties.

Not only could this test give a qualitative answer about potential carcinogenicity or mutagenicity, but, with regard to its in vivo characteristics, could furthermore give an indication of the carcinogenic and mutagenic potency and thus be of use in the risk evaluation of chemical substances; the relationship between the DNA synthesis inhibition and the underlying alkylation of guanine ‐0⁶ by different methylating agents is demonstrated.

As toxic effects decrease thymidine incorporation, too, means are discussed for distinguishing between “true”;, mutagenic, and “false”;, cytotoxic, DNA synthesis inhibition.

In conclusion, the implications of including the DSI‐test in a battery of mutagenicity tests are outlined.     

Protein synthesis and growth inOctopus vulgaris

Rates of protein synthesis in the whole body and tissues ofOctopus vulgaris collected in September 1985 and 1986 from the Bay of Naples were measured following a flooding dose injection of³H phenylalanine. There were stable phenylalanine free pool-specific radioactivities and linear incorporation of radiolabel into arm-tip protein from 10 to 30 min after the injection. In starved individuals there were no significant differences between the fractional rates of protein synthesis of the following tissues: ventricle, brain, branchial heart, arm tip, gill, stomach, arm, renal appendage and mantle. The mean value (± SE) for all the tissues was 3.02 ± 0.17% d^–1. In individuals fed varying amounts of crab, resulting in differing growth rates, there was a linear increase in fractional rates of whole-body protein synthesis with growth rate. A standard 148 g octopus growing at 3.0% d^–1 synthesised 0.54 g of protein, with 0.43 g of this protein retained as growth. The proportion of the total protein synthesis which was retained as growth increased with increasing growth rate; at a maximum growth rate of 6% d^–1, over 90% of the protein synthesised was retained as growth. The ventricle, arm tip, gill, arm and mantle also showed similar patterns of a linear increase in fractional rates of protein synthesis with increased growth rates. The RNA concentrations in the whole body and tissues increased with increasing growth rates, but the major change was an increase in the efficiency of translation. It is concluded that rapid growth rates inO. vulgaris are brought about by high rates of protein synthesis and high efficiencies of retention of synthesised protein and, therefore, little protein degradation.     

Light quality in relation to growth,photosynthetic rates and carbon metabolism in two species of marine plankton algae

The effect of light quality on growth, photosynthesis and carbon metabolism in two species of marine algae,Cyclotella nana (Hustedt) andDunaliella tertiolecta (Butcher), was examined. Relative growth constants forC. nana were 0.37, 0.29 and 0.25 in blue, white and green light, respectively. Corresponding constants were 0.41, 0.31 and 0.29 forD. tertiolecta. Photosynthetic rates in both species were higher in blue light and lower in green light compared with white light of the same intensity. More than 60% of¹⁴C assimilated byC. nana orD. tertiolecta grown in blue or green light was incorporated into the ethanol-insoluble fraction, compared with 10 to 30% in this fraction in white light. The relative importance of the various components within this fraction was independent of light quality. Although less¹⁴C was assimilated into the ethanol-soluble fraction in blue or green light, there was a relative increase in some amino acids and organic acids in this fraction and a decrease in sugars and sugar phosphates relative to white light of the same intensity. These differences were independent of light intensity, photosynthetic rate and cell density in the cultures.     

Temporal and spatial variation in bacterial production in the water column over a coral reef

Production and doubling times of the bacterial populations in the water around and over the reefs at Lizard Island, Great Barrier Reef were measured during summer and winter, 1982 and 1983. Bacterial productivity, determined from the rate of tritiated thymidine incorporation into DNA, was high over the reef flats and a Thalassia hemprichii sand flat (28 to 58 g Cl^-1 d^-1). Bacterial growth rates increased during the day and fell at night over the reef flats and seagrass bed. Growth rates were slower over the reef front and in open water. Doubling times ranged from about 2 d in the open water to about 3 h over the reef flat in summer. As numbers did not increase, grazing was probably intense on the reef flats. Growth rates were much slower in winter. The main source of organic nutrient used by the bacteria was probably mucus released following photosynthesis in the corals. The cyanobacterium Synechococcus sp. was sometimes very numerous, especially in summer when 2×10⁸ cells l^-1 were recorded in one water mass. The number of bacteria was also very high in summer, with values ranging from 1×10⁹ to 2.5×10⁹l^-1.     

Relationship between growth and biochemical indices in laboratory-reared juvenile Japanese flounder (Paralichthys olivaceus), and its application to wild fish

In order to estimate growth rates based on biochemical indices of the liver of wild Japanese flounder (Paralichthys olivaceus), juveniles were reared at six ration levels (0, 0.5, 2, 4, 6 and 8% body weight day⁻¹) in the laboratory for 14 days, and the relationship between their growth rates and biochemical indices (RNA/DNA, protein/DNA, triglyceride/DNA, phospholipid/DNA and cathepsin D activities) were determined. Positive and approximately linear relationships were seen between growth rates and the indices of RNA/DNA, protein/DNA and phospholipid/DNA. The triglyceride/DNA ratio decreased with increasing growth rates up to approximately 1% body weight day⁻¹, then increased linearly with increasing growth rates. There was no significant correlation between growth rates and cathepsin D activity, and the highest values were obtained in the starved fish. Compared with laboratory-reared specimens, wild specimens of similar sizes were found to have significantly larger livers. The RNA/DNA, protein/DNA and phospholipid/DNA ratios of wild specimens fell in a broad range between ration groups of reared juveniles. The protein/DNA ratios of wild specimens were low and outside the range of the reared juveniles at six ration levels. In contrast, the levels of cathepsin D activity of wild fish were highest compared to the reared fish. Estimated growth rates of wild fish from the RNA/DNA, protein/DNA and phospholipid/DNA regressions obtained from the rearing experiment were 1.66, −1.74 and 0.10% day⁻¹, respectively. Based on our results, the RNA/DNA index may be regarded as the most valid and reliable growth estimator. It is noted that the larger liver size, the lower liver protein/DNA ratio and the unexpectedly high level of cathepsin D activities of wild specimens found in this study may reflect the different metabolic conditions of fish reared in the laboratory compared to those collected in the field. Received: 29 February 2000 / Accepted: 26 August 2000     

Measuring RNA:DNA ratios in individual <Emphasis Type="Italic">Acartia tonsa</Emphasis> (Copepoda)

Acartia tonsa Dana is a dominant copepod in coastal waters and is therefore an important link in the food web between microplankton and higher trophic levels. RNA:DNA ratios have been used to describe growth and nutritional condition of field-collected copepods and to show strong correlation between RNA:DNA ratios and group egg production (EP). A method was developed using a sensitive, nucleic acid-specific fluorescent dye, and automated microplate fluorometer to measure DNA, RNA, and the RNA:DNA ratio of individual A. tonsa. DNA, RNA, and RNA:DNA ratios and EP were all significantly higher in copepods fed Thalassiosira sp. compared to starved copepods. There was a general trend toward an increase in RNA:DNA ratios with increase in EP, but due to the high degree of variation in both RNA:DNA ratios and EP among individual copepods there was no significant correlation between RNA:DNA ratios and EP. Significant differences in RNA:DNA ratios between fed (7.2) and starved (3.3) copepods were found after 2 days. This assay may be applied to other species of copepods sampled in the field to provide an index of the health of planktonic food webs.     

Biosynthesis of wax esters in oceanic crustaceans

Slices from the hepatopancreas of various oceanic curstaceans incorporated radioactivity into wax esters from ¹⁴C glucose and ¹⁴C aspartic acid to a lesser extent and from ¹⁴C palmitic acid to a much greater extent. Radioactivity was incorporated from ¹⁴C palmitic acid into both fatty acid and fatty alcohol moieties of wax esters, the percentage of total radioactivity present in alcohol moieties being greater in deep-living than in shallow-living species. Cell-free preparations from the hepatopancreas but not from muscle, supplemented with ATP and reduced pyridine nucleotides, incorporated radioactivity from 1-¹⁴C palmitoyl Coenzyme A into both fatty alcohol and fatty acid moieties of wax esters. Incorporation into fatty alcohol was NADPH- rather than NADH-specific. Preparations from deep-living species had a greater percentage of total radioactivity in the fatty alcohol moieties of wax esters than preparations from shallow-living species. We conclude that the level of wax esters in a given species is correlated with the rate at which the species biosynthesises these lipids de novo; deep-living species have higher rates of wax ester biosynthesis and higher levels of wax esters than shallow-living species. The results support the thesis that wax esters in oceanic crustaceans are derived largely from the animals' internal biosynthetic activities, presumably in response to particular biochemical and/or physiological requirements, rather than from their diets.     

RNA:DNA ratios and growth of herring (Clupea harengus) larvae reared in mesocosms

Autumn-spawned North Sea herring larvae (Clupea harengus L.) were released in two outdoor mesocosms of 2500 m³ (A) and 4000 m³ (B). The mesocosms were monitored for temperature, salinity, oxygen, chlorophyll a, zooplankton and herring larvae abundance. The density of suitable prey for first feeding larvae (mainly copepod nauplii) was initially low in Mesocosm A (<0.11^-1) compared to in Mesocosm B (>11^-1). Half-way through the experiment the situation was reversed, with higher densities of prey in Mesocosm A (>31^-1) as compared to Mesocosm B (～11^-1). The average temperature declined steadily in both mesocosms from 18°C at release to 11–12°C by the end of the experiment 60 d later. The RNA:DNA values of individual herring larvae were related to protein growth rates and temperature adjusted according to Buckley (1984). A corresponding DNA growth index (Gdi) was given as: Gdi=0.68 TEMP+3.05 RNA:DNA-9.92. The RNA:DNA based growth indices were significantly correlated with other somatic growth estimates. The average estimated protein growth rate in the two mesocosms followed the same temporal pattern as the somatic growth rate, but with a lag of 2 d or more. Residual analysis of the regression of ln RNA versus ln DNA also showed the same temporal pattern as the RNA:DNA ratios, but the shift in condition as estimated by this method occurred more in synchrony with the other somatic growth measures. Larvae in Mesocosm A had RNA:DNA values similar to the starvation control kept in the laboratory the first days after release, confirming that larvae in Mesocosm A initially were in poor nutritional condition. On the other hand, the majority of the herring from Mesocosm B were characterised as starving or in poor nutritional condition towards the end of the experiment. The assessment of growth and nutritional condition were in accordance with independent survival estimates which suggested that the majority of the total mortality occurred during the first 15 d in Mesocosm A and there-after in Mesocosm B.     

RNA:DNA ratios and growth of herring (Clupea harengus) larvae reared in mesocosms

Autumn-spawned North Sea herring larvae (Clupea harengus L.) were released in two outdoor mesocosms of 2500 m³ (A) and 4000 m³ (B). The mesocosms were monitored for temperature, salinity, oxygen, chlorophyll a, zooplankton and herring larvae abundance. The density of suitable prey for first feeding larvae (mainly copepod nauplii) was initially low in Mesocosm A (<0.11^-1) compared to in Mesocosm B (>11^-1). Half-way through the experiment the situation was reversed, with higher densities of prey in Mesocosm A (>31^-1) as compared to Mesocosm B (11^-1). The average temperature declined steadily in both mesocosms from 18°C at release to 11–12°C by the end of the experiment 60 d later. The RNA:DNA values of individual herring larvae were related to protein growth rates and temperature adjusted according to Buckley (1984). A corresponding DNA growth index (Gdi) was given as: Gdi=0.68 TEMP+3.05 RNA:DNA-9.92. The RNA:DNA based growth indices were significantly correlated with other somatic growth estimates. The average estimated protein growth rate in the two mesocosms followed the same temporal pattern as the somatic growth rate, but with a lag of 2 d or more. Residual analysis of the regression of ln RNA versus ln DNA also showed the same temporal pattern as the RNA:DNA ratios, but the shift in condition as estimated by this method occurred more in synchrony with the other somatic growth measures. Larvae in Mesocosm A had RNA:DNA values similar to the starvation control kept in the laboratory the first days after release, confirming that larvae in Mesocosm A initially were in poor nutritional condition. On the other hand, the majority of the herring from Mesocosm B were characterised as starving or in poor nutritional condition towards the end of the experiment. The assessment of growth and nutritional condition were in accordance with independent survival estimates which suggested that the majority of the total mortality occurred during the first 15 d in Mesocosm A and there-after in Mesocosm B.     

237Pu experiments with the plaice Pleuronectes platessa

Some experiments have been made on the metabolism of ²³⁷Pu — a high specific activity, gamma-emitting isotope of plutonium — by the plaice Pleuronectes platessa L. Very little of the isotope was incorporated into fish after 2 months exposure to labelled water. The oral retention from a variety of labelled foods was also very poor, and apart from the gut itself no incorporation of the isotope could, in fact, be demonstrated. Injected ²³⁷Pu was eliminated very slowly, and some evidence was obtained for an inverse relationship between the rate of ²³⁷Pu loss and the rate of growth of the fish. Growing fish incorporated a relatively larger fraction of the ²³⁷Pu body burden into skeletal material, and this was attained at the expense of the isotopic content of the liver. Very little ²³⁷Pu was incorporated into muscle.     

A method to determine in situ zooplankton grazing rates on natural particle assemblages

In situ zooplankton grazing rates on natural particle assemblages were stimated by measuring zooplankton uptake of labelled autotrophic (with Na¹⁴CO₃) and heterotrophic (with [methyl-³H]-thymidine) particulate matter in 1-h incubations in clear, Plexiglas, Haney chambers. The in situ grazing rates are in the same range as those measured for zooplankton in the laboratory using standard particle counting techniques. A negative selection coefficient for ³H-labelled particles indicated a lower filtration efficiency or avoidance of these particles by zooplankton.     

Products of photosynthesis in natural populations of marine phytoplankton from the Gulf of Maine

We have measured the photosynthetic assimilation of ¹⁴C-carbon dioxide into (1) ethanol-soluble, (2) hot-trichloroacetic acid (TCA)-soluble (polysaccharide), and (3) protein fractions of natural populations of marine phytoplankton. Diurnal studies showed a continuing incorporation of carbon-14 into the protein fraction during hours of darkness. This was accompanied by a concomitant decrease in the proportion assimilated into polysaccharide. When incorporation was measured under constant experimental conditions, the pattern of photosynthesis did not vary from one time of day to another. At one station approximately 12 km south of Boothbay Harbor, the proportion of carbon entering protein showed marked seasonal changes. During the winter, approximately 10 to 20% of the fixed carbon was incorporated into protein. During the summer the value increased to 22 to 35%. Between these times, a transient high value of 37 to 47% of the fixed carbon entering protein coincided with the spring bloom. The increases in proportion incorporated into protein were largely paralleled by equivalent decreases in the polysaccharide fraction. The proportion of carbon incorporated into protein during photosynthesis also increased markedly at reduced light intensities. This increase occurred both when populations were incubated in neutral-density filters and when incubated at increasing depths in the photic zone. There was little consistent and significant difference between the neutral-density filters and depth in the water column, suggesting a minimal role for light quality. The extent of the increased relative rate of protein synthesis at the lower light intensities depended on the nutritional state of the phytoplankton. For example, summer populations from water containing low concentrations of inorganic nutrients responded less dramatically to reduced light intensities than did populations from nutrient-rich waters.