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.     

Biosynthesis of lipds in zooplankton from Saanich Inlet,British Columbia,Canada

Lipid analyses were carried out on various species of zooplankton captured in Saanich Inlet, British Columbia, Canada, during September, 1972. The amphipod Cyphocaris challengeri had the highest level of lipid, consisting mainly of wax esters. The copepod Calanus pacificus had moderate amounts of lipid, with triglyceride as the major neutral lipid. The euphausiid Thysanoessa raschii contained mainly triglyceride in its moderate levels of neutral lipid, while wax esters and, to a lesser extent, triglycerides were present in the very small levels of neutral lipid in the chaetognath Sagitta elegans. The major fatty alcohols in wax esters of both Cyphocaris challengeri and Calanus pacificus were 20:1 and 22:1, although notable differences were found in the major fatty acids. Biosynthetic studies showed that phospholipids were labelled faster than neutral lipids in all species with both (U-¹⁴C) glucose and (1-¹⁴C) palmitic acid as precursors. Only species containing significant amounts of wax esters in their neutral lipids incorporated substantial amounts of radio-activity from (1-¹⁴C) palmitic acid into wax esters in (i) living animals, (ii) preparations containing fragments of tissue, (iii) cell-free systems. All species incorporated added fatty alcohols into wax esters in preparations containing tissue fragments and in cell-free preparations. Both the fatty acid and fatty alcohols of the wax esters of both Calanus pacificus and S. elegans were labelled from (1-¹⁴C) palmitic acid, consistent with de novo biosynthesis of the esters. (1-¹⁴C) hexadecanol was incorporated into wax esters almost entirely in the fatty alcohol moiety. It is concluded that all species examined formed was esters when presented with preformed fatty alcohols, but only those species that had wax esters as a major component of the neutral lipids were capable of de novo biosynthesis of the lipids.     

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.     

Uptake of inorganic carbon and internal carbon cycling in symbiont-bearing benthonic foraminifera

Incorporation rates of inorganic carbon and its distribution between the organic matter and the skeleton have been measured using ¹⁴C tracer techniques on two species of symbiont-bearing benthonic foraminifera in the Gulf of Elat: Amphistegina lobifera (a perforate species) and Amphisorus hemprichii (an imperforate species). Under constant experimental conditions, incorporation rates of the radiotracer become linear with time after several hours in A. hemprichii and after one day in A. lobifera. A. lobifera showed a lag time of 24 h for skeletal incorporation, whereas in A. hemprichii uptake into the skeleton started within 2 h. Pulse-chase incubations in radioactive seawater, followed by unlabelled incubations, demonstrate transfer of photosynthetically acquired ¹⁴C into the skeleton of A. lobifera. No such transfer was found in A. hemprichii. The total ¹⁴C uptake by A. lobifera increased during the first 24 h of cold chase incubation. This increase suggests the existence of an internal inorganic carbon pool that was lost (probably evaporated) during the analysis of pulse incubations. However, during the following chase incubations, the ¹⁴C in this pool was incorporated mainly into the skeleton and retained during analysis, causing the increase in the total uptake. No such increase was found in A. hemprichii. Additional ¹⁴C uptake experiments on other species of the genera Operculina, Heterostegina and Borelis suggest that the differences in pathways for incorporation of carbon between A. lobifera and A. hemprichii can be generalized to the perforate and imperforate foraminiferal groups. In perforate species, respired carbon originally taken up through photosynthesis is partly recycled into the skeleton. In imperforate species such a transfer has not been demonstrated. Perforate species seem to have a large internal inorganic carbon pool which serves mainly for calcification and possibly also for photosynthesis, while imperforate species may take up carbon for calcification directly from seawater or have a very small inorganic carbon pool.     

Removal of glycine from solution by the sea urchin Strongylocentrotus purpuratus

Radioautography and liquid-scintillation counting of juvenile sea urchins — Strongylocentrotus purpuratus (Stimpson) — exposed to ³H- and ¹⁴C-labeled glycine showed that the ³H is rapidly translocated throughout the animal and lost, while the ¹⁴C is slowly translocated into the interior of the urchin and incorporated into insoluble material.     

Compartmentation and turnover of organic carbon in the Staghorn coral Acropora formosa

Four colonies of Acropora formosa were incubate with Na₂ ¹⁴CO₃ for separate 2 h periods within a 24 h period, and then returned to the reef from which they were collected. Terminal branches were collected at intervals over the following 5 d and analysed for radioactivity associated with the skeleton and certain organic pools. Colonies incubated at night showed little or no loss of fixed radioactivity during the 5 d on the reef. However, 50–60% of photosynthetically-fixed ¹⁴C was lost from the terminal branches during the first 40 h on the reef. This loss of radioactivity probably resulted from release of mucus and dissolved organic carbon from the coral tissues. Most of the loss of photosynthetically-fixed ¹⁴C was due to decrease in the radioactivity of lipids (80% of the total ¹⁴C loss) and methanol-water soluble compounds. Determination of any sequencing in metabolic compartments was made difficult by the rapidity with which ¹⁴C dissappeared from most of the metabolic pools measured. ¹⁴C was incorporated into the skeleton throughout the 5 d on the reef, although the rate of incorporation was very low in colonies which had been incubated with Na₂ ¹⁴CO₃ at night.     

The pattern of carbon fixation in the marine unicellular alga Phaeodactylum tricornutum

The short- and long-term fixation of ¹⁴CO₂ by Phaeodactylum tricornutum was studied using methods of fractionation that allowed examination of all the products labelled with ¹⁴C. There was no doubt that the major pathway of CO₂ fixation was into 3-phosphoglycerate, but there was also significant incorporation by -carboxylation by means of phosphoenol-pyruvate carboxylase and transamination into aspartate. At short time intervals (10 sec), 90% of the radioactive products found were accounted for by 3-phosphoglycerate and aspartate. The lipids associated with the photosynthetic apparatus contained a high proportion of the ¹⁴C fixed, which at 60 sec was located mainly in the carbohydrate portion of the lipids. At 30 and 300 min, the chlorophylls, carotenoids and the long-chain fatty acids were heavily labelled with ¹⁴C. The monogalacto-diglycerides, the digalacto-diglycerides and the sulpholipids each had a characteristic long-chain fatty acid composition. The cell proteins and a reserve polysaccharide were also labelled with ¹⁴C at short time intervals and increased their radioactivity in a linear fashion up to the longest period studied, 300 min. The activity of the enzymes ribulose diphosphate carboxylase and phosphoenolpyruvate carboxylase was sufficient to account for the pattern of fixation found.     

Biodegradation kinetics of cymoxanil in aquatic system

A potential method to detoxify pesticides in aquatic system is using bioremediation. In this study, four microorganisms (Pseudomonas sp (EB11), Streptomyces sp. (EB12), Aspergillus niger (EB13) and Trichoderma viride (EB14) were isolated from cucumber leaves previously treated with cymoxanil using enrichment technique. These strains were evaluated for their potential to detoxify cymoxanil in aquatic system at the concentration level of 5×10^?4M. The effect of pH and temperature on the growth ability of the tested strains was also investigated by measuring the intracellular protein and mycelia dry weight for bacterial and fungal strains, respectively. Moreover, the remaining toxicity of cymoxanil after 28 days of incubation with tested strains was evaluated to confirm the complete removal of any toxic materials (cymoxanil and its metabolites). The results showed that the optimum pH for the growth of cymoxanil degrading strains (bacteria and fungi) was 7. A temperature of 30°C appears to be the optimum for the growth of either fungal or bacterial strains. Pseudomonas sp. (EB11) was the most effective strain in cymoxanil degradation followed Streptomyces sp (EB12), Trichoderma viride (EB14) and Aspergillus niger (EB13), with half-lives of 4.33, 9.5, 17.3 and 24.7 days, respectively. The degradation of cymoxanil by bacterial strains was much faster than fungal one. There is no remaining toxicity of cymoxanil detected in aqueous media previously treated with Pseudomonas sp. (EB11) for 28 days. The results suggest that bioremediation by Pseudomonas sp. (EB11) are promising for the detoxification of cymoxanil in aqueous media.     

The essential amino-acid requirements of the prawnPalaemon serratus. The growth of prawns on diets containing proteins of different amino-acid compositions

Two specimens ofPalaemon serratus Pennant (of about 4 g wet weight) were each given intrahaemocoelic injections of [U-¹⁴C] acetate. They were maintained under controlled conditions, and the release of¹⁴CO₂ and other labelled metabolites was followed for 6 days. The prawns were then sacrificed and fractionated into trichloroacetic acid soluble compounds, lipids, nucleic acids and a protein/chitin residue. Little or no¹⁴C had been incorporated into arginine, methionine, valine, threonine, isoleucine, leucine, lysine, histidine, phenylalanine and tryptophan. It was inferred thatP. serratus cannot synthesize these amino acids from ordinarily available materials, and that the prawns have an absolute dietary requirement for them. The glucosamine isolated from the protein/chitin residue contained more radioactivity than any of the other compounds examined. This implies a rapid turnover of glucosamine and chitin inP. serratus. The gain in weight of prawns fed on compound diets containing freeze-dried cod muscle was about 70% that of prawns fed on fresh mussel mantle. Predigestion of freeze-dried cod muscle with proteolytic enzymes did not significantly enhance its nutritional valve as assessed by increases in weight. Prawns fed two other compounded diets containing proteins of low nutritional value (gelatin and zein, respectively) grew at about 1/5 the rate of prawns fed fresh mussel mantle. Supplementation of the gelatin and zein in these diets with tryptophan, and with lysine plus tryptophan, respectively, failed to augment their nutritional value significantly.     

Carbon flow into the end-products of photosynthesis in short and long incubations of a natural phytoplankton population

During a cruise to the eastern Canadian Arctic (Northern Baffin Bay) in the summer of 1980, we took advantage of the 24-h photoperiod to conduct a 32-h time course experiment of ¹⁴C accumulation under natural solar radiation. The degree of non-linearity in the time course was judged against a time-dependent curve of radioactivity constructed by cumulatively adding the amount of ¹⁴C taken up in sequential short (2 h) incubations of plankton held in a replicate bottle but left unlabelled until removed for assay. Departure from linearity was due first to decreasing rates of ¹⁴C incorporation into polysaccharides and then into lipids. There was a close correspondence between ¹⁴C incorporation into proteins in the 32-h incubation and in the sequence of short incubations. These observations are consistent with patterns in utilization of photosynthetic end-products established from laboratory studies of unicellular algal cultures. Based on parallel or independent control experiments, it was judged that complicating factors such as diel light changes, nitrogenous nutrient exhaustion, bottle size effects or inhibitory conterminants in NaH¹⁴CO₃ stock solutions would not seriously affect our interpretation that non-linearity resulted from catabolic loss of radiocarbon.     

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.     

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.     

The composition and biosynthesis of lipids in Thysanoessa raschi from the Clyde Estuary,Scotland

The lipid composition and biosynthesising activity of Thysanoessa raschi collected from the Clyde Estuary, Scotland, in May 1981 were examined. Triacylglycerols were the major lipid class present, although 16.7% of the total lipid were wax esters in which phytol was the dominant fatty alcohol. The thoracic contents (hepatopancreas) of the krill were capable of biosynthesising lipids in vitro from various labelled substrates. Radioactivity from [1-¹⁴C] palmitic acid was incorporated into lipids in the order phospholipids>triacylglycerols>wax esters; the bulk of the radioactivity was present in all cases in the fatty acyl moieties of the lipids. [U-¹⁴C] glucose labelled lipids in the order phospholipids>triacylglycerols>free fatty acids> was esters; in the first two lipids the radioactivity was mainly in the glycerol moieties, whereas in was esters it was solely in the fatty acyl moieties. The extent of labelling of these lipids from [U-¹⁴C] alanine was less than that from [U-¹⁴C] glucose, but the pattern of labelling was generally similar. More than 90% of the radioactivity incorporated into total lipid from ³H₂O was present in free fatty acids from which it was calculated that the hepatopancreas of T. raschi can synthesise 2.5 g of fatty acid per hour at 15°C. This value is approximately three times lower than that previously determined for T. inermis from Balsfjorden, northern Norway. The results are discussed in terms of the sources of the dietary lipids of krill and the role of endogenous biosynthesis in contributing to its lipid reserves.     

Lipogenesis in the arctic euphausiid Thysanoessa inermis

Biosynthesis of lipids by Thysanoessa inermis collected from Balsfjorden, northern Norway, in May 1980, was examined in vitro. The highest concentration of lipid within the krill was in the hepatopancreas, and this organ was the most active in esterifying free fatty acids into wax esters. The hepatopancreas (i.e., thoracic contents) incorporated (¹⁴C) glucose, (¹⁴C) alanine and ³H₂O into wax esters, with the fatty alcohol moieties being labelled more than the fatty acids. (¹⁴C) fatty acid was incorporated preferentially into the fatty acid moieties of wax esters, this incorporation being markedly stimulated by free fatty alcohol. It is concluded that the fatty alcohols of wax esters are preferentially biosynthesized de novo from dietary protein and carbohydrates, whereas the fatty acids derive preferentially from dietary lipid. On the basis of ³H incorporated from ³H₂O, the hepatopancreas in a 50 mg II-group (2 yr old) individual of T. inermis is capable of biosynthesizing de novo, approximately 0.1 mg of lipid (as fatty acids) per day at 5°C.     

Coral illumination through an optic glass-fiber: incorporation of 14C photosynthates

The pathways of ¹⁴C incorporation into the three major compartments of the coral body were analysed in colonies of Stylophora pistillata. We used the optic glass-fiber method to carry out two sets of experiments: in the first, 11 different colonies were sampled immediately after incubation; in the second, 3 colonies were returned to the reef at the termination of incubation for a further period of 29 h. Within the tissue compartment, significantly more ¹⁴C labeled products were incorporated into illuminated tips or bases than into unilluminated sections. Tips located above illuminated bases contained amounts of ¹⁴C products similar to unilluminated tips. Within the organic matrix compartment, illumination of tip or base segments again resulted in increased amounts of ¹⁴C fixation, and again unilluminated tips located above the illuminated bases did not accumulate more ¹⁴C photosynthates than other tips on the same branches. The absence of detectable translocation was also confirmed after a post-incubation period of 29 h, and raises questions as to the validity of the widely accepted theory of upward translocation. Within the skeletal carbonate compartment, although the results were associated with a high coefficient of variation, significantly more ¹⁴C accumulated in the tips than in the bases. Illumination of tips or bases did not enhance ¹⁴C uptake. A light-independent carbon assimilation (dark fixation) is significant in S. pistillata within the three tested compartments (the tissue, the organic matrix of the skeleton, and the skeletal carbonate). It is suggested that the dark fixation process in corals in a result of accumulation of respiratory CO₂ and CO₂ from sea water as malate or other titratable acids during the night. During the day these acids are broken down, releasing free CO₂ for C₃ pathway photosynthesis.     

Movements of tiger sharks (Galeocerdo cuvier) in coastal Hawaiian waters

The giant clam Tridacna crocea harbors in the mantle tissue symbiotic microalgae commonly called zooxanthellae. Isolated zooxanthellae release glycerol into the medium in the presence of mantle tissue homogenate (MH), but it is not clear whether the cells do so in situ. In order to determine the photosynthetic products released by zooxanthellae in the mantle of the giant clam we traced photosynthetic fixation products from ¹³C- and ¹⁴C-bicarbonate both in the clam and in isolated zooxanthellae (IZ) in the presence or absence of MH. After 15 min incubation in the absence of MH the IZ released less than 0.6% of the fixed labeled carbon, mainly as glucose. The major intracellular photosynthates were neutral lipids, which constituted 20 to 40% of the total extractable ¹⁴C. In the presence of MH, the IZ released up to 5.6% of the total fixed ¹⁴C, mostly as glycerol, and the major intracellular photosynthate was glucose. In an intact clam incubated in sea water containing ¹⁴C-bicarbonate, 46 to 80% of the fixed ¹⁴C was translocated from the zooxanthellae to the host tissues. Most of the ¹⁴C in the hemolymph, in the isolated zooxanthellae and in intact mantle tissue (containing zooxanthellae) was recovered as glucose. No ¹⁴C-glycerol was detected in the mantle after 1 to 30 min incubation, and, even after 60 min, far less ¹⁴C-glycerol was synthesized than by IZ in the presence of MH. The possibility that in clam tissue glycerol is converted to glucose was examined by tracing the labeled carbon from ¹⁴C-glycerol injected into the adductor muscle. After 5 min incubation, no labeled glucose was found in the hemolymph, but after 60 min, some 20% was found as glucose. Thin slices containing zooxanthellae, cut from the surface of the mantle, fixed inorganic carbon supplied as NaH¹⁴CO₃ in the medium and mainly released ¹⁴C-glucose. The addition of MH to the surrounding medium did not affect the release rate or form of release product. When the slices were cut into smaller pieces, however, the ratio of glycerol to glucose in the release product increased. These results indicate that in the presence of MH the metabolism of isolated zooxan- thellae was different from that of zooxanthellae in the mantle. In the presence of MH, isolated zooxanthellae release mostly glycerol, whereas in the mantle they release glucose. Received: 18 February 1998 / Accepted: 4 December 1998     

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.     

Soil microbial diversity and C turnover modified by tillage and cropping in Laos tropical grassland

Agricultural practices should modify the diversity of soil microbes. However, the precise relationships between soil properties and microbial diversity are poorly known. Here, we study the effect of agricultural management on soil microbial diversity and C turnover in tropical grassland of north-eastern Laos. Three years after native grassland conversion into agricultural land, we compared soils from five land use management systems: one till versus two no-till rotational cropping systems, one no-till improved pasture and the natural grassland. Soils were incubated in microcosms during 64 days at optimum temperature and humidity. Bacterial and fungal diversity were evaluated by metagenomic 454-pyrosequencing of 16S and 18SrRNA genes, respectively. Changes in soil respiration patterns were evaluated by monitoring ¹²C- and ¹³C-CO₂ release after soil amendment with 13C-labelled wheat residues. Results show that residue mineralization increased with bacterial richness and diversity in the tilled treatment 7 days after soil amendment. Native soil organic C mineralization and priming effect increased with fungal richness and diversity in improved pasture and natural grassland. No-till cropping systems represented intermediate situations between tillage and pasture systems. Our findings evidence the potential of controlling soil microbial diversity by agricultural practices to improve soil biological properties. We suggest the promotion of no-till systems as a fair compromise between the need for agriculture intensification and soil ecological processes preservation.     

Assimilation of symbiont-derived photosynthates in some solitary and colonial radiolaria

Evidence for host assimilation of ¹⁴C-labeled symbiont photosynthates is presented from laboratory studies of the solitary radiolarian Thalassicolla nucleata and the colonial species Collosphaera huxleyi. The amount of ¹⁴C-labeled product assimilated in the central capsule of T. nucleata is directly related to the amount of ¹⁴C incorporated by the symbionts. In C. huxleyi central capsules, the percentage of ¹⁴C-label occurring in the water-soluble fraction is 38% and in the lipid-soluble fraction is 20%, the remainder being in insoluble products. Within the lipid-soluble fraction, a substantial percentage of the ¹⁴C activity is associated with the triglyceride and wax ester fractions. The significance of these findings is discussed in relation to the possible physiological role of symbionts in sustaining the host and stabilizing the host-symbiont association.     

Mantle protein excretion and calcification in the hardshell clam Mercenaria mercenaria. II. Protein synthesis and excretion by the isolated mantle

Sheets of mantle tissue from above the mantle line of Mercenaria mercenaria were incubated for 2, 6 and 20 h at 20°C in 50 Ci ³H-glycine in 50 ml artificial seawater. Incorporation of tritium into soluble proteins excreted by the mantle and into tissue proteins was followed. The excretion of soluble protein continued throughout the experiment; the proportion of incorporated label excreted reached 17% by 20 h. The initiation of excretion of labelled protein seemed to lag 60 to 70 min behind the initiation of protein synthesis. Protein synthesis by the mantle contributed to proteins of the extrapallium and mantle chambers and, thus, may be involved in synthesis and regulation of proteins involved in the shell-formation process.