Effects of host feeding and dissolved ammonium on cell division and nitrogen status of zooxanthellae in the hydroid Myrionema amboinense

There is a relationship between host feeding, nitrogen status and mitotic activity of zooxanthellae symbiotic with the marine hydroid Myrionema amboinense. Decreases in the mitotic index of zooxanthellae in starved M. amboinense, and in internal pool sizes of glutamine and glutamate, amino acids involved in ammonium assimilation via the glutamine synthetase-glutamate synthase (GS/GOGAT) pathway, were partially restored by addition of ammonium chloride to seawater in which hydroids were incubated. Levels of glutamine were more sensitive to host starvation than levels of glutamate, resulting in a decrease in the glutamine: glutamate molar ratio to that found in zooxanthellae cultured on nitrate. Hydroids starved for 5 d and then incubated in different concentrations of ammonium chloride showed a positive correlation between ammonium concentration and mitotic index of their symbiotic zooxanthellae. Host starvation caused a decrease in perturbation of levels of glutamine and glutamate during ammonium assimilation, as well as decreases in rates of assimilation of [¹⁴C]-leucine into TCA-insoluble protein, and in photosynthetic incorporation of [¹⁴C]-bicarbonate. These observations suggest that host starvation reduces nitrogen supply to the zooxanthellae, causing nitrogen stress to the symbionts and reduction in metabolic processes associated with nitrogen assimilation and photosynthesis as well as with cell division.     

Ontogenetic change in the diet ofAplodactylus punctatus (Pisces: Aplodactylidae): an ecophysiological explanation

Aplodactylus punctatus is a temperate berbivorous fish that changes from an omnivorous to a herbivorous diet and increases its ability ot assimilate algae as it grows. To investigate whether this dietary shift is related to size-specific differences in energetic demands imposed by metabolism and the amount of assimilated energy, oxygen consumption ( ) was determined experimentally in 12 specimens ranging in size from 62 to 545 g. increased allometrically with body size from 8.41 to 55.95 mg O₂ individual^-1 h^-1. Individual energetic requirements were 2.8 to 33.7 kJ d^-1. The assimilated energy was estimated, taking into consideration: (1) the energetic value of the most important alga in the diet (Lessonia trabeculata); (2) size-specific differences in assimilation rates for fish fed on this alga; (3) size-specific differences in throughput time and in the amount of food in a full gut. Comparison of the energy required and the assimilated energy revealed that fishes of < 22 to 29 cm total length had a negative energetic balance when consuming algae exclusively. This may explain the reliance of smallA. punctatus on more easily-digested invertebrates. The largest individuals can meet their energetic demands by consuming algae alone, apparently because of their higher assimilation capability. InA. punctatus, changing energetic requirements and capacities for algal assimilation may be responsible for the observed ontogenetic change in diet.     

Nitrogen fixation (acetylene reduction) associated with macroalgae in a coral-reef community in the Bahamas

N₂ fixation (C₂H₂ reduction) was associated with several species of macroalgae on a coral reef near Grand Bahama Island. The highest rates were associated with Microdictyon sp. (Chlorophyceae) and Dictyota sp. (Phaeophyceae). Extensive mats of filamentous blue-green algae, not heterotrophic bacteria, were the N₂ fixing agents: in experiments with samples of Microdictyon sp., the activity was lightdependent and not stimulated by organic compounds under either aerobic or anaerobic conditions. Assays in situ, at 20 m depth, and on shipboard, gave similar rates of N₂ fixation; the cyanophytes presumably have pigment adaptations to function in blue light. The maximum rate of N₂ fixation, associated with Microdictyon sp., was 3.8 g N fixed g dry weight^-1 h^-1. Coral-reef communities flourish in nutrientimpoverished waters, and therefore any input of nitrogen is probably important in stabilizing such ecosystems.     

An integrated river basin-coast-sea modelling scenario for nitrogen management in coastal waters

Linked river basin and coastal water models were applied to analyse the effects of an optimal nitrogen management scenario in the Oder/Odra river basin on water quality in the Oder (Szczecin) Lagoon and the Pomeranian Bay (Baltic Sea). This scenario would reduce nitrogen loads into the coastal waters by about 35%, a level which is similar to the load of the late 1960’s. During summer the primary production and algae biomass in the Oder estuary is limited by nitrogen, which makes a nitrogen management reasonable. The comparison of the late 1960’s and the mid 1990’s shows that an optimal nitrogen management has positive effects on coastal water quality and algae biomass. However, this realistic nitrogen reduction scenario would not ensure a good coastal water quality according to the European Water Framework Directive. A good water quality in the river will not be sufficient to ensure a good water quality in the lagoon. Nitrogen load reductions bear the risk of increased potentially toxic, blue-green algae blooms, especially in the Baltic coastal sea. However, to reach water quality improvements in lagoons and inner coastal waters, nitrogen cuts are necessary. A mere focus on phosphorus is not sufficient.     

Nitrogen fixation by blue-green algae associated with the siphonous green seaweed Codium decorticatum: effects on ammonium uptake

Nitrogen fixation (acetylene reduction) at rates of up to 1.2 g N₂ g dry wt^-1 h^-1 was measured for the siphonous green seaweed Codium decorticatum. No nitrogenase activity was detected in C. isthmocladum. The nitrogenase activity was light sensitive and was inhibited by the addition of DCMU and triphenyl tetrazolium chloride. Additions of glucose did not stimulate nitrogen fixation. Blue-green algae (Calothrix sp., Anabaena sp., and Phormidium sp.) were implicated as the organisms responsible for the nitrogenase activity. They occurred in a reduced microzone within the C. decorticatum thallus where nitrogen fixation was optimized. Nitrogen fixation did not affect the kinetic constants for ammonium uptake in C. decorticatum (K_s=12.0 M, V_max=13.4 mol NH₃ g dry wt^-1 h^-1) determined using the perturbation method. Nevertheless, C. decorticatum thalli which fixed nitrogen had internal dissolved nitrogen concentrations which were over 1.4 times higher than in non-fixing thalli. This suggests that if C. decorticatum does derive part of its nitrogen requirement from the blue-green algae which it harbors, the transfer does not involve competition between this process and the uptake of ambient ammonium.     

Primary site and initial products of ammonium assimilation in the symbiotic sea anemone Anemonia viridis

Invertebrates containing endosymbiotic dinoflagellate algae (zooxanthellae) retain excretory nitrogen, and many are able to take up ammonium from the surrounding seawater. However, the site of assimilation and role of nitrogen recycling between symbiont and host remains unclear. In the present study, ammonium uptake by the symbiotic sea anemone Anemonia viridis (Forskål) was examined by following the pathway of assimilation using ¹⁵N-enriched ammonium. Since zooxanthellae became enriched with ¹⁵N from ammonium at up to 17 times the rate of the host, they appear to be the primary site of assimilation. In the light, the rate of zooxanthellae enrichment at 20?M was twice that at 10?M, whereas the rate of host enrichment was not significantly affected by ammonium concentration. When anemones were incubated with [¹⁵N]ammonium in the dark, after 12?h without light the rate of enrichment was lowered in both zooxanthellae and host. However, while the enrichment of the host was significantly reduced when the light level was lowered from 300 to 150?μmol photons m^?2?s^?1, zooxanthellae enrichment was unchanged. Low molecular weight material from the zooxanthellae became enriched at 20 times the rate of that from the host, and enrichment was detected in the amino acids glutamate, glutamine, aspartate, alanine, glycine, phenylalanine, threonine, valine, tyrosine, and leucine from zooxanthellae. In the zooxanthellae, amino acids accounted for 65% of the total enrichment of low molecular weight material. Of the amino acids detected in zooxanthellae, over 90% of the enrichment was accounted for by glutamate, glutamine and aspartate. The enrichment of the amide group of glutamine was greater than that of the amine group of glutamate or glutamine, consistent with the glutamine synthetase/glutamine 2-oxoglutarate amidotransferase cycle as the mechanism of ammonium assimilation. To examine the flux of ¹⁵N from zooxanthellae to host, anemones were pulse-labelled with [¹⁵N]ammonium and then transferred to an unlabelled chase. Over a 2?h period there was no evidence for a flux of nitrogen from zooxanthellae to host. However, during the chase period, the enrichment of low molecular weight material declined and that of high molecular weight material increased in both zooxanthellae and host, indicating that protein was synthesized using ¹⁵N from ammonium in both components of the symbiosis. Again by using a pulse-chase system, it was found that glutamate was metabolised most rapidly by zooxanthellae, followed by (in order of decreasing rate of turnover) aspartate, alanine, glycine and valine (no data are available for glutamine). Unlike these amino acids, nitrogen was transferred to the essential amino acids phenylalanine and threonine, increasing their enrichment during the chase period. While recycled nitrogen is clearly important to this symbiosis, the mechanism by which it is cycled remains to be resolved.     

Indications from photosynthetic components that iron is a limiting nutrient in primary producers on coral reefs

Because iron is not available generally in oxygenated sea water, it may be a limiting factor in marine primary production. This hypothesis was tested in the context of Davies Reef, Latitude 18°50′S (one of the coral reefs in the central region of the Great Barrier Reef system). Samples were collected for study in the period August, 1980 to March, 1981. Sea water around the reef contained ≦2x10^-6 M Fe, surface sediments from the reef contained 66±26 (1 SD) ppm total Fe, and interstitial water near the surface contained ≧5x10^-7 M Fe. Thus, Fe constituted a trace component of the reef environment, but limited Fe should be available to algae associated with the sediments. Specific biochemical analyses to test the Fe status of benthic photosynthetic organisms were carried out with a common blue-green alga, Phormidium sp., and a ubiquitous symbiotic dinoflagellate, Gymnodinium microadriaticum (zooxanthellae). The blue-green alga contained the electron transport protein, flavodoxin, which is found only in Fe-deficient organisms. Supporting evidence for Fe stress in this organism included chlorosis in the presence of plentiful biliprotein, and very low extractable photosynthetic cytochrome, c-553. The latter observations were shown to be the result of Fe deficiency in laboratory cultures of a blue-green alga, Synechococcus sp. These cultures showed that production of flavodoxin is not a universal response of algae to Fe stress, but that lowered cellular concentrations of Fe-containing proteins involved in photosynthesis probably is universal. The zooxanthellae from a soft coral, Sinularia sp., had three-fold lower total Fe and ferredoxin (an electron transport protein), than the same alga from a clam, Tridacna maxima. Thus, some algae in symbiotic associations may also suffer Fe-deficiency. It was concluded that the degree and extent of Fe-stress in primary producers on a coral reef may influence growth rates, biomass, and distribution of species.     

Marine blue-green algae have a unique osmoregulatory system

Currently, blue-green algae are classified as either freshwater or marine depending on the ionic requirements of the strain, not on the type of habitat from which the strain was isolated. As a result many strains isolated from saline environments are classified as freshwater strains. New parameters were sought which might correlate better the physiology of marine strains with their habitat. This study reveals that blue-green algae isolated from the marine environment have a unique osmoregulatory system which distinguishes them as a coherent physiological group, distinct from blue-green algae isolated from non-marine habitats. The “marine” blue-green algae can be identified by their ability to synthesise and accumulate 2-O-α-D-glucopyranosylglycerol (glucosylglycerol), a major osmoregulatory compound, and by their related ability to grow in seawater-based medium with total maximum NaCl of 6 to 11% (w/v). These two properties allow a more rigorous definition of “marine” than the current method of classification.     

Trace Metal Uptake By Pichia Spartinae, an Endosymbiotic Yeast in the Salt Marsh Cord Grass Spartina Alterniflora

The ascosporogenous marine yeast Pichia spartinae is a dominant endosymbiont of the marsh grass Spartina alterniflora. Results of previous studies suggested that P. spartinae is involved in iron transport processes in the grass. of particular interest has been the mechanisms of metal uptake and metabolism by the yeast, and the ecological and plant biochemical significance of these processes. This investigation examined the uptake of iron and other metals (Zn, Cu, Cd, Ni, Pb, Cr) by P. spartinae, and provides data on possible mechanisms of this activity. the results suggest a) the yeast can assimilate divalent and trivalent forms of inorganic iron, as well as large organic-Fe(III) complexes, b) the uptake of inorganic trivalent iron under soluble iron-deficient conditions proceeded by a different mechanism than that of soluble Fe(II), with intracellular loadings of iron much increased under the former conditions; c) trivalent iron uptake is not mediated by hydroxamate siderophores at levels detectable by sensitive screening assays; d) the assimilation of some trace metals (Cu, Zn, Cd, Ni) is likely to be mediated by low molecular weight cysteine rich proteins, possibly metallothionein, and; e) siderophores from other fungi can provide iron for P. spartinae. the iron assimilation data suggested that multiple mechanisms are involved, and are influenced by the concentration and speciation of iron in the system. in general, iron assimilation mechanisms are comparable to those described for closely related yeasts, such as Saccharomyces cerevisiae. Among other things, these results indicated that future studies of trace metal mobilization and plant assimilation in salt marsh ecosystems must account for the activities of microbial symbionts associated with the plants.     

Physiology and chemical composition of nitrogen-fixing phytoplankton in the central North Pacific Ocean

The magnitude and physiological characteristics of biological nitrogen fixation have been studied in the oligotrophic waters of the North pacific gyre. The filamentous blue-green algae Trichodesmium spp. and Richelia intracellularis were the important nitrogen-fixing phytoplankton. Most of the nitrogen fixation occurs in the upper 40 m of the water column, with detectable fixation as deep as 90 m, which corresponds to about the 1 % light depth. There was no evidence of photoinhibition of nitrogen fixation, although CO₂ reduction was depressed slightly at the highest light levels. The rate of nitrogen fixation in the water column varied throughout the day, being highest in mid-morning and in late afternoon. Relatively high fixation rates were also found during periods of darkness. Elevated oxygen concentrations had a marked inhibitory effect on rates of nitrogen fixation, a pO₂ of 0.4 atm causing a 75% inhibition. Data from studies of nitrogen fixation and assimilation rates of ¹⁵N-labelled nitrate, ammonium, and urea indicate that nitrogen fixation furnished about 3% of the total daily fixed nitrogen requirement for phytoplankton growth. Studies with isolated colonies of Trichodesmium spp. indicated that 100% of their nitrogen requirement was met by nitrogen fixation. Chemical composition of the Trichodesmium colonies showed that the C:N ratio was 4.1 and that their phosphorus content relative to carbon or nitrogen was much lower than that of the total particulate material in the water column. Elevated ratios of carbon: adenosine triphosphate (ATP) also suggest that phosphorus deficiency may be limiting the growth of Trichodesmium. The magnitude of nitrogen fixation in the gyre is seasonally dependent, with high rates in late summer and autumn. At these times the water column is stratified, with phosphate and nitrate barely detectable in the upper 100 m. Our data suggest that during these months of stratification, biological fixation of nitrogen amounts to about 33 g-at N/m²/day.     

Physiological energetics of the intertidal sea anemone Anthopleura elegantissima

Energy budgets were calculated for individuals of the sea anemone Anthopleura elegantissima (Brandt), collected in 1981 and 1982 from Bodega Harbor, California, USA. Rates of ammonium excretion were measured in high-and low-intertidal, symbiotic and aposymbiotic sea anemones within 24 h of collection. Among symbiotic and aposymbiotic individuals, no differences in excretion rate were found on the basis of intertidal height. However, rates of ammonium excretion in aposymbiotic anemones (2.14 mol NH ₊ ⁴ g^-1 h^-1) were significantly higher than in symbiotic ones (0.288 mol NH ₊ ⁴ g^-1 h^-1). Rates of excretion were used with estimated rates of oxygen uptake to calculate nitrogen quotients (NQ). NQ and RQ values from the literature were used to calculate an oxyenthalpic equivalent [501 kJ (mol O₂)^-1 for R+U], and mass proportions of protein (54%), carbohydrate (44%) and lipid (2%) catabolized during routine metabolism in this species 24 h after feeding. Integrated energy budgets of these experimental anemones were calculated from data on ingestion, absorption and growth, and estimates of translocated energy from the symbiotic algae. Contribution of zooxanthellae to animal respiration based on translocation=90% and RQ=0.97 are 41 and 79% in high-and low-intertidal anemones, respectively. Calculated scope for growth is greater than directly measured growth in both high-and low-intertidal individuals. The deficit, estimated as 30% of assimilated energy in high-intertidal anemones, is attributed to unmeasured costs (specific dynamic effect) or production (mucus). Low-intertidal anemones lost mass during the experiment, implying that the magnitude of the deficit was greater in these anemones than in upper intertidal individuals. Anemones from both shore levels lost zooxanthellae during the experiment, which contributed to energy loss since the contribution of the zooxanthellae is greater in low-intertidal anemones. Scope for growth is preserved in high-intertidal anemones (29% of assimilated energy) because metabolic demands are lower due to aerial exposure, and prey capture rate is higher compared to lowshore anemones. Although possibly underestimated, lower scope for growth in low-shore anemones may result from continuous feeding and digestion processes that are less efficient than those of periodically feeding high-intertidal anemones.     

Role of glutamine synthetase in ammonia assimilation by symbiotic marine dinoflagellates (zooxanthellae)

The dinoflagellate symbionts (zooxanthellae) present in many reef corals aid in the survival of the symbiotic unit in nitrogen deficient tropical waters by providing additional routes of nitrogen uptake and metabolism. The enzymatic pathway of ammonia assimilation from seawater and the re-assimilation of coral ammonium waste by zooxanthellae was studied by examining the affinity of glutamine synthetase for one of its substrates, ammonia. Glutamine synthetase activity was measured in dinoflagellates of the species Symbiodinium microadriaticum found in symbiotic association with various marine coelenterates. Michaelis-Menten kinetics for the substrate ammonia were determined for freshly isolated dinoflagellates from Condylactis gigantea (apparent NH₃ K_m=33 M) and for cultured dinoflagellates from Zoanthus sociatus (apparent NH₃ K_m=60 M). On the basis of the low apparent K_ms for NH₃, it appears that ammonia assimilation by these symbiotic dinoflagellates occurs via the glutamine synthetase/glutamate synthase pathway. Additionally, the uptake of exogenous ammonium by an intact coelenterate-dinoflagellate symbiosis was strongly inhibited by 0.5 mM methionine sulfoximine, and inhibitor of glutamine synthetase.     

Essential amino acid synthesis and nitrogen recycling in an alga–invertebrate symbiosis

When aseptically-cultured sea anemones, Aiptasia pulchella, were incubated with ¹⁴C-labelled glucose, aspartate and glutamate, radioactivity was incorporated into animal protein. Radioactivity was recovered from all amino acids in the protein hydrolysates of A. pulchella bearing the symbiotic alga Symbiodinium sp., and from all but seven of the amino acids in A. pulchella experimentally deprived of their algae. These data suggest that these seven amino acids (histidine, isoleucine, leucine, lysine, phenylalanine, tyrosine and valine) may be synthesized by the symbiotic algae and translocated to the sea anemone's tissues; and that methionine and threonine, two amino acids traditionally considered as dietary essentials for animals, are synthesized by A. pulchella. Essential amino acid translocation from the symbiotic algae to the animal host is a core element in symbiotic nitrogen-recycling. Its nutritional value to the animal host is considered in the context of the amino acid biosynthetic capacity of the host. Received: 26 October 1998 / Accepted: 28 June 1999     

Daily photosynthesis,respiration, and carbon budgets in a tropical marine jellyfish (Mastigias sp.)

From measured diel photosynthesis and respiration rates, using oxygen electrodes, estimates of carbon flux between symbiotic algae (zooxanthellae) and host animal are presented for the marine scyphomedusan Mastigias sp. from a marine lake in Palau, Western Caroline Islands, during February and March 1982. The carbon budgets calculated for these lake medusae indicate that carbon fixed photosynthetically by zooxanthellae and made available to the host may satisfy up to 100% of the host's daily metabolic carbon demand (CZAR). The stable carbon isotope (¹³C) signature of the mesogleal carbon of lake Mastigias sp. was close to that of the zooxanthellae, supporting the interpretation that while these medusae may feed holozoically, some of their carbon comes from their symbionts. The diel photosynthesis, respiration, and preliminary estimates of carbon budgets of three individuals of another ecotype of Mastigias sp. collected from nearby oceanic lagoons are also given. Photosynthesis of lagoon medusae was generally greater than that for lake medusae of similar size, and lagoon medusae were phototrophic with respect to carbon, with commensurately greater CZAR values. Carbon translocated from the symbiotic algae also may contribute to the growth requirements of both lake and lagoon medusae. From carbon flux data, the lake jellyfish were estimated to contribute about 16% to the total primary productivity of their marine lake habitat.     

Symbiotic anemones can grow when starved: nitrogen budget for Anemonia viridis in ammonium-supplemented seawater

The ability of endosymbioses between anthozoans and dinoflagellate algae (zooxanthellae) to retain excretory nitrogen and take up ammonium from seawater has been well documented. However, the quantitative importance of these processes to the nitrogen budget of such symbioses is poorly understood. When starved symbiotic Anemonia viridis were incubated in a flow-through system in seawater supplemented with 20 μM ammonium for 91 d under a light regime of 12 h light at 150 μmol photons m⁻² s⁻¹ and 12 h darkness, they showed a mean net growth of 0.197% of their initial weight per day. Control anemones in unsupplemented seawater with an ammonium concentration of <1 μM lost weight by a mean of 0.263% of their initial weight per day. Attempts to construct a nitrogen budget showed that, over a 14 d period, ≃40% of the ammonium taken up could be accounted for by growth of zooxanthellae. It was assumed that the remainder was translocated from zooxanthellae to host. However, since the budget does not balance, only 60% of the growth of host tissue was accounted for by this translocation. The value for host excretory nitrogen which was recycled to the symbionts equalled that taken in by ammonium uptake from the supplemented seawater, indicating the importance of nitrogen retention to the symbiotic association. Received: 23 December 1997 / Accepted: 12 September 1998     

Low molecular-weight factor from Plesiastrea versipora (Scleractinia) that modifies release and glycerol metabolism of isolated symbiotic algae

When symbiotic dinoflagellate algae (Symbiodinium sp., isolated from the coral Plesiastrea versipora) were incubated with NaH¹⁴CO₃ in the light in seawater, they released 22.69±9.16 nmol carbon/10⁶ algae. Release of photosynthetically fixed carbon was stimulated more than six-fold for algae incubated in host-tissue homogenate (148.54±97.03 nmol C/10⁶ algae) and more than four-fold (102.00±49.16 nmol C/10⁶ algae) for algae incubated in a low molecular weight fraction (≤1 000 M _r ) prepared from host homogenate. Soluble released ¹⁴C-labelled products, as determined by chromatography and autoradiography, were the same when algae were incubated in either host homogenate or the low molecular weight fraction. After 4 h incubation in the light (300 mol photons m⁻² s⁻¹),␣intracellular␣glycerol increased in algae incubated with the low molecular weight fraction (an increase of 0.39 to␣0.67 nmol glycerol/10⁶ algae) compared with little or no increase in algae incubated in seawater (0 to 0.12 nmol glycerol/10⁶ algae). Partial inhibition of triglyceride synthesis (up to 51%) was also observed when algae were incubated in the low molecular weight fraction. All these effects are the same as those observed when algae were incubated in host homogenate. These data indicate that the “host release-factor” activity of P.␣versipora is a compound of low molecular weight. Received: 13 February 1997 / Accepted: 24 October 1997     

Assimilation of 210Po by the mussel Mytilus edulis from the alga Isochrysis galbana

Marine invertebrates are thought to accumulate ²¹⁰Po primarily from their food. In this study, a pulse-chase methodology was used to examine the assimilation and depuration of ²¹⁰Po by Mytilus edulis from the common marine alga Isochrysis galbana. The digestion of ²¹⁰Po from I. galbana occurred via a biphasic process, characteristic of a rapid (extracellular) and slow (intracellular) digestion typical of marine bivalves. The mantle/gill and foot have no known digestive role, yet their ²¹⁰Po specific activities increased after 24 h. It is proposed that this increase in ²¹⁰Po specific activity was related to ²¹⁰Po being incorporated into these tissues from ²¹⁰Po assimilated from I. galbana during extracellular digestion. It is proposed that the linear loss of ²¹⁰Po previously accumulated by control mussels was related to the continual state of renewal and replacement of cellular proteins, with ²¹⁰Po turnover and metabolism governed by protein turnover and metabolism. M. edulis' assimilation efficiency of ²¹⁰Po from the ²¹⁰Po-labelled alga was calculated to be 17.2 ± 2.1%, and thus similar to that of Ag, Cd, Co, Se and Zn by bivalves from other marine algae species. It is proposed that the assimilation efficiency of ²¹⁰Po is a function of protein assimilation. Received: 27 August 1998 / Accepted: 3 September 1999     

Transmission of symbiotic dinoflagellates through the sexual cycle of the host scyphozoan Linuche unguiculata

Intracellular symbiotic dinoflagellates are associated with the tropical scyphozoan Linuche unguiculata (Swartz, 1788) throughout all stages of the host's life cycle. During sexual reproduction, eggs are released in mucus strands that contain symbiotic dinoflagellates. Fertilization and development take place externally in the water column. Epifluorescence and transmission electron microscopy showed that unfertilized eggs did not contain intracellular algae, but that infection of the developing embryo was generally successful by the 128-cell stage (10 h after fertilization at 23° C). However, experiments with artificially provided Cellufluor-labeled algae demonstrated that older embryos and planulae could be infected by algae through at least 24 h post-fertilization, indicating that the L. unguiculata symbiosis represents a semi-closed system. This novel mode of symbiont acquisition results in most sexually-produced offspring becoming infected with maternally-transmitted algae during early development, but allows for acquisition of non-maternally-provided algae later in development. Most of the algal symbionts during the early stages of embryonic and larval development are located within ectodermal cells. This is in contrast to the other life-cycle stages of L. unguiculata (i.e., scyphistoma, medusa, ephyra), where symbionts are found within the gastrodermis of the host.     

Feeding behavior and acquisition of zooxanthellae by planula larvae of the sea anemone Anthopleura elegantissima

Symbiotic associations between cnidarians and photosynthetic dinoflagellates (i.e., zooxanthellae) are common in the marine environment. Many symbiotic cnidarians produce offspring that are initially nonsymbiotic. These new hosts must acquire symbiotic algae from environmental sources. We examined zooxanthella acquisition by laboratory-reared planula larvae of the temperate sea anemone Anthopleura elegantissima. Larvae ingested zooxanthellae while they were feeding. However, the signal that prompted larval feeding behavior did not originate from the symbiotic algae; the addition of algal cells to larval cultures never elicited a feeding response. In contrast, the addition of macerated animal tissue from several sources invariably generated a strong feeding response, which resulted in the larvae indiscriminately ingesting any particulate matter that was present, including zooxanthellae or other unicellular algae. Ingested zooxanthellae were incorporated into endodermal cells, where they remained undigested, while all other ingested material was digested or expelled within 24 h. Our results provide evidence that one source of zooxanthellae likely to serve as a route of infection in the natural environment is zooxanthella-laden mucus egested by anemones. This egested material fulfilled both of the criteria necessary for successful infection: it prompted larvae to begin feeding and provided an abundant supply of zooxanthellae that were ingested and taken up into endodermal cells of the new host.     

Nutritional effect of five species of marine algae on the growth,development, and survival of the brine shrimp Artemia salina

Five species of unicellular algae of the same age, cultured bacteria-free under standard growth conditions, were analyzed for chemical composition and fed to different size classes of Artemia salina. The green algae Chlamydomonas sphagnicolo, Dunaliella viridis, Platymonas elliptica and Chlorella conductrix had significantly higher percentages of protein and lipid than did the diatom Nitzschia closterium. Total ash value was highest in populations of N. closterium. Shrimp fed Chlamydomonas sphagnicolo cells assimilated highest percentages of organic matter, while those fed Chlorella conductrix had lowest assimilation rate. Respiration rates were inversely proportional to animal size (weight) and algal cell volume. Growth, survival, rate of sexual maturtion, and sex ratio were dependent on the growth and assimilation efficiencies obtained from each respective algal food. Shrimp fed Chlamydomonas sphagnicolo, D. viridis, or P. elliptica cells displayed highest growth and assimilation efficiencies.