Fecal discharge of zooxanthellae in the giant clam Tridacna derasa, with reference to their in situ growth rate

The population dynamics of zooxanthellae living in the mantle of a giant clam, Tridacna derasa, was studied. The giant clams with shell lengths of 5 to 6 cm which had been reared in the Palau Mariculture Demonstration Center, in the Republic of Palau, were transferred to aquaria on deck of the R.V. “Sohgen-maru” and kept in running sea water at 29 to 30 °C. Two clams were removed from the aquaria, and zooxanthellae in the mantle were isolated every 2 h for 24 h. Numbers of the zooxanthellae in or not in the cell division stage were counted for calculations of the zooxanthellae population in the mantle and their mitotic index (MI). The MI increased after sunset and reached the maximum values of 6.1 to 11.5% at 03:00 to 05:00 hrs. The specific growth rate, μ, estimated from the MI was 0.083 to 0.14 d⁻¹. Five clams were kept in each of 2 Plexiglas containers in the aquarium for collection of the discharged feces every 3 to 4 h. The discharged zooxanthellae in the feces were counted. The zooxanthellae discharged in 24 h were 0.38 to 1.46% of the total zooxanthella population in the mantle, and 2.7 to 16.9% of the newly formed zooxanthella population in a day. Increase of zooxanthella population in the mantle was estimated from clam shell growth rate and from the correlation between zooxanthella population and clam shell size. Daily increase of zooxanthella population in the mantle was estimated to be approximately 7.6 to 19% of the newly formed zooxanthella population. Therefore, the sum of zooxanthellae populations accounting for daily increase in the mantle and discharge in the feces was 11 to 36% of the newly formed population. About 64 to 89% of the newly formed cells were missing; some of these may have been digested by the clam. Received: 14 July 1996 / Accepted: 19 August 1996     

Phosphate acquisition in the giant clam-zooxanthellae symbiosis

The effect of phosphate on the giant clam Tridacna gigas and on its symbiotic dinoflagellate Symbiodinium sp. was compared with that on cultured Symbiodinium sp. originally isolated from the same clarn species. Incubation of whole clams in elevated phosphate (10 M) reduced their capacity for phosphate uptake, but the uptake capacity of the clam's zooxanthellae population was not influenced. In addition, there was no change in the zooxanthellae density and the N:P ratio, of these algae., On the other hand, cultured zooxanthellae were influenced by the phosphate regimen of their culture medium. Compared with controls (0 M P), addition of 10 M phosphate to the culture medium caused an increase of 100% in cell density and decreases of 50% in the N:P ratio, and 80% in the phosphate-uptake capacity of the zooxanthellae. Zooxanthellae freshly isolated from the clams exhibited properties similar to those of zooxanthellae cultured in the absence of phosphate. These results demonstrate that the zooxanthellae population of T. gigas have limited access to the inorganic phosphate in sea water and the phosphate reserves within the animal host.     

Nutrient enrichment and the ultrastructure of zooxanthellae from the giant clam Tridacna maxima

The separate and combined effects of ammonium (10M) and phosphate (2M) on the ultrastructure of zooxanthellae (Symbiodinium sp.) from giant clams, Tridacna maxima, were examined in the field. Nitrogen addition significantly changed the ultrastructure of the zooxanthellae inhabiting the clams. After 9 mo exposure, the cross-sectional area of zooxanthellae from N-treated clams was significantly lower than that from other treatments [N=39.3 m²; C=47.9 m²; P=43.2m²; N+P=44.5 m²; (P=0.001)]. There was also a significant decrease in the size of starch bodies, especially around the pyrenoid of the zooxanthellae from N and N+P treatments [N=1.2 m²; C=2.0 m²; P=1.8 m²; N+P=1.2 m²; (P=2.08E-11)]. This presumably occurs as a result of the mobilization of organic carbon stores in response to stimulated amino acid synthesis under enriched nutrient conditions. These data strongly suggest that the symbiotic zooxanthellae of clams are limited to some extent by the availability of inorganic nitrogen, and that relatively minor changes to the nutrient loading of the water column can have substantial effects on the biochemistry of symbioses such as that which exists between clams and zooxanthellae.     

Cellular and ultrastructural changes in the endoderm of the temperate sea anemone Anemonia viridis as a result of increased temperature

Exposure of the temperate sea anemone Anemonia viridis Forskål to increased seawater temperature (from 16 to 26°C) reduced the lysosomal latency of coelenterate tissues. Lysosomes in the mesenterial filaments of anemones were destabilised by increased temperature, with greater destabilisation in heat-shocked symbiotic anemones than in heat-shocked aposymbiotic anemones in the early stages of the experiment. Lysosomal enzyme activity in zooxanthellae from heat-shocked symbiotic anemones was associated with the algal membranes and the cytoplasm of degenerate algal cells. While the relationship between host coelenterate and symbiotic alga may confer many benefits under normal conditions, comparison of the responses of symbiotic and aposymbiotic anemones to heat shock suggests that there may be disadvantages for symbiotic anemones under stress.     

Effects of dissolved ammonium addition and host feeding with Artemia salina on photoacclimation of the hermatypic coral Stylophora pistillata

 Effects of nutrient treatments on photoacclimation of the hermatypic coral Stylophora pistillata (Esper) were studied. Studies on photoacclimation of colonies from different light regimes in the field were evaluated and used to design laboratory experiments. Coral colonies were collected in the Gulf of Eilat (Israel) from January to March 1993. Exterior branches of colonies from different depths (1 to 40 m) displayed different trends in production characteristics at reduced and very low levels of illumination. From 24 ± 3% to 12 ± 2% of incident surface photosynthetic active radiation (PAR_o), zooxanthella population density and chlorophyll a+c per 10⁶ zooxanthellae increased, a trend seen in the range of light levels optimal for coral growth (90 to 30% PAR_o). The P _max of CO₂ per 10⁶ zooxanthellae decreased, while P _max of CO₂ per 10³ polyps increased, indicating an increase in zooxanthella population density at low light levels. Proliferous zooxanthella frequency (PZF, a measure of zooxanthella division) declined significantly at light levels <18 ± 3% PAR_o. At the lowest levels of illumination (<5% PAR_o), zooxanthella population density decreased, as did the PZF; chl a+c per 10⁶ zooxanthellae was unchanged. In 28-d experiments, exterior coral branches from the upper surfaces of colonies from 3 m depth (65 ± 4% PAR_o) were incubated in aquaria under bright (80 to 90% PAR_o), reduced (20 to 30% PAR_o), and extremely low (2 to 4% PAR_o) light intensities. At each light intensity, the corals were maintained in three feeding treatments: sea water (SW); ammonium enriched SW (SW + N); SW with Artemia salina nauplii (SW + A). An increase in P _max of CO₂ per 10³ polyps was found in corals acclimated to reduced light (20 to 30% PAR_o) in nutrient-enriched SW, while in SW, where the increase in zooxanthella population density was smaller, it did not occur. Nutrient enrichments (SW + N at 2 to 4% PAR_o and SW + A at 20 to 30% PAR_o) increased zooxanthella population density, but had no effect on chl a+c per 10⁶ zooxanthellae. Acclimation for 14 d to reduced (10 to 20% PAR_o) and extremely low (1 to 3% PAR_o) light intensities shifted ¹⁴C photoassimilation into glycerol and other compounds (probably glycerides), rather than sugars. Both ammonium addition and feeding with Artemia salina nauplii resulted in an increase in photosynthetic assimilation of ¹⁴C into amino acids. We conclude that acclimation to reduced light consists of two processes: an increase in photosynthetic pigments and in zooxanthella population density. Both processes require nitrogen, the increase in zooxanthella population density needing more; this adaptation is therefore limited in nitrogen-poor sea water. Received: 19 June 1998 / Accepted: 13 June 2000     

Influence of light on algal symbionts of the deep water coral Leptoseris fragilis

Photoadaptations of zooxanthellae living within the deep water coral Leptoseris fragilis taken from the Gulf of Aqaba (Red Sea) were studied. Specimens-collected in summer 1988 between 110 and 120 m depth —were transplanted to 70 and 160 m. At each depth individuals were exposed in their natural growth position (oral side facing the surface) or in a reverse growth position (oral side facing the bottom). After 1 yr of exposure the corals were collected and the zooxanthellae were isolated. As a function of the availability of light with depth and growth position several algal parameters showed changes which are related to photoadaptations. The relatively low density of zooxanthellae of 0.15x10⁶ cellsxcm^-2 at a natural growth depth of 116 m decreased to 0.0034x10⁶ cellsxcm^-2 (2%) at 160 m in specimens growing with a natural orientation. In corals with a downward-facing oral surface at the same depth (160 m) only degenerated algae could be observed. With respect to depth dependence the volume of the algae decreased from 728 m³ at 116 m to 406 m³ at a depth of 160 m and the content of pigments increased. The augmentation of peridinin per cell was low (two times at 160 m compared to 116 m). Chlorophyll a and in particular chlorophyll c ₂ concentrations per cell were enhanced. Compared to natural amounts at 116 m, chl a was five times and chl c ₂ eight times higher at 160 m. At all depths the chl c ₂ content per cell was higher than for chl a. The formation of chl a/chl c ₂ complexes as light harvestor is discussed. Light harvesting, with chl c ₂ prevailing may be explained as a special type of chromatic adaptation of L. fragilis in a double sense: (1) in the habitat light short wavelengths predominate. This light can be directly absorbed with pigments such as chl a and chl c ₂. (2) Host pigments absorb visible violet light and transform these wavelengths, less suitable for photosynthesis, into longer ones by means of autofluorescence. The emitted longer wavelengths fit the absorption maxima of the algal pigments. Thus the host supports photosynthesis of his symbionts. Corals exposed at 160 m depth with a downward facing oral surface were alive after 1 yr and the host wavelength transforming pigment system was still present, but zooxanthellae were absent or degenerated. The light field at 160 m seems therefore to be critical: the combined photoadaptations of host and symbionts, allowing photosynthesis under barren light conditions, seem to be exhausted. In L. fragilis the photoadaptive strategies of host and symbionts cooperate harmoniously. In addition, the adaptations are interlocked with the particular light situation of the habitat with respect to light quantity and quality. The cooperation of physical and organismic parameters examplifies how evolution and, in particular, coevolution has led to optimal fitness.     

Effect of feeding regime and irradiance on the photophysiology of the symbiotic sea anemone Aiptasia pulchella

To determine how the animal and algal components of the symbiotic sea anemone Aiptasia pulchella respond to changes in food availability and culture irradiance, sea anemones from a single clone were maintained at four irradiance levels (320, 185, 115, and 45 E m^-2 s^-1) and either starved or fed for 5 wk. Changes in protein biomass of sea anemones maintained under these conditions were not related to the productivity of zooxanthellae, since the protein biomass of fed A. pulchella decreased with increase in irradiance and there was no difference in protein biomass among starved sea anemones at the four irradiance levels. Except for the starved high-light sea anemones, the density of symbiotic zooxanthellae was independent of culture irradiance within both starved and fed. A. pulchella. Starved sea anemones contained over twice the density of zooxanthellae as fed sea anemones. Within both starved and fed individuals, chlorophyll per zooxanthella increased with decreasing culture irradiance while algal size remained constant (in fed sea anemones) at about 8.80 m diameter. Chlorophyll a: c ₂ ratios of zooxanthellae increased with decreasing culture irradiance in zooxanthellae from starved sea anemones but remained constant in zooxanthellae from fed sea anemones. As estimated from mitotic index data, the in situ growth rates of zooxanthellae averaged 0.007 d^-1 and did not vary with irradiance or feeding regime. Photosynthesis-irradiance (P-I) responses of fed A. pulchella indicated an increase in photosynthetic efficiency with decreasing culture irradiance. But there was no consistent pattern in photosynthetic capacity with culture irradiance. Respiration rates of fed sea anemones also did not vary in relation to culture irradiance. The parameter I _k , defined as the irradiance at which light-saturated rates of photosynthesis are first attained, was the only parameter from the P-I curves which increased linearly with increasing culture irradiance. The daily ratio of net photosynthesis to respiration for A. pulchella ranged from 1.6 to 2.8 for sea anemones maintained at the three higher irradiances, but was negative for those maintained at 45 E m^-2 s^-1. Since the final protein biomass was greatest for sea anemones maintained at the lowest irradiance, these results indicate that sea anemone growth cannot be directly related to productivity of zooxanthellae in this symbiotic association.     

Host feeding and nutrient sufficiency for zooxanthellae in the sea anemone Aiptasia pallida

Nutrient sufficiency of zooxanthellae in the sea anemone Aiptasia pallida cultured in low nutrient seawater depends on the availability of particulate food to the host. Zooxanthellae in anemones unfed for 20 to 30 d exhibited the following characteristics of nutrient deficiency: cell division rates decreased; chlorophyll a content gradually decreased from 2 to <1 pg cell^–1; and C:N ratios increased from 7.5 to 16. Over a 3-mo period, algal populations in unfed anemones gradually decreased, indicating that zooxanthellae were lost faster than they were replaced by division. The mitotic index of zooxanthellae in unfed anemones was stimulated either by feeding the host or by the addition of inorganic N and P to the medium. Whether algae are nutrient-limited in hosts under field conditions has not been examined fully; however, C:N ratios in zooxanthellae from field-collected hosts are slightly higher (9.4 vs 7.5) than in hosts fed to repletion in laboratory cultures. This observation might indicate N limitation in the field.     

Uptake of dissolved inorganic nitrogen by the symbiotic clam Tridacna gigas and the coral Acropora sp.

Dissolved inorganic nitrogen flux was studied in the giant clam Tridacna gigas and the corals Acropora sp. and Tubastrea micrantha from the tropical reefs of Belau, Micronesia in 1983. T. micrantha, a nonsymbiotic coral, excreted ammonium. However, Tridacna gigas and Acropora sp., which contain symbiotic dinoflagellates (zooxanthellae) were able to take up both ammonium and nitrate. The requirement for a previous light exposure to sustain uptake by T. gigas is reported. The uptake kinetics of these symbioses are described and include the capacity of the zooxanthellae for surge uptake when given nutrient spikes.Contribution No. 417 of the Allan Hancock Foundation     

Adaptation of solitary corals and their zooxanthellae to low light and UV radiation

Adaptation of solitary corals, Fungia repanda and F. echinata, and their zooxanthellae to low light and ultraviolet light B (UV-B) was studied with respect to changes in their protein contents, photosynthetic pigment contents and the photosynthesis-irradiance (P-I) curves. The corals were collected from 1 to 50 m depths in the Republic of Belau (Paulau) in 1990 and 1991. The chlorophyll a content in a unit surface area of the coral did not change significantly with the depth of the habitat, whereas cellular chlorophyll a in the algae increased with the depth. Zooxanthellae density and protein content in a unit surface area of Fungia spp. decreased with the depth. Photosynthetic parameters normalized by a unit surface area of the Fungia spp., maximum gross photosynthetic rate (P _gmax area^-1) and dark respiration rate (R area^-1), were negatively correlated with the depth, while initial slope of the P-I curve () did not show significant correlation with the depth. Compensation light intensity (Ic) decreased with the depth. In isolated zooxanthellae, P _max chl a ^-1, and R chl a ^-1 decreased with the depth, while _{chl a} was constant. P _gmax cell^-1 and R cell^-1 did not change significantly but _cell increased with the depth. Ic decreased with the depth as in the intact corals. Reduction of protein content in a unit area of the coral from deeper habitat implies decrease of host animal tissues. Reduction of Ic can be explained by decrease of R area^-1, which may be due to the diminution of animal tissues. The photoadaptational response to low light intensity of intact Fungia spp. was found to be a combination of the photoadaptation of symbiotic algae and the decrease of host animal tissue. In order to study their adaptation to ultraviolet (UV) radiation, P-I curves of Fungia spp. and isolated zooxanthellae were analyzed before and after UV-B irradiation. 1 h UV-B irradiation showed no effect on the photosynthetic rate of the shallow water (1 m) corals, while it inhibited the photosynthesis of the deep water (30 m) corals and zooxanthellae isolated from both shallow and deep water corals. These results indicate that the host, Fungia spp., in shallow water have protective mechanism for intense UV-B in their habitat. These photoadaptational mechanisms seem to allow the Fungia spp. to have wide vertical distribution where light intensity spans more than two orders of magnitude.     

Floristic and physiological differences between the shallow and the deep nanophytoplankton community in the euphotic zone of the open tropical Atlantic revealed by HPLC analysis of pigments

Suspended matter sampled in 1982 in the North Equatorial Current, in the open Atlantic to the west of West Africa, was analyzed by high performance liquid chromatography. The pigment fingerprint of samples taken in the surface mixed layer was dominated by zeaxanthin and chlorophyll a, in agreement with observed dominance of coccoid cyanobacteria. Near the bottom of the euphotic zone the fingerprint was more complicated, with a sharp transition at the depth of the deep chlorophyll maximum layer to dominance of chlorophyll b, 19-hexanoyloxyfucoxanthin and an unknown fucoxanthin derivative in the lower part of this layer; this fingerprint suggests dominance of eukaryotes (green algae, Prymnesiophyceae and Chrysophyceae) at depth. Up to 90% of the chl a was contained in particles smaller than 8 m, and in the surface mixed layer even more than 50% in particles smaller than 1 m. The high concentration of zeaxanthin relative to chl a near the surface suggests adaptation of the cyanobacteria to exposure to high irradiance. Evidence of this adaptation was the very high specific phytoplankton growth rate between sunrise and sunset (=0.16 h^-1), measured by recording ¹⁴C incorporation into organic carbon and into chl a carbon after isolation of the latter by HPLC. The high concentration of chl b relative to chl a at depth was possibly caused by shade-adapted green algae containing more chl b than chl a. The specific growth rate of the deep shade community was low (<0.04 h^-1), yet net primary production, calculated on the basis of chl a increase during incubation, was greatest at depth.     

A new method for estimating phytoplankton growth rates and carbon biomass

A new method is described for the determination of phytoplankton growth rates and carbon biomass. This procedure is easy to apply and utilizes the labeling of chlorophyll a (chl a) with ¹⁴C. Pure chl a is isolated using two-way thin-layer chromatography, and the specific activity of chl a carbon is determined. Data from laboratory cultures indicate that the specific activity of chl a carbon becomes nearly equal to that of total phytoplankton carbon in incubations lasting 6 to 12 h and can be used to calculate phytoplankton growth rates and carbon biomass. Application of the method to the phytoplankton community in an eutrophic estuary in Hawaii indicates that the cells are growing with a doubling time of about 2 d and that about 85% of the particulate carbon consists of phytoplankton carbon.     

The role of chemosensory behavior of Symbiodinium microadriaticum,intermediate hosts,and host behavior in the infection of coelenterates and molluscs with zooxanthellae

Symbiotic dinoflagellates, Symbiodinium microadriaticum (=zooxanthellae), may gain access to aposymbiotic hosts (i.e., those lacking zooxanthellae) by chemosensory attraction of the motile algae by the potential host or via an intermediate host. Laboratory experiments showed that motile zooxanthellae were attracted to intact aposymbiotic host animals, but not to starved symbiotic hosts. Fed symbiotic hosts and brine shrimp (Artemia sp.) nauplii also attracted motile zooxanthellae. The attraction of these zooxanthellae was directly correlated with nitrogen levels in the seawater surrounding the hosts; thus ammonia and possibly nitrate could be atractants. Brine shrimp nauplii, acting as intermediate hosts actively ingested both motile and non-motile zooxanthellae. the ingested zooxanthellae tended to remain morphologically unaltered during and after passage through the gut of the brine shrimp. Capture and ingestion of brine shrimp containing zooxanthellae by aposymbiotic scyphistomae of the jellyfish Cassiopeia xamachana led to infection of the scyphistomae with zooxanthellae. Zooxanthellae isolated from 17 different species of coelenterates and molluscs could be transferred via brine shrimp to the endodermal cells of the scyphistomae. However only 10 of these isolates persisted to establish a permanent association with C. xamachana. Scyphistomae in suspensions of motile zooxanthellae responded by a classical coelenterate feeding response, which may facilitate ingestion of the potential symbionts and establishment of a symbiosis.     

Effects of feeding frequency and symbiosis with zooxanthellae on nitrogen metabolism and respiration of the coral Astrangia danae

Colonies of the temperate coral Astrangia danae occur naturally with and without zooxanthellae. Basal nitrogen excretion rates of nonsymbiotic colonies increased with increasing feeding frequency [average excretion rate was 635 ng-at N (mg-at tissue-N)^-1 h^-1]. Reduced excretion rates of symbiotic colonies were attributed to N uptake by the zooxanthellae. Nitrogen uptake rates of the zooxanthellae averaged 8 ng-at N (10⁶ cells)^-1 h^-1 in the dark and 21 ng-at N (10⁶ cells)^-1 h^-1 at 200 Ein m^-2 s^-1. At these rates the zooxanthellae could provide 54% of the daily basal N requirement of the coral if all of the recycled N was translocated. Basal respiration rates were 172 nmol O₂ cm^-2 h^-1 for starved colonies and 447 nmol O₂ cm^-2 h^-1 for colonies fed three times per week. There were no significant differences between respiration rates of symbiotic and nonsymbiotic colonies. N excretion and respiration rates of fed (symbiotic and nonsymbiotic) colonies increased greatly soon after feeding. N absorption efficiencies decreased with increasing feeding frequency. A N mass balance, constructed for hypothetical situations of nonsymbiotic and symbiotic (3×10⁶ zooxanthellae cm^-2) colonies, starved and fed 15 g-at N cm^-2wk^-1, showed that the presence of symbionts could double the N growth rate of feeding colonies, and reduce the turnover-time of starved ones, but could not provide all of the N requirements of starved colonies. Rates of secondary production, estimated from rates of photosynthesis and respiration were similar to those estimated for reef corals.     

Symbiotic zooxanthellae enhance boring and growth rates of the tropical sponge Anthosigmella varians forma varians

Several species of boring sponges harbor symbiotic zooxanthellae, and it is believed that the symbiont enhances boring activity of host sponges. This hypothesis was tested using manipulative field experiments to assess the effect of intracellular zooxanthella populations on boring rates of the tropical sponge Anthosigmella varians forma varians. Portions of sponge were attached to 60 calcium carbonate blocks of known weight. Three sets of 10 blocks were grown at high light levels and three sets of 10 blocks were grown at low light levels for 105 d in the Florida Keys, Florida, USA. Boring rates, growth rates (lateral growth and within-substratum tissue penetration), and zooxanthella populations were measured at the end of the experiment. Absolute rates of boring and growth of A. varians forma varians were significantly greater when zooxanthella densities were higher. Boring rate and tissue penetration related to final surface area of sponge attachment was also enhanced when zooxanthella densities were higher, suggesting that the symbiont plays a physiological role in the decalcification process. This is in contrast to the role that zooxanthellae play in coral hosts. Based on the results of this study, it appears that the presence of zooxanthellar symbionts has important ecological and life-history consequences for host sponges. Ability to laterally overgrow competitors will be correlated with the size and activity of zooxanthella populations. In addition, the fitness of host sponges will be enhanced by algal symbionts, since greater penetration within substrata will result in an increase in production of tissue that can be converted into storage, feeding and reproductive functions.     

UV-absorbing substances in zooxanthellate and azooxanthellate clams

The effects of UV-A and UV-B radiation on photosynthesis of zooxanthellae within the siphonal mantle of the giant clam, Tridacna crocea, and in isolation were studied. While UV-B irradiation (2.4 W m⁻², 20 min) completely suppressed photosynthesis of the isolated zooxanthellae, it had little effect on their photosynthetic ability if they were irradiated while within the siphonal mantle of the host tissue. Chemical analysis of the siphonal mantle of T. crocea showed the presence of significant amounts of mycosporine-like amino acids (MAAs), which absorb UV-A and -B light. However, no MAA was detected in the isolated zooxanthellae. MAAs were concentrated in the siphonal mantle and kidney tissues in comparison with other tissues. In the siphonal mantle, MAA concentrations were the highest in the outermost surface layer where most of the zooxanthella cells resided. This indicates that the zooxanthellae are protected from UV radiation by a screen of concentrated MAAs in the host clam. Aside from T. crocea, significant amounts of MAAs were found not only in other zooxanthellate clams, such as T. derasa, Hippopus hippopus, Colculum cardissa and Fragum unedo, but also in a closely related azooxanthellate clam, Vasticardium subrugosum. On the other hand, no MAA was detected in any of the zooxanthellae from these zooxanthellate clams. No MAA was detected in the tissues of a deep-sea bivalve, Calyptogena soyoae. Although MAAs seem to block strong UV radiation in the shallow-water clam, they are probably not essential for the clam's life in the dark. MAAs in shallow-water clams may be derived from food and accumulated in their tissues, especially in the siphonal mantle and kidney. Received: 29 November 1996 / Accepted: 13 January 1997     

Role of carbonic anhydrase in the supply of inorganic carbon to the giant clam—zooxanthellate symbiosis

The mechanism whereby inorganic carbon (C_i) is acquired by the symbiotic association between the giant clam (Tridacna derasa) and zooxanthellae (Symbiodinium sp.) has been investigated. C_i in the haemolymph of the clam is in equilibrium with the surrounding sea water. The photosynthesis rate exhibited by the intact clam varies as a function of the C_i concentration in the clam haemolymph. The gill tissue contains high carbonic anhydrase activity which may be important in adjusting the C_i equilibrium between haemolymph and sea water. Zooxanthellae (Symbiodinium sp.) isolated from the clam mantle prefer CO₂ to HCO ₃ ^- as a source of inorganic carbon. The zooxanthellae have low levels of carbonic anhydrase on the external surface of the cell; however, mantle extracts display high carbonic anhydrase activity. Carbonic anhydrase is absent from the mantle of aposymbiotic clams (T. gigas), indicating that this enzyme may be essential to the symbiosis. The enzyme is probably associated with the zooxanthellae tubes in the mantle. The results indicate that carbonic anhydrase plays an important role in the supply of carbon dioxide within the clam symbiosis.     

Effects of light of altered spectral composition on coral zooxanthellae associations and on zooxanthellae in vitro

Pocillopora damicornis (Linnaeus) and Montipora verrucosa (Lamarck) were collected from Hawaiian reefs. In two experiments (September 1979-January 1980: ca. 4 mo; August-October 1980; ca. 2 mo), these reef corals were grown under sunlight passed through filters producing light fields of similar quantum flux but different spectral composition. In vitro cultures of symbiotic zooxanthellae (Symbiodinium microadriaticum Freudenthal) from M. verrucosa were cultured under similar conditions for 15 d. Blue or white light promoted more coral skeletal growth than green or red light. In both coral species, blue light increased the total amount of chlorophyll a of the coral-zooxanthellae association. In the perforate species, M. verrucosa, the pigment concentration was elevated by an increase in the density of zooxanthellae, but the pigment concentrations per algal cell remained unchanged; in the non-perforate species, P. damicornis, it appears that pigment concentration was elevated by an increase in pigment per algal cell, and not by an increase in density of zooxanthellae. The sunloving reef-flat coral P. damicornis did not grow as rapidly as the shade-species M. verrucosa at the low quantum flux (about 10% sunlight) provided by the experimental treatments. The in vitro cultures of zooxanthellae from M. verrucosa exhibited growth rates in light of altered spectral quality that correlated with the responses of the host coral species: blue and white light supported significantly greater growth than green light, and red light resulted in the lowest growth rate.Contribution No. 678 of the Hawaii Institute of Marine Biology     

Enhancement of chlorophyll a production in Gulf Stream surface seawater by synthetic versus natural rain

Rainwater concentrations of either ammonium or nitrate were sufficient to stimulate chlorophyll a (chl a) production in bioassay experiments using Gulf Stream surface water collected off North Carolina during the summer of 1991. Previous studies primarily examined inshore waters and did not address the impact of rainwater ammonium. An increase in chl a occurred within 1 d of the addition of synthetic rainwater (2 or 5% rainwater, 98 or 95% seawater) containing up to 10 M ammonium; this increase was followed by a decrease in chl a the following day. A similar response to nitrate addition (5% addition of 20 M nitrate rain) was observed. In separate experiments, natural rainwater having nitrate and ammonium concentrations less than those in the experimental synthetic rain yielded a greater chl a response than synthetic rain when added at similar dilutions (0.5 to 5.0% rain). The maximum dissolved inorganic nitrogen concentration in the enriched seawater in these bioassays was 1.8 M; prior to enrichment the maximum was < 0.4 M. Bioassay experiments begun 2 d after a major storm event (sustained NE winds with gusts to 13 m s^-1 and ca. 390 mol m^-2 inorganic nitrogen deposition from rain) showed a chl a increase in response to addition of natural rainwater, but not to synthetic rainwater with similar dissolved inorganic nitrogen concentration. These results suggest that phytoplankton stimulants, in addition to nitrate and ammonium, exist in natural rain but not in the synthetic rain used in these experiments.