Methanotrophic symbiont location and fate of carbon incorporated from methane in a hydrocarbon seep mussel

Seep Mytilid Ia (SMIa), an undescribed mussel found at hydrocarbon seeps in the Gulf of Mexico, harbors intracellular methanotrophic symbionts. Two techniques were used to address the hypothesis that host digestion of symbionts is a significant mechanism of carbon transfer from symbiont to host in the SMIa association: lysosomal enzyme cytochemistry and ¹⁴C tissue autoradiography. Acid phosphatase activity was consistently localized in the Golgi apparatus and associated vesicles of gill cells, but was detected around bacteria in only three of approximately 50 bacteriocytes examined. These results indicate that the cellular equipment necessary for lysosomal digestion of symbionts is present in host bacteriocytes, but that acid phosphatase activity in symbiont vacuoles is rare at a given point in time. Tissue autoradiography was conducted with mussels collected in September 1992 to document carbon fixation by symbionts and follow the time course of transfer to host tissues. No asymbiotic host cell type showed a significant increase in relative grain density until at least 1 d after the end of incubation with ¹⁴C-methane. The ratio of label in the basal portion of bacteriocytes to total bacteriocyte label did not show a significant increase until 10 d after the end of the incubation period, indicating a slow increase of labeled carbon in the putative residual bodies, containing the remnants of lysosomal digestion. These results are consistent with the hypothesis that host digestion of symbionts is one route of nutrient acquisition in SMIa. Intracellular methanotrophic bacteria were found outside of the gill in SMIa juveniles, in mantle and foot epithelial tissues previously believed to be symbiont-free. These extra-gill symbionts and their host cells are morphologically similar to their gill counterparts and, like the gill symbionts, actively fix carbon from methane. Received: 29 March 1997 / Accepted: 12 May 1997     

Response of hydrothermal vent vestimentiferan <Emphasis Type="Italic">Riftia pachyptila</Emphasis> to differences in habitat chemistry

Vestimentiferan tubeworms, which rely on intracellular sulfide-oxidizing autotrophic bacteria for organic carbon, flourish at deep-sea hydrothermal vents despite the erratic nature of their habitat. To assess the degree to which differences in habitat chemistry (sulfide, pH/CO₂) might impact host and symbiont metabolic activity, Riftia pachyptila tubeworms were collected from habitats with low (H₂S < 0.0001 mM) and high (up to 0.7 mM) sulfide concentrations. The elemental sulfur content of the symbiont-containing trophosome organ was lower in specimens collected from the low-sulfide site. Symbiont abundance, RubisCO activity, and trophosome carbon fixation rates were not significantly different for individuals collected from low- versus high-sulfide habitats. Carbonic anhydrase activities were higher in the anterior gas exchange organs of R. pachyptila from the low-sulfide habitat. Despite large differences in habitat chemistry, symbiont abundance and autotrophic potential were consistent, while the host appears to tailor carbonic anhydrase activity to environmental CO₂ availability.     

Cytoenzymatic investigation of intracellular digestion in the symbiont-bearing hydrothermal bivalve <Emphasis Type="Italic">Bathymodiolus azoricus</Emphasis>

Invertebrates harbouring endosymbiotic chemoautotrophic bacteria are widely distributed in a variety of reducing marine habitats, including deep-sea hydrothermal vents. Bathymodiolids are dominants of the biomass at geochemically distinct vent sites of the Mid Atlantic Ridge (MAR) and thus are good candidates to study biological processes in response to site-specific conditions. To satisfy their nutritional requirements, these organisms depend to varying extent on two types of chemoautotrophic symbionts and on filterfeeding. The quantitative relationships of the nutritional modes are poorly understood. Using enzyme cytochemistry, electron microscopy and X-ray microanalysis, the structural and functional aspects of the cellular equipment necessary for lysosomal digestion was studied. We provide evidence for the following: (1) the basis of intracellular digestion of symbionts in Bathymodiolus azoricus from two geochemically distinct vent sites was not mainly in the large lysosomal bodies as previously thought (based on the membranous content resembling bacteria); (2) senescent bacteria are autolysed, possibly by bacterial acid phosphatase, that is more likely a cell cycling of the symbionts rather than an active lysosomal digestion by the host; (3) the consistent absence of hydrolases may indicate the improper use of the name “lysosome” for large vesicles at the base of the gill bacteriocytes (4) nutrient transfer in B. azoricus, therefore, may more likely be accomplished through leaking of metabolites from the symbiont to the host, not excluding lysosomal resorption of dead bacteria as an auxiliary strategy for organic molecule transfer; (5) evidence is provided for microvillar transfer of substances from the seawater that may indicate filter-feeding, in non-symbiotic ciliated gill cells of mussels from Lucky Strike; (6) two types of lysosomal vesicles can be distinguished in digestive cells based on their enzymatic content and their elemental composition.     

A multi-mutualist simulation: Applying biological market models to diverse mycorrhizal communities

We present a cellular automaton that simulates the interaction between a host tree and multiple potential mycorrhizal symbionts and generates testable hypotheses of how processes at the scale of individual root tips may explain mycorrhizal community composition. Existing theoretical biological market models imply that a single host is able to interact with and select from multiple symbionts to organize an optimal symbiont community. When evaluating the tree–symbiont interaction, two scales must be considered simultaneously: the scale of the entire host plant at which carbon utilization and nutrient demands operate, and the scale of the individual root tip, at which colonization and carbon-nutrient trade occurs. Three strategies that may be employed by the host tree for optimizing carbon use and nutrient acquisition through mycorrhizal symbiont communities are simulated: (1) carbon pool adjustment, in which the plant controls only the total amount of carbon to be distributed uniformly throughout the root system, (2) symbiont selection, wherein the plant opts either for or against the interaction at each fine root tip, and (3) selective carbon allocation, wherein the plant adjusts the amount of carbon allocated to each root tip based on the cost of nutrients. Strategies were tested over various nutrient availabilities (the amount of inorganically and organically bound nutrients). Success was defined on the basis of minimizing carbon expended for nutrient acquisition because this would allow more carbon to be utilized for growth and reproduction. In all cases, the symbiont selection and selective carbon allocation strategies were able to meet the nutritional requirements of the plant, but did not necessarily optimize carbon use. The carbon pool adjustment strategy is the only strategy that does not operate at the individual root tip scale, and the strategy was not successful when inorganic nutrients were scarce since there is no mechanism to exclude suboptimal symbionts. The combination of the symbiont selection strategy and the carbon pool adjustment resulted in optimal carbon use and nutrient acquisition under all environmental conditions but result in monospecific symbiont assemblages. On the other hand, the selective carbon allocation strategy is the only strategy that maintained successful, multi-symbiont communities. The simulations presented here thus imply clear hypotheses about the effect of nutrient availability on symbiont selection and mycorrhizal community richness and composition.     

The gill symbiont of the hydrothermal vent mussel Bathymodiolus thermophilus is a psychrophilic,chemoautotrophic, sulfur bacterium

Certain hydrothermal vent invertebrates, e.g. Riftia pachyptila and Calyptogena magnifica, are clearly established as harboring dense populations of chemoautotrophic sulfur bacteria in specialized tissues. By contrast, the physiological characteristics of the abundant intracellular gill symbiont of the vent mussel Bathymodiolus thermophilus have been questioned. The low activities of enzymes diagnostic for CO₂ fixation (Calvin cycle) and for sulfur-driven energy generation, as measured by other investigators, have been attributed to bacterial contamination of the gill surface. Based on research at the Galápagos Rift hydrothermal vents in 1988 and subsequent laboratory experiments, the current study confirms that the B. thermophilus symbiont is a psychrophile for which thiosulfate and sulfide stimulate CO₂ fixation. It strongly indicates that the symbiont is a chemoautotroph by establishing the following: (1) Sulfide and thiosulfate can stimulate CO₂ fixation by partially purified symbionts by up to 43-fold and 120-fold, respectively; (2) the ribulose-1,5-bisphosphate carboxylase/oxygenase activity of the symbiont is sufficient to account for its sulfide- or thiosulfate-stimulated CO₂ incorporation; (3) the symbiont's molar growth yield on thiosulfate, as judged by CO₂ incorporation, is indistinguishable from that of free-living chemoautotrophs. Due to the high protein-degrading activity of B. thermophilus gill lysate, it is also suggested that host lysis of symbionts plays a more important role in the nutrition of the vent mussel than in R. pachyptila or C. magnifica, for which no comparable protein-degrading activity was found.     

Algal symbiosis: A mathematical analysis

Host and algal symbion growth can be described by an iterative model which incorporates utilization efficiencies of host and symbiont. This model predicts that, with input of organic matter to the host and at very low host and algal utilization efficiences coupled with efficient recycling of nutrients between the host and symbionts, production of organic matter by the system can be increased by 2–3 orders of magnitude over that of a system comprised of only autotrophs and heterotrophs. Energy available for growth and respiration by the host is 1–2 orders of magnitude over that available to a heterotroph without symbionts. Algal symbiosis is highly advantageous in oligotrophic environments where radiant energy is abundant, growth-limiting nutrients are scarce and only concentrated in organic matter, and much energy must be expended to capture that organic matter.     

Plant secondary chemistry mediates the performance of a nutritional symbiont associated with a tree-killing herbivore

Many herbivores consume microbial food sources in addition to plant tissues for nutrition. Despite the ubiquity of herbivore-microbe feeding associations, few studies examine how host plant phenotypes affect microbial symbionts of herbivores. We tested the hypothesis that chemical polymorphism in a plant population mediates the performance of nutritional microbial symbionts. We surveyed the composition of ponderosa pine resin in northern Arizona, USA, for variation in six monoterpenes, and we approximated four chemical phenotypes. We reared populations of an herbivorous tree-killing beetle (Dendroctonus brevicomis) in ponderosa pine host material, controlling for three monoterpene compositions representing an alpha-pinene to delta-3-carene gradient. Beetles were reared in host material where the dominant monoterpene was alpha-pinene, delta-3-carene, or a phenotype that was intermediate between the two. We isolated nutritional fungal symbionts (Entomocorticium sp. B) from beetle populations reared in each phenotype and performed reciprocal growth experiments in media amended to represent four "average" monoterpene compositions. This allowed us to test the effects of natal host phenotype, chemical polymorphism, and the interaction between natal host phenotype and chemical polymorphism on a nutritional symbiont. Three important findings emerged: (1) fungal isolates grew 25-32% faster when acquired from beetles reared in the intermediate phenotype; (2) the mean growth rate of nutritional fungi varied up to 44% depending on which monoterpene composition media was amended with; and (3) fungal isolates uniformly performed best in the intermediate phenotype regardless of the chemical composition of their natal host. The performance of nutritional fungi related to both the chemical "history" of their associated herbivore and the chemical phenotypes they are exposed to. However, all fungal isolates appeared adapted to a common chemical phenotype. These experiments argue in favor of the hypothesis that chemical polymorphism in plant populations mediates growth of nutritional symbionts of herbivores. Intraspecific chemical polymorphism in plants contributes indirectly to the regulation of herbivore populations, and our experiments demonstrate that the ecological effects of plant secondary chemistry extend beyond the trophic scale of the herbivore-plant interaction.     

Bacterial symbiosis in Northeast Pacific Vestimentifera: a TEM study

A number of new vestimentiferan species occur at northeast Pacific hydrothermal vent sites. The trophosome and bacterial symbionts of three species, collected from the Juan de Fuca and Explorer Ridges between 1984 and 1986, were studied by transmission electron microscopy (TEM). As in Riftia pachyptila, trophosome tissue is organised into lobules each having an axial blood vessel, and intracellular bacterial symbionts are contained in membrane vacuoles. The bacteria have many cytoplasmic inclusions including tubular membrane systems, glycogen-like particles and poly--hydroxybutyrate (PHB) or sulfur bodies. Glycogen production may be quantitatively important to both the symbionts and the host. Glycogen-like granules appear to first accumulate in the bacterial cells and then be released into the bacteriocyte cytoplasm as bacteria are degraded. Although various stages of bacterial growth and degradation are observed, data are insufficient to verify any across-lobule progression of these processes. Morphological comparison of the symbionts reveals that similar symbionts are found in different vestimentiferan species and that one to two bacterial types exist within single individuals. Two possible models of trophosome function and nutrient exchange are discussed.Deceased     

Carbonic anhydrase in deep-sea chemoautotrophic symbioses

Carbonic anhydrase (CA) facilitates the rapid interconversion of carbon dioxide and bicarbonate. It is ubiquitous among organisms, and participates in numerous cellular processes. In several photoautotrophic marine organisms, CA facilitates uptake of inorganic carbon by catalyzing the reversible dehydration of bicarbonate to carbon dioxide, which diffuses more rapidly across cell membranes. In algal/invertebrate symbioses, this mechanism is correlated with significantly elevated CA activity in tissues where zooxanthellae are housed. Here, we determine whether elevated CA activity is also characteristic of tissues involved in inorganic carbon uptake by chemosynthetic invertebrates. Measurements of CA activity in several chemosynthetic clam and vestimentiferan species indicate that CA facilitates inorganic carbon uptake, with high activities present in clam gill tissue, and vestimentiferan plume and trophosome tissues. However levels of CA activity among species varied widely, from very low in mytilids to extraordinarily high in some vesicomyids and vestimentiferans. High CA activity is generally correlated with other adaptations for control of internal chemical environments where symbionts are housed, such as the presence of serum-borne sulfide-binding moieties. These findings suggest that a CA-based mechanism of carbon uptake may be one of a suite of adaptations to provide symbionts with essential metabolites.     

The influence of symbiont type on photosynthetic carbon flux in a model cnidarian–dinoflagellate symbiosis

We measured the relationship between symbiont diversity, nutritional potential, and symbiotic success in the cnidarian–dinoflagellate symbiosis, by infecting aposymbiotic (i.e. symbiont-free) specimens of the model sea anemone Aiptasia sp. with a range of Symbiodinium types. Four cultured heterologous Symbiodinium types (i.e. originally isolated from other host species) were used, plus both cultured and freshly isolated homologous zooxanthellae (i.e. from Aiptasia sp.). Rates of photosynthesis, respiration, and symbiont growth were measured during symbiosis establishment and used to estimate the contribution of the zooxanthellae to the animal’s respiratory carbon demands (CZAR). Anemones containing Symbiodinium B1 (both homologous and heterologous) tended to attain higher CZAR values and hence benefit most from their symbiotic partners. This was despite Symbiodinium B1 not achieving the highest cell densities, though it did grow more quickly during the earliest stages of the infection process. Rather, the heterologous Symbiodinium types A1.4, E2, and F5.1 attained the highest densities, with populations of E2 and F5.1 also exhibiting the highest photosynthetic rates. This apparent success was countered, however, by very high rates of symbiosis respiration that ultimately resulted in lower CZAR values. This study highlights the impact of symbiont type on the functionality and autotrophic potential of the symbiosis. Most interestingly, it suggests that certain heterologous symbionts may behave opportunistically, proliferating rapidly but in a manner that is energetically costly to the host. Such negative host–symbiont interactions may contribute to the host–symbiont specificity seen in cnidarian–dinoflagellate symbioses and potentially limit the potential for partner switching as an adaptive mechanism.     

There are many ways to be a mutualist: endophytic fungus reduces plant survival but increases population growth

One of the challenges to quantifying the costs and benefits of symbiosis is that symbionts can influence different components of host fitness. To improve understanding of the ecology of inherited symbionts, we developed general theory for a perennial host-hereditary symbiont interaction, in which symbionts can have independent and potentially opposing effects on host regeneration and survival. The model showed that negative effects on one component of fitness may be outweighed by positive effects on another, leading to a net positive impact of symbiosis on population growth. Model predictions depended on the availability of suitable patches, which influenced the relative contributions of survival vs. regeneration to host fitness. We then used experimental symbiont removal to quantify effects of a hereditary, fungal endophyte on a grass host. Endophyte presence strongly reduced host survival but increased regeneration. Application of the model revealed that negative effects on plant survival were overwhelmed by beneficial effects on regeneration, resulting in stable endophyte persistence at 100% frequency, consistent with field observations. Our work demonstrates the utility of a demographic perspective for predicting the dynamics of symbioses and supports the hypothesis that symbionts function as mutualists when host and symbiont fitness are coupled through vertical transmission.     

Phenotypic variations in the gills of the symbiont-containing bivalve Lucinoma aequizonata

The marine bivalve Lucinoma aequizonata (Lucinidae) maintains a population of sulfide-oxidizing chemoautotrophic bacteria in its gill tissue. These are housed in large numbers intracellularly in specialized host cells, termed bacteriocytes. In a natural population of L. aequizonata, striking variations of the gill colors occur, ranging from yellow to grey, brown and black. The aim of the present study was to investigate how this phenomenon relates to the physiology and numbers of the symbiont population. Our results show that in aquarium-maintained animals, black gills contained fewer numbers of bacteria as well as lower concentrations of sulfur and total protein. Nitrate respiration was stimulated by sulfide (but not by thiosulfate) 33-fold in homogenates of black gills and threefold in yellow gill homogenates. The total rates of sulfide-stimulated nitrate respiration were the same. Oxygen respiration could be measured in animals with yellow gills but not in animals with black gills. The cumulative data suggest that black-gilled clams maintained in the aquarium represent a starvation state. When collected from their natural habitat black gills contain the same number of bacteria as yellow gills. Also, no significant difference in glycogen concentrations of the host tissues was observed. Therefore, starvation is unlikely the cause of black gill color in a natural population. Alternative sources of nutrition to sulfur-based metabolism are discussed. Denaturing gradient gel electrophoresis (DGGE) performed on the different gill tissues, as well as on isolated symbionts, resulted in a single gill symbiont amplification product, the sequence of which is identical to published data. These findings provide molecular evidence that one dominant phylotype is present in the morphologically different gill tissues. Nevertheless, the presence of other phylotypes cannot formally be excluded. The implications of this study are that the gill of L. aequizonata is a highly dynamic organ which lends itself to more detailed studies regarding the molecular and cellular processes underlying nutrient transfer, regulation of bacterial numbers and host–symbiont communication. Received: 1 September 1999 / Accepted: 1 February 2000     

High genetic diversity of the symbiotic dinoflagellates in the coral Pocillopora meandrina from the South Pacific

Symbioses between dinoflagellates in the genus Symbiodinium (commonly referred to as zooxanthellae) and scleractinian corals are an essential feature for the maintenance of coral reefs. The fine-scale diversity and population structure of the zooxanthellae inhabiting the coral Pocillopora meandrina, a major reef building species in Polynesia, was examined. We used two polymorphic microsatellites to study seven populations from the South Pacific, whose host structuring has been previously investigated. The symbionts of P. meandrina showed high levels of diversity, with more than one zooxanthella genotype being identified in most of the host individuals. Genetic differentiation between symbiont populations was detected at a large scale (2,000 km) between the Tonga and the Society Archipelagos. Within the Society Archipelago, the two most remote populations (Tahiti and Bora-Bora; 200 km apart) were only weakly differentiated from each other. Statistical tests demonstrated that the symbiont genetic structure was not correlated with that of its host, suggesting that dispersal of the symbionts, whether they are transported within a host larva or free in the water, depends mainly on distance and water currents. In addition, the data suggests that hosts may acquire new symbionts after maternal transmission, possibly following a disturbance event. Lastly, the weak differentiation between symbiont populations of P. verrucosa and P. meandrina, both from Moorea, indicated that there was some host-symbiont fine-scale specificity detectable at the genetic resolution offered by microsatellites.     

A new record of the endosymbiont polychaete Veneriserva (Dorvilleidae), with description of a new sub-species, and relationships with its host Laetmonice producta (Polychaeta: Aphroditidae) in Southern Ocean waters (Antarctica)

A new record of the genus Veneriserva Rossi, 1984 (Polychaeta: Dorvilleidae) is reported, as an endosymbiont in the coelom of the polychaete Laetmonice producta Grube, 1877 (Aphroditidae) in the eastern Weddell Sea and off King George Island (Southern Ocean, Antarctica). The specimens studied were very similar to Veneriserva pygoclava Rossi, 1984; however, due to the greater morphological variability and larger dimensions of our specimens, as well as different host species and geographic locations, a new sub-species, V. pygoclava meridionalis, was erected. A total of 842 specimens of L. producta were examined, 163 of which hosted 209 symbionts (183 in the Weddell Sea samples and 26 in the King George Island samples). Symbiont prevalence was higher in the Weddell Sea samples, and increased with depth (max. 51% at stn 14, 850 m depth). Symbiont intensity was equal to one for 78% and to two for 19.6% of all hosts examined; a maximum of six symbionts per single host was observed. Mean symbiont density was equal to 0.36 and 0.07 for the Weddell Sea and King George Island host populations, respectively. A weak linear relationship was found between symbiont and host size. Eight symbiont specimens (all found at a single station, 850 m depth) were bearing eggs, ranging between 10 and 200 µm in diameter, while 13 specimens were observed in regeneration of the posterior part, suggesting the occurrence of both sexual and asexual reproduction. The way of feeding is still not clear; reduction of the jaw apparatus suggests a parasitic host-symbiont relationship, however, no evident damage was observed in the tissues of the host. These results point out that occurrence of polychaete endoparasites in large aphroditids may be a more frequent and widespread phenomenon than previously believed, and that more attention should be paid to this aspect also in temperate and tropical aphroditid species.     

Carbon metabolism and strobilation in Cassiopea andromedea (Cnidaria: Scyphozoa): Significance of endosymbiotic dinoflagellates

Scyphopolyps and scyphomedusae of Cassiopea andromeda Forskål (Cnidaria, Scyphozoa) containing dinoflagellate endosymbionts (zooxanthellae) were investigated for rates and pathways of carbon fixation. Photosynthesis by the algae, accounting for 80 and 15 mol C h^-1 on a dry weight basis in medusae and polyps, respectively, by far exceeds dark incorporation of inorganic carbon by the intact association. Photosynthetic carbon fixation is operated via C₃ pathway of carbon reduction. DCMU-treatment (1×10^-6 M and 1×10^-5 M) completely inhibits light-dependent carbon assimilation. Major photosynthates presumably involved in a metabolite flow from algal symbionts to animal tissue are glycerol and glucose. A total of 5–10% net algal photosynthate appears to be seleased in vivo to the host. This is probably less than the energy supply ultimately required for the nutrition of the polyps and medusae. The presence of zooxanthellae proved to be indispensable for strobilation in the scyphopolyps. However, photosynthesis by algal symbionts as well as photosynthate release is obviously not essential for the initiation of ephyrae as is shown by DCMU-treatment, culture in continous darkness, and aposymbiotic controls. It is therefore concluded that strobilation is supported, but not triggered by algal photosynthetic activity. The induction of strobilation thus seems to depend on a more complex system of regulation.     

Nemertean,copepod, and amphipod symbionts of the dimorphic ascidian Pyura stolonifera near Melbourne,Australia: specificities to host morphs,and factors affecting prevalences

On the central coast of Victoria. Australia, the dimorphic ascidian Pyura stolonifera (Heller, 1878) harbors three endosymbionts: the nemertean Gononemertes australiensis Gibson, 1974, the copepod Doropygus pulex (Thorell, 1859), and the amphipod Paraleucothoe novaehollandiae (Haswell, 1880). The specificities of these symbionts to two host colour morphs were studied during 1989 to 1991 as part of a multidisciplinary investigation aimed at determining whether the two morphs are genetically distinct. Distributional surveys revealed that nemerteans and copepods occur only in yellow and brown ascidians, respectively, and that amphipods live in both forms. These specificities held true not only when the two morphs were in allopatry, but also in sympatry. These observations, especially the sympatric data, suggest that the two host morphs might be genetically distinct. For example, the two morphs might have different genetically encoded internal milieus that favour the survival of nemerteans in yellow ascidians, and copepods in brown hosts. In transplant experiments, which involved moving ascidian morphs within and between habitats, the wrong symbionts never colonised the wrong hosts. These results, although consistent with the hypothesis of genetic maintenance of specificity, were deemed inconclusive because of the difficulty of establishing reliable controls (i.e. vacant hosts). The relationships between symbiont prevalences and several factors (season, year, site within host, host individual, host habitat, host size/age, host breeding condition, and co-occurrence of other symbiont species) were also analysed. Both simple (e.g. greater prevalences for large hosts) and complex (e.g. prevalence x season x gonad state of host) interactions were detected for all three symbiont species. These are among the very few quantitative analyses of factors affecting prevalences of ascidicolous nemerteans and amphipods. The present report identifies one of very few definite nemertean-ascidian symbioses. Since no differences in gross condition were ever noticed between occupied and vacant hosts, it is suggested that all three symbionts are commensals rather than parasites or mutuals.     

PSII activity and pigment dynamics of Symbiodinium in two Indo-Pacific corals exposed to short-term high-light stress

This study examined the capacity for photoprotection and repair of photo-inactivated photosystem II in the same Symbiodinium clade associated with two coexisting coral species during high-light stress in order to test for the modulation of the symbiont’s photobiological response by the coral host. After 4 days exposure to in situ irradiance, symbionts of the bleaching-sensitive Pocillopora damicornis showed rapid synthesis of photoprotective pigments (by 44 %) and strongly enhanced rates of xanthophyll cycling (by 446 %) while being insufficient to prevent photoinhibition (sustained loss in F _v/F _m at night) and loss of symbionts after 4 days. By contrast, Pavona decussata showed no significant changes in F _v/F _m, symbiont density or xanthophyll cycling. Given the association with the same Symbiodinium clade in both coral species, our findings suggest that symbionts in the two species examined may experience different in hospite light conditions as a result of different biometric properties of the coral host.     

Isolated bacteriocyte cell suspensions from the hydrothermal-vent tubeworm Riftia pachyptila,a potent tool for cellular physiology in a chemoautotrophic symbiosis

The hydrothermal-vent tubeworm Riftia pachyptila relies entirely on its intracellular chemoautotrophic symbionts to sustain its metabolism. The host must therefore provide them with inorganic metabolites, including carbon. This study describes a tool for studying cell processes occurring in a bacteria-containing cell by the dissociation of trophosome cell types. The physiological assays performed on cell preparations focused on carbon dioxide conversion and transport processes. Trophosome tissue was mechanically dissociated, resulting in cell suspensions enriched in small (7-20 µm) bacteriocytes, which were viable for several hours. In addition, medium-term cell cultures were also attempted. As a start to the understanding of the CO₂ metabolism of these cells, we were interested in evidence of carbonic anhydrase (CA) isoforms, ATPases and chloride exchangers. Variations in intracellular and extracellular pH, and in intracellular concentrations of sodium, potassium and chloride, were followed after addition of selective inhibitors. The data presented here suggest the occurrence of potential cytosolic and membrane-associated carbonic anhydrase isoforms in the bacteriocytes, proton-driven sodium-ATPases and a well represented anion transporter exchanging intracellular chloride against extracellular anions.     

A new bathymodioline mussel symbiosis at the Juan de Fuca hydrothermal vents

Until recently, the only major hydrothermal vent biogeographic province not known to include bathymodioline mussels was the spreading centers of the northeast Pacific, but deep-sea dives using DSV Alvin on the Endeavor segment of the Juan de Fuca Ridge (47°56N 129°06W; ∼2,200 m depth) in August 1999 yielded the only recorded bathymodioline mytilids from these northeastern Pacific vents. One specimen in good condition was evaluated for its relatedness to other deep-sea bathymodioline mussels and for the occurrence of chemoautotrophic and/or methanotrophic symbionts in the gills. Phylogenetic analyses of the host cytochrome oxidase I gene show this mussel shares evolutionary alliances with hydrothermal vent and cold seep mussels from the genus Bathymodiolus, and is distinct from other known species of deep-sea bathymodiolines, suggesting this mussel is a newly discovered species. Ultrastructural analyses of gill tissue revealed the presence of coccoid bacteria that lacked the intracellular membranes observed in methanotrophic symbionts. The bacteria may be extracellular but poor condition of the fixed tissue complicated conclusions regarding symbiont location. A single gamma-proteobacterial 16S rRNA sequence was amplified from gill tissue and directly sequenced from gill tissue. This sequence clusters with other mussel chemoautotrophic symbiont 16S rRNA sequences, which suggests a chemoautotrophic, rather than methanotrophic, symbiosis in this mussel. Stable carbon (δ¹³C = −26.6%) and nitrogen (δ¹⁵N = +5.19%) isotope ratios were also consistent with those reported for other chemoautotroph-mussel symbioses. Despite the apparent rarity of these mussels at the Juan de Fuca vent sites, this finding extends the range of the bathymodioline mussels to all hydrothermal vent biogeographic provinces studied to date.     

Temporal and spatial infection dynamics indicate recognition events in the early hours of a dinoflagellate/coral symbiosis

The obligate symbiotic relationship between dinoflagellates, Symbiodinium spp. and reef building corals is re-established each host generation. The solitary coral Fungia scutaria Lamarck 1801 harbors a single algal strain, Symbiodinium ITS2 type C1f (homologous strain) during adulthood. Previous studies have shown that distinct algal ITS2 types in clade C correlate with F. scutaria—Symbiodinium specificity during the onset of symbiosis in the larval stage. The present study examined the early specificity events in the onset of symbiosis between F. scutaria larvae and Symbiodinium spp., by looking at the temporal and spatial infection dynamics of larvae challenged with different symbiont types. The results show that specificity at the onset of symbiosis was mediated by recognition events during the initial symbiont—host physical contact before phagocytosis, and by subsequent cellular events after the symbionts were incorporated into host cells. Moreover, homologous and heterologous Symbiodinium sp. strains did not exhibit the same pattern of localization within larvae. When larvae were infected with homologous symbionts (C1f), ~70% of the total acquired algae were found in the equatorial area of the larvae, between the oral and aboral ends, 21 h after inoculation. In contrast, no spatial difference in algal localization was observed in larvae infected with heterologous symbionts. This result provides evidence of functional differences among gastrodermal cells, during development of the larvae. The cells in the larval equator function as nutritive phagocytes, and also appear to function as a region of enhanced symbiont acquisition in F. scutaria.