Bacteria In Gills Research Articles

Host-Microbiome Interactions in a Changing Sea: The Gill Microbiome of an Invasive Oyster under Drastic Temperature Changes.

The gill tissue of bivalve mollusks hosts rich symbiotic microbial communities that may contribute to host health. Spondylus spinosus is an invasive Lessepsian oyster in the Eastern Mediterranean Sea that has become highly abundant while constantly expanding its range northwestward. Using 16S rRNA gene amplicon sequencing, we examined how temperature affects S. spinosus oysters and their gill microbiota in a series of experiments: exposing them to the current annual seawater temperature range, to the colder temperature of the Western Mediterranean Sea, and to the elevated temperature as predicted under global warming scenarios. The bacterial genus Endozoicomonas dominated the communities of the S. spinosus, mainly upon exposure to winter-like (16 °C) temperatures. Exposure to the elevated seawater temperature resulted in a significant change in the bacterial communities, while the oysters maintained normal functioning, suggesting that the oyster may survive a seawater warming scenario. Exposure to 11 °C led to the health deterioration of the oysters, the emergence of opportunistic pathogens, such as Arcobacter, Vibrio, Colwelliaceae, and Pseudoalteromonas, and a decline in the relative abundance of Endozoicomonas, suggesting that S. spinosus might not survive Western Mediterranean Sea winters. Both the host and its gill bacteria are thus greatly affected by temperature, which could consequently restrict the range of expansion of this and other invasive oysters.

Association between melanin deposits in gill tissue and microbiome across different hatchery reared Atlantic salmon.

To investigate the relationship between microbial community profiles and gill pathology during a production cycle of Atlantic salmon in two commercial hatcheries. Relationships between gill histology, environmental conditions, and microbiome were determined using high-throughput data, including 16S rDNA amplicon sequencing data, histopathology data, and water quality parameters. Hatchery A used riverine water and operated a mixed system of recirculation aquaculture system (RAS) and flowthrough. Hatchery B was used bore water and operated a RAS. Melanin deposits, hyperplastic, and inflammatory lesions were observed histologically in the gills. A higher prevalence of melanin deposits was detected and correlated to a change in beta diversity of bacterial communities in early time points (fingerling and parr stages). High abundance of Sphaerotilus sp.,Pseudomonas sp.,Nitrospira sp.,Exiguobacterium sp.,Deinococcus sp.,and Comamonas sp. was correlated with a high prevalence of melanin in filaments. Bacterial diversity increased as the fish cohort transitioned from RAS to flowthrough in hatchery A. Under commercial conditions, the commensal community of gill bacteria was related to melanin prevalence.

Genomic and Environmental Factors Shape the Active Gill Bacterial Community of an Amazonian Teleost Holobiont.

Fish bacterial communities provide functions critical for their host's survival in contrasting environments. These communities are sensitive to environmental-specific factors (i.e., physicochemical parameters, bacterioplankton), and host-specific factors (i.e., host genetic background). The relative contribution of these factors shaping Amazonian fish bacterial communities is largely unknown. Here, we investigated this topic by analyzing the gill bacterial communities of 240 wild flag cichlids (Mesonauta festivus) from 4 different populations (genetic clusters) distributed across 12 sites in 2 contrasting water types (ion-poor/acidic black water and ion-rich/circumneutral white water). Transcriptionally active gill bacterial communities were characterized by a 16S rRNA metabarcoding approach carried on RNA extractions. They were analyzed using comprehensive data sets from the hosts genetic background (Genotyping-By-Sequencing), the bacterioplankton (16S rRNA) and a set of 34 environmental parameters. Results show that the taxonomic structure of 16S rRNA gene transcripts libraries were significantly different between the 4 genetic clusters and also between the 2 water types. However, results suggest that the contribution of the host's genetic background was relatively weak in comparison to the environment-related factors in structuring the relative abundance of different active gill bacteria species. This finding was also confirmed by a mixed-effects modeling analysis, which indicated that the dissimilarity between the taxonomic structure of bacterioplanktonic communities possessed the best explicative power regarding the dissimilarity between gill bacterial communities' structure, while pairwise fixation indexes (FST) from the hosts' genetic data only had a weak explicative power. We discuss these results in terms of bacterial community assembly processes and flag cichlid fish ecology. IMPORTANCE Host-associated microbial communities respond to factors specific to the host physiology, genetic backgrounds, and life history. However, these communities also show different degrees of sensitivity to environment-dependent factors, such as abiotic physico-chemical parameters and ecological interactions. The relative importance of host- versus environment-associated factors in shaping teleost bacterial communities is still understudied and is paramount for their conservation and aquaculture. Here, we studied the relative importance of host- and environment-associated factors structuring teleost bacterial communities using gill samples from a wild Amazonian teleost model (Mesonauta festivus) sampled in contrasting habitats along a 1500 km section of the Amazonian basin, thus ensuring high genetic diversity. Results showed that the contribution of the host's genetic background was weak compared to environment-related bacterioplanktonic communities in shaping gill bacterial assemblages, thereby suggesting that our understanding of teleost microbiome assembly could benefit from further studies focused on the ecological interplay between host-associated and free-living communities.

First Field-Based Evidence That the Seagrass-Lucinid Mutualism Can Mitigate Sulfide Stress in Seagrasses

Seagrass meadows form vital ecological components of coastal zones worldwide, but are rapidly declining. Large-scale seagrass diebacks have been related to accumulation of toxic sulfide in the sediment, a phenomenon predicted to occur more frequently in the near future due to ongoing global warming and increasing organic loading of coastal systems worldwide. Recently, a facultative mutualism between seagrasses and lucinid bivalves with endosymbiotic sulfide-consuming gill bacteria was discovered that may prevent toxic sulfide accumulation in seagrass sediments. Yet, direct field-based evidence for the importance of this mutualism in alleviating sulfide stress in seagrasses is currently lacking, as well as how its role may change when sediment sulfide levels increase due to environmental change. Here, we investigated the sulfide detoxification function of this seagrass-lucinid mutualism and its resilience to organic-loading induced sulfide stress in a temperate lagoon system (Thau lagoon, France), using a correlative field survey and a full factorial field experiment. The field survey revealed a strong positive correlation between seagrass above-ground biomass and lucinid densities, and pore water sulfide concentrations close to zero at all sites. Furthermore, the field experiment revealed that addition of organic matter (starch mixed with sucrose) increased sedimentary sulfide intrusion into seagrass (Zostera noltei) leaves (a proxy for sulfide stress), while experimentally enhanced lucinid densities reduced sulfide intrusion, regardless of addition of organic matter. Moreover, addition of organic matter reduced seagrass rhizome biomass and increased pore water sulfide levels, lucinid tissue sulfur content, lucinid condition (expressed as flesh/shell dry weight ratio), and total lucinid biomass, while enhancement of lucinid densities reduced lucinid condition. These results provide the first field-based evidence that lucinid bivalves and their sulfide-oxidizing gill symbionts mitigate sulfide stress in seagrasses, and suggests that the dependence of seagrass on this seagrass-lucinid mutualism will increase under conditions of enhanced sediment sulfide production, as predicted for the near future. Therefore, we suggest that awareness of the ecological importance of the seagrass-lucinid mutualism may be instrumental for designing new measures for improving long-term restoration success and seagrass resilience to global change.

Bacterial spoilage profiles in the gills of Pacific oysters (Crassostrea gigas) and Eastern oysters (C. virginica) during refrigerated storage

Microorganisms harbored in oyster gills are potentially related to the spoilage and safety of oyster during storage. In this study, the microbial activities and pH changes of the gills of the two species, Crassostrea gigas and C. virginica, harvested from three different sites were determined and sensory evaluation was conducted during refrigerated storage. The bacteria in gills with an initial aerobic plate count (APC) of 3.1–4.5 log CFU/g rose remarkably to 7.8–8.8 log CFU/g after 8-days of storage. The APC of Enterobacteriaceae increased from 2.5 to 3.6 log CFU/g to 4.5–4.8 log CFU/g, and that of lactic acid bacteria (LAB) fluctuated in the range of 1.4–3.0 log CFU/g during the whole storage period. The results of sensory analysis indicated that the oysters had 8-days of shelf-life and that the gill presented the fastest deterioration rate. The pH of all samples showed a decrease in the early stages followed by an increased after 4-days of storage. The dynamic changes in microbial profiles were depicted to characterize gill spoilage by Illumina Miseq sequencing to characterize gill spoilage. The results revealed that oysters harvested at different sites showed common bacterial profiles containing Arcobacter, Spirochaeta, Pseudoalteromonas, Marinomonas, Fusobacterium, Psychrobacter, Psychromonas, and Oceanisphaera when spoiled, especially, among which Psychrobacter and Psychromonas (psychrotrophic genus) were represented as the most important gill spoiled bacteria during refrigerated storage, and Arcobacter with pathogenic potential was the dominated bacteria in all spoiled oysters. The consumption quality and safety of refrigerated oysters could be greatly improved by targeted control of bacteria in oyster gills according to the results the present study provided.

Isolation and identification of prevalent bacterial pathogens from an exotic fish Tilapia zillii and Oreochromis mossambicus

Aim: Fishes are susceptible to a wide variety of bacterial pathogens which are part of the natural microflora of the environment. The aim of the present study was to isolate the pathogenic bacteria in gills, liver, gut, and skin of Tilapia zillii and Oreochromis mossambicus from the lake near Perumbakkam in Chennai. Materials and Methods: The experimental fish T. zillii and O. mossambicus were collected from large lake 2 km length which starts from Perumbakkam through Vengaivasal Chithalapakkam in Chennai. Results and Discussion: Based on the differentiation and characterization of isolates on specific culture media, human bacterial pathogen such as Escherichia coli, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Enterococcus faecalis, and Shigella dysenteriae was isolated. In this study, the total plate count in the tissues of T. zillii ranged between 7.14 × 105 and 23.2 105 cfu/g. Similarly, in O. mossambicus, the total cell count in the tissue ranged between 6.21 × 105 and 21.16 × 105, respectively. The isolation of enteric bacteria in fish serves as indicator organisms of fecal contamination and/or water pollution. Their presences are also hazardous to humans. Conclusion: The association of pathogenic bacteria among the fishes suggests that if fish are handled or prepared improperly it could lead to food safety hazards for consumers.

A facultative mutualistic feedback enhances the stability of tropical intertidal seagrass beds

Marine foundation species such as corals, seagrasses, salt marsh plants, and mangrove trees are increasingly found to engage in mutualistic interactions. Because mutualisms by their very nature generate a positive feedback between the species, subtle environmental impacts on one of the species involved may trigger mutualism breakdown, potentially leading to ecosystem regime shifts. Using an empirically parameterized model, we investigate a facultative mutualism between seagrass and lucinid bivalves with endosymbiotic sulfide-oxidizing gill bacteria in a tropical intertidal ecosystem. Model predictions for our system show that, by alleviating the build-up of toxic sulfide, this mutualism maintains an otherwise intrinsically unstable seagrass ecosystem. However, an increase in seagrass mortality above natural levels, due to e.g. desiccation stress, triggers mutualism breakdown. This pushes the system in collapse-and-recovery dynamics (‘slow-fast cycles’) characterized by long-term persistent states of bare and seagrass-dominated, with rapid transitions in between. Model results were consistent with remote sensing analyses that suggest feedback-mediated state shifts induced by desiccation. Overall, our combined theoretical and empirical results illustrate the potential of mutualistic feedbacks to stabilize ecosystems, but also reveal an important drawback as small environmental changes may trigger shifts. We therefore suggest that mutualisms should be considered for marine conservation and restoration of seagrass beds.

Symbiotic Bacteria in Gills and Guts of Chinese Mitten Crab (Eriocheir sinensis) Differ from the Free-Living Bacteria in Water.

Aquatic animals have a close relationship with water, but differences in their symbiotic bacteria and the bacterial composition in water remains unclear. Wild or domestic Chinese mitten crabs (Eriocheir sinensis) and the water in which they live were collected from four sampling sites in Jiangsu and Shanghai, China. Bacterial composition in water, gills or guts of E. sinensis, were compared by high-throughput sequencing using 16S rRNA genes. Analysis of >660,000 sequences indicated that bacterial diversity was higher in water than in gills or guts. Tenericutes and Proteobacteria were dominant phyla in guts, while Actinobacteria, Proteobacteria and Bacteroidetes were dominant in gills and water. Non-metric multidimensional scaling analysis indicated that microbiota from gills, guts or water clearly separated into three groups, suggesting that crabs harbor a more specific microbial community than the water in which they live. The dominant OTUs in crab gut were related to Mycoplasmataceae, which were low in abundance in gills, showing that, like mammals, crabs have body-site specific microbiota. OTUs related to Ilumatobacter and Albimonas, which are commonly present in sediment and seawater, were dominant in gills but almost absent from the sampled water. Considering E. sinensis are bottom-dwelling crustacean and they mate in saline water or seawater, behavior and life cycle of crabs may play an important role in shaping the symbiotic bacterial pattern. This study revealed the relationship between the symbiotic bacteria of Chinese mitten crab and their habitat, affording information on the assembly factors of commensal bacteria in aquatic animals.

A new intracellular bacterium, Candidatus Similichlamydia labri sp. nov. (Chlamydiaceae) producing epitheliocysts in ballan wrasse, Labrus bergylta (Pisces, Labridae).

Certain wrasse species (Labridae) are used as cleaner fish in salmon farms on the Norwegian coast, reducing salmon louse intensities. The pathogen repertoire of wrasse in Norway is poorly known, and the objective of the present study is to describe a novel intracellular bacterium detected in Norwegian Labrus bergylta. Histological examination of gill tissues from ballan wrasse, L. bergylta, revealed epitheliocysts occurring basally to the secondary lamellae in the interlamellar epithelium. Ultrastructurally, these had bacteria-filled inclusions with thickened membranes and radiating ray-like structures (actinae). 16S rRNA gene sequences from the gill bacteria showed the highest (97.1 %) similarity to Candidatus Similichlamydia latridicola from the gills of the latrid marine fish Latris lineata in Australia and 94.9 % similarity to Candidatus Actinochlamydia clariae, causing epitheliocystis in the freshwater catfish Clarias gariepinus in Uganda. A total of 47 gill samples from L. bergylta from Western Norway were screened by real time RT-PCR with an assay targeting Candidatus Actinochlamydiaceae 16S rRNA. Prevalence was 100 %. We propose the name Candidatus Similichlamydia labri sp. nov. for this new agent producing gill epitheliocysts in L. bergylta.

Gill bacteria enable a novel digestive strategy in a wood-feeding mollusk.

Bacteria play many important roles in animal digestive systems, including the provision of enzymes critical to digestion. Typically, complex communities of bacteria reside in the gut lumen in direct contact with the ingested materials they help to digest. Here, we demonstrate a previously undescribed digestive strategy in the wood-eating marine bivalve Bankia setacea, wherein digestive bacteria are housed in a location remote from the gut. These bivalves, commonly known as shipworms, lack a resident microbiota in the gut compartment where wood is digested but harbor endosymbiotic bacteria within specialized cells in their gills. We show that this comparatively simple bacterial community produces wood-degrading enzymes that are selectively translocated from gill to gut. These enzymes, which include just a small subset of the predicted wood-degrading enzymes encoded in the endosymbiont genomes, accumulate in the gut to the near exclusion of other endosymbiont-made proteins. This strategy of remote enzyme production provides the shipworm with a mechanism to capture liberated sugars from wood without competition from an endogenous gut microbiota. Because only those proteins required for wood digestion are translocated to the gut, this newly described system reveals which of many possible enzymes and enzyme combinations are minimally required for wood degradation. Thus, although it has historically had negative impacts on human welfare, the shipworm digestive process now has the potential to have a positive impact on industries that convert wood and other plant biomass to renewable fuels, fine chemicals, food, feeds, textiles, and paper products.

Molecular detection of prokaryote and protozoan parasites in the commercial bivalve Ruditapes decussatus from southern Portugal

A parasite screening combining histological and molecular techniques was performed on healthy grooved carpet shell clams collected from a commercial shellfish bed in Southern Portugal. The study included the first attempt to develop molecular techniques to detect and identify Rickettsia/Chlamydia-like bacteria and gill ciliates infecting this high-value bivalve. Although the animals failed to reveal significant pathologies relatable to infectious agents, both techniques detected low-moderate levels of infection by bacteria and protozoans, the latter including the apicomplexan Perkinsus olseni, the ciliate Boveria subcylindrica and a yet unclassified haplosporidian. Infections by P. olseni and bacteria were the most frequent and most disseminated within the two surveyed organs, gills and digestive glands. Gill and digestive gland bacteria belonged to distinct groups, the former more related to Rickettsiales and the latter to Chlamydiales. However, while gill bacteria could be clearly allocated within the Spongiobacter/Endocoizomonas group, thus likely belonging to Oceanospirillales, no evident taxonomic position could be attributed to digestive gland bacteria, most possibly consisting of a new species of parasite. In face of the current findings, symbiotic or commensal relationships between these bacteria and the bivalve should not be excluded. The results showed that healthy clams act as a repository of multiple agents of infection, which is of particular importance to a species that is known to be particularly sensitive to parasitism. The PCR techniques developed for the detection of Boveria sp. and bacteria were proven efficient and were appended to the methods for Perkinsus sp. and haplosporidians described elsewhere.

Phylogenetic Characterization of Episymbiotic Bacteria Hosted by a Hydrothermal Vent Limpet (Lepetodrilidae, Vetigastropoda)

Marine invertebrates hosting chemosynthetic bacterial symbionts are known from multiple phyla and represent remarkable diversity in form and function. The deep-sea hydrothermal vent limpet Lepetodrilus fucensis from the Juan de Fuca Ridge complex hosts a gill symbiosis of particular interest because it displays a morphology unique among molluscs: filamentous bacteria are found partially embedded in the host's gill epithelium and extend into the fluids circulating across the lamellae. Our objective was to investigate the phylogenetic affiliation of the limpet's primary gill symbionts for comparison with previously characterized bacteria. Comparative 16S rRNA sequence analysis identified one γ- and three ε-Proteobacteria as candidate symbionts. We used fluorescence in situ hybridization (FISH) to test which of these four candidates occur with the limpet's symbiotic gill bacteria. The γ-proteobacterial probes consistently hybridized to the entire area where symbiotic bacteria were found, but fluorescence signal from the ε-proteobacterial probes was generally absent. Amplification of the γ-proteobacterial 16S rRNA gene using a specific forward primer yielded a sequence similar to that of limpets collected from different ridge sections. In total, direct amplification or FISH identified a single γ-proteobacterial lineage from the gills of 23 specimens from vents separated by a distance up to about 200 km and collected over the course of 2 years, suggesting a highly specific and widespread symbiosis. Thus, we report the first filamentous γ-proteobacterial gill symbiont hosted by a mollusc.

Persistence, morphology, and nutritional state of a gastropod hosted bacterial symbiosis in different levels of hydrothermal vent flux

The limpet, Lepetodrilus fucensis McLean, is found in prominent stacks around hydrothermal vents on the Juan de Fuca Ridge. L. fucensis hosts a filamentous episymbiont on its gill lamellae that may be ingested directly by the gill epithelium. To assess the persistence of this symbiosis I used microscopy to examine the gills of L. fucensis from sites representing its geographic range and different habitats. The symbiosis is present on all the specimens examined in this study, including both sexes and a range of juvenile and adult sizes. Next, I aimed to determine if patterns in bacterial abundance, host condition, and gill morphology support the hypotheses that the bacteria are chemoautotrophic and provide limpets with a food resource. To do so, I compared specimens from high and low flux locations at multiple vents. My results support the above hypotheses: (1) gill bacteria are significantly less abundant in low flux where the concentrations of reduced chemicals (for chemoautotrophy) are negligible, (2) low flux specimens have remarkably poor tissue condition, and (3) the lamellae of high flux limpets have greater surface area: the blood space and bacteria-hosting epithelium are deeper and have more folds than low flux lamellae, modifications that support higher symbiont abundances. I next asked if the morphology of the lamellae could change. To test this, I moved high flux limpets away from a vent and after 1 year the lamellar depth and shape of the transplanted specimens resembled low flux gills. Last, I was interested in whether bacterial digestion by the gill epithelium is a significant feeding mechanism. As bacteria-like cells are rarely apparent in lysosomes of the gill epithelium, I predicted that lysosome number would be unrelated to bacterial abundance. My data support this prediction, suggesting that digestion of bacteria by the gill epithelium probably contributes only minimally to the limpet’s nutrition. Overall, the persistence and morphology of the L. fucensis gill symbiosis relates to the intensity of vent flux and indicates that specimens from a variety of habitats may be necessary to characterize the morphological variability of gill-hosted symbioses in other molluscs.

Influence of salmonid gill bacteria on development and severity of amoebic gill disease

The relationship between salmonid gill bacteria and Neoparamoeba sp., the aetiological agent of amoebic gill disease (AGD) was determined in vivo. Fish were divided into 4 groups and were subjected to following experimental infections: Group 1, amoebae only; Group 2, Staphylococcus sp. and amoebae; Group 3, Winogradskyella sp. and amoebae; Group 4, no treatment (control). Fish (Groups 1, 2 and 3) were exposed to potassium permanganate to remove the natural gill microflora prior to either bacterial or amoebae exposure. AGD severity was quantified by histological analysis of gill sections to determine the percentage of lesioned filaments and the number of affected lamellae within each lesion. All amoebae infected groups developed AGD, with fish in Group 3 showing significantly more filaments with lesions than other groups. Typically lesion size averaged between 2 to 4 interlamellar units in all AGD infected groups. The results suggest that the ability of Neoparamoeba sp. to infect filaments and cause lesions might be enhanced in the presence of Winogradskyella sp. The possibility is proposed that the prevalence of more severe AGD is due to the occurrence of Winogradskyella sp. at high concentrations on the gills.

Utilization of methane and carbon dioxide by symbiotrophic bacteria in gills of Mytilidae (Bathymodiolus) from the Rainbow and Logachev hydrothermal fields on the Mid-Atlantic Ridge

Bivalve mollusks Bathymodiolus asoricus and Bathymodiolus puteoserpentis collected from the Rainbow and Logachev hydrothermal fields during dives of Mir 1 and Mir 2 deep-sea manned submersibles were studied. Rates of methane oxidation and carbon dioxide assimilation in mussel gill tissue were determined by radiolabel analysis. During oxidation of 14Ch4, radiocarbon was detected in significant quantities not only in carbon dioxide but also in dissolved organic matter, most notably 14C-formate and 14C-acetate, occurring in a 2:1 ratio. Activities of hexulose-phosphate synthase, phosphoribulokinase, and ribulose 1,5-bisphosphate carboxylase were shown in the soluble fraction of gill tissue cells. At the same time, no activity of hydroxypyruvate reductase--the key enzyme of the serine pathway of C1-assimilation--was detected. The results of PCR amplification using genetic probes for membrane-bound methane monooxygenase (pmoA) and methanol dehydrogenase (mxaF) attest to the presence of the genes of these enzymes in the total DNA extracted from gill samples. However, no appropriate PCR responses were obtained with the mmoX primer system, which is a marker for soluble methane monooxygenase. All samples studied showed amplification with primers for the genera Methylobacter and Methylosphaera. At the same time, no genes specific to the genera Methylomonas, Methylococcus, Methylomicrobium, or Methylosinus and Methylocystis were detected. Electron microscopic examinations revealed the presence of two groups of endosymbiotic bacteria in the mussel gill tissue. The first group was represented by large cells possessing a complex system of cytoplasmic membranes, typical of methanotrophs of morphotype I. The other type of endosymbionts, having much smaller cells and lacking intracellular membrane structures, is likely to be constituted by sulfur bacteria.

AMS 14C Measurements of Dissolved Inorganic Carbon in Pore Waters from a Deep-Sea “Cold Seep” Giant Clam Community Off Hatsushima Island, Sagami Bay, Japan

We collected pore waters using an in situ pore water-squeezer for a submersible Shinkai 2000 at six depths beneath the sediment surface within a deep-sea “cold seep” giant clam community off Hatsushima Island, Sagami Bay, Japan. A box core sample was also collected ca. 4.5 km east of the community and pore waters were separated. Dissolved inorganic carbon (DIC) was extracted and purified in a vacuum line and 14C concentration was determined with a Tandetron accelerator mass spectrometer at Nagoya University after conversion to graphite targets using a batch Fe-catalytic hydrogen reduction method. ∆14C values decreased with increasing depth to −938‰ at the sulfate concentration minimum. This indicates that methane used for the active reduction of sulfate and formation of hydrogen sulfide, which is used by symbiotic chemoautotrophic bacteria in gills of the giant clams, is almost dead and is likely supplied from the deep. ∆14C values of DIC vary linearly with δ13C values along a mixing line between that in the bottom water and that produced by the oxidation of dead methane. The δ13C value of DIC oxidized from dead methane is estimated to be ca. −45‰.

Virus-like particles in bacteria symbiotic in bivalve gills

Thyasira gouldi,an arctic bivalve mollusc of the family Thyasiridae, has symbiotic, probably chemoautotrophic, bacteria living in its gills. In specimens collected from Loch Etive in Scotland in 1989, after the local population had undergone an unexplained decline, the gill bacteria contained numerous virus-like particles 60 nm in diameter. The bacteria are not lysed by the virus, and the particles are not present outside the bacteria except in phagocytic vacuoles of the gill cells where the bacteria have been digested, leaving the virus particles unchanged. Similar particles have been discovered in bacterial symbionts ofThyasira flexuosa,but in much smaller numbers.

Chemoautotrophic metabolism of intracellular gill bacteria in the marine bivalve Spisula subtruncata (da Costa)

Spisula substruncata, a mactrid bivalve living in well-oxygenated sublittoral sandy bottoms, contains two types of intracellular bacteria in its gills. Chemoautotrophic metabolism of these bacteria was tested in different ways. Homogenates of gill tissues showed a slight ribulose — 1.5 — bisphosphate carboxylase activity. Fixation of 14C-labelled bicarbonate in S. subtruncata was higher than in Venus gallina, a clam without bacteria living in the same environment. Other experiments were performed with and without two reduced compounds (sodium thiosulphate and ammonium chloride) at different incubation times (45 to 300 min). With the two compounds, the levels of fixed carbon per g of gill tissue increased significantly after 60 min incubation. A possible ammonia utilization by gill bacteria is demonstrated and may be explained by the fact that the most abundant dissolved reduced compound available for chemoautotrophs is ammonia. The values obtained in this association were lower than those recorded for other bivalves with bacterial cells in their gills. These low values may be explained by the relatively small number of the two types of bacteria observed in the gills of S. subtruncata.

Observations on Australian Solemyidae

Abstract Two species of Australian Solemyidae are examined. Gross structure, the condition of the gut, the histological organization of the gills, and the presence of intracellular gill bacteria are noted. Solemya velesiana (Iredale) is a small, intertidal protobranch with large ctenidia, and a functional gut. The species oxidizes sulphide and intracellular gill symbionts appear to contribute to its economy. Museum specimens of a species assigned to Solemya australis Lamarck also have large ctenidia with intracellular bacteria. The gut of this species is much reduced and the organism thus must be more dependent on sulphide-oxidizing symbiosis.