Fish are not abundant at hydrothermal vents due to the toxicity of venting fluids. Those that are present usually roam the periphery of the vent field or visit occasionally to feed on the abundance of life supported by chemosynthesis. In the past decade, dense aggregations of a newly described flatfish, Symphurus n.sp, have been observed in association with hydrothermal vents in the western Pacific hydrothermal vent biogeographic province. In this thesis I provide evidence that Symphurus n.sp is a vent obligate and consider the ramifications that this association with hydrothermal vents may have for its distribution, population characteristics, behaviour and diet. Symphurus n.sp has a widespread but disjunct distribution throughout the western Pacific hydrothermal vent biogeographic province. Symphurus n.sp appears to be restricted to hydrothermally active, shallow, sulphur rich seamounts. Symphurus n.sp occurs on unconsolidated volcanoclastic ash and solid sulphur crusts and in close association with molten elemental sulphur. The obvious affinity that this species has for native sulphur is unusual and remains unexplained. Unlike most vent-associated fish, Symphurus n.sp occurs in close contact with point source venting and its distribution extends to the periphery of vent fields but not beyond. The density of flatfish on these seamounts surpasses density estimates of flatfish nursery grounds on the continental shelf. On Daikoku Seamount (Mariana Volcanic Arc), mean flatfish abundances were 100 and 66 individuals m-2 in 2005 and 2006 respectively. The prey items that support such high densities of flatfish vary over spatial scales. Differing prey, in turn, results in differing foraging modes. On Nikko Seamount (Mariana Volcanic Arc), Symphurus n.sp is a “sit and wait” predator that feeds exclusively on a vent endemic shrimp, Opaepele loihi. On other seamounts, Symphurus n.sp is an opportunistic forager that preys mostly on polychaetes and small crustaceans. By counting annuli on otoliths I constructed growth curves and determined that growth rates differ between seamounts. This difference in growth rates is likely due to differences in their diet and foraging strategies. Symphurus n.sp may be allocating more energy to growth when less energy is required to forage. Furthermore, size distributions also differ between populations, likely due to variability in growth rates as well as differences in strong recruitment years.

We investigated oceanic dissolved inorganic nitrogen (N) dynamics, focussing on processes removing bio-available N and ultimately affecting primary productivity, in sulfidic hydrothermal vent fluids discharging from the subsurface on the Juan de Fuca Ridge (northeast Pacific Ocean) and in anoxic bottom waters of Saanich Inlet, a British Columbia fjord, using a combination of geochemical and molecular microbial ecology techniques. During episodes of mixing with oxygenated sea-water, both systems can switch from anoxic to oxic conditions. Strong inter-site variations in the concentrations and δ15N of ammonium (NH4+) in high-T fluids suggested different N sources (deep-sea nitrate (NO3-) versus organic sediments) for hydrothermally discharged NH4+. Increase in the isotopic composition of NO3- (δ15N and δ18O), concomitant with decreased [NO3-], indicated NO3- assimilation or denitrification in the subsurface. NO3- isotope anomalies, i.e. deviations from the 15N:18O isotopic enrichment of 1:1 in marine environments, were observed and confirmed the occurrence of NO3- regeneration in vent fluids. Denitrification was the dominant N-loss pathway, suggesting that bacterial denitrification out-competes anaerobic NH4+ oxidation (anammox) in diffuse hydrothermal vent waters. The diversity of denitrifying bacteria encoding the nirS-form of nitrite reductase was low in vent fluids. Quantitative polymerase chain reaction (qPCR) analysis revealed that denitrifiers accounted for up to 38% (nirk-encoding γ-proteobacteria of the SUP05 cluster) and 8% (nirS-encoding bacteria) of the total bacterial abundance. Furthermore, nirS gene operational taxonomic units from two vent fields clustered into different groups in the phylogenetic tree, suggesting a link between denitrifying bacterial community membership and small-scale geographic isolation and/or fluid physico-chemical properties. Significant correlations existed between fixed N-loss rates and in-situ dissolved inorganic N deficits in the fluids. Based on our rate measurements, and on published data on hydrothermal fluid fluxes and residence times, we estimated that up to ~10 Tg N yr−1 could be removed globally in the subsurface biosphere. In Saanich Inlet, a gradual increase in both the δ15N and δ18O of NO3- associated with a decrease in [NO3-] and an increase in biological excess N2, was observed after bottom water renewal events in fall 2008, following NO3- consumption by denitrifiers in an expanding suboxic zone. N-to-O negative NO3- isotope anomalies were observed in surface and bottom waters, confirming the occurrence of NO3- regeneration and/or external NO3- input. Closed and open-system-model derived NO3- isotope effects in anoxic bottom waters were lower (as low as ~11‰) than the ~25‰ for water column denitrification reported in other studies, suggesting that ~50% of the denitrification could occur, with a highly suppressed isotope effect, in the sediments of the Inlet. / Graduate

Thesis (M.S.)--Miami University, Dept. of Geology, 2004. / Title from first page of PDF document. Includes bibliographical references (p. 126-129).

Bathymodiolus azoricus est un bivalve endémique des sources hydrothermales de la dorsale Médio Atlantique qui présente une forte capacité à accumuler différents métaux dans ses tissus, la qualifiant comme espèce modèle en écotoxicologie. L’objectif de ce travail a consisté à décrire les mécanismes moléculaires de tolérance et de détoxication des métaux, que ce soit au sein des populations naturelles ou durant des expositions expérimentales, par le biais d’approches chimiques (quantification des métaux) et transcriptomiques (PCR quantitative et puces à ADN), afin de comprendre comment les facteurs environnementaux pouvaient influencer l’expression des gènes de B. azoricus et d’identifier des biomarqueurs potentiels utiles pour des études écotoxicologiques en environnement extrême. Les mesures de bioaccumulation de différents métaux (Cd, Cu, Fe, Pb,…) sur des individus provenant de sites aux caractéristiques contrastées ont révélé des patrons spécifiques aux populations et aux tissus examinés chez cette espèce, suggérant des phénomènes d’organotropisme. Nos résultats suggèrent également que les bactéries symbiotiques des branchies pourraient participer à la tolérance aux métaux et aux processus de détoxication. Des tendances comparables sont observées pour l’expression relative de certains gènes candidats impliqués dans la réponse au stress métallique, par exemple les métallothionéines, les superoxyde dismutases, les ferritines et les phytochélatines. Les réponses distinctes observées pour certaines populations ou certains tissus traduisent des différences d’état physiologique qui ne seraient pas directement reliées avec l’accumulation de métaux. En outre, l’analyse des puces à ADN à une échelle globale nous a permis d’identifier les réseaux métaboliques principaux pour chaque population et chaque tissu. / Bathymodiolus azoricus is an endemic bivalve from hydrothermal vents in the Mid-Atlantic ridge, which is known to accumulate different types of metals in various tissues at high concentrations, and has therefore been granted model species for exotoxicology. The objective of this work is to describe the mechanisms of metal tolerance and detoxification as they occur in natural populations and during experimental exposures, using both chemical (metal quantification) and transcriptomic (qPCR and microarrays) approaches to understand how environmental factors influence gene expression response in B. azoricus and to identify potential biomarkers useful for ecotoxicological studies in extreme environments. Bioaccumulation of different metals (Cd, Cu, Fe, Pb,…) was measured on individuals from contrasted vent sites, revealing specific population patterns and tissue differences for this species that could be related to processes of organotropism. Our results also suggest that the symbiotic bacteria in gills may be involved in metal tolerance and detoxification. Similar variation trends are observed in the relative expression of candidate genes involved in response to metal stress, e.g. metallothioneins, superoxide dismutase, ferritin, and phytochelatin, revealing contrasted responses at population and tissue level, and reflecting differences in physiological status not directly correlated with the accumulation of metals. In addition, global scale microarray analysis allowed us to identify the principal biological pathways representative of each population and tissues.

L'importance des symbioses dans l'évolution du vivant est désormais admise et les associations symbiotiques sont observées dans une grande diversité d'habitats. Notre étude porte sur une symbiose au sein d'un écosystème réduit, les sources hydrothermales de l'océan profond. Bathymodiolus azoricus est un bivalve hydrothermal vivant le long de la ride Médio-Atlantique, qui héberge dans des cellules branchiales spécialisées, deux types de γ-protéobactéries différentes : des méthanotrophes (MOX) et des sulfo-oxydantes (SOX). Ces dernières sont capables d'oxyder les composés réduits présents dans le fluide hydrothermal fournissant ainsi énergie et/ou source de carbone à leur hôte. Cette double endosymbiose est plastique ainsi, l'abondance relative du type de symbionte hébergé (SOX vs. MOX) varie en fonction des concentrations en composés réduits présent dans le milieu (H2S, CH4). L'objectif de ce travail de thèse est d'identifier les acteurs moléculaires impliqués dans l'acquisition, le maintien et la régulation des bactéries symbiotiques. Pour ce faire, une analyse OMICs globale (protéomique -nano LC-MS/MS- et transcriptomique -micro-array-) a été mise en ¿uvre sur des individus symbiotiques issus de population naturelle (site hydrothermal Lucky Strike, -1700m) et sur des individus ayant expérimentalement perdu ou maintenu leurs symbiotes. Suite à cette approche globale et exploratoire, une approche plus spécifique a été menée sur des familles de protéines impliquées dans des processus immunitaire et/ou d'interactions hôte/symbiotes. Cette thèse apporte un éclaircissement sur les mécanismes régissant les relations et la communication hôte/symbiote. / Hydrothermal vents are located on the mid-ocean ridges, and are characterized by challenging physico-chemical conditions. Despite these conditions dense hydrothermal communities develop down around hydrothermal fluid emissions. The presence of marine invertebrates relies on their capacity to cope with these challenging factors, and, for those forming most of the biomass, on their ability to live in symbiosis with chemoautotrophic bacteria. Bathymodiolus azoricus is one of these symbiotic species that harbors two types of γ-proteobacteria, a sulfide-oxidizing bacterium (SOX) (using the oxidation of H2S as the source of energy and CO2 as source of carbon) and a methane-oxidizing bacterium (MOX) (that uses the oxidation of CH4 as both a source of energy and carbon). These bacteria are located in specific epithelial cells in the gill tissue of the mussel. The proportion and number of these symbiont types (SOX vs. MOX) in B.azoricus can change in response to environmental conditions, and especially on the relative concentration of reduced compounds. The aim of our study is to understand the molecular mechanisms of acquisition, regulation and maintenance of the symbiotic charge in B .azoricus gills. We therefore, performed a global OMICs analysis (proteomics –nano LC-MS/MS and transcriptomics- micro-array) on mussels from natural population (Lucky Strike, -1700m) and on mussels that experimentally loose or maintain their symbiotic rate. This exploratory approach was followed by a more specific approach on family of proteins involved in immunity process and/or in host/symbiont interactions. This PhD provides hypotheses on the mechanisms governing the relationship and communication between host and symbionts.

Thesis (Ph.D.)--Joint Program in Oceanography (Massachusetts Institute of Technology, Dept. of Earth, Atmospheric, and Planetary Sciences and the Woods Hole Oceanographic Institution), 2000. / Vita. / Includes bibliographical references (leaves 179-187). / A spectral camera (ALISS - Ambient Light Imaging and Spectral System) was used to image ambient light from high-temperature vents at 9°N East Pacific Rise and the Juan de Fuca Ridge during 1997 and 1998 Alvin dive cruises. ALISS is a low-light digital camera with custom-designed optics. A set of nine lenses, each covered by an individual bandpass filter (50 and 100 nm nominal bandwidths), allows vents to be imaged in nine wavelength bands simultaneously spanning the range of 400-1000 nm. Thus, both spatial and spectral information are obtained. ALISS was used to image three types of vents: black smokers, flange pools, and beehives. The primary source of light is thermal radiation due to the high temperature of the hydrothermal fluid (~350°C). This light is dominant at wavelengths greater than 700 nm. At flange pools, where the fluid is relatively stable, only thermal radiation is present. Black smokers and beehives, however, are subject to mixing with ambient seawater (2°C) leading to mineral precipitation. Data from these types of vents show the existence of non-thermal, temporally varying light in the 400-700 nm region. This light is probably caused by mechanisms related to mixing and precipitation, such as chemiluminescence, crystalloluminescence and triboluminescence. / by Sheri N. White. / Ph.D.

Despite over four decades of research on deep-sea hydrothermal vent ecosystems, major gaps remain in our understanding of these systems. Knowledge of microeukaryote diversity, abundance, and involvement in ecosystem function lags far behind that of prokaryotes, and contributions of the non-endosymbiotic microbiome in faunal assemblages to ecosystem processes and overall hydrothermal vent microbial diversity are not known. This research addresses these gaps using high-throughput sequencing of 16S/18S rRNA genes and metagenomes from vent and surrounding non-vent habitats encompassing diffuse hydrothermal fluids, plumes, deep seawater, and microbes in assemblages of the foundation tubeworm species Ridgeia piscesae. Co-occurrence/covariance is a central method used, first, between prokaryotes with known extreme habitat preferences and microeukaryotes to infer potential endemism in the latter, and then between microbes and fauna in R. piscesae assemblages to infer interspecies interactions. Microeukaryote distribution and abundance suggest potential vent endemic microeukarya are infrequently encountered, potentially in low abundance, and belong to novel lineages of Rhizaria and Stramenopila. Potential endemism is inferred for relatives of known apusomonads, excavates, and some clades of Syndiniales. R. piscesae assemblages are shown to be hotspots of microbial taxonomic richness and exhibit a robust temperature-driven distinction in assemblage composition above and below ~25°C spanning micro, meio and macro size classes and microbial domains (Bacteria, Archaea, and micro-Eukarya). Likely interacting faunal and microbial taxa among R. piscesae assemblages are identified as ‘core communities’, which included eight macro- and meiofaunal taxa and members of the Bacteroidetes and Epsilonbacteraeota in highT communities (>25°C) and more meiofaunal species in addition to Alpha- and Gammaproteobacteria, and Actinobacteria in lowT communities (<25°C). Core communities were used to guide metagenomic investigations of microbial functional potential. Exploratory metagenomic analysis required development of new methods to deal with compositional data. ‘Enrichment leanings’ were developed to prioritize in-depth functional comparisons between sample types, which revealed clades within core community microbes with differing functional potential between highT and lowT assemblages and between assemblages and fluids. The balance of autotrophy-heterotrophy genes and patterns of genes for different carbon, nitrogen, and sulfur-cycling processes were tested as potential metrics of community-level function but did not distinguish assemblages by highT/lowT designations. This research brings us closer to understanding hydrothermal vent ecosystem function and suggests sizeable continued discovery potential. / Graduate / 2022-06-08

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21830号 / 農博第2343号 / 新制||農||1068(附属図書館) / 学位論文||H31||N5202(農学部図書室) / 京都大学大学院農学研究科応用生物科学専攻 / (主査)教授 佐藤 健司, 教授 澤山 茂樹, 准教授 豊原 治彦 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

The Juan de Fuca Ridge, located 400 km off the coast of Washington State, is home to unstable and unpredictable hydrothermal vent sites where chemosynthetic communities flourish. In 2007 the manned submersible ALVIN retrieved a Ridgeia piscesae tubeworm community in its entirety from the side of the Tower sulfide edifice from the Endeavor segment of the Juan de Fuca Ridge (47 55.416720 N, 129 6.487020 W, at a depth of 2269 m) using the Bushmaster Jr. collection device. The collection was analyzed for community structure and the data collected were compared to that from several other hydrothermal vent communities. It was determined that substrate composition is a factor that heavily influences community structure. The data were then compared to the community succession model developed by Sarrazin et. al. in 1997 and 1999 (Sarrazin et. al. 1997, Sarrazin and Juniper 1999). The Tower community was found to expand the model as a new community succession classification; that of community iii low flow. The Tower community was then analyzed for diversity, structure and tubeworm morphology in conjunction with two other communities from differing substrata. The Ridgeia piscesae tubeworms were found to be of the "long skinny" morphotype, one that was previously thought to only reside on basaltic substrate. The Tower community has similar species richness and higher species evenness than those from basaltic substrate, but similar richness and lower evenness that those from sulfide. This community type combines the characteristics of those from both substrata, resulting in a community with diversity and structure that is an intermediary between sulfide and basaltic substrates. / Biology

Metagenomics has revealed that the marine microbial biosphere is immensely more diverse than originally considered, and is an almost untapped reservoir for the potential discovery of microbial natural products. Despite numerous advances in culturing, biosynthetic engineering and genomic-based screening efforts to uncover much of this diversity in relatively accessible environments, a high rediscovery rate has resulted in the investigation of unique, relatively unexplored ecosystems harboring phylogenetically diverse communities of marine organisms. The focus of this research was to establish a culture repository of microorganisms collected from the Red Sea and from deep-sea hydrothermal vents, and to assess their biosynthetic potential for the production of new chemical scaffolds. Cultivation of marine cyanobacteria from the Red Sea has led to the identification of five new cyclic depsipeptides, apratoxin H, grassypeptolides D and E, Ibu-epidemethoxylyngbyastin 3 and leptochelin, the latter possessing a unique chemical scaffold capable of binding metals. A collection of deep-sea hydrothermal vent sediment and microbial mat samples led to the isolation of 64 unique bacterial strains, with eight assigned as members of the order Actinomycetales. Importantly, these isolates, along with a collection of deep-vent invertebrates and microbes, have led to the development of methods for the collection, culturing and biological screening of organisms from this extreme environment for future natural products research. / Graduation date: 2013