Genus Beggiatoa Research Articles

Mechanism of Intracellular Elemental Sulfur Oxidation in Beggiatoa leptomitoformis, Where Persulfide Dioxygenase Plays a Key Role.

Representatives of the colorless sulfur bacteria of the genus Beggiatoa use reduced sulfur compounds in the processes of lithotrophic growth, which is accompanied by the storage of intracellular sulfur. However, it is still unknown how the transformation of intracellular sulfur occurs in Beggiatoa representatives. Annotation of the genome of Beggiatoa leptomitoformis D-402 did not identify any genes for the oxidation or reduction of elemental sulfur. By searching BLASTP, two putative persulfide dioxygenase (PDO) homologs were found in the genome of B. leptomitoformis. In some heterotrophic prokaryotes, PDO is involved in the oxidation of sulfane sulfur. According to HPLC-MS/MS, the revealed protein was reliably detected in a culture sample grown only in the presence of endogenous sulfur and CO2. The recombinant protein from B. leptomitoformis was active in the presence of glutathione persulfide. The crystal structure of recombinant PDO exhibited consistency with known structures of type I PDO. Thus, it was shown that B. leptomitoformis uses PDO to oxidize endogenous sulfur. Additionally, on the basis of HPLC-MS/MS, RT-qPCR, and the study of PDO reaction products, we predicted the interrelation of PDO and Sox-system function in the oxidation of endogenous sulfur in B. leptomitoformis and the connection of this process with energy metabolism.

Lipoxygenase in a giant sulfur bacterium: an evolutionary solution for size and complexity?

Discovery of Thiomargarita magnifica - an exceptionally large giant sulfur bacterium - urges us to pay additional attention to the giant sulfur bacteria and to revisit our recent bioinformatic finding of lipoxygenases in the representatives of the genus Beggiatoa. These close relatives of Thiomargarita magnifica meet the similar size requirements by forming multicellular structures. We hypothesize that their lipoxygenases are a part of the oxylipin signaling system that provides high level of cell-to-cell signaling complexity which, in turn, enables them to reach large sizes.

Lipoxygenase in a Giant Sulfur Bacterium: An Evolutionary Solution for Size and Complexity?

Discovery of Thiomargarita magnifica - an exceptionally large giant sulfur bacterium - urges us to pay additional attention to the giant sulfur bacteria and to revisit our recent bioinformatic finding of lipoxygenases in the representatives of the genus Beggiatoa. These close relatives of Thiomargarita magnifica meet the similar size requirements by forming multicellular structures. We hypothesize that their lipoxygenases are a part of the oxylipin signaling system that provides high level of cell-to-cell signaling complexity which, in turn, enables them to reach large sizes.

Microbial Sulfide Filter along a Benthic Redox Gradient in the Eastern Gotland Basin, Baltic Sea

The sediment-water interface is an important site for material exchange in marine systems and harbor unique microbial habitats. The flux of nutrients, metals, and greenhouse gases at this interface may be severely dampened by the activity of microorganisms and abiotic redox processes, leading to the “benthic filter” concept. In this study, we investigate the spatial variability, mechanisms and quantitative importance of a microbially-dominated benthic filter for dissolved sulfide in the Eastern Gotland Basin (Baltic Sea) that is located along a dynamic redox gradient between 65 and 173 m water depth. In August-September 2013, high resolution (0.25 mm minimum) vertical microprofiles of redox-sensitive species were measured in surface sediments with solid-state gold-amalgam voltammetric microelectrodes. The highest sulfide consumption (2.73–3.38 mmol m−2 day−1) occurred within the top 5 mm in sediments beneath a pelagic hypoxic transition zone (HTZ, 80–120 m water depth) covered by conspicuous white bacterial mats of genus Beggiatoa. A distinct voltammetric signal for polysulfides, a transient sulfur oxidation intermediate, was consistently observed within the mats. In sediments under anoxic waters (>140 m depth), signals for Fe(II) and aqueous FeS appeared below a subsurface maximum in dissolved sulfide, indicating a Fe(II) flux originating from older sediments presumably deposited during the freshwater Ancylus Lake that preceded the modern Baltic Sea. Our results point to a dynamic benthic sulfur cycling in Gotland Basin where benthic sulfide accumulation is moderated by microbial sulfide oxidation at the sediment surface and FeS precipitation in deeper sediment layers. Upscaling our fluxes to the Baltic Proper; we find that up to 70% of the sulfide flux (2281 kton yr−1) toward the sediment-seawater interface in the entire basin can be consumed at the microbial mats under the HTZ (80–120 m water depth) while only about 30% the sulfide flux effuses to the bottom waters (>120 m depth). This newly described benthic filter for the Gotland Basin must play a major role in limiting the accumulation of sulfide in and around the deep basins of the Baltic Sea.

Beggiatoa leptomitoformis sp. nov., the first freshwater member of the genus capable of chemolithoautotrophic growth.

A strain of filamentous sulfur bacteria was isolated from freshwater spring contaminated with residential and agricultural wastewater in Moscow region, Russia. According to the results of phylogenetic analysis, strain D-402T belonged to the genus Beggiatoa within the family Beggiatoaceae of the class Gammaproteobacteria. Within the genus Beggiatoa, strain D-402T was most closely related to Beggiatoa alba strains. Strain D-402T had a DNA G+C content 42.1 mol%. The DNA-DNA hybridization value between strain D-402T and Beggiatoa alba strain B15LD was 33 %. Predominant fatty acids were C18 : 1 (46.1 and 53.3 %), C16 : 0 (15.5 and 16.2 %) and C16 : 1 (32.9 and 25.4 %) for strains D-402T and B15LD, respectively. In contrast to known representatives of Beggiatoa, strain D-402T was capable of chemolithoautotrophic growth with sulfide and thiosulfate as electron donors. Oxidation of sulfide and thiosulfate was accompanied by deposition of sulfur globules within the cells. Strain D-402T was capable of heterotrophic growth. The strain was capable of using different organic compounds, sulfur compounds and hydrogen as electron donors. Based on these observations, strain D-402T is considered as a representative of a species Beggiatoa leptomitoformis sp. nov. of the genus Beggiatoa. The type strain is D-402T (=DSM 14946T=UNIQEM U 779T).

Temporal fluctuations in the trophic role of large benthic sulfur bacteria in mangrove sediment

Filamentous sulfur bacteria of the genus Beggiatoa form large mats covering the sediment in the shallow waters of a Guadeloupean mangrove (French West Indies). The abundance of these bacteria varies over the year and their trophic role may, therefore, also vary. We investigated this variation by conducting a survey examining the stable isotopic compositions of four grazers and four food sources during nine sampling sessions in three different periods of the year. We analyzed bulk isotopic compositions for each component except for the bacterial and diatom communities, for which we carried out a compound-specific 13C analysis of phospholipid-derived fatty acids (PLFAs). Correlations between isotopic compositions revealed a predominance of diatoms in the diet of nematodes and the important role of detritus and bacteria in the diet of the polychaete Ceratocephale sp. None of the grazers had an isotopic composition correlated with that of Beggiatoa suggesting that sulfur bacteria were not a predominant part of the diet of any grazer. Beggiatoa has a large central vacuole, resulting in a very low carbon content-to-volume ratio, potentially accounting for its low level of attractiveness to grazers. Mangrove sediments are habitats rich in organic carbon, in which, the addition of a food source, such as Beggiatoa, would have a limited effect on the structure of the food web over the course of the year.

Barite encrustation of benthic sulfur‐oxidizing bacteria at a marine cold seep

Crusts and chimneys composed of authigenic barite are found at methane seeps and hydrothermal vents that expel fluids rich in barium. Microbial processes have not previously been associated with barite precipitation in marine cold seep settings. Here, we report on the precipitation of barite on filaments of sulfide-oxidizing bacteria at a brine seep in the Gulf of Mexico. Barite-mineralized bacterial filaments in the interiors of authigenic barite crusts resemble filamentous sulfide-oxidizing bacteria of the genus Beggiatoa. Clone library and iTag amplicon sequencing of the 16S rRNA gene show that the barite crusts that host these filaments also preserve DNA of Candidatus Maribeggiatoa, as well as sulfate-reducing bacteria. Isotopic analyses show that the sulfur and oxygen isotope compositions of barite have lower δ(34)S and δ(18)O values than many other marine barite crusts, which is consistent with barite precipitation in an environment in which sulfide oxidation was occurring. Laboratory experiments employing isolates of sulfide-oxidizing bacteria from Gulf of Mexico seep sediments showed that under low sulfate conditions, such as those encountered in brine fluids, sulfate generated by sulfide-oxidizing bacteria fosters rapid barite precipitation localized on cell biomass, leading to the encrustation of bacteria in a manner reminiscent of our observations of barite-mineralized Beggiatoa in the Gulf of Mexico. The precipitation of barite directly on filaments of sulfide-oxidizing bacteria, and not on other benthic substrates, suggests that sulfide oxidation plays a role in barite formation at certain marine brine seeps where sulfide is oxidized to sulfate in contact with barium-rich fluids, either prior to, or during, the mixing of those fluids with sulfate-containing seawater in the vicinity of the sediment/water interface. As with many other geochemical interfaces that foster mineral precipitation, both biological and abiological processes likely contribute to the precipitation of barite at marine brine seeps such as the one studied here.

Spatial heterogeneity and underlying geochemistry of phylogenetically diverse orange and white Beggiatoa mats in Guaymas Basin hydrothermal sediments

Sulfide-oxidizing bacteria of the genus Beggiatoa are found in conspicuous, colorful mats on the seafloor above active hydrothermal seeps at Guaymas Basin. Guaymas Beggiatoa filaments fall into discrete size classes representing at least five separate 16S rRNA phylotypes, and appear either white, yellow, or orange. During two R/V Atlantis cruises to Guaymas Basin, 78 temperature profiles were taken near and within 15 different orange and white Beggiatoa mats by the Alvin submersible to investigate spatial relationships between mat color and hydrothermal fluid seeps, as indicated by elevated temperatures. The surface temperatures from 78 profiles are similar to each other (on average 8–12°C, warmer than bare sediments at 3–4°C), indicating that Guaymas Basin Beggiatoa spp., although relying on the hydrothermal system for energy and carbon sources, live within a relatively cool temperature range. Temperatures from 40cm below orange Beggiatoa versus white Beggiatoa are the same, at 84°C averaged across all mat systems. However, within a single mat system, temperatures are higher beneath the predominantly orange center of the mat than beneath the white mat periphery. Push core transects across the orange-to-white color change of three Beggiatoa mats showed stronger upward compression of isotherms and metabolic zones beneath the orange mat center than beneath white mat periphery. Hydrothermal temperature gradients push the microbial processes generating carbon and energy sources for Beggiatoa mats towards the sediment surface. The resulting steep gradients of hydrothermal electron donors and carbon sources to the sediment surface, rather than the in situ temperature by itself, control the relative positioning of orange and white filaments within a Guaymas Basin Beggiatoa mat. Given the wide spectrum of temperature and hydrothermal flux regimes between different mats, the orange/white pattern represents a relative preference or even a competitive balance among different Beggiatoa types that establishes itself within each hydrothermal hot spot.

Unusual polyphosphate inclusions observed in a marine Beggiatoa strain

Sulfide-oxidizing bacteria of the genus Beggiatoa are known to accumulate phosphate intracellularly as polyphosphate but little is known about the structure and properties of these inclusions. Application of different staining techniques revealed the presence of unusually large polyphosphate inclusions in the marine Beggiatoa strain 35Flor. The inclusions showed a co-occurrence of polyphosphate, calcium and magnesium when analyzed by scanning electron microscopy and energy dispersive X-ray analysis. Similar to polyphosphate-enriched acidocalcisomes of prokaryotes and eukaryotes, the polyphosphate inclusions in Beggiatoa strain 35Flor are enclosed by a lipid layer and store cations. However, they are not notably acidic. 16S rRNA gene sequence-based phylogenetic reconstruction showed an affiliation of Beggiatoa strain 35Flor to a monophyletic branch, comprising other narrow vacuolated and non-vacuolated Beggiatoa species. The polyphosphate inclusions represent a new type of membrane surrounded storage compartment within the genus Beggiatoa, distinct from the mostly nitrate-storing vacuoles known from other marine sulfide-oxidizing bacteria of the family Beggiatoaceae.

Filamentous sulfur bacteria, Beggiatoa spp., in arctic marine sediments (Svalbard, 79°N)

Fjord sediments on the west coast of the arctic archipelago Svalbard were surveyed to understand whether large filamentous sulfur bacteria of the genus Beggiatoa thrive at seawater temperatures permanently near freezing. Two sediments had abundant populations of Beggiatoa, while at six sites, only sporadic occurrences were observed. We conclude that Beggiatoa, although previously unnoticed, are widespread in these arctic fjord sediments. Beggiatoa ranged in diameter from 2 to 52 microm and, by those tested, stored nitrate in vacuoles at up to 260 mM. The 16S rRNA gene sequence of a 20-microm-wide filament is closely associated with other large, marine, nitrate-storing Beggiatoa. The Beggiatoa mostly occurred in the upper 2-5 cm of oxidized surface sediment between oxygen and the deeper sulfidic zone. In spite of a very low or an undetectable sulfide concentration, sulfate reduction provided abundant H(2)S in this zone. The total living biomass of Beggiatoa filaments at one study site varied over 3 years between 1.13 and 3.36 g m(-2). Because of their large size, Beggiatoa accounted for up to 15% of the total prokaryotic biomass, even though the filament counts at this site were rather low, comprising <1/10,000 of the bacterial numbers on a cell basis.

Comparative analysis of Beggiatoa from hypersaline and marine environments

The main criterion to classify a microorganism as belonging to the genus Beggiatoa is its morphology. All multicellular, colorless, gliding bacterial filaments containing sulfur globules described so far belong to this genus. At the ultrastructural level, they show also a very complex cell envelope structure. Here we describe uncultured vacuolated and non-vacuolated bacteria from two different environments showing all characteristics necessary to assign a bacterium to the genus Beggiatoa. We also intended to investigate whether narrow and vacuolate Beggiatoa do differ morphologically as much as they do phylogenetically. Both large, vacuolated trichomes and narrow filaments devoid of vacuoles were observed. We confirmed the identity of the narrow filaments by 16S rRNA phylogenetic analysis. The diameters of the trichomes ranged from 2.4 to 34 μm, and their lengths ranged from 10 μm to over 30 mm. Narrow trichomes moved by gliding at 3.0 μm/s; large filaments moved at 1.5 μm/s. Periplasmic sulfur inclusions were observed in both types of filaments, whereas phosphorus-rich bodies were found only in narrow trichomes. On the other hand, nitrate vacuoles were observed only in large trichomes. Ultra-thin section transmission electron microscopy showed differences between the cell ultrastructure of narrow (non-vacuolated) and large (vacuolated) Beggiatoa. We observed that cell envelopes from narrow Beggiatoa consist of five layers, whereas cell envelopes from large trichomes contain four layers.

Methane seepage along the Hikurangi Margin, New Zealand: Overview of studies in 2006 and 2007 and new evidence from visual, bathymetric and hydroacoustic investigations

This paper is an introduction to and an overview of papers presented in the Special Issue of Marine Geology “Methane seeps at the Hikurangi Margin, New Zealand”. In 2006 and 2007, three research cruises to the Hikurangi Margin at the east coast of New Zealand's North Island were dedicated to studying methane seepage and gas hydrates in an area where early reports suggested they were widespread. Two cruises were carried out on RV TANGAROA and one on RV SONNE using the complete spectrum of state-of-the-art equipment for geophysics (seismic, sidescan, controlled source electromagnetics, ocean bottom seismometers and hydrophones, singlebeam and multibeam), seafloor observations (towed camera systems, ROV), sediment and biological sampling (TV-guided multi-corer, gravity-corer, grab, epibenthic sled), deployment of in-situ observatories (landers) as well as water column sampling and oceanographic studies (CTD, moorings). The scientific disciplines involved ranged from geology, geophysics, petrography, geochemistry, to oceanography, biology and microbiology. These cruises confirmed that a significant part of the Hikurangi Margin has been active with locally intense methane seepage at present and in the past, with the widespread occurrence of dead seep faunas and knoll-forming carbonate precipitations offshore and on the adjacent land. A close link to seismically detected fluid systems and the outcropping of the base of the gas hydrate stability zone can be found at some places. Pore fluid and free gas release were found to be linked to tides. Currents as well as density layers modulate the methane distribution in the water column. The paper introduces the six working areas on the Hikurangi Margin, and compiles all seep locations based on newly processed multibeam and multibeam backscatter data, water column hydroacoustic and visual data that are combined with results presented elsewhere in this Special Issue. In total, 32 new seep sites were detected that commonly show chemoherm-type carbonates or carbonate cemented sediment with fissures and cracks in which calyptogenid clams and bathymodiolid mussels together with sibloglinid tube worms live. White bacterial mats of the genus Beggiatoa and dark gray beds of heterotrophic ampharetid polychaetes typically occur at active sites. Bubble release has frequently been observed visually as well as hydroacoustically (flares) and geochemical analyses show that biogenic methane is released. All seep sites, bubbling or not, were inside the gas hydrate stability zone. Gas hydrate itself was recovered at three sites from the seafloor surface or 2.5 m core depth as fist-sized chunks or centimeter thick veins. The strong carbonate cementation that in some cases forms 50 m high knolls as well as some very large areas being paved with clam shells indicates very strong and long lasting seep activity in the past. This activity seems to be less at present but nevertheless makes the Hikurangi Margin an ideal place for methane-related seep studies in the SW-Pacific.

Microbial community composition of black band disease on the coral host Siderastrea siderea from three regions of the wider Caribbean

Microbial communities associated with black band disease (BBD) on colonies of the reef building coral Siderastrea siderea from reefs in 3 regions of the wider Caribbean were studied using 16S rRNA gene-targeted amplification, cloning, and sequencing. Samples were collected from 7 BBD-infected S. siderea colonies on 2 reefs near Lee Stocking Island, Bahamas, one reef in the Florida Keys, Florida, USA, and 2 reefs in St. Croix, US Virgin Islands. Genomic DNA extracted from BBD samples was PCR-amplified with universal bacterial primers (27F and 1492R). Seven clone libraries were constructed and 411 sequences were retrieved. All of the clone libraries were domi- nated by Alphaproteobacteria and contained sequences associated with bacteria of the sulfur cycle, including the first report of the molecular detection of sequences related to the sulfide-oxidizing genus Beggiatoa in BBD field samples. Additionally, all clone libraries had sequence types of bacte- ria associated with toxin producing dinoflagellates. These sequences were most abundant in a sewage impacted reef site in St. Croix, which also had the highest prevalence of BBD-infected colonies. Non-metric multidimensional scaling and cluster analysis showed clustering of BBD micro- bial communities by relative level of anthropogenic impact. The results support the hypothesis that BBD is composed of variable members of distinct physiological and toxin associated bacterial groups and that the microbial community variation is associated with environmental differences such as anthropogenic impact. We propose that with degrading water quality (i.e. increasing nutrients) cer- tain proteobacteria thrive and increase BBD virulence.

Regulation of Metabolic and Electron Transport Pathways in the Freshwater Bacterium Beggiatoa leptomitiformis D-402

The biomass yield of freshwater filamentous sulfur bacteria of the genus Beggiatoa, when grown lithoheterotrophically or mixotrophically, has been shown to increase 2 to 2.5 times under microaerobic conditions (0.12 mg/l oxygen) as compared to aerobic conditions (9 mg/l oxygen). The activity of the glyoxylate cycle key enzymes have been found to increase two to three times under microaerobic conditions (at an O2 concentration of 2 mg/l), and the activities of the sulfur metabolism enzymes increased three to five times (at an O2 concentration of 0.1-0.5 mg/l). It has also been found that, under microaerobic conditions, thiosulfate was almost completely oxidized to sulfate by the bacteria, without accumulation of intermediate metabolites. At the same time, a 2- to 15-fold decrease in the activities of the tricarboxylic acid cycle enzymes involved in the reduction of NAD and FAD was observed. Reorganization of the respiratory chain after changes in aeration and type of nutrition was also observed. It has been found that, in cells grown heterotrophically, the terminal part of the respiratory chain contained an aa3-type oxidase, whereas, during mixotrophic, lithoheterotrophic, and autotrophic growth, aa3-type oxidase synthesis was inhibited, and the synthesis of a cbb3-type oxidase, which is induced under microaerobic conditions, was activated. The gene of the catalytic subunit CcoN of the cbb3-type oxidase was sequenced and proved to be highly homologous to the corresponding genes of other proteobacteria.

Dynamics of nematodes in a high organic loading rotating biological contactors

Nematode diversity and dynamics of a full-scale rotating biological contactor plant (RBC) has been studied. Analysis of biofilm composition showed a well-established zoning of microfauna among the three RBC sections analysed. Nematodes appeared to be the dominant group within the larger microfauna populations with average abundances between 200 and 300 ind/mg or 8000 and 17 000 ind/cm 2. The most abundant nematode species were Diplogasteritus nudicapitatus and Paroigolaimella coprophages and, to a lesser extent, Paroigolaimella bernensis and Steinernema intermedia. The relationship between nematodes and filamentous bacteria (specifically the genus Beggiatoa) was the most significant biotic relationship found, and to a lesser extent, nematodes with ciliates. The relationship between the abundance of nematode species and the physical–chemical variables suggests that nematodes may be good indicators of low pollutant load levels in the entry of the RBC system. Finally, the results indicate that nematodes may have a relevant role for a good biofilm development.

Purification and physicochemical properties of malate dehydrogenase from bacteria of the genus Beggiatoa.

Homogeneous malate dehydrogenase (MDH) with a specific activity of 20-24 units per mg protein was purified from the sulfur bacterium Beggiatoa leptomitiformis strain D-402 grown organotrophically and lithotrophically and from the organotrophic bacterium Beggiatoa alba. MDHs from the B. leptomitiformis strain D-402 grown under organotrophic conditions and from B. alba are homodimers with the subunit molecular weight of 40 kD. Tetrameric MDH is formed in B. leptomitiformis strain D-402 grown under lithotrophic conditions. The dimeric and tetrameric forms of MDH from B. leptomitiformis D-402 display some differences in kinetic properties.

Dependence of Malate Dehydrogenase Structure on the Type of Metabolism in Freshwater Filamentous Colorless Sulfur Bacteria of the Genus Beggiatoa

Major pathways of carbon metabolism were studied in strains D-402 and D-405 of freshwater colorless sulfur bacteria of the genus Beggiatoa grown organotrophically and mixotrophically. The bacteria were found to possess all the enzymes of the tricarboxylic acid (TCA) and glyoxylate cycles. When organotrophic growth changed to mixotrophic growth, the activity of the TCA cycle enzymes decreased 2- to 3-fold, but the activity of enzymes of the glyoxylate cycle increased threefold. It follows that, in the oxidation of thiosulfate, organic compounds no longer play the leading part in the energy metabolism, and most of electrons that enter the electron transport chain (ETC) derive from inorganic sulfur compounds. A connection was established between the structure and kinetic characteristics of malate dehydrogenase—an enzyme of the TCA and glyoxylate cycles—and the type of carbon metabolism in the strains studied. Malate dehydrogenase in organotrophically grown cells of strains D-402 and D-405 is dimeric, whereas in strain D-402 grown mixotrophically it is tetrameric.

Microbiological processes of the carbon and sulfur cycle in cold methane seeps in the North Atlantic

Functioning of microbial communities in surface sediments of the Haakon Mosby underwater mud volcano (lat. 72 degrees N) and in gas seepage fields of the Vestnesa Ridge was investigated using Mir-1 and Mir-2 deep-sea submersibles during the 40th expedition of the research vessel Academician Mstislav Keldysh. Large areas of sedimentary deposits of the Haakon Mosby mud volcano (HMMV) and pockmarks of the Vestnesa Ridge (VR) are covered with bacterial mats 0.1 to 0.5 cm thick. The microbial community making up bacterial mats of the HMMV was predominated by large filamentous bacteria with filaments measuring up to 100 microns in length and 2 to 8 microns in width. The occurrence of rosettes allowed the observed filamentous bacteria to be referred to the morphologically similar genera Leucothrix or Thiothrix. Three morphological types of filamentous bacteria were identified in bacterial mats covering VR pockmarks. Filaments of type one are morphologically similar with representatives of the genera Thioploca or Desmanthos. Type two filaments had numerous inclusions of sulfur and resembled representatives of the genus Thiothrix. The third morphological type was constituted by single filaments made up of tightly connected disk-like cells and can, apparently, be assigned to the genus Beggiatoa. The rates of methane oxidation (up to 1570 microliters C/(dm3 day)) and sulfate reduction (up to 17 mg S/(dm3 day)) measured in surface sediments of HMMV and VR were close to the maximum rates of these processes observed in badly polluted regions of the northwestern shelf of the Black Sea. High rates of microbiological processes correlated with the high number of bacteria. The rate of methane production in sediments studied was notably lower and ranged from 0.1 to 3.5 microliters CH4/(dm3 day). Large areas of the HMMV caldera were populated by pogonophoras, represented by the two species, Sclerolinum sp. and Oligobrachia sp. The mass development of Sclerolinum sp. in the HMMV caldera was by the activity of aerobic methane-oxidizing bacteria localized inside the cells of these animals. Bacterial cells were also found in the trophosome tissue of Oligobrachia sp., but in cells of these bacteria, we did not observe the membrane structures typical of methanotrophs. The localization pattern of pogonophoras on the surface of reduced sediments suggests that the predominant bacteria in Oligobrachia tissues are sulfur-oxidizing endosymbionts.

Occurence and regulation of Calvin cycle enzymes in non-autotrophic Beggiatoa strains

To date only marine strains of the genus Beggiatoa have been proven to have the capacity for lithoautotrophic growth. The two best characterized freshwater strains (B18LD and OH-75-2a) — previously tested with no evidence of autotrophic potential — were reinvestigated in the current study. Heterotrophically grown cells from exponential or early stationary phase showed extremely low ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activities (0.002–0.2 nmol CO2·mg prot-1 min-1). After acetate limitation terminated growth both strains demonstrated increased RuBisCO activity with a maximum achieved 1–3 days into stationary phase. For a single growth cycle, activity could increase by more than 100-fold (strain OH-75-2a), but the highest activity achieved was less than 5% of that observed in a lithoautotrophic control. Phosphoribulokinase activity was approximately an order of magnitude greater than the corresponding RuBisCO activity and showed a similar pattern of regulation. A gene probe derived from the RuBisCO large subunit gene of Anacystis nidulans hybridized strongly with DNA from strain OH-75-2a and the autotrophic marine control but not with DNA from strain B18LD.

Characterization of Vitreoscilla beggiatoides and Vitreoscilla filiformis sp. nov., nom. rev., and Comparison with Vitreoscilla stercoraria and Beggiatoa alba

A group of Beggiatoa-like organisms which do not deposit sulfur from sulfide but which were isolated from the same locations as sulfide-oxidizing beggiatoas should be considered as a species within the genus Vitreoscilla. These organisms fit the general description of Vitreoscilla beggiatoides as originally described by E. G. Pringsheim, and strain B23SS (= ATCC 43189) is herein designated as the neotype strain. The guanine-plus-cytosine content of the deoxyribonucleic acid of strain B23SS was 42 mol%. A few strains that were formerly considered as Beggiatoa spp. have been shown to lack the ability to accumulate sulfur in inclusions when exposed to hydrogen sulfide. Because rigorous tests were not previously carried out to determine whether these organisms really deposited sulfur, it is probable that they were never capable of doing so. These strains, which form a distinct group of microorganisms, now should be considered to be a Vitreoscilla species. This group of organisms is most characteristic of the species V. filiformis as originally described by E. G. Pringsheim. Because strains had not been characterized since the original description, V. filiformis was not included in the Approved List of Bacterial Names. This name has, therefore, been revived, and strain L1401-2 (= ATCC 43190), a strain originally isolated by E. G. Pringsheim, is designated as the neotype strain. The guanine-plus-cytosine content of the deoxyribonucleic acid of these strains ranged from 59 to 63 mol%. These two groups of organisms are compared to V. stercoraria, the best-characterized Vitreoscilla strain, and to Beggiatoa alba, the best-characterized and type species of the genus Beggiatoa.