A coastal marine sulfide-oxidizing autotrophic bacterium produces hydrophilic filamentous sulfur as a novel metabolic end product. Phylogenetic analysis placed the organism in the genus Arcobacter in the epsilon subdivision of the Proteobacteria. This motile vibrioid organism can be considered difficult to grow, preferring to grow under microaerophilic conditions in flowing systems in which a sulfide-oxygen gradient has been established. Purified cell cultures were maintained by using this approach. Essentially all 4,6-diamidino-2-phenylindole dihydrochloride-stained cells in a flowing reactor system hybridized with Arcobacter-specific probes as well as with a probe specific for the sequence obtained from reactor-grown cells. The proposed provisional name for the coastal isolate is "Candidatus Arcobacter sulfidicus." For cells cultured in a flowing reactor system, the sulfide optimum was higher than and the CO 2 fixation activity was as high as or higher than those reported for other sulfur oxidizers, such as Thiomicrospira spp. Cells associated with filamentous sulfur material demonstrated nitrogen fixation capability. No ribulose 1,5-bisphosphate carboxylase/oxygenase could be detected on the basis of radioisotopic activity or by Western blotting techniques, suggesting an alternative pathway of CO 2 fixation. The process of microbial filamentous sulfur formation has been documented in a number of marine environments where both sulfide and oxygen are available. Filamentous sulfur formation by "Candidatus Arcobacter sulfidicus" or similar strains may be an ecologically important process, contributing significantly to primary production in such environments.In the marine environment, hydrogen sulfide is a ubiquitous end product of anaerobic processes of organic matter remineralization (5,23,29,30). At ridge crest sites on the ocean floor, it is produced from the geothermal transformation of sulfate and elemental sulfur leaching via seawater-basaltic rock interaction (26,40,62). When brought into contact with the aerobic biosphere, hydrogen sulfide becomes an energy-yielding substrate for chemosynthetic colorless sulfur-oxidizing bacteria. Members of this group include free-living rods or ovoids of the genera Thiobacillus, Thiomonas, Acidiphilium, Thiomicrospira, and Thiovulum (31,32,34,42) as well as the morphologically conspicuous gliding and nongliding filamentous forms of the genera Beggiatoa, Thioploca, and Thiothrix (19,47,48,69). These organisms are characterized by their ability to catalyze the oxidation of sulfide and its chemically and biologically mediated partial oxidation products (polysulfides, S n 2Ϫ ; elemental sulfur, S 0 ; sulfane monosulfonic acids, HSS n O 3 2Ϫ ]; and polythionates, S n O 6 2Ϫ ) (14, 33, 66) coupled to the fixation of carbon dioxide to organic carbon by utilizing the same CalvinBassham-Benson cycle enzymes as those used by oxygenic phototrophs.Because of the differential rates of oxidation of hydrogen sulfide and derived oxidation products, intermediates may substantially accu...

Based on 16S rRNA gene surveys, bacteria of the subdivision of proteobacteria have been identified to be important members of microbial communities in a variety of environments, and quite a few have been demonstrated to grow autotrophically. However, no information exists on what pathway of autotrophic carbon fixation these bacteria might use. In this study, Thiomicrospira denitrificans and Candidatus Arcobacter sulfidicus, two chemolithoautotrophic sulfur oxidizers of the subdivision of proteobacteria, were examined for activities of the key enzymes of the known autotrophic CO 2 fixation pathways. Both organisms contained activities of the key enzymes of the reductive tricarboxylic acid cycle, ATP citrate lyase, 2-oxoglutarate: ferredoxin oxidoreductase, and pyruvate:ferredoxin oxidoreductase. Furthermore, no activities of key enzymes of other CO 2 fixation pathways, such as the Calvin cycle, the reductive acetyl coenzyme A pathway, and the 3-hydroxypropionate cycle, could be detected. In addition to the key enzymes, the activities of the other enzymes involved in the reductive tricarboxylic acid cycle could be measured. Sections of the genes encoding the ␣-and ␤-subunits of ATP citrate lyase could be amplified from both organisms. These findings represent the first direct evidence for the operation of the reductive tricarboxylic acid cycle for autotrophic CO 2 fixation in -proteobacteria. Since -proteobacteria closely related to these two organisms are important in many habitats, such as hydrothermal vents, oxic-sulfidic interfaces, or oilfields, these results suggest that autotrophic CO 2 fixation via the reductive tricarboxylic acid cycle might be more important than previously considered.Almost all major groups of prokaryotes include representatives that are able to grow autotrophically (33). These organisms play an essential role in ecosystems by providing a continuous supply of organic carbon for heterotrophs. The Calvin-Benson-Bassham cycle (Calvin cycle) represents the most important extant autotrophic carbon fixation pathway (43, 50). Despite its global significance, it is restricted to organisms with high-energy yield from a chemotrophic or phototrophic lifestyle. Microorganisms present in extreme environments, e.g., high temperature or anaerobic or acidic conditions, generally utilize different CO 2 fixation pathways (17, 33). At present, there are three alternative pathways known: the reductive tricarboxylic acid (TCA) cycle, the reductive acetyl coenzyme A (CoA) pathway, and the 3-hydroxypropionate cycle (4, 33).It has been proposed that the first autotrophic pathway was akin to either the reductive TCA cycle or the reductive acetylCoA pathway (11,17,35,45,58). The reductive TCA cycle has the characteristics of an autocatalytic cycle and leads to a complex cyclic reaction network from which other anabolic pathways could have evolved (11, 58): e.g., the oxidative TCA cycle (8,45). Based upon biochemical and isotopic analyses, the reductive TCA cycle appears to operate in phylogenetically diverse aut...

Studies were conducted in opposing gradients of oxygen and sulfide in microslide capillaries to (i) characterize the chemical microenvironment preferred by Candidatus Arcobacter sulfidicus, a highly motile, sulfur-oxidizing bacterium that produces sulfur in filamentous form, and (ii) to develop a model describing the mechanism of filamentous-sulfur formation. The highly motile microorganisms are microaerophilic, with swarms effectively aggregating within oxic-anoxic interfaces by exhibiting a chemotactic response. The position of the band was found to be largely independent of the sulfide concentration as it always formed at the oxic-anoxic interface. Flux calculations based on steady state gradients of oxygen and sulfide indicate that sulfide is incompletely oxidized to sulfur, in line with the formation of filamentous sulfur by these organisms. It is proposed that Candidatus Arcobacter sulfidicus effectively competes with other sulfur-oxidizing bacteria in the environment by being able to tolerate higher concentrations of hydrogen sulfide (1-2 mM) and by possessing the ability to grow at very low oxygen concentrations (1-10 muM). The formation of mat-like structures from filamentous sulfur appears to be a population mediated effort allowing these organisms to effectively colonize environments characterized by high sulfide, low oxygen and dynamic fluid movement.

Epidemiologic studies of farm children are of international interest because farm children are less often atopic, have less allergic disease, and often have less asthma than do nonfarm children—findings consistent with the hygiene hypothesis. We studied a cohort of rural Iowa children to determine the association between farm and other environmental risk factors with four asthma outcomes: doctor-diagnosed asthma, doctor-diagnosed asthma/medication for wheeze, current wheeze, and cough with exercise. Doctor-diagnosed asthma prevalence was 12%, but at least one of these four health outcomes was found in more than a third of the cohort. Multivariable models of the four health outcomes found independent associations between male sex (three asthma outcomes), age (three asthma outcomes), a personal history of allergies (four asthma outcomes), family history of allergic disease (two asthma outcomes), premature birth (one asthma outcome), early respiratory infection (three asthma outcomes), high-risk birth (two asthma outcomes), and farm exposure to raising swine and adding antibiotics to feed (two asthma outcomes). The high prevalence of rural childhood asthma and asthma symptoms underscores the need for asthma screening programs and improved asthma diagnosis and treatment. The high prevalence of asthma health outcomes among farm children living on farms that raise swine (44.1%, p = 0.01) and raise swine and add antibiotics to feed (55.8%, p = 0.013), despite lower rates of atopy and personal histories of allergy, suggests the need for awareness and prevention measures and more population-based studies to further assess environmental and genetic determinants of asthma among farm children.

The effects of light and anaerobiosis on germination of marine dinoflagellate resting cysts were examined. Germination of all species was completely inhibited during 7 weeks of anaerobic incubation, although the cysts remained viable. Light requirements were more variable. Germination rates in the light varied 20-fold between species; dark rates varied by a similar factor but were generally much lower. One species (Gonyaulax polyedra) required light to germinate, three germinated faster in the light than in the dark (Gonyaulax tamarensis, Scrippsiella sp., and Gonyaulax verior), and one germinated at comparable rates in the light and in the dark (Gonyaulax rugosum). Brief exposure to light at the beginning of the experiments made it impossible to say whether germination is possible in constant darkness. It is clear, however, that prolonged light exposure is a requirement only for G. polyedra, although it can significantly accelerate germination of the other species. Germination inhibition by darkness and anaerobiosis helps to explain the subsurface accumulation of dinoflagellate cysts in marine sediments and the persistence of benthic cyst populations at times when temperatures favor germination. These constraints are sufficiently common that many cysts will never germinate once they reach bottom sediments.

At deep-sea hydrothermal vents, a large source of potential chemical energy is created when reducing vent fluid and oxidizing seawater mix. In this environment, chemolithoautotrophic microbes catalyze exergonic redox reactions which in turn provide the energy needed to fuel their growth and the fixation of CO 2 into biomass. In addition to producing new organic matter, this process also consumes compounds contained both in vent fluid and entrained seawater (e.g. H 2 , NO 3-). Despite their biogeochemical importance, such reactions have remained difficult to quantify due to methodological limitations. To address this knowledge gap, this study reports a novel application of isobaric gas-tight fluid samplers for conducting incubations of hydrothermal vent fluids at in situ temperature and pressure. Eighteen ~24 h incubations were carried out, representing seven distinct conditions that

A highly motile chemoautotrophic strain of hydrogen sulfide-oxidizing bacteria from coastal seawater produces solid sulfur filaments of dimensions 0.5 to 2.0 micrometers by 20 to 500 micrometers. Filamentous sulfur is rapidly produced by direct excretion by a vibrioid organism, and the newly produced filaments are thickened by the deposition of sulfur by other members of the population. Microscopic observations of the flocculent discharge material collected from diffuse-flow hydrothermal vents (gON, East Pacific Rise) revealed that the material from this source is composed largely of filamentous sulfur of morphology nearly identical to that obtained in the model laboratory system.

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