The anaerobic zones of Lakes Cis6 and Vilar (Banyoles karstic area, NE Spain) had mass developments of purple sulfur bacteria during summer 1982. In Lake Vilar, Chromatium spp. was dominant (up to 92% of the microbial biovolume). In Lake Ciso, the predominant microorganisms were Chromatium spp. (up to 7 1%) and another purple sulfur bacterium forming aggregates (20%).The bacterial layer could be divided according to the physiological state of the cells into a top part of maximal specific activity, a peak of maximal abundance, and a bottom part of inactive cells. The bacteria in the peak were predominantly limited by light; sulfide, phosphate, and acetate were not limiting in the middle of the day. The light limitation started at the depth having the maximal concentration of cells; the top of the layer appeared to be sulfide-limited. Specific contents of photopigments, elemental sulfur, and reserve polymers decreased from the top to the bottom of the bacterial layer. These phenomena point to the crucial role of light in the development of layers of phototrophic bacteria in stratified lakes.
Two kinds of predatory bacteria have been observed and characterized by light and electron microscopy in samples from freshwater sulfurous lakes in northeastern Spain. The first bacterium, named Vampirococcus, is Gram-negative and ovoidal (0.6 jam wide). An anaerobic epibiont, it adheres to the surface of phototrophic bacteria (Chromatium spp.) by specific attachment structures and, as it grows and divides by fission, destroys its prey. An important in situ predatory role can be inferred for Vampirococcus from direct counts in natural samples. The second bacterium, named Daptobacter, is a Gram-negative, facultatively anaerobic straight rod (0.5 x 1.5 ,um) with a single polar flagellum, which collides, penetrates, and grows inside the cytoplasm of its prey (several genera of Chromatiaceae). Considering also the well-known case of Bdellovibrio, a Gram-negative, aerobic curved rod that penetrates and divides in the periplasmic space of many chemotrophic Gram-negative bacteria, there are three types of predatory prokaryotes presently known (epibiotic, cytoplasmic, and periplasmic). Thus, we conclude that antagonistic relationships such as primary consumption, predation, and scavenging had already evolved in microbial ecosystems prior to the appearance of eukaryotes. Furthermore, because they represent methods by which prokaryotes can penetrate other prokaryotes in the absence of phagocytosis, these associations can be considered preadaptations for the origin of intracellular organelles.Although symbiotic bacteria have been extensively studied and their evolutionary importance in the origin of eukaryotic cells has been recognized (1, 2), predatory behavior in bacteria is known only for Bdellovibrio (3, 4) and Vampirovibrio (5,6). Antagonistic relationships among large organisms are considered to be properties of ecosystems and integrated into ecological theory (7); however, such behavior (e.g., primary consumption, predation, and scavenging) attributed only to animals and plants (8) MATERIALS AND METHODS Studies were conducted in Lake Estanya (420 02' N, 0O 32' E) and Lake Cis6 (420 08' N, 20 45' E) in northeastern Spain.Both lakes are sinkholes formed in karstic areas, rich in calcium sulfate as gypsum and anhydrite. They receive most of their water inputs through seepage. The water conductivity, about 1800 gS cm-l for Lake Estanya and 1300 ;LS-cm-'for Lake Cis6, is high, primarily as a consequence of dissolved salts as sulfates (siemens are reciprocal ohms; S = 1/fl). From 7 to 10 mM sulfate is present in solution in the hypolimnia of both lakes. Lake Estanya, figure-eight shaped, has two basins 12 and 20 m deep, respectively. They are separated by a 2-m-deep sill (10). Lake Cis6, an almost semispherical basin, is 9 m deep and 25 m in average diameter at the surface. Because of high production of hydrogen sulfide in the sediments, it is completely anoxic during mixing (11). Details of lake ecology and methods of study have been published (12)(13)(14). In both lakes light penetrates down to the thermocline,...
A photosynthetic microbial mat was investigated in a large pond of a Mediterranean saltern (Salins-de-Giraud, Camargue, France) having water salinity from 70 per thousand to 150 per thousand (w/v). Analysis of characteristic biomarkers (e.g., major microbial fatty acids, hydrocarbons, alcohols and alkenones) revealed that cyanobacteria were the major component of the pond, in addition to diatoms and other algae. Functional bacterial groups involved in the sulfur cycle could be correlated to these biomarkers, i.e. sulfate-reducing, sulfur-oxidizing and anoxygenic phototrophic bacteria. In the first 0.5 mm of the mat, a high rate of photosynthesis showed the activity of oxygenic phototrophs in the surface layer. Ten different cyanobacterial populations were detected with confocal laser scanning microscopy: six filamentous species, with Microcoleus chthonoplastes and Halomicronema excentricum as dominant (73% of total counts); and four unicellular types affiliated to Microcystis, Chroococcus, Gloeocapsa, and Synechocystis (27% of total counts). Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments confirmed the presence of Microcoleus, Oscillatoria, and Leptolyngbya strains (Halomicronema was not detected here) and revealed additional presence of Phormidium, Pleurocapsa and Calotrix types. Spectral scalar irradiance measurements did not reveal a particular zonation of cyanobacteria, purple or green bacteria in the first millimeter of the mat. Terminal-restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA gene fragments of bacteria depicted the community composition and a fine-scale depth-distribution of at least five different populations of anoxygenic phototrophs and at least three types of sulfate-reducing bacteria along the microgradients of oxygen and light inside the microbial mat.
Accurate studies of the pigment composition and isolation in pure cultures of Chlorobiaceae from samples of eight Spanish lakes show that there are two main coexisting groups of green and brown Chlorobium spp. represented respectively by Chlorobium limicola and Chlorobium phaeobacteroides. Laboratory experiments with pure and mixed cultures of the isolated strains show that light quality plays a selective role on the species composition among Chlorobiaceae. This selection depends on the pigment composition which determines the in vivo absorption spectrum of the cells as well as on their ability to adjust the intracellular concentration of light-harvesting pigments to the spectral distribution and energy of light. Correlation analysis performed with field data resulted in significant, but low, correlation coefficients. Nevertheless, they were consistent with laboratory data showing that brown Chlorobiaceae were dominant in deep layers in meromictic lakes, whereas green Chlorobiaceae dominated in layers nearer the surface or underneath plates of Chromatiaceae. The combination of laboratory and field observations stress the role of biological light filtering in determining the species composition among Chlorobiaceae in lakes.
The microenvironment and community composition of microbial mats developing on beaches in Scapa Flow (Orkney Islands) were investigated. Analysis of characteristic biomarkers (major fatty acids, hydrocarbons, alcohols, and alkenones) revealed the presence of different groups of bacteria and microalgae in mats from Waulkmill and Swanbister beach, including diatoms, Haptophyceae, cyanobacteria, and sulfate-reducing bacteria. These analyses also indicated the presence of methanogens, especially in Swanbister beach mats, and therefore a possible role of methanogenesis for the carbon cycle of these sediments. High amounts of algal lipids and slightly higher numbers (genera, abundances) of cyanobacteria were found in Waulkmill Bay mats. However, overall only a few genera and low numbers of unicellular and filamentous cyanobacteria were present in mats from Waulkmill and Swanbister beach, as deduced from CLSM (confocal laser scanning microscopy) analysis. Spectral scalar irradiance measurements with fiber-optic microprobes indicated a pronounced heterogeneity concerning zonation and density of mainly anoxygenic phototrophs in Swanbister Bay mats. By microsensor and T-RFLP (terminal restriction fragment length polymorphism) analysis in Swanbister beach mats, the depth distribution of different populations of purple and sulfate-reducing bacteria could be related to the microenvironmental conditions. Oxygen, but also sulfide and other (inorganic and organic) sulfur compounds, seems to play an important role in the stratification and diversity of these two major bacterial groups involved in sulfur cycling in Swanbister beach mats.
The spatio-temporal distribution of phototrophic communities of the hypersaline photosynthetic Camarguc microbial mat (Salins-de-Giraud, France) was investigated over a diel cycle by combining microscopic and molecular approaches. Microcoleus chthonoplastes and Halomicronema excentricum, the dominant cyanobacteria of this oxyphotrophic community, were observed with confocal laser scanning microscopy to determine their biomass profiles. Both bacteria have similar vertical distributions, varying from a homogenous distribution through the mat during the night, to a specific localization in the upper oxic zone of 1.5 mm during the day. Terminal restriction fragment length polymorphism of PCR-amplified pufM gene fragments revealed three groups of anoxyphototrophic populations, which varied according to the two opposite periods of the diel cycle under study. They were either specifically detected in only one period, or homogenously distributed through the mat in all periods, or located in specific zones of the mat depending on the period considered. Oxygen concentrations, pH and biomass of the major filamentous cyanobacteria were the determinative factors in the distribution of these anoxyphototrophs across the mat. Thus, vertical migration, cell-cell aggregate formation and metabolic switches were the most evident defence of the photosynthetic populations against the adverse effects of sulfide and oxygen fluxes during a diel cycle.
Recent studies have shown that the cyanobacterium Microcoleus chthonoplastes forms a consortium with heterotrophic bacteria present within the cyanobacterial sheath. These studies also show that this consortium is able to grow in the presence of crude oil, degrading aliphatic heterocyclic organo-sulfur compounds as well as alkylated monocyclic and polycyclic aromatic hydrocarbons. In this work, we characterize this oil-degrading consortium through the analysis of the 16S rRNA gene sequences. We performed the study in cultures of Microcoleus grown in mineral medium and in cultures of the cyanobacterium grown in mineral medium supplemented with crude oil. The results indicate that most of the clones found in the polluted culture correspond to well-known oil-degrading and nitrogen-fixing microorganisms, and belong to different phylogenetic groups, such as the Alpha, Beta, and Gamma subclasses of Proteobacteria, and the Cytophaga/Flavobacteria/Bacteroides group. The control is dominated by one predominant organism (88% of the clones) closely affiliated to Pseudoxanthomonas mexicana (similarity of 99.8%). The presence of organisms closely related to well-known nitrogen fixers such as Rhizobium and Agrobacterium suggests that at least some of the cyanobacteria-associated heterotrophic bacteria are responsible for nitrogen fixation and degradation of hydrocarbon compounds inside the polysaccharidic sheath, whereas Microcoleus provides a habitat and a source of oxygen and organic matter.
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