Experimental investigation of insect deposition in lentic environments and implications for formation of Konservat Lagerstätten

Terrestrial insects are often remarkably well preserved in lacustrine Konservat Lagerstätten. However, the assumption that carcasses should sink fast through the water column seems contradictory as this scenario is unlikely due to excessive buoyancy and surface tension. The mechanisms that promote rapid and permanent emplacement onto the sediment surface (RPESS) of such terrestrial animal remains are not fully understood. Here we use taphonomic experiments to show that floating in water, growth of microbial biofilms and reception of rapid sediment load promote RPESS of terrestrial insect remains in lentic water bodies. Our results show that the optimum conditions for RPESS occur when terrestrial insects enter a lentic water body in articulation, experience brief decay in association with growth of microbes, then are buried rapidly by airborne volcanic ash. These results provide a model for preservation of articulated terrestrial insects and emphasize the importance of microbial activity and volcanism for insect preservation in lacustrine Konservat Lagerstätten.     

A microbial model for the Lower Devonian stromatactis mud mounds of the Montagne Noire (France)

Summary Lower Devonian mud mounds and stromatactis fabrics are exceptionally well exposed in quarry walls and industrially sawed blocks in the Montagne Noire in southern France. Interlayered red biomicrites and white to grey sparitic calcites form mounds up to 70 m high. The red biomicrites contain predominantly bryozoans, sponges and echinoderms. The sparitic layers show typical features of stromatactis fabrics, as outlined byBathurst (1982). We recognize two types of stromatactis fabrics: (1) Stromatactis type A: exentsive cavity systems filled by multiple cement generations, which are interpreted to be related to microbial mats, and (2) Stromatactis type B: smaller patches of blocky spar which are mainly diagenetic in origin, but show characteristic features of stromatactis. Type A is far more important in terms of rock volume. The cyclic interlayering of red biomicrites and sparitic layers is supposed to result from frequent changes in the composition of the mound biota. The bryozoan/sponge community was displaced by short term propagations of microbial mats during times of extremely low sedimentation. Sedimentation and thus the biotic community was probably determined by high frequency (6th order) sea level changes. Despite these changes, mound growth continued, because once established the ecological advantage over the surroundings was maintained by both communities alternating with each other. The microbial mats and the cavities they left after their decay were important for the stabilization of the mounds, the latter allowing for enormous quantities of dissolved carbonate to be transported and precipitated. We anticipate a close interrelation between mound formation and stromatactis formation, and we believe that it is not incidential that both, mud mounds and stromatactis, are mainly restricted to the same interval, namely the Paleozoic.     

A new eocrinoid from the Guanshan Biota (Cambrian Series 2, Stage 4), with implication of the development of different attachment modes in early Cambrian

Eocrinoids are scarce in the Guanshan Biota (Cambrian Series 2, Stage 4), Yunnan Province, southwest China. Here, we introduce a new gogiid: Kunmingeocrinus cupuliformis n. gen. n. sp. which is characterized by a short stalk and a well-developed attachment disc. Preservation may indicate a weakly biomineralized body for the new taxon. Morphological similarities between the new taxon and other eocrinoids with attachment discs from Cambrian Lagerstätten of Guizhou Province (Series 2, Stage 4 and Miaolingian, Wuliuan) might suggest a similar mode of life. The eocrinoids from the Guanshan Biota possibly utilized different attachment modes.     

Suspected microbial-induced sedimentary structures (MISS) in Furongian (Upper Cambrian; Jiangshanian,Sunwaptan) strata of the Upper Mississippi Valley

The Furongian (Upper Cambrian; Jiangshanian and Sunwaptan) Tunnel City Group (Lone Rock Formation and Mazomanie Formation), exposed in Wisconsin and Minnesota, represents a shallow-marine clastic environment during a time of exceptionally high sea level. Lithofacies from shoreface to transitional-offshore settings document deposition in a wave- and storm-dominated sea. Flooding of the cratonic interior was associated with formation of a condensed section and the extensive development of microbial mats. Biolamination, mat fragments, wrinkle structures, and syneresis cracks are preserved in various sandstone facies of the Lone Rock Formation, as is evidence for the cohesive behavior of sand. These microbial-induced sedimentary structures (MISS) provide unique signals of biological–physical processes that physical structures alone cannot mimic. The MISS are associated with a trilobite extinction event in the Steptoean–Sunwaptan boundary interval. This may support recent claims that Phanerozoic microbial mats were opportunistic disaster forms that flourished during periods of faunal turnover. Further investigation of stratigraphic, taphonomic, and other potential biases, however, is needed to fully test this hypothesis.     

Microbial mats and physicochemistry in a saltern in the Bretagne (France) and in a laboratory scale saltern model

Abstract A saltern near La Baule (Bretagne, France) was remodeled in a programmable temperature and humidity controlled walk-in environmental chamber resembling the characteristics of the original saltern. The saltern showed different types of microbial mats predominantly composed of algae, oxy- and anoxyphotobacteria, and associated chemoorganotrophic bacteria, fungi and animals. Well-developed microbial mats were found up to a salinity of 10% during the three or four months in summer when salinity gradients and NaCl precipitation were established. The main phototrophic organisms were diatoms, the cyanobacteria Aphanothece, Microcoleus, Spirulina , and Oscillatoria , and Chromatiaceae. At higher salinity, Halobacterium sp., diatoms, and Dunaliella were dominant. Typical microbial mats and saltern-typical invertebrate, algal and bacterial species also developed in the saltern model, building up a stable community. The ionic composition of the brines and physicochemical parameters were similar to those determined for the original saltern. Different photosynthetic organisms, e.g. a filamentous purple bacterium and a hypersaline Chloroflexus -like organism, could be enriched within the microbial mats by changing the light regime.     

Late Devonian marine anoxia challenged by benthic cyanobacterial mats

Mass occurrence of benthic cyanobacterial mats in a sequence of Late Devonian black shales and bituminous limestones of the Holy Cross Mts. (central Poland), enclosing the famous Kellwasser and Hangenberg extinction horizons, is reported. The microbiota forming the mats is compared with some modern benthic chroococcalean cyanobacteria. Similarly to their extant counterparts, the Devonian cyanobacteria must had been phototrophic and oxygenic aerobes which could, however, tolerate slightly sulfidic conditions characterizing the near-bottom waters of the Late Devonian epicontinental sea. The cyanobacterial mats successfully colonized the oxygen-deficient and H(2) S-enriched seabed otherwise unfavorable for most other benthic biota. The redox state of this sluggish Late Devonian sea, ascribed previously mostly to anoxic or euxinic conditions, is reassessed as probably pulsating between anoxic, dysoxic, and weakly oxic conditions. The redox state was dependent on the rate of oxygen production by the cyanobacterial mats, the intensity of H(2) S emissions from the decaying mat biomass, and the rate of planktonic production.     

Microbial mats in tidal channels at San Carlos,Baja California Sur,Mexico

Microbial communities of stratified phototrophic bacteria in laminated intertidal sediments north of Estuary El Puente, near San Carlos, Baja California Sur, Mexico,‐were studied. This study describes the macroscopic and microscopic characteristics of the mats, including their annual growth. The mats were located in and along meandering mangrove‐lined tidal channels. Their thickness ranged from 0.5 to 25 cm. Square‐meter areas of polygonal mats were detected in several ponds infiltrated by sea water. The principal microbial community of the upper surface of various morphotypes of microbial mats was identified as cyanobacteria belonging to the genera Microcoleus, Lyngbya, Phormidium, and Oscillatoria. Other cyanobacte‐rial genera such as Pseudanabaena, Spirulina, Synechococcus, and Gloeocapsa, as well as many unidentified diatoms, were also present but at lower population densities. The second inward reddish layers of the microbial mats contained similar cyano‐bacterial genera plus anoxygenic phototrophic bacteria belonging to the genera Chloroflexus, Thiocapsa, Chromatium, Prosthecochloris, Rhodopseudomonas, and Chlorobium, as well as several unidentified bacteria. In situ measurements on the growth of the mats, from intermittent tide sites, showed an annual buildup of two layers: green and reddish. These layers corresponded to a vertical growth of 1.4 ± 0.27 mm/year. Permanently submerged mats did not show vertical growth during the same period of time.     

Three-dimensional anatomy of the Tully monster casts doubt on its presumed vertebrate affinities

Tullimonstrum gregarium, also known as the Tully monster, is a well-known phylogenetic enigma, fossils of which have been found only in the Mazon Creek Lagerstätte. The affinities of Tullimonstrum have been debated since its discovery in 1966, because its peculiar morphology with stalked eyes and a proboscis cannot easily be compared with any known animal morphotypes. Recently, the possibility that Tullimonstrum was a vertebrate has attracted much attention, and it has been postulated that Tullimonstrum might fill a gap in the fossil record of early vertebrates, providing important insights into vertebrate evolutionary history. With the hope of resolving this debate, we collected 3D surface data from 153 specimens of Tullimonstrum using a high-resolution laser 3D scanner and conducted x-ray micro-computed tomographic (μCT) analysis of stylets in the proboscis. Our investigation of the resulting comprehensive 3D morphological dataset revealed that structures previously regarded as myomeres, tri-lobed brain, tectal cartilages and fin rays are not comparable with those of vertebrates. These results raise further doubts about its vertebrate affinities, and suggest that Tullimonstrum may have been either a non-vertebrate chordate or a protostome.     

A Microbial Mat of a Large Sulfur Bacterium Preserved in a Miocene Methane-Seep Limestone

A Miocene methane-seep limestone from the Romagna Apennine (Pietralunga, Italy) was found to contain an extraordinarily well-preserved microbial mat consisting of filamentous fossils. Individual filaments of the lithified Pietralunga mat are 50 to 80 μ m in diameter and resemble the sulfide-oxidizing bacterium Beggiatoa. Mats of sulfur bacteria are common around modern methane-seeps, but have not yet been reported from ancient seep limestones. This is thought to be related to the conditions prevailing in metabolically active mats of sulfur bacteria that do not favor carbonate formation. The preservation of the Pietralunga mat was most likely caused by a sudden change from oxidizing to anoxic conditions, leading to the rapid carbonate precipitation induced by anaerobic oxidation of methane. Lipid biomarkers specific for archaea and sulfate-reducing bacteria linked with the anaerobic oxidation of methane co-occur with compounds derived from methanotrophic bacteria and ciliates. These findings confirm a close proximity of oxic and anoxic conditions, as required for the growth of sulfide-oxidizing bacteria in the methane-based ecosystem. The lack of earlier reports on fossilized thiotrophic mats in seep limestones is most likely related to the rarity of environmental changes rapid enough to preserve the filaments rather than to a lower frequency of thiotrophic mats around methane-seeps in the geological past.     

Ecosystems on ice: the microbial ecology of Markham Ice Shelf in the high Arctic

Microbial communities occur throughout the cryosphere in a diverse range of ice-dominated habitats including snow, sea ice, glaciers, permafrost, and ice clouds. In each of these environments, organisms must be capable of surviving freeze-thaw cycles, persistent low temperatures for growth, extremes of solar radiation, and prolonged dormancy. These constraints may have been especially important during global cooling events in the past, including the Precambrian glaciations. One analogue of these early Earth conditions is the thick, landfast sea ice that occurs today at certain locations in the Arctic and Antarctic. These ice shelves contain liquid water for a brief period each summer, and support luxuriant microbial mat communities. Our recent studies of these mats on the Markham Ice Shelf (Canadian high Arctic) by high performance liquid chromatography (HPLC) showed that they contain high concentrations of chlorophylls a and b, and several carotenoids notably lutein, echinenone and beta-carotene. The largest peaks in the HPLC chromatograms were two UV-screening compounds known to be produced by cyanobacteria, scytonemin, and its decomposition product scytonemin-red. Microscopic analyses of the mats showed that they were dominated by the chlorophyte genera cf. Chlorosarcinopsis, Pleurastrum, Palmellopsis, and Bracteococcus, and cyanobacteria of the genera Nostoc, Phormidium, Leptolyngbya, and Gloeocapsa. From point transects and localized sampling we estimated a total standing stock on this ice shelf of up to 11,200 tonnes of organic matter. These observations underscore the ability of microbial communities to flourish despite the severe constraints imposed by the cryo-ecosystem environment.     

Sedimentology,taphonomy, and palaeoecology of a laminated plattenkalk from the Kimmeridgian of the northern Franconian Alb (southern Germany)

At Wattendorf in the northern Franconian Alb, southern Germany, centimetre- to decimetre-thick packages of finely laminated limestones (plattenkalk) occur intercalated between well bedded graded grainstones and rudstones that blanket a relief produced by now dolomitized microbialite-sponge reefs. These beds reach their greatest thickness in depressions between topographic highs and thin towards, and finally disappear on, the crests. The early Late Kimmeridgian graded packstone–bindstone alternations represent the earliest plattenkalk occurrence in southern Germany. The undisturbed lamination of the sediment strongly points to oxygen-free conditions on the seafloor and within the sediment, inimical to higher forms of life. The plattenkalk contains a diverse biota of benthic and nektonic organisms. Excavation of a 13 cm thick plattenkalk unit across an area of 80 m² produced 3500 fossils, which, with the exception of the bivalve Aulacomyella, exhibit a random stratigraphic distribution. Two-thirds of the individuals had a benthic mode of life attached to hard substrate. This seems to contradict the evidence of oxygen-free conditions on the sea floor, such as undisturbed lamination, presence of articulated skeletons, and preservation of soft parts. However, palaeoecological and taphonomic analyses indicate that the benthic faunal elements are allochthonous having settled out of suspension and thus must have been derived from hard substrate areas of neighbouring topographic highs. Solely the bivalve Aulacomyella, which occurs concentrated on a single bedding plane, may have colonised the plattenkalk depression during brief periods of oxygenation. Alternatively, a pseudoplanktonic mode of life, attached to floating sea weed, is envisaged for the bivalve. The formation of the plattenkalk is related to local and global factors: a drop in relative sea level caused the death of the microbialite-sponge reefs. Restricted circulation in depressions between the crests of the former reefs led to anoxic conditions and the formation of finely laminated sediments stabilised by microbial mats. The small depressions received sediment and skeletal elements of benthic organisms that were swept off neighbouring topographic highs by storms and that entered the depressions partly as turbidity currents, partly as suspension clouds. A rise in the relative sea level finally terminated the restricted circulation in the depressions and closed the taphonomic window that led to the preservation of the plattenkalk biota.     

Fossil liberation: a model to explain high biodiversity in the Triassic Cassian Formation

With 1429 animal species, the Triassic Cassian Formation in the Dolomites, Southern Alps (Italy), yields the highest species richness reported from any spatially constrained pre-Quaternary formation known to science. The high preserved diversity is partly attributable to a high primary diversity governed by the tropical setting, increasing alpha diversity, and the breadth of habitats spurring beta diversity. More important is the excellent preservation of fossils and the ease with which they can be extracted from the poorly lithified sediments. We propose the term ‘liberation Lagerstätten’ to capture this preservational window. In contrast to conservation Lagerstätten, liberation Lagerstätten like the Cassian Formation originate from normal marine conditions but low-grade diagenesis. Molluscs contribute substantially to species richness, comprising 67% of all invertebrate species in the Cassian Formation. The gastropod dominance (39% of all species) is nearly as great as in Recent tropical settings, contradicting the concept of a substantial Cenozoic rise.     

Microscopic examination of distribution and phenotypic properties of phylogenetically diverse Chloroflexaceae-related bacteria in hot spring microbial mats

We investigated the diversity, distribution, and phenotypes of uncultivated Chloroflexaceae-related bacteria in photosynthetic microbial mats of an alkaline hot spring (Mushroom Spring, Yellowstone National Park). By applying a directed PCR approach, molecular cloning, and sequence analysis of 16S rRNA genes, an unexpectedly large phylogenetic diversity among these bacteria was detected. Oligonucleotide probes were designed to target 16S rRNAs from organisms affiliated with the genus Chloroflexus or with the type C cluster, a group of previously discovered Chloroflexaceae relatives of this mat community. The application of peroxidase-labeled probes in conjunction with tyramide signal amplification enabled the identification of these organisms within the microbial mats by fluorescence in situ hybridization (FISH) and the investigation of their morphology, abundance, and small-scale distribution. FISH was combined with oxygen microelectrode measurements, microscope spectrometry, and microautoradiography to examine their microenvironment, pigmentation, and carbon source usage. Abundant type C-related, filamentous bacteria were found to flourish within the cyanobacterium-dominated, highly oxygenated top layers and to predominate numerically in deeper orange-colored zones of the investigated microbial mats, correlating with the distribution of bacteriochlorophyll a. Chloroflexus sp. filaments were rare at 60 degrees C but were more abundant at 70 degrees C, where they were confined to the upper millimeter of the mat. Both type C organisms and Chloroflexus spp. were observed to assimilate radiolabeled acetate under in situ conditions.     

Identification and characterization of Rhodopseudomonas spp., a purple, non-sulfur bacterium from microbial mats

A species of facultative photo-organotrophic, purple, non-sulfur bacterium was isolated from mixed-species microbial mats, characterized and examined for metal tolerance and bioremediation potential. Contributing mats were natural consortia of microbes, dominated by cyanobacteria and containing several species of bacteria arranged in a laminar structure, stabilized within a gel matrix. Constructed microbial mats were used for bioremediation of heavy metals and organic chemical pollutants. Purple, non-sulfur bacteria are characteristically found in lower strata of intact mats, but their contributing function in mats survival and function by mediating the chemical environment has not been explored. The gram-negative rod-shaped bacterium, reported here, produced a dark red culture under phototrophic conditions, reproduced by budding and formed a lamellar intracytoplasmic membrane (ICM) system parallel to cytoplasmic membrane, which contained bacteriochlorophyll a and carotenoids. This strain was found to have multiple metal resistances and to be effective in the reductive removal of Cr(VI) and the degradation of 2,4,6-trichlorophenol. Based on the results obtained from morphology, nutrient requirements, major bacteriochlorophyll content, GC content, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) profile and 16S-rDNA phylogenetic analysis, this member of the microbial mats may be identified as a new strain of the genus Rhodopseudomonas.     

Biogeochemical processes in the saline meromictic Lake Kaiike, Japan: implications from molecular isotopic evidences of photosynthetic pigments

Stable carbon and nitrogen isotopic compositions were determined for individual photosynthetic pigments isolated and purified from the saline meromictic Lake Kaiike, Japan, to investigate species-independent biogeochemical processes of photoautotrophs in the natural environment. In the anoxic monimolimnion and benthic microbial mats, the carbon isotopic compositions of BChls e and isorenieratene related to brown-coloured strains of green sulfur bacteria are substantially ( approximately 10 per thousand) depleted in (13)C relative to those found in the chemocline. In conjunction with 16S rDNA evidence reported previously, it strongly suggests that Pelodyctyon luteolum inhabited and photosynthesized in the anoxic monimolimnion and benthic microbial mats by using (13)C-depleted regenerated CO(2). By contrast, both Chl a and BChl a in the monimolimnion and microbial mats have similar isotopic compositions as they do in the chemocline, implying that the source organisms live only in the chemocline. In the chemocline, the nitrogen isotopic compositions of BChl e homologues ranges from -7.7 to-6.5 per thousand, whereas that of BChl a is -2.1 per thousand. These isotopic compositions suggest that green sulfur bacteria Chlorobium phaeovibrioides would conduct nitrogen fixation in the chemocline, whereas purple sulfur bacteria Halochromatium sp. and cyanobacteria Synechococcus sp. may assimilate nitrite.     

Biogeochemistry and community composition of iron- and sulfur-precipitating microbial mats at the Chefren mud volcano (Nile Deep Sea Fan, Eastern Mediterranean)

In this study we determined the composition and biogeochemistry of novel, brightly colored, white and orange microbial mats at the surface of a brine seep at the outer rim of the Chefren mud volcano. These mats were interspersed with one another, but their underlying sediment biogeochemistries differed considerably. Microscopy revealed that the white mats were granules composed of elemental S filaments, similar to those produced by the sulfide-oxidizing epsilonproteobacterium "Candidatus Arcobacter sulfidicus." Fluorescence in situ hybridization indicated that microorganisms targeted by a "Ca. Arcobacter sulfidicus"-specific oligonucleotide probe constituted up to 24% of the total the cells within these mats. Several 16S rRNA gene sequences from organisms closely related to "Ca. Arcobacter sulfidicus" were identified. In contrast, the orange mat consisted mostly of bright orange flakes composed of empty Fe(III) (hydr)oxide-coated microbial sheaths, similar to those produced by the neutrophilic Fe(II)-oxidizing betaproteobacterium Leptothrix ochracea. None of the 16S rRNA gene sequences obtained from these samples were closely related to sequences of known neutrophilic aerobic Fe(II)-oxidizing bacteria. The sediments below both types of mats showed relatively high sulfate reduction rates (300 nmol x cm(-3) x day(-1)) partially fueled by the anaerobic oxidation of methane (10 to 20 nmol x cm(-3) x day(-1)). Free sulfide produced below the white mat was depleted by sulfide oxidation within the mat itself. Below the orange mat free Fe(II) reached the surface layer and was depleted in part by microbial Fe(II) oxidation. Both mats and the sediments underneath them hosted very diverse microbial communities and contained mineral precipitates, most likely due to differences in fluid flow patterns.     

Diazotrophic microbial community of coastal microbial mats of the southern North Sea

The diazotrophic community in microbial mats growing along the shore of the North Sea barrier island Schiermonnikoog (The Netherlands) was studied using microscopy, lipid biomarkers, stable carbon (δ(13) C(TOC) ) and nitrogen (δ(15) N) isotopes as well as by constructing and analyzing 16S rRNA gene libraries. Depending on their position on the littoral gradient, two types of mats were identified, which showed distinct differences regarding the structure, development and composition of the microbial community. Intertidal microbial mats showed a low species diversity with filamentous non-heterocystous Cyanobacteria providing the main mat structure. In contrast, supratidal microbial mats showed a distinct vertical zonation and a high degree of species diversity. Morphotypes of non-heterocystous Cyanobacteria were recognized as the main structural component in these mats. In addition, unicellular Cyanobacteria were frequently observed, whereas filamentous heterocystous Cyanobacteria occurred only in low numbers. Besides the apparent visual dominance of cyanobacterial morphotpyes, 16S rRNA gene libraries indicated that both microbial mat types also included members of the Proteobacteria and the Cytophaga-Flavobacterium-Bacteroides group as well as diatoms. Bulk δ(15) N isotopes of the microbial mats ranged from +6.1‰ in the lower intertidal to -1.2‰ in the supratidal zone, indicating a shift from predominantly nitrate utilization to nitrogen fixation along the littoral gradient. This conclusion was supported by the presence of heterocyst glycolipids, representing lipid biomarkers for nitrogen-fixing heterocystous Cyanobacteria, in supratidal but not in intertidal microbial mats. The availability of combined nitrogen species might thus be a key factor in controlling and regulating the distribution of the diazotrophic microbial community of Schiermonnikoog.     

Evidence for seafloor microbial mats and associated metazoan lifestyles in Lower Cambrian phosphorites of Southwest China

The increase in the depth and intensity of bioturbation through the Proterozoic-­Phanerozoic transition changed the substrates on which marine benthos lived from being relatively firm with a sharp sediment-water interface to having a high water content and blurry sediment-water interface. Additionally, microbial mats, once dominant on normal marine Proterozoic seafloors, were relegated to stressed settings lacking intense metazoan activity. This change in substrates has been termed the 'agronomic revolution', and its impact on benthic metazoans has been termed the 'Cambrian substrate revolution'. The shallow marine phosphorites of the Lower Cambrian Meishucun Formation of southwest China contain evidence suggestive of the presence of seafloor microbial mats. This evidence includes abundant and distinctive red-colored bedding planes enriched in heavy iron minerals and mica, interpreted as resulting from mat-decay mineralization and mica trapping by microbial mats. The radular grazing trace fossil Radulichnus is also found in this formation, indicating a firm, microbial mat-bound substrate. These radular scratches are always preserved with circular impressions around 10 cm in diameter, possibly the fossils of soft-bodied organisms. The first relatively intense bioturbation in this region is found in this formation and is dominated by horizontal Thalassinoides burrows, which could represent undermat mining behavior. The evidence for the presence of microbial mats in the Lower Cambrian Meishucun Formation, and for metazoan lifestyles associated with such mat-bound seafloors, reveals that normal marine environments dominated by typical Proterozoic-style soft substrates still existed during the Cambrian substrate revolution.     

Microscopic Examination of Distribution and Phenotypic Properties of Phylogenetically Diverse Chloroflexaceae-Related Bacteria in Hot Spring Microbial Mats†

We investigated the diversity, distribution, and phenotypes of uncultivated Chloroflexaceae-related bacteria in photosynthetic microbial mats of an alkaline hot spring (Mushroom Spring, Yellowstone National Park). By applying a directed PCR approach, molecular cloning, and sequence analysis of 16S rRNA genes, an unexpectedly large phylogenetic diversity among these bacteria was detected. Oligonucleotide probes were designed to target 16S rRNAs from organisms affiliated with the genus Chloroflexus or with the type C cluster, a group of previously discovered Chloroflexaceae relatives of this mat community. The application of peroxidase-labeled probes in conjunction with tyramide signal amplification enabled the identification of these organisms within the microbial mats by fluorescence in situ hybridization (FISH) and the investigation of their morphology, abundance, and small-scale distribution. FISH was combined with oxygen microelectrode measurements, microscope spectrometry, and microautoradiography to examine their microenvironment, pigmentation, and carbon source usage. Abundant type C-related, filamentous bacteria were found to flourish within the cyanobacterium-dominated, highly oxygenated top layers and to predominate numerically in deeper orange-colored zones of the investigated microbial mats, correlating with the distribution of bacteriochlorophyll a. Chloroflexus sp. filaments were rare at 60°C but were more abundant at 70°C, where they were confined to the upper millimeter of the mat. Both type C organisms and Chloroflexus spp. were observed to assimilate radiolabeled acetate under in situ conditions.