Biogeochemistry of hypersaline microbial mats illustrates the dynamics of modern microbial ecosystems and the early evolution of the biosphere

Photosynthetic microbial mats are remarkably complete self-sustaining ecosystems at the millimeter scale, yet they have substantially affected environmental processes on a planetary scale. These mats may be direct descendents of the most ancient biological communities in which even oxygenic photosynthesis might have developed. Photosynthetic mats are excellent natural laboratories to help us to learn how microbial populations associate to control dynamic biogeochemical gradients.     

Bacterial and archaeal profiling of hypersaline microbial mats and endoevaporites,under natural conditions and methanogenic microcosm experiments

Extremophiles - Bacterial and archaeal community structure of five microbial communities, developing at different salinities in Baja California Sur, Mexico, were characterized by 16S rRNA...     

Phototrophs in high iron microbial mats: microstructure of mats in iron-depositing hot springs

Chocolate Pots Hot Springs in Yellowstone National Park are high in ferrous iron, silica and bicarbonate. The springs are contributing to the active development of an iron formation. The microstructure of photosynthetic microbial mats in these springs was studied with conventional optical microscopy, confocal laser scanning microscopy and transmission electron microscopy. The dominant mats at the highest temperatures (48-54 degrees C) were composed of Synechococcus and Chloroflexus or Pseudanabaena and Mastigocladus. At lower temperatures (36-45 degrees C), a narrow Oscillatoria dominated olive green cyanobacterial mats covering most of the iron deposit. Vertically oriented cyanobacterial filaments were abundant in the top 0.5 mm of the mats. Mineral deposits accumulated beneath this surface layer. The filamentous microstructure and gliding motility may contribute to binding the iron minerals. These activities and heavy mineral encrustation of cyanobacteria may contribute to the growth of the iron deposit. Chocolate Pots Hot Springs provide a model for studying the potential role of photosynthetic prokaryotes in the origin of Precambrian iron formations.     

Archaean metabolic evolution of microbial mats

Microbial mats of coexisting bacteria and archaea date back to the early Archaean: many of the major steps in early evolution probably took place within them. The earliest mats may have formed as biofilms of cooperative chemolithotrophs in hyperthermophile settings, with microbial exploitation of diversifying niches. Anoxygenic photosynthesis using bacteriochlorophyll could have allowed mats, including green gliding bacteria, to colonize anaerobic shallow-water mesothermophile habitats. Exploitation of the Calvin–Benson cycle by purple bacteria allowed diversification of microbial mats, with some organisms in more aerobic habitats, while green sulphur bacteria specialized in anaerobic niches. Cyanobacterial evolution led to more complex mats and plankton, allowing widespread colonization of the globe and the creation of further aerobic habitat. Microbial mat structure may reflect this evolutionary development in broad terms, with anaerobic lower levels occupied by archaeal and bacterial respirers, fermenters and green bacteria, while the higher levels contain aerobic purple bacteria and are dominated by cyanobacteria. A possible origin of eukaryotes is from a fusion of symbiotic partners living across a redox boundary in a mat. The geological record of Archaean mats may be present as isotopic fingerprints: with the presence of cyanobacteria, mats may have had a nearly modern structure as early as 3.5 Ga ago (1 Ga = 10⁹ years).     

Biogeochemistry and microbial ecology of methane oxidation in anoxic environments: a review

Evidence supporting a key role for anaerobic methane oxidation in the global methane cycle is reviewed. Emphasis is on recent microbiological advances. The driving force for research on this process continues to be the fact that microbial communities intercept and consume methane from anoxic environments, methane that would otherwise enter the atmosphere. Anaerobic methane oxidation is biogeochemically important because methane is a potent greenhouse gas in the atmosphere and is abundant in anoxic environments. Geochemical evidence for this process has been observed in numerous marine sediments along the continental margins, in methane seeps and vents, around methane hydrate deposits, and in anoxic waters. The anaerobic oxidation of methane is performed by at least two phylogenetically distinct groups of archaea, the ANME-1 and ANME-2. These archaea are frequently observed as consortia with sulfate-reducing bacteria, and the metabolism of these consortia presumably involves a syntrophic association based on interspecies electron transfer. The archaeal member of a consortium apparently oxidizes methane and shuttles reduced compounds to the sulfate-reducing bacteria. Despite recent advances in understanding anaerobic methane oxidation, uncertainties still remain regarding the nature and necessity of the syntrophic association, the biochemical pathway of methane oxidation, and the interaction of the process with the local chemical and physical environment. This review will consider the microbial ecology and biogeochemistry of anaerobic methane oxidation with a special emphasis on the interactions between the responsible organisms and their environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Bioremediation of oil by marine microbial mats

Cyanobacterial mats developing in oil-contaminated sabkhas along the African coasts of the Gulf of Suez and in the pristine Solar Lake, Sinai, were collected for laboratory studies. Samples of both mats showed efficient degradation of crude oil in the light, followed by development of an intense bloom of Phormidium spp. and Oscillatoria spp. Isolated cyanobacterial strains, however, did not degrade crude oil in axenic cultures. Strains of sulfate-reducing bacteria and aerobic heterotrophs were capable of degrading model compounds of aliphatic and aromatic hydrocarbons. Results indicate that degradation of oil was done primarily by aerobic heterotrophic bacteria. The oxygenic photosynthesis of oil-insensitive cyanobacteria supplied the molecular oxygen for the efficient aerobic metabolism of organisms, such as Marinobacter sp. The diurnal shifts in environmental conditions at the mat surface, from highly oxic conditions in the light to anaerobic sulfide-rich habitat in the dark, may allow the combined aerobic and anaerobic degradation of crude oil at the mat surface. Hence, coastal cyanobacterial mats may be used for the degradation of coastline oil spills. Oxygen microelectrodes detected a significant inhibition of photosynthetic activity subsequent to oil addition. This prevailed for a few hours and then rapidly recovered. In addition, shifts in bacterial community structure following exposure to oil were determined by denaturing gradient gel electrophoresis of PCR-amplified fractions of 16S rRNA from eubacteria, cyanobacteria and sulfate-reducing bacteria. Since the mats used for the present study were obtained from oil-contaminated environments, they were believed to be preequilibrated for petroleum remediation. The mesocosm system at Eilat provided a unique opportunity to study petroleum degradation by mats formed under different salinities (up to 21%). These mats, dominated by cyanobacteria, can serve as close analogues to the sabkhas contaminated during the Gulf War in Kuwait and Saudi Arabia. Electronic Publication     

Effect of a temperature gradient on Sphagnum fallax and its associated living microbial communities: a study under controlled conditions

Microbial communities living in Sphagnum are known to constitute early indicators of ecosystem disturbances, but little is known about their response (including their trophic relationships) to climate change. A microcosm experiment was designed to test the effects of a temperature gradient (15, 20, and 25°C) on microbial communities including different trophic groups (primary producers, decomposers, and unicellular predators) in Sphagnum segments (0-3 cm and 3-6 cm of the capitulum). Relationships between microbial communities and abiotic factors (pH, conductivity, temperature, and polyphenols) were also studied. The density and the biomass of testate amoebae in Sphagnum upper segments increased and their community structure changed in heated treatments. The biomass of testate amoebae was linked to the biomass of bacteria and to the total biomass of other groups added and, thus, suggests that indirect effects on the food web structure occurred. Redundancy analysis revealed that microbial assemblages differed strongly in Sphagnum upper segments along a temperature gradient in relation to abiotic factors. The sensitivity of these assemblages made them interesting indicators of climate change. Phenolic compounds represented an important explicative factor in microbial assemblages and outlined the potential direct and (or) indirect effects of phenolics on microbial communities.     

Evolution of a microbial nitrilase gene family: a comparative and environmental genomics study

Background  

Completed genomes and environmental genomic sequences are bringing a significant contribution to understanding the evolution of gene families, microbial metabolism and community eco-physiology. Here, we used comparative genomics and phylogenetic analyses in conjunction with enzymatic data to probe the evolution and functions of a microbial nitrilase gene family. Nitrilases are relatively rare in bacterial genomes, their biological function being unclear.     

Artificial soil microcosms: a tool for studying microbial autecology under controlled conditions

A novel artificial microcosm containing all the essential chemical components of soil, but with reduced heterogeneity and biological complexity, has been developed. Its utility for supporting realistic microbial populations was demonstrated and an example of how competing bacteria can be studied is illustrated.     

Effects of environmental conditions on microbial proteolysis in a pork myofibril model system

P.M. KENNEALLY, N.G. FRANSEN, H. GRAU, E.E. O'NEILL & E.K. ARENDT.1999.A number of bacterial strains used for meat fermentations were screened for proteolytic activity. A strain of Micrococcus which was found to be proteolytic was evaluated for the effects of environmental conditions on its proteolytic activity against pork myofibrillar proteins using response surface methodology. Three strains of micrococci were also tested for the ability to produce free amino acids from pork myofibrils. Analysis of the effects of environmental conditions showed that proteolytic activity would be minimal under conditions normally found in fermented sausages, thereby suggesting that proteolysis in these products is largely due to endogenous meat enzymes. The three strains of micrococci were shown to produce free amino acids from pork myofibrils, thereby demonstrating the presence of peptidase activity in these strains.     

Annual growth layers as proxies of past growth conditions for benthic microbial mats in a perennially ice-covered Antarctic lake

Perennial microbial mats can be the dominant autotrophic community in Antarctic lakes. Their seasonal growth results in clearly discernible annual growth layering. We examined features of live microbial mats from a range of depths in Lake Hoare, Antarctica, that are likely to be preserved in these layers to determine their potential as proxies of past growth performance. Cyanobacteria dominated the mat for all but the deepest depth sampled. Changes in areal concentrations of phycobilin pigments, organic matter and extracellular polysaccharide and in species composition did not correspond to changes in various water column properties, but showed a linear relationship with irradiance. Carbonate accumulation in the mats correlated with biomass markers and may be inferred as an index of mat performance. We examined the carbonate content of annual layers laid down from 1958–1959 to 1994–1995 in sediment cores from 12 m depth. The carbonate content in the layer showed a significant correlation with the mean summer air temperature. These data suggest a link between air temperature and microbial mat growth performance, and suggest that it is mediated via irradiance. Laminated microbial mats in Antarctic lakes have the potential to act as fine-resolution records of environmental conditions in the recent past, although interpretation is complex.     

Leaf temperatures under controlled environmental conditions

A wind tunnel and an external microwave energy source formed the basis of an experimental technique to investigate many complex functional relationships between climatic parameters and leaf temperatures. The results of preliminary experimental plant-environment interactions are compared with published theoretical results obtained by solving the energy balance equation for a single leaf.     

Methods of assessing microbial activity and inhibition under anaerobic conditions: a literature review

This work reviews the existing methodologies for assessing microbial activity and inhibition under anaerobic conditions. The anaerobic digestion process consists of several metabolic steps–the Anaerobic Digestion Model No. 1 (ADM1) has attempted to describe these steps in the form of a mathematical model with the intention of providing a reference base for all further efforts in the modelling of anaerobic processes. The existence of a reference point for modelling has highlighted the fact that there is a lack of coherence between the many different methodologies for experimentally assessing anaerobic activity and inhibition.A working group of the International Water Association was recently founded to harmonise the existing methodologies with the ultimate intention of developing a unified reference procedure– a primary objective of the group will be the establishment of a standard terminology in the field of anaerobic digestion, activity and inhibition assessment. Secondly, it will compare the existing methodologies and develop standard protocols for assessing the kinetic parameters (e.g. maximum uptake rate, half-saturation constant) of anaerobic processes that may be entered directly into ADM1 and its successors.This paper revises and enlarges a contribution presented by the authors at the workshop Harmonisation of anaerobic biodegradation, activity and inhibition assays (Ligthart & Nieman 2002, Proc. workshop held in Orta (Italy) June 7–8, 2002) and aims to promote a clear understanding of the currently established methodology.Numerous methods have been developed over the past 30 years, since Van den Berg et al. (1974, Biotechnol Bioeng 16(11)– 1459–1469) measured methanogenic activity, by using a manometric device equipped with a photoelectric sensor to quantify the gas production. Methanogenesis is often the rate limiting step of the entire process and since the quantification of gas flowrate is relatively easy to perform, most of the methods reported in literature monitor the production of biogas. These methods can be termed volumetric or manometric methods, as the volume of biogas produced or the pressure increase due to gas production inside a close vessel are assessed, respectively. However, this same concept can be employed to assess activity or inhibition of individual metabolic steps preceding the methanogenic one, providing that they are rate limiting for the whole process. The reliability of activity assessment through gas measurement has been proven to be strongly dependent on the equilibrium between liquid and gas phase in a closed vessel. This can be influenced by many factors, e.g. the amount and characteristics of the test substrate; the concentration of the biomass; the gas-to-liquid ratio– all these aspects will need to be addressed in the standard procedure. Other direct or indirect methods, targeting physico-chemical or microbiological parameter exist and have been investigated by many authors. Besides the interest for research purposes, the definition of reference methods to assess activity and inhibition can be of great interest for engineers, both phy. Specific reference procedures might be needed for particular applications, e.g. the (kinetic) study of rate limiting microbial steps and might require ad-hoc methodologies to be devised. A microbiological technique such as FISH, coupled with microsensors have been reported to have a great potential in the near future.Passed away on April 7th 2003.     

Cyanobacterial composition of microbial mats from an Australian thermal spring: a polyphasic evaluation

Cyanobacterial composition of microbial mats from an alkaline thermal spring issuing at 43-71 degrees C from tropical north-eastern Australia are described using a polyphasic approach. Eight genera and 10 species from three cyanobacterial orders were identified based on morphological characters. These represented taxa previously known as thermophilic from other continents. Ultrastructural analysis of the tower mats revealed two filamentous morphotypes contributed the majority of the biomass. Both types had ultrastructural characteristics of the family Pseudanabaenaceae. DNA extracts were made from sections of the tentaculiform towers and the microbial community analysed by 16S cyanobacteria-specific PCR and denaturing-gradient gel electrophoresis. Five significant bands were identified and sequenced. Two bands clustered closely with Oscillatoria amphigranulata isolated from New Zealand hot springs; one unique phylotype had only moderate similarity to a range of Leptolyngbya species; and one phylotype was closely related to a number of Geitlerinema species. Generally the approaches yielded complementary information, however the results suggest that species designation based on morphological and ultrastructural criteria alone often fails to recognize their true phylogenetic position. Conversely some molecular techniques may fail to detect rare taxa suggesting that the widest possible suite of techniques be applied when conducting analyses of cyanobacterial diversity of natural populations. This is the first polyphasic evaluation of thermophilic cyanobacterial communities from the Australian continent.     

Contribution of microbial mats to sedimentary surface structures

Summary This paper summarizes studies of sedimentary surface structures in which microbial mats play a role. Intertidal/supratidal transitions of tidal flats of the North Sea coast, and shallow hypersaline water bodies of salterns (Bretagne, Canary and Balearic Islands), and Gavish Sabkha (Sinai) reveal a multitude of sedimentary surface structures which can be grouped and primary biologically controlled structures. Physically controlled surface structures include shrinkage cracks, erosion marks, deformation structures caused by water friction, gas pressure and mineral encrustation. Shrinkage cracks in microbial mats reveal the following features: (i) horizontally arranged cauliflower pattern that differs from the usually orthogonally regular crack morphology in clay, (ii) rounded edges and pillow-like thickening along the crack edges, caused by the growth of mats into the cracks. Criteria of erosion are pocket-like depressions and ripple marks on the thus exposed non-stabilized sand, and residual stacks of microbial mats. Deformation structures are due to water friction causing flotation of loosely attached microbial mats which fold and tear. Gas migration from deeper layers causes domal upheaval, protuberance structures, folds and “fairy rings”. Protuberance structures are caused by the rupture of gas domes and rapid escape of the enclosed gas. The sudden drop of pressure forces sediment to well up from below through the gas channels and to fill the internal hollow spaces of the domes. “Fairy rings” are horizontal ringshaped structures. Their center is the exit point of gas bubbles which escape from the substrate into the shallow water. The bubbles generate concentric waves which cause displacement of fine muddy sediments at the sediment-water interface Such gradual displacement guides mat-constructing microbes to grow concentrically. The “fairy rings” are crowned by pinnacle structures of bacterial and diatom origin. Pinnacles, “fairy rings” and pillow-like coatings of crack margins are biogenic structures which have to be genetically separated from purely physically controlled structures.     

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.     

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.