16S rDNA diversity of cultured and uncultured prokaryotes of a mat sample from Lake Fryxell, McMurdo Dry Valleys, Antarctica

The prokaryotic diversity of aerobic and anaerobic bacterial isolates and of bacterial and archaeal 16S rDNA clones was determined for a microbial mat sample from the moated region of Lake Fryxell, McMurdo Dry Valleys, Antarctica. Among the anaerobic bacteria, members of Clostridium estertheticum and some other psychrotolerant strains dominated whereas methanogens and other Archaea were lacking. Isolates highly related to Flavobacterium hibernum, Janthiniobacterium lividum, and Arthrobacter flavus were among the aerobic bacteria most frequently isolated. Assessment of more than 350 partial 16S rDNA clone sequences of libraries generated by Bacteria- and Archaea-specific PCR primers revealed a rich spectrum of bacterial diversity but only two different archaeal clone sequences. Among the Bacteria, representative sequences belonged to the class Proteobacteria, order Verrucomicrobiales, class Actinobacteria, Clostridium/Bacillus subphylum of Gram-positives, and the Cytophaga-Flavobacterium-Bacteroides phylum. The clones formed about 70 higher taxonomy groups (<98% sequence similarity) and 133 potential species, i.e., groups of clones sharing greater than 98% similarity. Only rarely were clone sequences found to be highly related to Lake Fryxell isolates and to strains of described species. Subsequent analysis of ten sequencing batches of 36 individual clones indicated that the diversity might be still higher than had been assessed.     

Heterotrophic bacterial diversity in aquatic microbial mat communities from Antarctica

Heterotrophic bacteria isolated from five aquatic microbial mat samples from different locations in continental Antarctica and the Antarctic Peninsula were compared to assess their biodiversity. A total of 2,225 isolates obtained on different media and at different temperatures were included. After an initial grouping by whole-genome fingerprinting, partial 16S rRNA gene sequence analysis was used for further identification. These results were compared with previously published data obtained with the same methodology from terrestrial and aquatic microbial mat samples from two additional Antarctic regions. The phylotypes recovered in all these samples belonged to five major phyla, Actinobacteria, Bacteroidetes, Proteobacteria, Firmicutes and Deinococcus-Thermus, and included several potentially new taxa. Ordination analyses were performed in order to explore the variance in the diversity of the samples at genus level. Habitat type (terrestrial vs. aquatic) and specific conductivity in the lacustrine systems significantly explained the variation in bacterial community structure. Comparison of the phylotypes with sequences from public databases showed that a considerable proportion (36.9%) is currently known only from Antarctica. This suggests that in Antarctica, both cosmopolitan taxa and taxa with limited dispersal and a history of long-term isolated evolution occur.     

Cold-active acetogenic bacteria from surficial sediments of perennially ice-covered Lake Fryxell, Antarctica

RibR is a minor cryptic flavokinase (EC 2.7.1.26) of the Gram-positive bacterium Bacillus subtilis with an unknown cellular function. The flavokinase activity appears to be localized to the N-terminal domain of the protein. Using the yeast three-hybrid system, it was shown that RibR specifically interacts in vivo with the nontranslated wild-type leader of the mRNA of the riboflavin biosynthetic operon. This interaction is lost partially when a leader containing known cis-acting deregulatory mutations in the so-called RFN element is tested. The RFN element is a sequence within the rib-leader mRNA reported to serve as a receptor for an FMN-dependent 'riboswitch'. In RibR itself, interaction was localized to the carboxy-terminate part of the protein, a segment of unknown function that does not show similarity to other proteins in the public databases. Analysis of a ribR-defective strain revealed a mild deregulation with respect to flavin (riboflavin, FMN and FAD) biosynthesis. The results indicate that the RNA-binding protein RibR may be involved in the regulation of the rib genes.     

Culture-independent analysis of bacterial diversity in a child-care facility

Background  

Child-care facilities appear to provide daily opportunities for exposure and transmission of bacteria and viruses. However, almost nothing is known about the diversity of microbial contamination in daycare facilities or its public health implications. Recent culture-independent molecular studies of bacterial diversity in indoor environments have revealed an astonishing diversity of microorganisms, including opportunistic pathogens and many uncultured bacteria. In this study, we used culture and culture-independent methods to determine the viability and diversity of bacteria in a child-care center over a six-month period.     

Geochemistry of aquatic humic substances in the Lake Fryxell Basin, Antarctica

Dissolved organic carbon (DOC) in Lake Fryxell, 10 streams flowing into the lake, and the moat surrounding the lake was studied to determine the influence of sources and biogeochemical processes on its distribution and chemical nature. Lake Fryxell is an amictic, permanently ice-covered lake in the McMurdo Dry Valleys which contains benthic and planktonic microbial populations, but receives essentially no input of organic material from the ahumic soils of the watershed. Biological activity in the water column does not appear to influence the DOC depth profile, which is similar to the profiles for conservative inorganic constituents. DOC values for the streams varied with biomass in the stream channel, and ranged from 0.2 to 9.7 mg C/L. Fulvic acids in the streams were a lower percentage of the total DOC than in the lake. These samples contain recent carbon and appear to be simpler mixtures of compounds than the lake samples, indicating that they have undergone less humification. The fulvic acids from just above the sediments of the lake have a high sulfur content and are highly aliphatic. The main transformations occurring as these fractions diffuse upward in the water column are 1) loss of sulfur groups through the oxycline and 2) decrease in aliphatic carbon and increase in the heterogeneity of aliphatic moieties. The fraction of modem¹⁴C content of the lake fulvic acids range from a minimum of 0.68 (approximately 3000 years old) at 15m depth to 0.997 (recent material) just under the ice. The major processes controlling the DOC in the lake appear to be: 1) The transport of organic matter by the inflow streams resulting in the addition of recent organic material to the moat and upper waters of the lake; 2) The diffusion of organic matter composed of relict organic material and organic carbon resulting from the degradation of algae and bacteria from the bottom waters or sediments of the lake into overlying glacial melt water, 3) The addition of recent organic matter to the bottom waters of the lake from the moat.     

Temperature and nutrient induced responses of Lake Fryxell sulfate-reducing prokaryotes and description of Desulfovibrio lacusfryxellense, sp. nov., a pervasive, cold-active, sulfate-reducing bacterium from Lake Fryxell, Antarctica

The effects of temperature and carbon substrate availability on the stimulation of sulfate reduction by indigenous populations of sulfate-reducing prokaryotes (SRP) in permanently ice-covered Lake Fryxell, Antarctica were investigated. Psychrophilic and halotolerant, lactate-degrading SRP showed significant metabolic activity throughout all sampled depths of the water column, suggesting that such organisms, possibly of marine origin, may be key contributors to carbon and sulfur cycling in Lake Fryxell. Planktonic and benthic strains of lactate-oxidizing sulfate-reducing bacteria (SRB) were isolated from samples of various depths of the anoxic water column and from surficial sediments. Phylogenetic analyses of 16S rRNA gene sequences placed the Fryxell sulfate-reducer (FSR) strains within the Deltaproteobacteria and showed them to be most closely related to the Arctic marine species of SRB Desulfovibrio frigidus and Desulfovibrio ferrireducens. Based on phylogenetic and phenotypic differences between the Antarctic FSR strains and related species of the genus Desulfovibrio, strain FSRs^T (=DSM 23315^T =ATCC BAA-2083^T) is proposed as the type strain of a novel species of cold-active SRB, Desulfovibrio lacusfryxellense, sp. nov.     

Biodiversity of methanogenic and other archaea in the permanently frozen Lake Fryxell, Antarctica

Archaea were detected in molecular diversity studies of the permanently frozen Lake Fryxell, Antarctica. Two clusters of methanogens were detected in the sediments, and another cluster of possibly methanotrophic Euryarchaeota was detected in the anoxic water column just above the sediments. One crenarchaeote was detected in water just below the oxycline. The Archaea present in Lake Fryxell are likely involved in the major biogeochemical cycles that occur there.     

Isolation, characterization, and ecology of cold-active, chemolithotrophic, sulfur-oxidizing bacteria from perennially ice-covered Lake Fryxell, Antarctica

Novel strains of obligately chemolithoautotrophic, sulfur-oxidizing bacteria have been isolated from various depths of Lake Fryxell, Antarctica. Physiological, morphological, and phylogenetic analyses showed these strains to be related to mesophilic Thiobacillus species, such as T. thioparus. However, the psychrotolerant Antarctic isolates showed an adaptation to cold temperatures and thus should be active in the nearly freezing waters of the lake. Enumeration by most-probable-number analysis in an oxic, thiosulfate-containing medium revealed that the sulfur-oxidizing chemolithotroph population peaks precisely at the oxycline (9.5 m), although viable cells exist well into the anoxic, sulfidic waters of the lake. The sulfur-oxidizing bacteria described here likely play a key role in the biogeochemical cycling of carbon and sulfur in Lake Fryxell.     

Isolation, Characterization, and Ecology of Cold-Active, Chemolithotrophic, Sulfur-Oxidizing Bacteria from Perennially Ice-Covered Lake Fryxell, Antarctica

Novel strains of obligately chemolithoautotrophic, sulfur-oxidizing bacteria have been isolated from various depths of Lake Fryxell, Antarctica. Physiological, morphological, and phylogenetic analyses showed these strains to be related to mesophilic Thiobacillus species, such as T. thioparus. However, the psychrotolerant Antarctic isolates showed an adaptation to cold temperatures and thus should be active in the nearly freezing waters of the lake. Enumeration by most-probable-number analysis in an oxic, thiosulfate-containing medium revealed that the sulfur-oxidizing chemolithotroph population peaks precisely at the oxycline (9.5 m), although viable cells exist well into the anoxic, sulfidic waters of the lake. The sulfur-oxidizing bacteria described here likely play a key role in the biogeochemical cycling of carbon and sulfur in Lake Fryxell.     

Biodiversity of Methanogenic and Other Archaea in the Permanently Frozen Lake Fryxell, Antarctica

Archaea were detected in molecular diversity studies of the permanently frozen Lake Fryxell, Antarctica. Two clusters of methanogens were detected in the sediments, and another cluster of possibly methanotrophic Euryarchaeota was detected in the anoxic water column just above the sediments. One crenarchaeote was detected in water just below the oxycline. The Archaea present in Lake Fryxell are likely involved in the major biogeochemical cycles that occur there.     

Cyanobacterial diversity in natural and artificial microbial mats of Lake Fryxell (McMurdo Dry Valleys,Antarctica): a morphological and molecular approach

Currently, there is no consensus concerning the geographic distribution and extent of endemism in Antarctic cyanobacteria. In this paper we describe the phenotypic and genotypic diversity of cyanobacteria in a field microbial mat sample from Lake Fryxell and in an artificial cold-adapted sample cultured in a benthic gradient chamber (BGC) by using an inoculum from the same mat. Light microscopy and molecular tools, including 16S rRNA gene clone libraries, denaturing gradient gel electrophoresis, and sequencing, were used. For the first time in the study of cyanobacterial diversity of environmental samples, internal transcribed spacer (ITS) sequences were retrieved and analyzed to complement the information obtained from the 16S rRNA gene. Microscopy allowed eight morphotypes to be identified, only one of which is likely to be an Antarctic endemic morphotype. Molecular analysis, however, revealed an entirely different pattern. A much higher number of phylotypes (15 phylotypes) was found, but no sequences from Nodularia and Hydrocoryne, as observed by microscopy, were retrieved. The 16S rRNA gene sequences determined in this study were distributed in 11 phylogenetic lineages, 3 of which were exclusively Antarctic and 2 of which were novel. Collectively, these Antarctic sequences together with all the other polar sequences were distributed in 22 lineages, 9 of which were exclusively Antarctic, including the 2 novel lineages observed in this study. The cultured BGC mat had lower diversity than the field mat. However, the two samples shared three morphotypes and three phylotypes. Moreover, the BGC mat allowed enrichment of one additional phylotype. ITS sequence analysis revealed a complex signal that was difficult to interpret. Finally, this study provided evidence of molecular diversity of cyanobacteria in Antarctica that is much greater than the diversity currently known based on traditional microscopic analysis. Furthermore, Antarctic endemic species were more abundant than was estimated on the basis of morphological features. Decisive arguments concerning the global geographic distribution of cyanobacteria should therefore incorporate data obtained with the molecular tools described here.     

Diversity and distribution of sulfate-reducing bacteria in permanently frozen Lake Fryxell, McMurdo Dry Valleys, Antarctica

The permanently frozen freshwater Lake Fryxell, located in the Dry Valleys of Antarctica, exhibits an ideal geochemistry for microbial sulfate reduction. To investigate the population of sulfate-reducing bacteria in Lake Fryxell, both 16S rRNA gene and metabolic primer sets targeting the dsrA gene for the dissimilatory sulfite reductase alpha subunit were employed to analyze environmental DNA obtained from the water column and sediments of Lake Fryxell. In addition, enrichment cultures of sulfate-reducing bacteria established at 4 degrees C from Lake Fryxell water were also screened using the dsrA primer set. The sequence information obtained showed that a diverse group of sulfate-reducing prokaryotes of the domain Bacteria inhabit Lake Fryxell. With one exception, the enrichment culture sequences were not represented within the environmental sequences. Sequence data were compared with the geochemical profile of Lake Fryxell to identify possible connections between the diversity of sulfate-reducing bacteria and limnological conditions. Several clone groups were highly localized with respect to lake depth and, therefore, experienced specific physiochemical conditions. However, all sulfate-reducing bacteria inhabiting Lake Fryxell must function under the constantly cold conditions characteristic of this extreme environment.     

Diversity and Distribution of Sulfate-Reducing Bacteria in Permanently Frozen Lake Fryxell, McMurdo Dry Valleys, Antarctica

The permanently frozen freshwater Lake Fryxell, located in the Dry Valleys of Antarctica, exhibits an ideal geochemistry for microbial sulfate reduction. To investigate the population of sulfate-reducing bacteria in Lake Fryxell, both 16S rRNA gene and metabolic primer sets targeting the dsrA gene for the dissimilatory sulfite reductase alpha subunit were employed to analyze environmental DNA obtained from the water column and sediments of Lake Fryxell. In addition, enrichment cultures of sulfate-reducing bacteria established at 4°C from Lake Fryxell water were also screened using the dsrA primer set. The sequence information obtained showed that a diverse group of sulfate-reducing prokaryotes of the domain Bacteria inhabit Lake Fryxell. With one exception, the enrichment culture sequences were not represented within the environmental sequences. Sequence data were compared with the geochemical profile of Lake Fryxell to identify possible connections between the diversity of sulfate-reducing bacteria and limnological conditions. Several clone groups were highly localized with respect to lake depth and, therefore, experienced specific physiochemical conditions. However, all sulfate-reducing bacteria inhabiting Lake Fryxell must function under the constantly cold conditions characteristic of this extreme environment.     

A gas vesiculate planktonic strain of the purple non-sulfur bacterium Rhodoferax antarcticus isolated from Lake Fryxell, Dry Valleys, Antarctica

A moderately psychrophilic purple non-sulfur bacterium, Rhodoferax antarcticus strain Fryx1, is described. Strain Fryx1 was isolated from the water column under the ice of the permanently frozen Lake Fryxell, Antarctica. Cells of Fryx1 are long thin rods and contain gas vesicles, the first report of such structures in purple non-sulfur bacteria. Gas vesicles are clustered at 2–4 sites per cell. Surprisingly, the 16S rRNA gene sequence of strain Fryx1 is nearly identical to that of Rfx. antarcticus strain AB, a short, vibrio-shaped phototroph isolated from an Antarctic microbial mat. Although showing physiological parallels, strains AB and Fryx1 differ distinctly in their morphology and absorption spectra. DNA–DNA hybridization shows that the genomes of strains AB and Fryx1 are highly related, yet distinct. We conclude that although strains AB and Fryx1 may indeed be the same species, their ecologies are quite different. Unlike strain AB, strain Fryx1 has adapted to a planktonic existence in the nearly freezing water column of Lake Fryxell.Dedicated to Prof. Dr. Hans Günter Schlegel on the occasion of his 80th birthday.     

The microbial plankton of Lake Fryxell,southern Victoria Land,Antarctica during the summers of 1992 and 1994

 Samples collected from Lake Fryxell, southern Victoria Land, Antarctica in January 1992 and 1994 were analysed for the abundance of bacterioplankton and the diversity and abundance of protistan plankton. At the times of sampling, 14 ciliate species and 10 species of autotrophic flagellate were recorded. The samples contained two species of rotifer (Philodina spp.), which formed the first record of planktonic metazoans in the Dry Valley lakes of this region of Antarctica. Bacterial concentrations ranged between 1.0 and 3.8×10⁸ l^-1 in the upper oxic waters increasing to 20×10⁸ l^-1 in the anoxic waters. Heterotrophic flagellates decreased in abundance down the oxygenated water column, disappearing completely at 9 m, and ranged between 0.28 and 7.39×10⁵ l^-1 in abundance. Autotrophic flagellates were much more abundant exhibiting a number of distinct peaks down the water column (1.89–25.3×10⁸ l^-1). The ciliated protozoa were very abundant (up to 7720 l^-1) in relation to flagellate and bacterial numbers, typical of oligotrophic lakes world-wide. The distribution of the protistan plankton showed marked zonation, probably in response to the differing salinity and temperature gradients in the water column. Possible trophic interactions are discussed and comparisons with other continental Antarctic lakes made. Received: 29 November 1995/Accepted: 18 February 1996     

Culture-independent analysis of gut bacteria: the pig gastrointestinal tract microbiota revisited

The phylogenetic diversity of the intestinal bacterial community in pigs was studied by comparative 16S ribosomal DNA (rDNA) sequence analysis. Samples were collected from a total of 24 pigs representing a variety of diets, ages, and herd health status. A library comprising 4,270 cloned 16S rDNA sequences obtained directly by PCR from 52 samples of either the ileum, the cecum, or the colon was constructed. In total, 375 phylotypes were identified using a 97% similarity criterion. Three hundred nine of the phylotypes (83%) had a <97% sequence similarity to any sequences in the database and may represent yet-uncharacterized bacterial genera or species. The phylotypes were affiliated with 13 major phylogenetic lineages. Three hundred four phylotypes (81%) belonged to the low-G+C gram-positive division, and 42 phylotypes (11.2%) were affiliated with the Bacteroides and Prevotella group. Four clusters of phylotypes branching off deeply within the low-G+C gram-positive bacteria and one in the Mycoplasma without any cultured representatives were found. The coverage of all the samples was 97.2%. The relative abundance of the clones approximated a lognormal distribution; however, the phylotypes detected and their abundance varied between two libraries from the same sample. The results document that the intestinal microbial community is very complex and that the majority of the bacterial species colonizing the gastrointestinal tract in pigs have not been characterized.     

Vertical distribution of organic constituents in an Antarctic lake: Lake Fryxell

Vertical distribution of organic constituents, i.e. total organic carbon (TOC), hydrocarbons, fatty acids and hydroxy acids in water and sediment samples from Lake Fryxell (77° 35 S, 163° 15 E) of southern Victoria Land, Antarctica were studied to elucidate their features in relation to stratification of the lake waters and likely distribution of microorganisms. The TOC content of the surface water (5.0 m; just below the ice cover of 4.50 m thickness) was 1.4 mg l^–1. It increased markedly with depth and attained a maximum value of 21.7 mg C l^–1 at a depth of 17.5 m, but decreased to the bottom (13.3 mg C l^–1). The high TOC content of the anoxic bottom layers (> 15 m) is attributable to the concentration of refractory organic substances over long periods following the degradation of labile organic constituents. Hydrocarbons were not found in the water column, but the major constituent of the bottom sediment was n-C_{29 : 2} alkene. Total concentrations of fatty acids in the oxic layers ( 10 m) were highest at 10.0 m and much higher than those in the anoxic layers (> 10 m), probably reflecting the phytoplankton population. The content of branched (iso and anteiso) fatty acids and 3-hydroxy acids in the anoxic layers were much greater than those in the oxic layers which would seem to reflect the distribution of bacterial abundance. The differences of organic composition between the water column and sediments imply that sinking dead organisms were quickly degraded in the lake bottom. Also, the composition of microorganisms in the water column must be very different from that in the sediments.     

Hydrocarbon contamination changes the bacterial diversity of soil from around Scott Base, Antarctica

A combination of culture-independent and culturing methods was used to determine the impacts of hydrocarbon contamination on the diversity of bacterial communities in coastal soil from Ross Island, Antarctica. While numbers of culturable aerobic heterotrophic microbes were 1-2 orders of magnitude higher in the hydrocarbon-contaminated soil than control soil, the populations were less diverse. Members of the divisions Fibrobacter/Acidobacterium, Cytophaga/Flavobacterium/Bacteroides, Deinococcus/Thermus, and Low G+C gram positive occurred almost exclusively in control soils whereas the contaminated soils were dominated by Proteobacteria; specifically, members of the genera Pseudomonas, Sphingomonas and Variovorax, some of which degrade hydrocarbons. Members of the Actinobacteria were found in both soils.     

Analyses of soil bacterial diversity of the Schirmacher Oasis,Antarctica

Schirmacher Oasis, Antarctica, is a region with relatively large exposed area and consisted of many freshwater lakes. Nevertheless, only a few studies were done on the bacterial diversity of this region. Hence, this project was undertaken to determine the bacterial community in soil samples collected from the Schirmacher Oasis using the denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA fragments. A total of 79 partial 16S rDNA sequences were obtained from the excised DGGE bands, which corresponded to 63 different operational taxonomic units (OTUs) representing bacteria from seven different phyla. The most dominant phyla in descending order were Acidobacteria, Proteobacteria, Bacteroidetes, and Actinobacteria, Planctomycetes, Cyanobacteria and BRC1. There were 5.4 % of unclassified bacteria which cannot be grouped into any of the existing phyla. Eighty-seven percent of the OTUs had highest similarity with the uncultured bacteria from the NCBI GenBank database. Thirty-two percent of the OTUs were similar to bacteria reported in other parts of the Antarctica, while the others were related to bacteria found elsewhere outside the Antarctic.