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Cold desert soil microbiomes thrive despite severe moisture and nutrient limitations. In Eastern Antarctic soils, bacterial primary production is supported by trace gas oxidation and the light-independent RuBisCO form IE. This study aims to determine if atmospheric chemosynthesis is widespread within Antarctic, Arctic and Tibetan cold deserts, to identify the breadth of trace gas chemosynthetic taxa and to further characterize the genetic determinants of this process. H2 oxidation was ubiquitous, far exceeding rates reported to fulfill the maintenance needs of similarly structured edaphic microbiomes. Atmospheric chemosynthesis occurred globally, contributing significantly (p Chloroflexota, Firmicutes, Deinococcota and Verrucomicrobiota genomes. We identify a novel group of high-affinity [NiFe]-hydrogenases, group 1m, through phylogenetics, gene structure analysis and homology modeling, and reveal substantial genetic diversity within RuBisCO form IE (rbcL1E), and high-affinity 1h and 1l [NiFe]-hydrogenase groups. We conclude that atmospheric chemosynthesis is a globally-distributed phenomenon, extending throughout cold deserts, with significant implications for the global carbon cycle and bacterial survival within environmental reservoirs.

Snow algae are predicted to expand in polar regions due to climate warming, which can accelerate snowmelt by reducing albedo. Green snow frequently occurs near penguin colonies, and red snow distributes widely along ocean shores. However, the mechanisms underpinning the assemblage of algae and heterotrophs in colored snow remain poorly characterized. We investigated algal, bacterial, and fungal communities and their interactions in red and green snows in the Antarctic Peninsula using a high-throughput sequencing method. We found distinct algal community structure in red and green snows, and the relative abundance of dominant taxa varied, potentially due to nutrient status differences. Contrastingly, red and green snows exhibited similar heterotrophic communities (bacteria and fungi), whereas the relative abundance of fungal pathogens was substantially higher in red snow by 3.8-fold. Red snow exhibited a higher network complexity, indicated by a higher number of nodes and edges. Red snow exhibited a higher proportion of negative correlations among heterotrophs (62.2% vs 3.4%) and stronger network stability, suggesting the red-snow network is more resistant to external disturbance. Our study revealed that the red snow microbiome exhibits a more stable microbial network than the green snow microbiome.

Glaciers represent important biomes of Earth and are recognized as key species pools for downstream aquatic environments. Worldwide, rapidly receding glaciers are driving shifts in hydrology, species distributions and threatening microbial diversity in glacier-fed aquatic ecosystems. However, the impact of glacier surface snow-originating taxa on the microbial diversity in downstream aquatic environments has been little explored. To elucidate the contribution of glacier surface snow-originating taxa to bacterial diversity in downstream aquatic environments, we collected samples from glacier surface snows, downstream streams and lakes along three glacier-fed hydrologic continuums on the Tibetan Plateau. Our results showed that glacier stream acts as recipients and vectors of bacteria originating from the glacier environments. The contributions of glacier surface snow-originating taxa to downstream bacterial communities decrease from the streams to lakes, which was consistently observed in three geographically separated glacier-fed ecosystems. Our results also revealed that some rare snow-originating bacteria can thrive along the hydrologic continuums and become dominant in downstream habitats. Finally, our results indicated that the dispersal patterns of bacterial communities are largely determined by mass effects and increasingly subjected to local sorting of species along the glacier-fed hydrologic continuums. Collectively, this study provides insights into the fate of bacterial assemblages in glacier surface snow following snow melt and how bacterial communities in aquatic environments are affected by the influx of glacier snow-originating bacteria. This article is protected by copyright. All rights reserved.

Candidatus phylum Eremiobacterota (formerly WPS-2) is an as-yet-uncultured bacterial clade that takes its name from Ca. Eremiobacter, an Antarctic soil aerobe proposed to be capable of a novel form of chemolithoautotrophy termed atmospheric chemosynthesis, that uses the energy derived from atmospheric H(2)-oxidation to fix CO(2) through the Calvin-Benson-Bassham (CBB) cycle via type 1E RuBisCO. To elucidate the phylogenetic affiliation and metabolic capacities of Ca. Eremiobacterota, we analysed 63 public metagenome-assembled genomes (MAGs) and nine new MAGs generated from Antarctic soil metagenomes. These MAGs represent both recognized classes within Ca. Eremiobacterota, namely Ca. Eremiobacteria and UBP9. Ca. Eremiobacteria are inferred to be facultatively acidophilic with a preference for peptides and amino acids as nutrient sources. Epifluorescence microscopy revealed Ca. Eremiobacteria cells from Antarctica desert soil to be coccoid in shape. Two orders are recognized within class Ca. Eremiobacteria: Ca. Eremiobacterales and Ca. Baltobacterales. The latter are metabolically versatile, with individual members having genes required for trace gas driven autotrophy, anoxygenic photosynthesis, CO oxidation, and anaerobic respiration. UBP9, here renamed Ca. Xenobia class. nov., are inferred to be obligate heterotrophs with acidophilic adaptations, but individual members having highly divergent metabolic capacities compared to Ca. Eremiobacteria, especially with regard to respiration and central carbon metabolism. We conclude Ca. Eremiobacterota to be an ecologically versatile phylum with the potential to thrive under an array of “extreme” environmental conditions.

Background: Bacteria in low abundance represent the majority of Earth’s biodiversity and perform vital ecological functions, but little is known about their biogeography nor the ecological processes that drive their community assembly in terrestrial ecosystems. Here, we investigated the community compositions and phylogenies of rare (relative abundance < 0.1%) and abundant (> 1%) soil bacteria along a transect containing three alpine grassland types (meadow, steppe, and desert) on the Tibetan Plateau. Results: Our results revealed similar biogeographical patterns of rare and abundant bacteria, with their community compositions and phylogenies shifting gradually along the transect. The similar patterns, however, were driven by contrasting community assembly processes, with rare subcommunity being more heavily influenced by stochasticity (72%) than abundant (57%). The composition of abundant subcommunity (80%) was better explained by local (including soil and vegetation factors), geospatial and climatic factors than that of rare subcommunity (41%), while the phylogeny of the rare one (36%) was better explained than that of the abundant one (29%). Variation partitioning analysis indicated that pure local factors consistently explained a higher proportion of the community composition than geospatial factors in both rare (12.3% and 8.7%, respectively) and abundant bacteria (18.3% and 14.1%, respectively). In contrast, the phylogeny of rare subcommunity was explained by local and geospatial factors equally (11.5% and 11.9%, respectively), while that of abundant subcommunity was more explained by geospatial (22.1%) than local factors (11.3%). Furthermore, our results revealed a tighter connection between the community phylogeny and composition in rare than in abundant bacteria. Conclusions: Our results revealed consistent biogeographical patterns of rare and abundant bacteria in grassland soils, but their assembly processes were distinct. We further demonstrated that rare subcommunity was less predictable than the abundant subcommunity by environmental and geospatial factors. Rare and abundant bacteria responded differentially to factors, which was attributed to the distinct life strategies. Our study provides novel insights into the assembly processes and biographical patterns of rare and abundant bacteria in terrestrial ecosystems.

Global warming leads to deglaciations in high-elevation regions, which exposes deglaciated soils to microbial colonization. Disparity in year-to-year successional patterns of bacterial community and influencing factors in freshly deglaciated soils remain unclear.We explored the abundance of bacterial 16S rRNA gene and community succession in deglaciated soils along a 14-year chronosequence after deglaciation using qPCR and Illumina sequencing on the Tibetan Plateau. The results showed that the abundance of bacterial 16S rRNA gene gradually increased with increasing deglaciation age. Soil bacterial community succession was clustered into three deglaciation stages, which were the early (zero-year old), transitional (1–7 years old) and late (8–14 years old) stages. A significantly abrupt bacterial community succession occurred from the early to the transitional stage (P < 0.01), while a mild succession (P = 0.078) occurred from the transitional to the late stage. The bacterial community at the early and transitional stages were dominated by Proteobacteria, while the late stage was dominated by Actinobacteria. Less abundant ( < 10%) Acidobacteria, Gemmatimonadetes, Verrucomicrobia, Chloroflexi, Planctomycetes, unclassified bacteria dominantly occurred in the transition and late stage and Cyanobacteria in the early stage. Total organic carbon (24.7%), post deglaciation age (21%), pH (16.5%) and moisture (10.1%) significantly contributed (P < 0.05) to the variation of bacterial community succession. Our findings provided a new insight that short time-scale chronosequence is a good model to study yearly resolution of microbial community succession.

Soil microbiomes within oligotrophic cold deserts are extraordinarily diverse. Increasingly, oligotrophic sites with low levels of phototrophic primary producers are reported, leading researchers to question their carbon and energy sources. A novel microbial carbon fixation process termed atmospheric chemosynthesis recently filled this gap as it was shown to be supporting primary production at two Eastern Antarctic deserts. Atmospheric chemosynthesis uses energy liberated from the oxidation of atmospheric hydrogen to drive the Calvin-Benson-Bassham (CBB) cycle through a new chemotrophic form of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), designated IE. Here, we propose that the genetic determinants of this process; RuBisCO type IE (rbcL1E) and high affinity group 1h-[NiFe]-hydrogenase (hhyL) are widespread across cold desert soils and that this process is linked to dry and nutrient-poor environments. We used quantitative PCR (qPCR) to quantify these genes in 122 soil microbiomes across the three poles; spanning the Tibetan Plateau, 10 Antarctic and three high Arctic sites. Both genes were ubiquitous, being present at variable abundances in all 122 soils examined (rbcL1E, 6.25 × 103–1.66 × 109 copies/g soil; hhyL, 6.84 × 103–5.07 × 108 copies/g soil). For the Antarctic and Arctic sites, random forest and correlation analysis against 26 measured soil physicochemical parameters revealed that rbcL1E and hhyL genes were associated with lower soil moisture, carbon and nitrogen content. While further studies are required to quantify the rates of trace gas carbon fixation and the organisms involved, we highlight the global potential of desert soil microbiomes to be supported by this new minimalistic mode of carbon fixation, particularly throughout dry oligotrophic environments, which encompass more than 35% of the Earth’s surface.

Biochar and mineral-enriched biochar (MEB) have been used as soil amendments to improve soil fertility, sequester carbon and mitigate greenhouse gas emissions. Such beneficial outcomes could be partially mediated by soil bacteria, however little is known about how they directly interact with biochar or MEB. We therefore analyzed the diversity and functions of bacterial communities on the surfaces of one biochar and two different MEBs after a 140-day incubation in soil. The results show that the biochar and the MEBs harbor distinct bacterial communities to the bulk soil. Communities on biochar and MEBs were dominated by a novel Gammaproteobacterium. Genome reconstruction combined with electron microscopy and high-resolution elemental analysis revealed that the bacterium generates energy from the oxidation of iron that is present on the surface. Two other bacteria belonging to the genus Thiobacillus and a novel group within the Oxalbacteraceae were enriched only on the MEBs and they had the genetic capacity for thiosulfate oxidation. All three surface-enriched bacteria also had the capacity to fix carbon dioxide, either in a potentially strictly autotrophic or mixotrophic manner. Our results show the dominance of chemolithotrophic processes on the surface of biochar and MEB that can contribute to carbon sequestration in soil.

Salinity is one of the most important environmental factors influencing bacterial plankton communities in lake waters, while its influence on bacterial interactions has been less explored. Here, we investigated the influence of salinity on the bacterial diversity, interactions and community structure in Tibetan Plateau lakes. Our results revealed that saline lakes (salinity between 0.5 and 50 g/L) harboured similar or even higher bacterial diversity compared with freshwater lakes (< 0.5 g/L), while hyper-saline lakes (> 50 g/L) exhibited the lowest diversity. Network analysis demonstrated that hyper-saline lakes exhibited the highest network complexity, with higher total correlation numbers (particularly the negative correlations), but lower network module numbers than freshwater and saline lakes. Furthermore, salinity dominantly explained the bacterial community structure variations in saline lakes, while those in freshwater and hyper-saline lakes were predominately explained by water temperature and geospatial distance, respectively. The core operational taxonomic units (OTUs), which were ubiquitously present in all lakes, were less sensitive to enhancing salinity than the indicative OTUs whose presence was dependent on lake type. Our findings offer a new understanding of how salinity influences bacterial community in plateau lakes.

Glaciers harbour diverse microbes and autotrophic microbes play a key role in sustaining the glacial ecosystems by providing organic carbon. The succession of glacier-originated autotrophic microbes and their effects on downstream aquatic ecosystems remain unknown. We herein investigated the shift of autotrophic microbial communities in waters (not biofilms) along a glacier meltwater transect consisting of a glacier terminus outflow (subglacial), a glacial stream, two glacier-fed lakes (upper and lower) and their outflow on the Tibetan Plateau. The autotrophic community was characterized by cbbL gene using qPCR, T-RFLP and clone library/sequencing methods. The results demonstrated that form IC and ID autotrophic microbes exhibited a much higher abundance than form IAB in all waters along the transect. Form IAB autotrophic abundance in waters gradually decreased, while the form IC exhibited a substantial increase in the upper lake waters, and ID exhibited a substantial increase in the lower lake waters. The water form IC autotrophic community structure exhibited a distinguished shift from the glacier terminus outflow to the stream, while the form ID showed a dramatic shift from the stream to the lower lake. Our results revealed the succession patterns of glacier-originated autotrophic microbial communities and possible effects on downstream aquatic ecosystems.

Landscape heterogeneity impacts community assembly in animals and plants, but it is not clear if this ecological concept extends to microbes. To examine this question, we chose to investigate polar soil environments from the Antarctic and Arctic, where microbes often form the major component of biomass. We examined soil environments that ranged in connectivity from relatively well-connected slopes to patchy, fragmented landforms that comprised isolated frost boils. We found landscape connectedness to have a significant correlation with microbial community structure and connectivity, as measured by co-occurrence networks. Soils from within fragmented landforms appeared to exhibit less local environmental heterogeneity, harboured more similar communities, but fewer biological associations than connected landforms. This effect was observed at both poles, despite the geographical distances and ecological differences between them. We suggest that microbial communities inhabiting well-connected landscape elements respond consistently to regional-scale gradients in biotic and edaphic factors. Conversely, the repeated freeze thaw cycles that characterize fragmented landscapes create barriers within the landscape and act to homogenize the soil environment within individual frost boils and consequently the microbial communities. We propose that lower microbial connectivity in the fragmented landforms is a function of smaller patch size and continual disturbances following soil mixing.

Mitchell Peninsula is located towards the East of the Windmill Islands in eastern Antarctica. It is an ice-free polar desert, and knowledge of its soil microbial taxonomic composition is limited. In this study, we investigated the soil microbial taxonomic composition using multiplex 454 pyrosequencing targeting the bacterial 16S rRNA and the fungal ITS genes; and the bacterial and fungal abundances were estimated using quantitative PCR. In total, 40 bacterial and five fungal phyla were identified comprising 111 bacterial and 22 fungal classes, respectively. Mitchell Peninsula soil exhibited a unique bacterial taxonomic profile. In contrast to the usual dominance of Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes in polar and temperate soils, Mitchell Peninsula was rich in the poorly studied Chloroflexi (31.7 %), candidate divisions WPS-2 (8.1 %) and AD3 (5.1 %), while the commonly observed Bacteroidetes and Firmicutes were present in relative abundances below 1 %. The fungal community consisted of Ascomycota (77 %) and Basidiomycota (10 %), and was dominated by the lichenous fungal class Lecanoromycetes (46.4 %). Network analysis revealed the presence of several microbial clusters that each potentially occupied a different environmental niche, and fewer numbers of correlations were identified between bacteria within each cluster compared with the lichen community, where extensive community dynamics may be present. As Mitchell Peninsula exhibits a unique microbial taxonomic composition, not previously observed in any reported polar or temperate ecosystem, we believe it is a potential microbial biodiversity “hotspot”, which warrants further investigation to examine the role of the dominance of these uncharacterised candidate divisions in this extreme ecosystem.

A Gram-stain-negative, rod-shaped, non-motile bacterium, designated D4N98T, was isolated from a desert soil near Glendambo, Australia. The taxonomic position of strain D4N98T was investigated using a polyphasic approach. Strain D4N98T shared 97 % 16S rRNA gene sequence similarity with the only reference strain of the genus Izhakiella ( Izhakiella capsodis N6PO6T), and less than 96 % similarity with other species of genera in the family Enterobacteriaceae with validly published names. Phylogenetic analysis, based on 16S rRNA, rpoB, atpD, gyrB genes and a concatenated sequence comprising 37 single copy marker genes, as well as chemotaxonomic data (major polar lipids: phosphatidyl ethanolamine, phosphatidylglycerol, diphosphatidylglycerol, a neutral lipid, and a glycolipid. Major fatty acids (>10 %): C16 : 0, C17 : 0 cyclo, summed feature 2 (C14 : 0 3-OH and/or iso-C16 : 1 I), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) support the affiliation of this strain to the genus Izhakiella . The results of in silico DNA–DNA hybridisation plus physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain D4N98T from the other species of the genus Izhakiella with validly published names. Therefore, strain D4N98T represents a novel species, for which the name Izhakiella australiensis sp. nov. is proposed. The type strain is D4N98T (LMG 30066T=DSM 105030T).

Microbial activities in Arctic and Antarctic soils are of particular interest due to uncertainty surrounding the fate of the enormous polar soil organic matter (SOM) pools and the potential to lose unique and vulnerable micro-organisms from these ecosystems. We quantified richness, evenness and taxonomic composition of both fungi and bacteria in 223 Arctic and Antarctic soil samples across 8 locations to test the global applicability of hypotheses concerning edaphic drivers of soil microbial communities that have been primarily developed from studies of bacteria in temperate and tropical systems. We externally validated our model's conclusions with an independent dataset comprising 33 Arctic heath samples. We also explored if our system was responding to large scale climatic or biogeographical processes that we had not measured by evaluating model stability for one location, Mitchell Pennisula, that had been extensively sampled. Soil Fertility (defined as organic matter, nitrogen and chloride content) was the most important edaphic property associated with measures of α-diversity such as microbial richness and evenness (especially for fungi), whereas pH was primarily associated with measures of β-diversity such as phylogenetic structure and diversity (especially for bacteria). Surprisingly, phosphorus emerged as consistently the second most important driver of all facets of microbial community structure for both fungi and bacteria. Despite the clear importance of edaphic factors in controlling microbial communities, our analyses also indicated that fungal/bacterial interactions play a major, but causally unclear, role in structuring the soil microbial communities of which they are a part.

No systems have been reported for genetic manipulation of cold-adapted Archaea. Halorubrum lacusprofundi is an important member of Deep Lake, Antarctica (~10% of the population), and is amendable to laboratory cultivation. Here we report the development of a shuttle-vector and targeted gene-knockout system for this species. To investigate the function of acetamidase/formamidase genes, a class of genes not experimentally studied in Archaea, the acetamidase gene, amd3, was disrupted. The wild-type grew on acetamide as a sole source of carbon and nitrogen, but the mutant did not. Acetamidase/formamidase genes were found to form three distinct clades within a broad distribution of Archaea and Bacteria. Genes were present within lineages characterized by aerobic growth in low nutrient environments (e.g. haloarchaea, Starkeya) but absent from lineages containing anaerobes or facultative anaerobes (e.g. methanogens, Epsilonproteobacteria) or parasites of animals and plants (e.g. Chlamydiae). While acetamide is not a well characterized natural substrate, the build-up of plastic pollutants in the environment provides a potential source of introduced acetamide. In view of the extent and pattern of distribution of acetamidase/formamidase sequences within Archaea and Bacteria, we speculate that acetamide from plastics may promote the selection of amd/fmd genes in an increasing number of environmental microorganisms.

Browning Peninsula is an ice-free polar desert situated in the Windmill Islands, Eastern Antarctica. The entire site is described as a barren landscape, comprised of frost boils with soils dominated by microbial life. In this study, we explored the microbial diversity and edaphic drivers of community structure across this site using traditional cultivation methods, a novel approach the soil substrate membrane system (SSMS), and culture-independent 454-tag pyrosequencing. The measured soil environmental and microphysical factors of chlorine, phosphate, aspect and elevation were found to be significant drivers of the bacterial community, while none of the soil parameters analyzed were significantly correlated to the fungal community. Overall, Browning Peninsula soil harbored a distinctive microbial community in comparison to other Antarctic soils comprised of a unique bacterial diversity and extremely limited fungal diversity. Tag pyrosequencing data revealed the bacterial community to be dominated by Actinobacteria (36%), followed by Chloroflexi (18%), Cyanobacteria (14%), and Proteobacteria (10%). For fungi, Ascomycota (97%) dominated the soil microbiome, followed by Basidiomycota. As expected the diversity recovered from culture-based techniques was lower than that detected using tag sequencing. However, in the SSMS enrichments, that mimic the natural conditions for cultivating oligophilic “k-selected” bacteria, a larger proportion of rare bacterial taxa (15%), such as Blastococcus, Devosia, Herbaspirillum, Propionibacterium and Methylocella and fungal (11%) taxa, such as Nigrospora, Exophiala, Hortaea, and Penidiella were recovered at the genus level. At phylum level, a comparison of OTU's showed that the SSMS shared 21% of Acidobacteria, 11% of Actinobacteria and 10% of Proteobacteria OTU's with soil. For fungi, the shared OTUs was 4% (Basidiomycota) and

The candidate division TM7 is ubiquitous and yet uncultured phylum of the Bacteria that encompasses a commonly environmental associated clade, TM7-1, and a ‘host-associated’ clade, TM7-3. However, as members of the TM7 phylum have not been cultured, little is known about what differs between these two clades. We hypothesized that these clades would have different environmental niches. To test this, we used a large-scale global soil dataset, encompassing 223 soil samples, their environmental parameters and associated bacterial 16S rRNA gene sequence data. We correlated chemical, physical and biological parameters of each soil with the relative abundance of the two major classes of the phylum to deduce factors that influence the groups’ seemingly ubiquitous nature. The two classes of the phylum (TM7-1 and TM7-3) were indeed distinct from each other in their habitat requirements. A key determinant of each class’ prevalence appears to be the pH of the soil. The class TM7-1 displays a facultative anaerobic nature with correlations to more acidic soils with total iron, silicon, titanium and copper indicating a potential for siderophore production. However, the TM7-3 class shows a more classical oligotrophic, heterotroph nature with a preference for more alkaline soils, and a probable pathogenic role with correlations to extractable iron, sodium and phosphate. In addition, the TM7-3 was abundant in diesel contaminated soils highlighting a resilient nature along with a possible carbon source. In addition to this both classes had unique co-occurrence relationships with other bacterial phyla. In particular, both groups had opposing correlations to the Gemmatimonadetes phylum, with the TM7-3 class seemingly being outcompeted by this phylum to result in a negative correlation. These ecological controls allow the characteristics of a TM7 phylum preferred niche to be defined and give insight into possible avenues for cultivation of this previously uncultured group.

Despite decreasing costs, generating large-scale, well-replicated and multivariate microbial ecology investigations with sequencing remains an expensive and time-consuming option. As a result, many microbial ecology investigations continue to suffer from a lack of appropriate replication. We evaluated two fingerprinting approaches – terminal restriction fragment length polymorphism (T-RFLP) and automated ribosomal intergenic spacer analysis (ARISA) against 454 pyrosequencing, by applying them to 225 polar soil samples from East Antarctica and the high Arctic. By incorporating local and global spatial scales into the dataset, our aim was to determine whether various approaches differed in their ability and hence utility, to identify ecological patterns. Through the reduction in the 454 sequencing data to the most dominant OTUs, we revealed that a surprisingly small proportion of abundant OTUs (< 0.25%) was driving the biological patterns observed. Overall, ARISA and T-RFLP had a similar capacity as sequencing to separate samples according to distance at a local scale, and to correlate environmental variables with microbial community structure. Pyrosequencing had a greater resolution at the global scale but all methods were capable of significantly differentiating the polar sites. We conclude fingerprinting remains a legitimate approach to generating large datasets as well as a cost-effective rapid method to identify samples for elucidating taxonomic information or diversity estimates with sequencing methods.

The phylum candidatus Saccharibacteria formerly known as Candidate Division TM7 is a highly ubiquitous phylum with 16S rRNA gene sequences reported in soils, sediments, wastewater and animals, as well as a host of clinical environments. Here, the application of two taxon-specific primers on environmental and human-associated samples using bar-coded tag pyrosequencing revealed two new clades for this phylum to exist and we propose that the division consists of 2 monophyletic and 2 polyphyletic clades. Investigation into TM7 ecology revealed that a high proportion (58%) of phylotypes were sample specific, few were widely distributed and of those most widely distributed all belonged to subdivision 3. Additionally, 50% of the most relatively abundant phylotypes observed were also subdivision 3 members. Community analysis showed that despite the presence of a high proportion of unique phylotypes, specific groups of samples still harbor similar TM7 communities with samples clustering together. The lack of relatively abundant phylotypes from subdivisions 1, 2 and 4 and the presence of very few cosmopolitan members' highlights not only the site specific nature of this phylum but provides insight into why the majority of studies into TM7 have been biased towards subdivision 3.