Dominant Phylotypes Research Articles

Sex differences in gut microbiota in patients with major depressive disorder.

ObjectiveOur previous studies found that disturbances in gut microbiota might have a causative role in the onset of major depressive disorder (MDD). The aim of this study was to investigate whether there were sex differences in gut microbiota in patients with MDD.Patients and methodsFirst-episode drug-naïve MDD patients and healthy controls were included. 16S rRNA gene sequences extracted from the fecal samples of the included subjects were analyzed. Principal-coordinate analysis and partial least squares-discriminant analysis were used to assess whether there were sex-specific gut microbiota. A random forest algorithm was used to identify the differential operational taxonomic units. Linear discriminant-analysis effect size was further used to identify the dominant sex-specific phylotypes responsible for the differences between MDD patients and healthy controls.ResultsIn total, 57 and 74 differential operational taxonomic units responsible for separating female and male MDD patients from their healthy counterparts were identified. Compared with their healthy counterparts, increased Actinobacteria and decreased Bacteroidetes levels were found in female and male MDD patients, respectively. The most differentially abundant bacterial taxa in female and male MDD patients belonged to phyla Actinobacteria and Bacteroidia, respectively. Meanwhile, female and male MDD patients had different dominant phylotypes.ConclusionThese results demonstrated that there were sex differences in gut microbiota in patients with MDD. The suitability of Actinobacteria and Bacteroidia as the sex-specific biomarkers for diagnosing MDD should be further explored.

Gender Based Compositional Fluctuations in Gut Microbiota in Hypertension

Background: Emerging evidence suggests that human gut microbiota plays a critical role in maintaining physiological homeostasis, and gut dysbiosis is involved directly or indirectly in cardiovascular diseases including hypertension. Aim: The aim of the present study was to investigate whether there were sex-based compositional differences in gut microbiota of patients with hypertension. Methods: Fecal samples were collected from male and female hypertensive patients, and healthy individuals from Xi’an, Shaanxi province of China. Touchdown PCR and denaturing gradient gel electrophoresis (PCR-DGGE) with primers specifically targeting V3 region of 16S rRNA, and real-time quantitative PCR (qPCR) were performed to characterize all the samples. High-throughput sequencing of the V3–V4 regions was performed on Illumina HiSeq 2500. Results: Diversity and richness indices for the gut microbiome were compared among the four groups. The relative abundance of Firmicutes was found increased in hypertensive groups (both female and male patients) while that of Bacteroidetes was decreased in male hypertensive patients only, as compared to the healthy male subjects. The most differentially abundant bacterial taxa belonged to the genera Prevotella and Megasphaera in female hypertensive patients and Megamonas in male hypertensive patients. The female and male hypertensive patients also showed different dominant phylotypes. Conclusion: These results demonstrated that there were sex-based differences in gut microbial composition of patients with hypertension. Our findings indicate Prevotella, Megasphaera and Megamonas as the potential sex-specific biomarkers of hypertension, and need to be further evaluated.

Overall bacterial community composition and abundance of nitrifiers and denitrifiers in a typical macrotidal estuary

Coupled nitrogen cycling processes can alleviate the negative effects of eutrophication caused by excessive nitrogen load in estuarine ecosystems. The abundance and diversity of nitrifiers and denitrifiers across different environmental gradients were examined in the sediment of Hangzhou Bay. Quantitative PCR and Pearson's correlation analyses suggested that the bacterial ammonia-oxidizers (AOB) were the dominant phylotypes capable of ammonia oxidation, while the nirS-encoding denitrifiers predominated in the denitrification process. Simultaneously, nitrite and pH were found to be the two major factors influencing amoA and nir gene abundances, and the distribution of bacterial communities. Moreover, the ratio of nirS/AOB amoA gene abundance showed negative correlation with nitrite concentration. Fluorescence in situ hybridization further demonstrated that AOB and acetate-denitrifying cells were closely connected and formed obvious aggregates in the sediment. Together, all these results provided us a preliminary insight for coupled nitrification-denitrification processes in the sediment of Hangzhou Bay.

Fibrobacter communities in the gastrointestinal tracts of diverse hindgut-fermenting herbivores are distinct from those of the rumen.

The genus Fibrobacter contains cellulolytic bacteria originally isolated from the rumen. Culture-independent investigations have since identified Fibrobacter populations in the gastrointestinal tracts of numerous hindgut-fermenting herbivores, but their physiology is poorly characterized due to few representative axenic cultures. To test the hypothesis that novel Fibrobacter diversity exists in hindgut fermenters, we performed culturing and 16S rRNA gene amplicon sequencing on samples collected from phylogenetically diverse herbivorous hosts. Using a unique approach for recovering axenic Fibrobacter cultures, we isolated 45 novel strains from 11 different hosts. Full-length 16S rRNA gene sequencing of these isolates identified nine discrete phylotypes (cutoff = 0.03%) among them, including several that were only isolated from hindgut-fermenting hosts, and four previously unrepresented by axenic cultures. Our phylogenetic analysis indicated that six of the phylotypes are more closely related to previously described subspecies of Fibrobacter succinogenes, while the remaining three were more closely related to F. intestinalis. Culture-independent bacterial community profiling confirmed that most isolates were representative of numerically dominant phylotypes in their respective samples and strengthened the association of certain phylotypes with either ruminants or hindgut-fermenters. Despite considerable phylogenetic diversity observed among the Fibrobacter strains isolated here, phenotypic characterization suggests a conserved specialization for growth on cellulose.

Effect of seasonality and physicochemical parameters on bacterial communities in two hot spring microbial mats from Araró, Mexico

In this study, we explored the diversity of culturable bacterial communities residing in hot springs from Araró, México, and analysed the effect of seasonality and related changes in physicochemical parameters of spring water. Two hot springs with unique features, Tina and Bonita, were analysed. Seventy-nine unique 16S rRNA gene phylotypes were detected, belonging to the bacterial phyla Firmicutes, Proteobacteria, and Actinobacteria. A group of dominant phylotypes of the genus Bacillus was recovered in 3 out of 4 of the sampling seasons. Another group of phylotypes was recovered in 2 samplings, while the remaining groups were detected in only 1 season. Ecological indexes for species richness and evenness showed moderate to low diversity in both hot springs, and a Sørensen analysis revealed that the 2 communities shared 64% of their bacterial phylotypes. Physicochemical parameters measured every season showed slight variations, except for temperature and arsenic content. Fluctuations in bacterial composition in the Tina hot spring were correlated mainly with salt content, while diversity in the Bonita hot spring was significantly correlated with temperature, pH, and arsenic content.

Geographical structure of endosymbiotic bacteria hosted by Bathymodiolus mussels at eastern Pacific hydrothermal vents

BackgroundChemolithoautotrophic primary production sustains dense invertebrate communities at deep-sea hydrothermal vents and hydrocarbon seeps. Symbiotic bacteria that oxidize dissolved sulfur, methane, and hydrogen gases nourish bathymodiolin mussels that thrive in these environments worldwide. The mussel symbionts are newly acquired in each generation via infection by free-living forms. This study examined geographical subdivision of the thiotrophic endosymbionts hosted by Bathymodiolus mussels living along the eastern Pacific hydrothermal vents. High-throughput sequencing data of 16S ribosomal RNA encoding gene and fragments of six protein-coding genes of symbionts were examined in the samples collected from nine vent localities at the East Pacific Rise, Galápagos Rift, and Pacific-Antarctic Ridge.ResultsBoth of the parapatric sister-species, B. thermophilus and B. antarcticus, hosted the same numerically dominant phylotype of thiotrophic Gammaproteobacteria. However, sequences from six protein-coding genes revealed highly divergent symbiont lineages living north and south of the Easter Microplate and hosted by these two Bathymodiolus mussel species. High heterogeneity of symbiont haplotypes among host individuals sampled from the same location suggested that stochasticity associated with initial infections was amplified as symbionts proliferated within the host individuals. The mussel species presently contact one another and hybridize along the Easter Microplate, but the northern and southern symbionts appear to be completely isolated. Vicariance associated with orogeny of the Easter Microplate region, 2.5–5.3 million years ago, may have initiated isolation of the symbiont and host populations. Estimates of synonymous substitution rates for the protein-coding bacterial genes examined in this study were 0.77–1.62%/nucleotide/million years.ConclusionsOur present study reports the most comprehensive population genetic analyses of the chemosynthetic endosymbiotic bacteria based on high-throughput genetic data and extensive geographical sampling to date, and demonstrates the role of the geographical features, the Easter Microplate and geographical distance, in the intraspecific divergence of this bacterial species along the mid-ocean ridge axes in the eastern Pacific. Altogether, our results provide insights into extrinsic and intrinsic factors affecting the dispersal and evolution of chemosynthetic symbiotic partners in the hydrothermal vents along the eastern Pacific Ocean.

Conserved nature of Helicoverpa armigera gut bacterial flora on different host plants and in vitro interactions with PI proteins advocates role in host digestive physiology

Abstract Helicoverpa armigera is anxious insect pest of agricultural crops. Array of defensive molecules in host plants and extensive use of chemical insecticides are unable to cease the attack incidences. Gut bacterial communities are found to contribute in various physiological activities in most of the arthropods. In the current study the bacterial communities were isolated from gut of H. armigera feeding on three host plants (Pigeonpea, Chickpea and Cotton) by culture dependent and culture independent methods. Predominant bacterial communities were identified by terminal restriction fragment length polymorphism (TRFLP). Three dominant phylotypes namely proteobacteria, actinobacteria and firmicutes were identified by TRFLP and found to conserve on different host plant selected. Five Bacillus species namely Bacillus sp. JR14, Bacillus sp. YP1, Bacillus safensis CG1, Bacillus subtillis KAVK2 and Bacillus megaterium 47N were purified by culture dependent method and identified by 16S rRNA sequencing. Among all identified Bacillus, Bacillus sp. YP1 strain was found to be potent protease producer as assisted by dot-blot assay and in vitro solution assays. The in vitro interactions of these proteases with host plant PIs were studied by reverse zymography and gel X-ray contact print (GXCP) analysis. Reduction in activity of PIs and degradation pattern of PI bands on gels in presence of trypsin and protease extract of Bacillus sp. YP1 indicates inactivation of PIs. Thus, conserved nature and in vitro response to PI proteins advocates role of gut bacterial flora in H. armigera digestive physiology.

Acceptable symbiont cell size differs among cnidarian species and may limit symbiont diversity

Reef-building corals form symbiotic relationships with dinoflagellates of the genus Symbiodinium. Symbiodinium are genetically and physiologically diverse, and corals may be able to adapt to different environments by altering their dominant Symbiodinium phylotype. Notably, each coral species associates only with specific Symbiodinium phylotypes, and consequently the diversity of symbionts available to the host is limited by the species specificity. Currently, it is widely presumed that species specificity is determined by the combination of cell-surface molecules on the host and symbiont. Here we show experimental evidence supporting a new model to explain at least part of the specificity in coral–Symbiodinium symbiosis. Using the laboratory model Aiptasia–Symbiodinium system, we found that symbiont infectivity is related to cell size; larger Symbiodinium phylotypes are less likely to establish a symbiotic relationship with the host Aiptasia. This size dependency is further supported by experiments where symbionts were replaced by artificial fluorescent microspheres. Finally, experiments using two different coral species demonstrate that our size-dependent-infection model can be expanded to coral–Symbiodinium symbiosis, with the acceptability of large-sized Symbiodinium phylotypes differing between two coral species. Thus the selectivity of the host for symbiont cell size can affect the diversity of symbionts in corals.

Comparison of Prokaryotic Diversity in Cold, Oligotrophic Remote Lakes of Chilean Patagonia

The prokaryotic abundance and diversity in three cold, oligotrophic Patagonian lakes (Témpanos, Las Torres and Mercedes) in the northern region Aysén (Chile) were compared in winter and summer using 16S rRNA fluorescence in situ hybridization and PCR-denaturing gradient gel electrophoresis technique. Prokaryotic abundances, numerically dominated by Bacteria, were quite similar in the three lakes, but higher in sediments than in waters, and they were also higher in summer than in winter. The relative contribution of Archaea was greater in waters than in sediments, and in winter rather than in summer. Despite the phylogenetic analysis indicated that most sequences were affiliated to a few taxonomic groups, mainly referred to Proteobacteria (consisting of Beta-, Alpha- and Gammaproteobacteria) and Euryarchaeota (mainly related to uncultured methanogens), their relative abundances differed in each sample, resulting in different bacterial and archaeal assemblages. In winter, the abundance of the dominant bacterial phylotypes were mainly regulated by the increasing levels of total organic carbon in waters. Archaeal abundance and richness appeared mostly influenced by pH in winter and total nitrogen content in summer. The prokaryotic community composition at Témpanos lake, located most northerly and closer to a glacier, greatly differed in respect to the other two lakes. In this lake was detected the highest bacterial diversity, being Betaproteobacteria the most abundant group, whereas Alphaproteobacteria were distinctive of Mercedes. Archaeal community associated with sediments was mainly represent by members related to the order of Methanosarcinales at Mercedes and Las Torres lakes, and by Crenarchaeota at Témpanos lake. Our results indicate that the proximity to the glacier and the seasonality shape the composition of the prokaryotic communities in these remote lakes. These results may be used as baseline information to follow the microbial community responses to potential global changes and to anthropogenic impacts.

Species Coexistence in Nitrifying Chemostats: A Model of Microbial Interactions

In a previous study, the two nitrifying functions (ammonia oxidizing bacteria (AOB) or nitrite-oxidizing bacteria (NOB)) of a nitrification reactor—operated continuously over 525 days with varying inputs—were assigned using a mathematical modeling approach together with the monitoring of bacterial phylotypes. Based on these theoretical identifications, we develop here a chemostat model that does not explicitly include only the resources’ dynamics (different forms of soluble nitrogen) but also explicitly takes into account microbial inter- and intra-species interactions for the four dominant phylotypes detected in the chemostat. A comparison of the models obtained with and without interactions has shown that such interactions permit the coexistence of two competing ammonium-oxidizing bacteria and two competing nitrite-oxidizing bacteria in competition for ammonium and nitrite, respectively. These interactions are analyzed and discussed.

Hydrocarbon degraders establish at the costs of microbial richness, abundance and keystone taxa after crude oil contamination in permafrost environments.

Oil spills from pipeline ruptures are a major source of terrestrial petroleum pollution in cold regions. However, our knowledge of the bacterial response to crude oil contamination in cold regions remains to be further expanded, especially in terms of community shifts and potential development of hydrocarbon degraders. In this study we investigated changes of microbial diversity, population size and keystone taxa in permafrost soils at four different sites along the China-Russia crude oil pipeline prior to and after perturbation with crude oil. We found that crude oil caused a decrease of cell numbers together with a reduction of the species richness and shifts in the dominant phylotypes, while bacterial community diversity was highly site-specific after exposure to crude oil, reflecting different environmental conditions. Keystone taxa that strongly co-occurred were found to form networks based on trophic interactions, that is co-metabolism regarding degradation of hydrocarbons (in contaminated samples) or syntrophic carbon cycling (in uncontaminated samples). With this study we demonstrate that after severe crude oil contamination a rapid establishment of endemic hydrocarbon degrading communities takes place under favorable temperature conditions. Therefore, both endemism and trophic correlations of bacterial degraders need to be considered in order to develop effective cleanup strategies.

Microbial Community Structure of Subalpine Snow in the Sierra Nevada, California

ABSTRCTMounting evidence suggests that Earth’s cryosphere harbors diverse and active microbial communities. However, our understanding of microbial composition and diversity in seasonal snowpack of montane ecosystems remains limited. We sequenced the 16S rRNA gene to determine microbial structure (composition and diversity) of snow at two depths (0–15 and 15–30 cm) of a subalpine site in the Southern Sierra Critical Zone Observatory, California, U.S.A. Proteobacteria dominated both depths (~72% of sequences), and this phylum was composed mostly of bacteria within the Rhodospirillales order. Cyanobacteria were almost exclusively present in the upper snow layer, while Actinobacteria and Firmicutes were more abundant in the deep snow layer. Many of the most abundant phylotypes were Acetobacteraceae. Phylotype NCR4874, which comprised 22%–32% of the sequences, was most closely related to the N2-fixing bacteria Asaia siamensis, suggesting that N2 fixation may be an important process within the Sierra snowpack. In addition, just under half (45%) of the numerically dominant phylotypes shared >98% similarity with sequences recovered from other cold environments. Our results suggest that microbial communities of subalpine Sierra Nevada snowpack are diverse, with both snow depths harboring distinct but overlapping communities consisting largely of cold-adapted bacteria.

Community structure, population dynamics and diversity of fungi in a full-scale membrane bioreactor (MBR) for urban wastewater treatment

Community structure, population dynamics and diversity of fungi were monitored in a full-scale membrane bioreactor (MBR) operated throughout four experimental phases (Summer 2009, Autumn 2009, Summer 2010 and Winter, 2012) under different conditions, using the 18S-rRNA gene and the intergenic transcribed spacer (ITS2-region) as molecular markers, and a combination of temperature-gradient gel electrophoresis and 454-pyrosequencing. Both total and metabolically-active fungal populations were fingerprinted, by amplification of molecular markers from community DNA and retrotranscribed RNA, respectively. Fingerprinting and 454-pyrosequencing evidenced that the MBR sheltered a dynamic fungal community composed of a low number of species, in accordance with the knowledge of fungal diversity in freshwater environments, and displaying a medium-high level of functional organization with few numerically dominant phylotypes. Population shifts were experienced in strong correlation with the changes of environmental variables and operation parameters, with pH contributing the highest level of explanation. Phylotypes assigned to nine different fungal Phyla were detected, although the community was mainly composed of Ascomycota, Basidiomycota and Chytridiomycota/Blastocladiomycota. Prevailing fungal phylotypes were affiliated to Saccharomycetes and Chytridiomycetes/Blastocladiomycetes, which displayed antagonistic trends in their relative abundance throughout the experimental period. Fungi identified in the activated sludge were closely related to genera of relevance for the degradation of organic matter and trace-organic contaminants, as well as genera of dimorphic fungi potentially able to produce plant operational issues such as foaming or biofouling. Phylotypes closely related to genera of human and plant pathogenic fungi were also detected.

Basin-scale distribution of prokaryotic phylotypes in the epipelagic layer of the Central South Pacific Ocean during austral summer

In the present study, we used catalyzed reporter deposition-fluorescence in situ hybridization to quantify the abundance of five bacterial (Alphaproteobacteria, SAR11, Gammaproteobacteria, SAR86, and Bacteroidetes) and two archaeal (Crenarchaeota and Euryarchaeota) phylotypes in the epipelagic layer (0–200 m) of the Central South Pacific Ocean along 170°W from 0° to 40°S. We found that the distribution patterns of these phylotypes differed from each other. All phylotypes except Gammaproteobacteria were particularly abundant at the surface water of the equatorial region, whereas Gammaproteobacteria was relatively abundant in the area from the southern part of the South Pacific Ocean. SAR11, affiliated with Alphaproteobacteria was the dominant phylotype at all depths, throughout the study area. The abundance of SAR11 significantly increased with chlorophyll a concentration, suggesting that phytoplankton could affect their distribution pattern. There was a positive correlation between Bacteroidetes abundance and water temperature, suggesting that the temperature gradient could be a critical factor determining their distribution in the South Pacific Ocean. Crenarchaeota and Euryarchaeota were more abundant at the equatorial region than in other study areas. Euryarchaeota abundance significantly decreased with depth, and increased with chlorophyll a concentration. This suggests that there was ecological interaction between Euryarchaeota and phytoplankton in the equatorial surface. Our data indicate that distinct hydrographic properties such as seawater temperature, salinity, and the concentrations of chlorophyll a and nutrients can principally control the basin-scale distribution of different prokaryotic phylotypes in the epipelagic layer of the Central South Pacific Ocean.

Microbial communities within the water column of freshwater Lake Radok, East Antarctica: predominant 16S rDNA phylotypes and bacterial cultures

Antarctic lake ecosystems provide a rare opportunity to study the evolution and adaptation of microorganisms to extreme conditions, as well as to discover new species useful for biotechnological applications. Four water samples were collected from various layers of the water column of freshwater Lake Radok in East Antarctica. Two regions (v3-v5 and v4-v8) of the 16S rRNA gene were amplified by PCR and sequenced. Twenty dominant phylotypes were detected representing five bacterial phyla (Actinobacteria, α, β and δ Proteobacteria, Bacteroidetes, Planctomycetes, OD1) and two eukaryotic divisions (stramenopiles and green algae). Of these, 16 phylotypes demonstrated ≤98 % identity to the nearest taxa in GenBank and were therefore classified as new unknown species. Seven phylotypes demonstrated ≤90 % identity and thus remained unidentified. Actinobacteria was the most abundant phylum with 157 clones (41 % of the total number) representing 5 phylotypes. A species complex (3 clades from acI-A subgroup) of Candidatus Planktophila limnetica was prevalent in all layers. Representatives of the OD1 phylum and δ-proteobacteria were discovered by sequencing of the v3-v5 region of 16S rRNA, while Planctomycetes, β-proteobacteria and mtDNA of stramenopiles were discovered by sequencing of the v4-v8 region. This highlights the necessity of sequencing at least two 16S rRNA gene regions to gain more data on microbial community characterization. In general, despite the uniformity in the physical and chemical properties in the water column, a prominent stratification of microbial groups was observed, at the levels of both divisions and phylotypes.

Variations in Bacterial Community in a Temperate Lake Associated with an Agricultural Watershed.

Terrestrially derived carbon and nutrients are washed into lakes, providing nutritional drivers for both microbial heterotrophy and phototrophy. Changes in the quantity and diversity of carbon and nutrients exported from watersheds in response to alterations in long-term land use have led to a need for evaluation of the linkage between watershed-exported carbon and nutrients and bacterial community structure in watershed associated lakes. To learn more about these interactions, we investigated Muskrat Lake in Michigan, which has a well-defined moderately sized watershed dominated by agriculture. We measured the water chemistry, characterized the dissolved organic carbon, and determined the structure of the bacterial communities at the inlet and center of this lake (five depths per site) over the summer and fall of 2008. The lake had temporal and rain event-based fluctuations in water chemistry, as well as temporal and rain event-dependent shifts in bacterial communities as measured by terminal restriction fragment length polymorphism. Agricultural watershed inputs were observed in the lake during and after rain events. Terminal restriction fragment length polymorphism and 454 pyrosequencing of the bacterial communities indicated that there were differences over time and that the dominant phylotypes shifted between summer and late fall. Some populations (e.g., Polynucleobacter and Mycobacterium) increased during fall, while others (e.g., Gemmatimonas) diminished. Redundancy and partitioning analyses showed that water chemistry is highly correlated with variations in the bacterial community of the lake, which explained 34% of the variations in the bacterial community. Dissolved organic carbon had the greatest effects on variations in the Muskrat Lake bacterial community (2%). The results of this study provide information that will enable a better understanding of the interaction between the bacterial community of lakes and changes in chemical properties as a result of nutrient importation from the surrounding watershed.

Rumen and Cecum Microbiomes in Reindeer (Rangifer tarandus tarandus) Are Changed in Response to a Lichen Diet and May Affect Enteric Methane Emissions.

Reindeer (Rangifer tarandus tarandus) are large Holarctic herbivores whose heterogeneous diet has led to the development of a unique gastrointestinal microbiota, essential for the digestion of arctic flora, which may include a large proportion of lichens during winter. Lichens are rich in plant secondary metabolites, which may affect members of the gut microbial consortium, such as the methane-producing methanogenic archaea. Little is known about the effect of lichen consumption on the rumen and cecum microbiotas and how this may affect methanogenesis in reindeer. Here, we examined the effects of dietary lichens on the reindeer gut microbiota, especially methanogens. Samples from the rumen and cecum were collected from two groups of reindeer, fed either lichens (Ld: n = 4), or a standard pelleted feed (Pd: n = 3). Microbial densities (methanogens, bacteria and protozoa) were quantified using quantitative real-time PCR and methanogen and bacterial diversities were determined by 454 pyrosequencing of the 16S rRNA genes.In general, the density of methanogens were not significantly affected (p>0.05) by the intake of lichens. Methanobrevibacter constituted the main archaeal genus (>95% of reads), with Mbr. thaueri CW as the dominant species in both groups of reindeer. Bacteria belonging to the uncharacterized Ruminococcaceae and the genus Prevotella were the dominant phylotypes in the rumen and cecum, in both diets (ranging between 16–38% total sequences). Bacteria belonging to the genus Ruminococcus (3.5% to 0.6%; p = 0.001) and uncharacterized phylotypes within the order Bacteroidales (8.4% to 1.3%; p = 0.027), were significantly decreased in the rumen of lichen-fed reindeer, but not in the cecum (p = 0.2 and p = 0.087, respectively). UniFrac-based analyses showed archaeal and bacterial libraries were significantly different between diets, in both the cecum and the rumen (vegan::Adonis: pseudo-F<0.05). Based upon previous literature, we suggest that the altered methanogen and bacterial profiles may account for expected lower methane emissions from lichen-fed reindeer.

Improved metagenome assemblies and taxonomic binning using long-read circular consensus sequence data.

DNA assembly is a core methodological step in metagenomic pipelines used to study the structure and function within microbial communities. Here we investigate the utility of Pacific Biosciences long and high accuracy circular consensus sequencing (CCS) reads for metagenomic projects. We compared the application and performance of both PacBio CCS and Illumina HiSeq data with assembly and taxonomic binning algorithms using metagenomic samples representing a complex microbial community. Eight SMRT cells produced approximately 94 Mb of CCS reads from a biogas reactor microbiome sample that averaged 1319 nt in length and 99.7% accuracy. CCS data assembly generated a comparative number of large contigs greater than 1 kb, to those assembled from a ~190x larger HiSeq dataset (~18 Gb) produced from the same sample (i.e approximately 62% of total contigs). Hybrid assemblies using PacBio CCS and HiSeq contigs produced improvements in assembly statistics, including an increase in the average contig length and number of large contigs. The incorporation of CCS data produced significant enhancements in taxonomic binning and genome reconstruction of two dominant phylotypes, which assembled and binned poorly using HiSeq data alone. Collectively these results illustrate the value of PacBio CCS reads in certain metagenomics applications.

Different genotypes of Silene vulgaris (Moench) Garcke grown on chromium-contaminated soils influence root organic acid composition and rhizosphere bacterial communities.

Plant-microbe interactions are considered to be important processes determining the efficiency of phytoremediation of heavy metal-contaminated soils. However, relatively little is known about how these interactions are influenced by chromium (Cr) contamination. The effect of Cr stress on metal uptake, root organic acid composition, and rhizosphere bacterial communities was studied using two genotypes of the metallophyte Silene vulgaris, which have shown different tolerance to Cr(VI). The results indicated that root biomass and shoot biomass were not significantly influenced by Cr treatment, but metal uptake in shoots and roots was significantly impacted by the genotype. Principal component analyses (PCA) showed that variation in organic acids oxalic, citric, malic, formic, lactic, acetic, and succinic differed between genotypes. Changes in root organic acid contents in response to Cr revealed a significant increase of oxalic acid in genotype SV-21. The denaturing gradient gel electrophoresis (DGGE) cluster analysis showed that the community structure (determined by PCR-DGGE) was affected by plant genotype and, to a lesser extent, by Cr contamination, the first being the most influential factor shaping the rhizosphere microbiome. Under Cr pollution, a shift in the relative abundance of specific taxa was found and dominant phylotypes were identified as Variovorax in SV-21 and Chitinophaga niastensis, Pontibacter sp., and Ramlibacter sp. in SV-38. These results provided the basis for further studies aimed at the combined use of plants and soil microorganisms in the remediation of Cr-polluted soils.

Oil removal and effects of spilled oil on active microbial communities in close to salt-saturation brines

Abiotic and biotic processes associated with the degradation of a light petroleum in brines close to the salt-saturation (~31%) and the effect of labile organic matter (LOM) supply (casaminoacids/citrate; 0.2 and 0.1% w/v, respectively) were followed during an incubation of 30days. After 4-week incubation at 40°C under light/dark cycles, a 24% of abiotic degradation was observed in untreated brines. The stimulation of native brines community with LOM addition allowed an additional 12.8% oil attenuation due to biodegradation processes. Successional changes in the active microbial community structure due to the oil contamination (16S rRNA DGGE approach) showed the selection of one phylotype affiliated to Salinibacter and the disappearance of Haloquadratum walsbyi in untreated brines. In LOM-amended microcosms, phylotypes related to Salinibacter, Haloarcula, Haloterrigena and Halorhabdus were selected. An effect of hydrocarbon contamination was only observed in the bacterial community with the inhibition of two dominant proteobacterial phylotypes. This study further confirms that short-term and moderate oil biodegradation is possible in LOM-stimulated brines. Biodegradation should be much more reduced under in situ conditions. Self-cleaning capacities of close to saturation hypersaline lakes appears, therefore very limited compared to non-extreme haline environments.