Josephine Bay Paul Center in Comparative Molecular Biology and Evolution
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The Josephine Bay Paul Center for Comparative Molecular Biology and Evolution explores the evolution and interaction of genomes of diverse organisms that play significant roles in environmental biology and human health. This dynamic research program integrates the powerful tools of genome science, molecular phylogenetics, and molecular ecology to advance our understanding of how living organisms are related to each other, to provide the tools to quantify and assess biodiversity, and to identify genes and underlying mechanisms of biomedical importance. Projects span all evolutionary time scales, ranging from deep phylogenetic divergence of ancient eukaryotic and prokaryotic lineages, to ecological analyses of how members of diverse communities contribute and respond to environmental change.
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ArticleA comprehensive update to the Mycobacterium tuberculosis H37Rv reference genome(Nature Research, 2022-11-18) Chitale, Poonam ; Lemenze, Alexander D. ; Fogarty, Emily C. ; Shah, Avi ; Grady, Courtney ; Odom-Mabey, Aubrey R. ; Johnson, W. Evan ; Yang, Jason H. ; Eren, A. Murat ; Brosch, Roland ; Kumar, Pradeep ; Alland, DavidH37Rv is the most widely used Mycobacterium tuberculosis strain, and its genome is globally used as the M. tuberculosis reference sequence. Here, we present Bact-Builder, a pipeline that uses consensus building to generate complete and accurate bacterial genome sequences and apply it to three independently cultured and sequenced H37Rv aliquots of a single laboratory stock. Two of the 4,417,942 base-pair long H37Rv assemblies are 100% identical, with the third differing by a single nucleotide. Compared to the existing H37Rv reference, the new sequence contains ~6.4 kb additional base pairs, encoding ten new regions that include insertions in PE/PPE genes and new paralogs of esxN and esxJ, which are differentially expressed compared to the reference genes. New sequencing and de novo assemblies with Bact-Builder confirm that all 10 regions, plus small additional polymorphisms, are also present in the commonly used H37Rv strains NR123, TMC102, and H37Rv1998. Thus, Bact-Builder shows promise as an improved method to perform accurate and reproducible de novo assemblies of bacterial genomes, and our work provides important updates to the primary M. tuberculosis reference genome.
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ArticleA phylogenomic approach to resolving interrelationships of polyclad flatworms, with implications for life-history evolution(The Royal Society, 2023-03-29) Goodheart, Jessica A. ; Collins, Allen G. ; Cummings, Michael P. ; Egger, Bernhard ; Rawlinson, Kate A.Platyhelminthes (flatworms) are a diverse invertebrate phylum useful for exploring life-history evolution. Within Platyhelminthes, only two clades develop through a larval stage: free-living polyclads and parasitic neodermatans. Neodermatan larvae are considered evolutionarily derived, whereas polyclad larvae are hypothesized to be ancestral due to ciliary band similarities among polyclad and other spiralian larvae. However, larval evolution has been challenging to investigate within polyclads due to low support for deeper phylogenetic relationships. To investigate polyclad life-history evolution, we generated transcriptomic data for 21 species of polyclads to build a well-supported phylogeny for the group. The resulting tree provides strong support for deeper nodes, and we recover a new monophyletic clade of early branching cotyleans. We then used ancestral state reconstructions to investigate ancestral modes of development within Polycladida and more broadly within flatworms. In polyclads, we were unable to reconstruct the ancestral state of deeper nodes with significant support because early branching clades show diverse modes of development. This suggests a complex history of larval evolution in polyclads that likely includes multiple losses and/or multiple gains. However, our ancestral state reconstruction across a previously published platyhelminth phylogeny supports a direct developing prorhynchid/polyclad ancestor, which suggests that a larval stage in the life cycle evolved along the polyclad stem lineage or within polyclads.
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ArticleAccuracy and quality of massively parallel DNA pyrosequencing(BioMed Central, 2007-07-20) Huse, Susan M. ; Huber, Julie A. ; Morrison, Hilary G. ; Sogin, Mitchell L. ; Mark Welch, David B.Massively parallel pyrosequencing systems have increased the efficiency of DNA sequencing, although the published per-base accuracy of a Roche GS20 is only 96%. In genome projects, highly redundant consensus assemblies can compensate for sequencing errors. In contrast, studies of microbial diversity that catalogue differences between PCR amplicons of ribosomal RNA genes (rDNA) or other conserved gene families cannot take advantage of consensus assemblies to detect and minimize incorrect base calls. We performed an empirical study of the per-base error rate for the Roche GS20 system using sequences of the V6 hypervariable region from cloned microbial ribosomal DNA (tag sequencing). We calculated a 99.5% accuracy rate in unassembled sequences, and identified several factors that can be used to remove a small percentage of low-quality reads, improving the accuracy to 99.75% or better. By using objective criteria to eliminate low quality data, the quality of individual GS20 sequence reads in molecular ecological applications can surpass the accuracy of traditional capillary methods.
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PreprintAdaptive evolution of voltage-gated sodium channels : the first 800 million years( 2012-04) Zakon, Harold H.Voltage-gated Na+-permeable (Nav) channels form the basis for electrical excitability in animals. Nav channels evolved from Ca2+ channels and were present in the common ancestor of choanoflagellates and animals although this channel was likely permeable to both Na+ and Ca2+. Thus, like many other neuronal channels and receptors, Nav channels predated neurons. Invertebrates possess two Nav channels (Nav1, Nav2), whereas vertebrate Nav channels are of the Nav1 family. Approximately 500 MYA in early chordates Nav channels evolved a motif that allowed them to cluster at axon initial segments, 50MY later with the evolution of myelin, Nav channels “capitalized” on this property and clustered at nodes of Ranvier. The enhancement of conduction velocity along with the evolution of jaws likely made early gnathostomes fierce predators and the dominant vertebrates in the ocean. Later in vertebrate evolution, the Nav channel gene family expanded in parallel in tetrapods and teleosts (~9-10 genes in amniotes, 8 in teleosts). This expansion occurred during or after the late Devonian extinction when teleosts and tetrapods each diversified in their respective habitats and coincided with an increase in the number of telencephalic nuclei in both groups. The expansion of Nav channels may have allowed for more sophisticated neural computation and tailoring of Nav channel kinetics with potassium channel kinetics to enhance energy savings. Nav channels show adaptive sequence evolution for increasing diversity in communication signals (electric fish), in protection against lethal Nav channel toxins (snakes, newts, pufferfish, insects), and in specialized habitats (naked mole rats).
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ArticleAlkenone producers inferred from well-preserved 18S rDNA in Greenland lake sediments(American Geophysical Union, 2006-09-19) D'Andrea, William J. ; Lage, Melissa ; Martiny, Jennifer B. H. ; Laatsch, Abby D. ; Amaral-Zettler, Linda A. ; Sogin, Mitchell L. ; Huang, YongsongThe 18S ribosomal DNA (rDNA) sequences of haptophyte algae were successfully amplified using the polymerase chain reaction (PCR) from water filtrate, surface sediments, and a late-Holocene sediment sample (∼1000 years old) from a group of lakes in the Søndre Strømfjord region of west Greenland. The DNA of the algal primary producer is extremely well preserved in the laminated lake sediments which have been deposited in cold (1°–2°C), anoxic, and sulphidic bottom water. Phylogenetic analyses of the Greenland haptophyte rDNA sequences suggest that alkenones in the Greenland lake sediments are produced by haptophyte algae of the class Prymnesiophyceae. The 18S rDNA sequences from the Greenland samples cluster within a distinct phylotype, differing from both marine haptophytes and from those reported previously from Ace Lake, Antarctica. The similarity of haptophyte rDNA sequences among all samples in this study suggests a single alkenone-based temperature calibration may be applied to these lakes for at least the past 1000 years. These sedimentary archives hold great promise for high-resolution, alkenone-based paleotemperature reconstruction of southern west Greenland, a region sensitive to atmospheric-oceanic climate phenomena such as the North Atlantic Oscillation (NAO).
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ArticleAnalysis of expressed sequence tags of the cyclically parthenogenetic rotifer Brachionus plicatilis(Public Library of Science, 2007-08-01) Suga, Koushirou ; Mark Welch, David B. ; Tanaka, Yukari ; Sakakura, Yoshitaka ; Hagiwara, AtsushiRotifers are among the most common non-arthropod animals and are the most experimentally tractable members of the basal assemblage of metazoan phyla known as Gnathifera. The monogonont rotifer Brachionus plicatilis is a developing model system for ecotoxicology, aquatic ecology, cryptic speciation, and the evolution of sex, and is an important food source for finfish aquaculture. However, basic knowledge of the genome and transcriptome of any rotifer species has been lacking. We generated and partially sequenced a cDNA library from B. plicatilis and constructed a database of over 2300 expressed sequence tags corresponding to more than 450 transcripts. About 20% of the transcripts had no significant similarity to database sequences by BLAST; most of these contained open reading frames of significant length but few had recognized Pfam motifs. Sixteen transcripts accounted for 25% of the ESTs; four of these had no significant similarity to BLAST or Pfam databases. Putative up- and downstream untranslated regions are relatively short and AT rich. In contrast to bdelloid rotifers, there was no evidence of a conserved trans-spliced leader sequence among the transcripts and most genes were single-copy. Despite the small size of this EST project it revealed several important features of the rotifer transcriptome and of individual monogonont genes. Because there is little genomic data for Gnathifera, the transcripts we found with no known function may represent genes that are species-, class-, phylum- or even superphylum-specific; the fact that some are among the most highly expressed indicates their importance. The absence of trans-spliced leader exons in this monogonont species contrasts with their abundance in bdelloid rotifers and indicates that the presence of this phenomenon can vary at the subphylum level. Our EST database provides a relatively large quantity of transcript-level data for B. plicatilis, and more generally of rotifers and other gnathiferan phyla, and can be browsed and searched at gmod.mbl.edu.
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ArticleAnalysis of lung microbiota in bronchoalveolar lavage, protected brush and sputum samples from subjects with mild-to-moderate cystic fibrosis Lung Disease(Public Library of Science, 2016-03-04) Hogan, Deborah A. ; Willger, Sven D. ; Dolben, Emily L. ; Hampton, Thomas H. ; Stanton, Bruce A. ; Morrison, Hilary G. ; Sogin, Mitchell L. ; Czum, Julianna ; Ashare, AlixIndividuals with cystic fibrosis (CF) often acquire chronic lung infections that lead to irreversible damage. We sought to examine regional variation in the microbial communities in the lungs of individuals with mild-to-moderate CF lung disease, to examine the relationship between the local microbiota and local damage, and to determine the relationships between microbiota in samples taken directly from the lung and the microbiota in spontaneously expectorated sputum. In this initial study, nine stable, adult CF patients with an FEV1>50% underwent regional sampling of different lobes of the right lung by bronchoalveolar lavage (BAL) and protected brush (PB) sampling of mucus plugs. Sputum samples were obtained from six of the nine subjects immediately prior to the procedure. Microbial community analysis was performed on DNA extracted from these samples and the extent of damage in each lobe was quantified from a recent CT scan. The extent of damage observed in regions of the right lung did not correlate with specific microbial genera, levels of community diversity or composition, or bacterial genome copies per ml of BAL fluid. In all subjects, BAL fluid from different regions of the lung contained similar microbial communities. In eight out of nine subjects, PB samples from different regions of the lung were also similar in microbial community composition, and were similar to microbial communities in BAL fluid from the same lobe. Microbial communities in PB samples were more diverse than those in BAL samples, suggesting enrichment of some taxa in mucus plugs. To our knowledge, this study is the first to examine the microbiota in different regions of the CF lung in clinically stable individuals with mild-to-moderate CF-related lung disease.
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ArticleAnalysis, optimization and verification of Illumina-generated 16S rRNA gene amplicon surveys(Public Library of Science, 2014-04-10) Nelson, Michael C. ; Morrison, Hilary G. ; Benjamino, Jacquelynn ; Grim, Sharon L. ; Graf, JoergThe exploration of microbial communities by sequencing 16S rRNA genes has expanded with low-cost, high-throughput sequencing instruments. Illumina-based 16S rRNA gene sequencing has recently gained popularity over 454 pyrosequencing due to its lower costs, higher accuracy and greater throughput. Although recent reports suggest that Illumina and 454 pyrosequencing provide similar beta diversity measures, it remains to be demonstrated that pre-existing 454 pyrosequencing workflows can transfer directly from 454 to Illumina MiSeq sequencing by simply changing the sequencing adapters of the primers. In this study, we modified 454 pyrosequencing primers targeting the V4-V5 hyper-variable regions of the 16S rRNA gene to be compatible with Illumina sequencers. Microbial communities from cows, humans, leeches, mice, sewage, and termites and a mock community were analyzed by 454 and MiSeq sequencing of the V4-V5 region and MiSeq sequencing of the V4 region. Our analysis revealed that reference-based OTU clustering alone introduced biases compared to de novo clustering, preventing certain taxa from being observed in some samples. Based on this we devised and recommend an analysis pipeline that includes read merging, contaminant filtering, and reference-based clustering followed by de novo OTU clustering, which produces diversity measures consistent with de novo OTU clustering analysis. Low levels of dataset contamination with Illumina sequencing were discovered that could affect analyses that require highly sensitive approaches. While moving to Illumina-based sequencing platforms promises to provide deeper insights into the breadth and function of microbial diversity, our results show that care must be taken to ensure that sequencing and processing artifacts do not obscure true microbial diversity.
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PreprintAncient mitochondrial pseudogenes reveal hybridization between distant lineages in the evolution of the Rupicapra genus( 2017-07) Pérez, Trinidad ; Rodriguez, Fernando ; Fernández, M. ; Albornoz, Jesús ; Domínguez, AnaMitochondrial pseudogenes (numts) inserted in the nuclear genome are frequently found in population studies. Its presence is commonly connected with problems and errors when they are confounded with true mitochondrial sequences. In the opposite side, numts can provide valuable phylogenetic information when they are copies of ancient mitochondrial lineages. We show that Rupicapra individuals of different geographic origin from the Cantabrian Mountains to the Apennines and the Caucasus share a nuclear COI fragment. The numt copies are monophyletic, and their pattern of differentiation shows two outstanding features: a long evolution as differentiated true mitochondrial lineage, and a recent integration and spread through the chamois populations. The COI pseudogene is much older than the present day mitochondrial clades of Rupicapra and occupies a basal position within the Rupicapra-Ammotragus- Arabitragus node. Joint analysis of this numt and a cytb pseudogene with a similar pattern of evolution places the source mitochondrial lineage as a sister branch that separated from the Ammotragus-Arabitragus lineage 6 million years ago (Mya). The occurrence of this sequence in the nucleus of chamois suggests hybridization between highly divergent lineages. The integration event seems to be very recent, more recent than the split of the present day mtDNA lineages of Rupicapra (1.9 Mya). This observation invites to think of the spread across the genus by horizontal transfer through recent male-biased dispersal.
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ArticleAnvi’o : an advanced analysis and visualization platform for ‘omics data(PeerJ, 2015-10-08) Eren, A. Murat ; Esen, Ozcan C. ; Quince, Christopher ; Vineis, Joseph H. ; Morrison, Hilary G. ; Sogin, Mitchell L. ; Delmont, Tom O.Advances in high-throughput sequencing and ‘omics technologies are revolutionizing studies of naturally occurring microbial communities. Comprehensive investigations of microbial lifestyles require the ability to interactively organize and visualize genetic information and to incorporate subtle differences that enable greater resolution of complex data. Here we introduce anvi’o, an advanced analysis and visualization platform that offers automated and human-guided characterization of microbial genomes in metagenomic assemblies, with interactive interfaces that can link ‘omics data from multiple sources into a single, intuitive display. Its extensible visualization approach distills multiple dimensions of information about each contig, offering a dynamic and unified work environment for data exploration, manipulation, and reporting. Using anvi’o, we re-analyzed publicly available datasets and explored temporal genomic changes within naturally occurring microbial populations through de novo characterization of single nucleotide variations, and linked cultivar and single-cell genomes with metagenomic and metatranscriptomic data. Anvi’o is an open-source platform that empowers researchers without extensive bioinformatics skills to perform and communicate in-depth analyses on large ‘omics datasets.
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PreprintAquarium microbiome response to ninety-percent system water change : clues to microbiome management( 2015-04) Van Bonn, William ; LaPointe, Allen ; Gibbons, Sean M. ; Frazier, Angel ; Hampton-Marcell, Jarrad T. ; Gilbert, Jack A.The bacterial community composition and structure of water from an established teleost fish system was examined before, during and after a major water change to explore the impact of such a water-change disturbance on the stability of the aquarium water microbiome. The diversity and evenness of the bacterial community significantly increased following the 90% water replacement. While the change in bacterial community structure was significant, it was slight, and was also weakly correlated with changes in physicochemical parameters. Interestingly there was a significant shift in the correlative network relationships between operational taxonomic units from before to after the water replacement. We suggest this shift in network structure is due to the turnover of many taxa during the course of water replacement. These observations will inform future studies into manipulation of the microbiome by changing system environmental parameter values to optimize resident animal health.
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PreprintArchaea and bacteria with surprising microdiversity show shifts in dominance over 1,000-year time scales in hydrothermal chimneys( 2009-12-06) Brazelton, William J. ; Ludwig, Kristin A. ; Sogin, Mitchell L. ; Andreishcheva, Ekaterina N. ; Kelley, Deborah S. ; Shen, Chuan-Chou ; Edwards, R. Lawrence ; Baross, John A.The Lost City Hydrothermal Field, an ultramafic-hosted system located 15 km west of the Mid-Atlantic Ridge, has experienced at least 30,000 years of hydrothermal activity. Previous studies have shown that its carbonate chimneys form by mixing of ~90ºC, pH 9-11 hydrothermal fluids and cold seawater. Flow of methane and hydrogen-rich hydrothermal fluids in the porous interior chimney walls supports archaeal biofilm communities dominated by a single phylotype of Methanosarcinales. In this study, we have extensively sampled the carbonate-hosted archaeal and bacterial communities by obtaining sequences of >200,000 amplicons of the 16S rRNA V6 region and correlated the results with isotopic (230Th) ages of the chimneys over a 1200 year period. Rare sequences in young chimneys were often more abundant in older chimneys, indicating that members of the rare biosphere can become dominant members of the ecosystem when environmental conditions change. These results suggest that a long history of selection over many cycles of chimney growth has resulted in numerous closely related species at Lost City, each of which is pre-adapted to a particular set of re-occurring environmental conditions. Due to the unique characteristics of the Lost City Hydrothermal Field, these data offer an unprecedented opportunity to study the dynamics of a microbial ecosystem's rare biosphere over a thousand-year time scale.
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ArticleASPIRE : the Amundsen Sea Polynya International Research Expedition(The Oceanography Society, 2012-09) Yager, Patricia L. ; Sherrell, Robert M. ; Stammerjohn, Sharon E. ; Alderkamp, Anne-Carlijn ; Schofield, Oscar M. E. ; Abrahamsen, E. Povl ; Arrigo, Kevin R. ; Bertilsson, Stefan ; Garay, D. Lollie ; Guerrero, Raul ; Lowry, Kate E. ; Moksnes, Per-Olav ; Ndungu, Kuria ; Post, Anton F. ; Randall-Goodwin, Evan ; Riemann, Lasse ; Severmann, Silke ; Thatje, Sven ; van Dijken, Gert L. ; Wilson, StephanieIn search of an explanation for some of the greenest waters ever seen in coastal Antarctica and their possible link to some of the fastest melting glaciers and declining summer sea ice, the Amundsen Sea Polynya International Research Expedition (ASPIRE) challenged the capabilities of the US Antarctic Program and RVIB Nathaniel B. Palmer during Austral summer 2010–2011. We were well rewarded by both an extraordinary research platform and a truly remarkable oceanic setting. Here we provide further insights into the key questions that motivated our sampling approach during ASPIRE and present some preliminary findings, while highlighting the value of the Palmer for accomplishing complex, multifaceted oceanographic research in such a challenging environment.
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ArticleAssociation of cesarean delivery and formula supplementation with the intestinal microbiome of 6-week-old infants(American Medical Association, 2016-01-11) Madan, Juliette C. ; Hoen, Anne G. ; Lundgren, Sara N. ; Farzan, Shohreh F. ; Cottingham, Kathryn L. ; Morrison, Hilary G. ; Sogin, Mitchell L. ; Li, Hongzhe ; Moore, Jason H. ; Karagas, Margaret R.The intestinal microbiome plays a critical role in infant development, and delivery mode and feeding method (breast milk vs formula) are determinants of its composition. However, the importance of delivery mode beyond the first days of life is unknown, and studies of associations between infant feeding and microbiome composition have been generally limited to comparisons between exclusively breastfed and formula-fed infants, with little consideration given to combination feeding of both breast milk and formula.
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ArticleAxial Seamount(Oceanography Society, 2010-03) Chadwick, William W. ; Butterfield, David A. ; Embley, Robert W. ; Tunnicliffe, Verena ; Huber, Julie A. ; Nooner, Scott L. ; Clague, David A.Axial Seamount is a hotspot volcano superimposed on the Juan de Fuca Ridge (JdFR) in the Northeast Pacific Ocean. Due to its robust magma supply, it rises ~ 800 m above the rest of JdFR and has a large elongate summit caldera with two rift zones that parallel and overlap with adjacent segments of the spreading center.
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ArticleBacterial biogeography across the Amazon river-ocean continuum(Frontiers Media, 2017-05-23) Doherty, Mary ; Yager, Patricia L. ; Moran, Mary Ann ; Coles, Victoria J. ; Fortunato, Caroline S. ; Krusche, Alex V. ; Medeiros, Patricia M. ; Payet, Jérôme P. ; Richey, Jeffrey E. ; Satinsky, Brandon ; Sawakuchi, Henrique O. ; Ward, Nicholas D. ; Crump, Byron C.Spatial and temporal patterns in microbial biodiversity across the Amazon river-ocean continuum were investigated along ∼675 km of the lower Amazon River mainstem, in the Tapajós River tributary, and in the plume and coastal ocean during low and high river discharge using amplicon sequencing of 16S rRNA genes in whole water and size-fractionated samples (0.2–2.0 μm and >2.0 μm). River communities varied among tributaries, but mainstem communities were spatially homogeneous and tracked seasonal changes in river discharge and co-varying factors. Co-occurrence network analysis identified strongly interconnected river assemblages during high (May) and low (December) discharge periods, and weakly interconnected transitional assemblages in September, suggesting that this system supports two seasonal microbial communities linked to river discharge. In contrast, plume communities showed little seasonal differences and instead varied spatially tracking salinity. However, salinity explained only a small fraction of community variability, and plume communities in blooms of diatom-diazotroph assemblages were strikingly different than those in other high salinity plume samples. This suggests that while salinity physically structures plumes through buoyancy and mixing, the composition of plume-specific communities is controlled by other factors including nutrients, phytoplankton community composition, and dissolved organic matter chemistry. Co-occurrence networks identified interconnected assemblages associated with the highly productive low salinity near-shore region, diatom-diazotroph blooms, and the plume edge region, and weakly interconnected assemblages in high salinity regions. This suggests that the plume supports a transitional community influenced by immigration of ocean bacteria from the plume edge, and by species sorting as these communities adapt to local environmental conditions. Few studies have explored patterns of microbial diversity in tropical rivers and coastal oceans. Comparison of Amazon continuum microbial communities to those from temperate and arctic systems suggest that river discharge and salinity are master variables structuring a range of environmental conditions that control bacterial communities across the river-ocean continuum.
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ArticleBacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis(BioMed Central, 2016-04-19) Zozaya, Marcela ; Ferris, Michael J. ; Siren, Julia D. ; Lillis, Rebecca ; Myers, Leann ; Nsuami, M. Jacques ; Eren, A. Murat ; Brown, Jonathan ; Taylor, Christopher M. ; Martin, David H.The epidemiology of bacterial vaginosis (BV) suggests it is sexually transmissible, yet no transmissible agent has been identified. It is probable that BV-associated bacterial communities are transferred from male to female partners during intercourse; however, the microbiota of sexual partners has not been well-studied. Pyrosequencing analysis of PCR-amplified 16S rDNA was used to examine BV-associated bacteria in monogamous couples with and without BV using vaginal, male urethral, and penile skin specimens. The penile skin and urethral microbiota of male partners of women with BV was significantly more similar to the vaginal microbiota of their female partner compared to the vaginal microbiota of non-partner women with BV. This was not the case for male partners of women with normal vaginal microbiota. Specific BV-associated species were concordant in women with BV and their male partners. In monogamous heterosexual couples in which the woman has BV, the significantly higher similarity between the vaginal microbiota and the penile skin and urethral microbiota of the male partner, supports the hypothesis that sexual exchange of BV-associated bacterial taxa is common.
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ArticleBacterial diversity and community composition from seasurface to subseafloor(Nature Publishing Group, 2015-10-02) Walsh, Emily A. ; Kirkpatrick, John B. ; Rutherford, Scott D. ; Smith, David C. ; Sogin, Mitchell L. ; D'Hondt, StevenWe investigated compositional relationships between bacterial communities in the water column and those in deep-sea sediment at three environmentally distinct Pacific sites (two in the Equatorial Pacific and one in the North Pacific Gyre). Through pyrosequencing of the v4–v6 hypervariable regions of the 16S ribosomal RNA gene, we characterized 450 104 pyrotags representing 29 814 operational taxonomic units (OTUs, 97% similarity). Hierarchical clustering and non-metric multidimensional scaling partition the samples into four broad groups, regardless of geographic location: a photic-zone community, a subphotic community, a shallow sedimentary community and a subseafloor sedimentary community (greater than or equal to1.5 meters below seafloor). Abundance-weighted community compositions of water-column samples exhibit a similar trend with depth at all sites, with successive epipelagic, mesopelagic, bathypelagic and abyssopelagic communities. Taxonomic richness is generally highest in the water-column O2 minimum zone and lowest in the subseafloor sediment. OTUs represented by abundant tags in the subseafloor sediment are often present but represented by few tags in the water column, and represented by moderately abundant tags in the shallow sediment. In contrast, OTUs represented by abundant tags in the water are generally absent from the subseafloor sediment. These results are consistent with (i) dispersal of marine sedimentary bacteria via the ocean, and (ii) selection of the subseafloor sedimentary community from within the community present in shallow sediment.
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ArticleBacterial group II introns in a deep-sea hydrothermal vent environment(American Society for Microbiology, 2002-12) Podar, Mircea ; Mullineaux, Lauren S. ; Huang, Hon-Ren ; Perlman, Philip S. ; Sogin, Mitchell L.Group II introns are catalytic RNAs and mobile retrotransposable elements known to be present in the genomes of some nonmarine bacteria and eukaryotic organelles. Here we report the discovery of group II introns in a bacterial mat sample collected from a deep-sea hydrothermal vent near 9°N on the East Pacific Rise. One of the introns was shown to self-splice in vitro. This is the first example of marine bacterial introns from molecular population structure studies of microorganisms that live in the proximity of hydrothermal vents. These types of mobile genetic elements may prove useful in improving our understanding of bacterial genome evolution and may serve as valuable markers in comparative studies of bacterial communities.
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PreprintBidirectional incompatibility among divergent Wolbachia and incompatibility level differences among closely related Wolbachia in Nasonia( 2007-03-07) Bordenstein, Seth R. ; Werren, John H.Most insect groups harbor obligate bacterial symbionts from the alphaproteobacterial genus Wolbachia. These bacteria alter insect reproduction in ways that enhance their cytoplasmic transmission. One of the most common alterations is cytoplasmic incompatibility (CI) - a post-fertilization modification of the paternal genome that renders embryos inviable or unable to complete diploid development in crosses between infected males and uninfected females or infected females harboring a different strain. The parasitic wasp species complex Nasonia (N. vitripennis, N. longicornis, and N. giraulti) harbor at least six different Wolbachia that cause cytoplasmic incompatibility. Each species have double infections with a representative from both the A and B Wolbachia subgroups. CI relationships of the A and B Wolbachia of N. longicornis with those of N. giraulti and N. vitripennis are investigated here. We demonstrate that all pairwise crosses between the divergent A strains are bidirectionally incompatible. We were unable to characterize incompatibility between the B Wolbachia, but we establish that the B strain of N. longicornis induces no or very weak CI in comparison to the closely related B strain in N. giraulti that expresses complete CI. Taken together with previous studies, we show that independent acquisition of divergent A Wolbachia has resulted in three mutually incompatible strains, while codivergence of B Wolbachia in N. longicornis and N. giraulti is associated with differences in CI level. Understanding the diversity and evolution of new incompatibility strains will contribute to a fuller understanding of Wolbachia invasion dynamics and Wolbachia-assisted speciation in certain groups of insects.