Search

Many bacteria and other organisms carry out fermentations forming acetate. These fermentations have broad importance for foods, agriculture, and industry. They also are important for bacteria themselves because they often generate ATP. Here, we found a biochemical pathway for forming acetate and synthesizing ATP that was unknown in fermentative bacteria. We found that the bacterium Cutibacterium granulosum formed acetate during fermentation of glucose. It did not use phosphotransacetylase or acetate kinase, enzymes found in nearly all acetate-forming bacteria. Instead, it used a pathway involving two different enzymes. The first enzyme, succinyl coenzyme A (succinyl-CoA):acetate CoA-transferase (SCACT), forms acetate from acetyl-CoA. The second enzyme, succinyl-CoA synthetase (SCS), synthesizes ATP. We identified the genes encoding these enzymes, and they were homologs of SCACT and SCS genes found in other bacteria. The pathway resembles one described in eukaryotes, but it uses bacterial, not eukaryotic, gene homologs. To find other instances of the pathway, we analyzed sequences of all biochemically characterized homologs of SCACT and SCS (103 enzymes from 64 publications). Homologs with similar enzymatic activity had similar sequences, enabling a large-scale search for them in genomes. We searched nearly 600 genomes of bacteria known to form acetate, and we found that 6% encoded homologs with SCACT and SCS activity. This included >30 species belonging to 5 different phyla, showing that a diverse range of bacteria encode the SCACT/SCS pathway. This work suggests the SCACT/SCS pathway is important for acetate formation in many branches of the tree of life. IMPORTANCE Pathways for forming acetate during fermentation have been studied for over 80 years. In that time, several pathways in a range of organisms, from bacteria to animals, have been described. However, one pathway (involving succinyl-CoA:acetate CoA-transferase and succinyl-CoA synthetase) has not been reported in prokaryotes. Here, we discovered enzymes for this pathway in the fermentative bacterium Cutibacterium granulosum. We also found >30 other fermentative bacteria that encode this pathway, demonstrating that it could be common. This pathway represents a new way for bacteria to form acetate from acetyl-CoA and synthesize ATP via substrate-level phosphorylation. It could be a target for controlling yield of acetate during fermentation, with relevance for foods, agriculture, and industry.

The cpdB gene is pro-virulent in avian pathogenic Escherichia coli and in Salmonella enterica, where it encodes a periplasmic protein named CpdB. It is structurally related to cell wall-anchored proteins, CdnP and SntA, encoded by the also pro-virulent cdnP and sntA genes of Streptococcus agalactiae and Streptococcus suis, respectively. CdnP and SntA effects are due to extrabacterial hydrolysis of cyclic-di-AMP, and to complement action interference. The mechanism of CpdB pro-virulence is unknown, although the protein from non-pathogenic E. coli hydrolyzes cyclic dinucleotides. Considering that the pro-virulence of streptococcal CpdB-like proteins is mediated by c-di-AMP hydrolysis, S. enterica CpdB activity was tested as a phosphohydrolase of 3′-nucleotides, 2′,3′-cyclic mononucleotides, linear and cyclic dinucleotides, and cyclic tetra- and hexanucleotides. The results help to understand cpdB pro-virulence in S. enterica and are compared with E. coli CpdB and S. suis SntA, including the activity of the latter on cyclic-tetra- and hexanucleotides reported here for the first time. On the other hand, since CpdB-like proteins are relevant to host-pathogen interactions, the presence of cpdB-like genes was probed in eubacterial taxa by TblastN analysis. The non-homogeneous genomic distribution revealed taxa with cpdB-like genes present or absent, identifying eubacteria and plasmids where they can be relevant. [ABSTRACT FROM AUTHOR]

ABSTRACT Modulation of the intracellular cyclic di-GMP (c-di-GMP) pool is central to the formation of structured bacterial communities. Genome annotations predict the presence of dozens of conserved c-di-GMP catalytic enzymes in many bacterial species, but the functionality and regulatory control of the vast majority remain underexplored. Here, we begin to fill this gap by utilizing an experimental evolution system in Pseudomonas fluorescens Pf0-1, which repeatedly produces a unique social behavior through bidirectional transitions between two distinct phenotypes converging on c-di-GMP modulation. Parallel evolution of 33 lineages captured 147 unique mutations among 191 evolved isolates in genes that are empirically demonstrated, bioinformatically predicted, or previously unknown to impact the intracellular pool of c-di-GMP. Quantitative chemistry confirmed that each mutation causing the phenotypic shift either amplifies or reduces c-di-GMP production. We identify missense or in-frame deletion mutations in numerous diguanylate cyclase genes that largely fall outside the conserved catalytic domain. We also describe a novel relationship between a regulatory component of branched-chain amino acid biosynthesis and c-di-GMP production, and predict functions of several other unexpected proteins that clearly impact c-di-GMP production. Sequential mutations that continuously disrupt or recover c-di-GMP production across discrete functional elements suggest a complex and underappreciated interconnectivity within the c-di-GMP regulome of P. fluorescens. IMPORTANCE Microbial communities comprise densely packed cells where competition for space and resources is fierce. Aging colonies of Pseudomonas fluorescens are known to repeatedly produce mutants with two distinct phenotypes that physically work together to spread away from the overcrowded population. We demonstrate that the mutants with one phenotype produce high levels of cyclic di-GMP (c-di-GMP) and those with the second phenotype produce low levels. C-di-GMP is an intracellular signaling molecule which regulates many bacterial traits that cause tremendous clinical and environmental problems. Here, we analyze 147 experimentally selected mutations, which manifest either of the two phenotypes, to identify key residues in diverse proteins that force or shut down c-di-GMP production. Our data indicate that the intracellular pool of c-di-GMP is modulated through the catalytic activities of many independent c-di-GMP enzymes, which appear to be in tune with several proteins with no known links to c-di-GMP modulation.

ABSTRACT Phages and prophages are one of the principal modulators of microbial populations. However, much of their diversity is still poorly understood. Here, we extracted 33,624 prophages from 13,713 complete prokaryotic genomes to explore the prophage diversity and their relationships with their host. Our results reveal that prophages were present in 75% of the genomes studied. In addition, Enterobacterales were significantly enriched in prophages. We also found that pathogens are a significant reservoir of prophages. Finally, we determined that the prophage relatedness and the range of genomic hosts were delimited by the evolutionary relationships of their hosts. On a broader level, we got insights into the prophage population, identified in thousands of publicly available prokaryotic genomes, by comparing the prophage distribution and relatedness between them and their hosts. IMPORTANCE Phages and prophages play an essential role in controlling their host populations either by modulating the host abundance or providing them with genes that benefit the host. The constant growth in next-generation sequencing technology has caused the development of powerful computational tools to identify phages and prophages with high precision. Making it possible to explore the prophage populations integrated into host genomes on a large scale. However, it is still a new and under-explored area, and efforts are still required to identify prophage populations to understand their dynamics with their hosts.

Background The species checklists for Bhutan published in the Global Biodiversity Information Facility (GBIF) were originally collated for the publication ‘Biodiversity Statistics of Bhutan 2017, A Preliminary Baseline’ (BSB 2017). This document was published in 2019 and is the first comprehensive overview of Bhutan's species richness, recording more than 11,000 species across all five kingdoms. Collation of biodiversity checklists has been limited in Bhutan. Thus, this paper and its associated data provide an overview of all species from the kingdoms Plantae, Animalia, Fungi, Chromista and Eubacteria in the country that have been recorded in scientific publications. New information The checklists showcase 11,175 species from the kingdoms Plantae, Animalia, Fungi, Chromista and Eubacteria. Research conducted into identifying species from the kingdoms Protista and Archaebacteria revealed zero records. These checklists include at least 33 species new to science and 566 species new to Bhutan, discovered between 2009 and 2017. Unidentified species are not taken into account in this publication. [ABSTRACT FROM AUTHOR]

Background: Blood-feeding arthropods support a diverse array of symbiotic microbes, some of which facilitate host growth and development whereas others are detrimental to vectorborne pathogens. We found a common core constituency among the microbiota of 16 different arthropod blood-sucking disease vectors, including Bacillaceae, Rickettsiaceae, Anaplasmataceae, Sphingomonadaceae, Enterobacteriaceae, Pseudomonadaceae, Moraxellaceae and Staphylococcaceae. By comparing 21 genomes of common bacterial symbionts in blood-feeding vectors versus non-blooding insects, we found that certain enteric bacteria benefit their hosts by upregulating numerous genes coding for essential nutrients. Bacteria of blood-sucking vectors expressed significantly more genes (p < 0.001) coding for these essential nutrients than those of non-blooding insects. Moreover, compared to endosymbionts, the genomes of enteric bacteria also contained significantly more genes (p < 0.001) that code for the synthesis of essential amino acids and proteins that detoxify reactive oxygen species. In contrast, microbes in non-blood-feeding insects expressed few gene families coding for these nutrient categories. We also discuss specific midgut bacteria essential for the normal development of pathogens (e.g., Leishmania) versus others that were detrimental (e.g., bacterial toxins in mosquitoes lethal to Plasmodium spp.). [ABSTRACT FROM AUTHOR]

Arylamines constitute a large group of industrial chemicals detoxified by certain bacteria through conjugation reactions catalyzed by N-acetyltransferase (NAT) enzymes. NAT homologs, mostly from pathogenic bacteria, have been the subject of individual studies that do not lend themselves to direct comparisons. By implementing a practicable pipeline, we carried out a comparative investigation of 15 NAT homologs from 10 bacteria, mainly bacilli, streptomycetes, and one alphaproteobacterium. The new homologs were characterized for their sequence, phylogeny, predicted structural features, substrate specificity, thermal stability, and interaction with components of the enzymatic reaction. Bacillus NATs demonstrated the characteristics of xenobiotic metabolizing N-acetyltransferases, with the majority of homologs generating high activities. Nonpathogenic bacilli are thus proposed as suitable mediators of arylamine bioremediation. Of the Streptomyces homologs, the NAT2 isoenzyme of S. venezuelae efficiently transformed highly toxic arylamines, while the remaining homologs were inactive or generated low activities, suggesting that xenobiotic metabolism may not be their primary role. The functional divergence of Streptomyces NATs was consistent with their observed sequence, phylogenetic, and structural variability. These and previous findings support classification of microbial NATs into three groups. The first includes xenobiotic metabolizing enzymes with dual acetyl/propionyl coenzyme A (CoA) selectivity. Homologs of the second group are more rarely encountered, acting as malonyltransferases mediating specialized ecological interactions. Homologs of the third group effectively lack acyltransferase activity, and their study may represent an interesting research area. Comparative NAT enzyme screens from a broad microbial spectrum may guide rational selection of homologs likely to share similar biological functions, allowing their combined investigation and use in biotechnological applications. [ABSTRACT FROM AUTHOR]

Many bacteria and other organisms carry out fermentations forming acetate. These fermentations have broad importance for foods, agriculture, and industry. They also are important for bacteria themselves because they often generate ATP. Here, we found a biochemical pathway for forming acetate and synthesizing ATP that was unknown in fermentative bacteria. We found that the bacterium Cutibacterium granulosum formed acetate during fermentation of glucose. It did not use phosphotransacetylase or acetate kinase, enzymes found in nearly all acetate-forming bacteria. Instead, it used a pathway involving two different enzymes. The first enzyme, succinyl coenzyme A (succinyl-CoA):acetate CoA-transferase (SCACT), forms acetate from acetyl-CoA. The second enzyme, succinyl-CoA synthetase (SCS), synthesizes ATP. We identified the genes encoding these enzymes, and they were homologs of SCACT and SCS genes found in other bacteria. The pathway resembles one described in eukaryotes, but it uses bacterial, not eukaryotic, gene homologs. To find other instances of the pathway, we analyzed sequences of all biochemically characterized homologs of SCACT and SCS (103 enzymes from 64 publications). Homologs with similar enzymatic activity had similar sequences, enabling a largescale search for them in genomes. We searched nearly 600 genomes of bacteria known to form acetate, and we found that 6% encoded homologs with SCACT and SCS activity. This included .30 species belonging to 5 different phyla, showing that a diverse range of bacteria encode the SCACT/SCS pathway. This work suggests the SCACT/SCS pathway is important for acetate formation in many branches of the tree of life. [ABSTRACT FROM AUTHOR]

Here, I would like to overview an overlooked part of the nomenclatural history of the bacterial domain of life. The usage of the word Bacteria was quite wide before Ferdinand Cohn's famous 1872 work to which it is now ascribed. This word applied to microorganisms appeared 27 years before this (in 1845). Also, the name Eubacteria appeared three years earlier than is thought now (in 1883). Both arose thanks to persons other than Ferdinand Cohn and Joseph Schröter to whom they are now attributed. [ABSTRACT FROM AUTHOR]

A method for identifying mixed infections, dysbioses, and inflammatory processes from specific markers (fatty acids, aldehydes, and sterols) by chromatographic mass spectrometry was used to study the microecological status of patients eligible for liver transplantation. Heterogeneous microorganisms (aerobes, anaerobes, actinobacteria, fungi, and viruses) were quantified from molecular markers in an experiment within 3 hours after the samples were sent to the laboratory. The examinees were found to have excessive growth of gram-positive anaerobes (Clostridia, eubacteria), actinobacteria of the genera Streptomyces, Nocardia, and Candida yeasts. Next were staphylococci, streptococci, and gram-negative microorganisms of the group Moraxella/Acinetobacter, Pseudomonas aeruginosa, Helicobacter pylory, and Fusobacterium/Haemophylus. Antibiotics and probiotics were chosen to correct dysbiosis and infection, by taking into account the data available in the literature. The speed and accuracy of the procedure ensure real-time monitoring of a treatment process under control of mass spectrometry.

Background: Blood-feeding arthropods support a diverse array of symbiotic microbes, some of which facilitate host growth and development whereas others are detrimental to vector-borne pathogens. We found a common core constituency among the microbiota of 16 different arthropod blood-sucking disease vectors, including Bacillaceae, Rickettsiaceae, Anaplasmataceae, Sphingomonadaceae, Enterobacteriaceae, Pseudomonadaceae, Moraxellaceae and Staphylococcaceae. By comparing 21 genomes of common bacterial symbionts in blood-feeding vectors versus non-blooding insects, we found that certain enteric bacteria benefit their hosts by upregulating numerous genes coding for essential nutrients. Bacteria of blood-sucking vectors expressed significantly more genes (p < 0.001) coding for these essential nutrients than those of non-blooding insects. Moreover, compared to endosymbionts, the genomes of enteric bacteria also contained significantly more genes (p < 0.001) that code for the synthesis of essential amino acids and proteins that detoxify reactive oxygen species. In contrast, microbes in non-blood-feeding insects expressed few gene families coding for these nutrient categories. We also discuss specific midgut bacteria essential for the normal development of pathogens (e.g., Leishmania) versus others that were detrimental (e.g., bacterial toxins in mosquitoes lethal to Plasmodium spp.).

Abstrak: Tujuan dari penelitian ini adalah untuk mengungkap miskonsepsi siswa kelas X pada konsep Archaebacteria dan Eubacteria menggunakan tes diagnostik two-tier multiple choice di SMAN 9 Bandar Lampung. Penelitian ini termasuk kedalam penelitian kualitatif deskriptif. Populasi dalam penelitian ini adalah siswa kelas X MIA. Sampel dalam penelitian ini sebanyak 54 orang. Data tes diagnostik dikumpulkan menggunakan two-tier multiple choice untuk mengidentifikasi pemahaman konsep siswa kedalam paham konsep, miskonsepsi, menebak dan tidak paham konsep. Berdasarkan hasil penelitian yang diperoleh dari tes diagnostik two-tier multiple choice terhadap siswa kelas X MIA1 - X MIA6 SMA Negeri di Bandar Lampung bahwa miskonsepsi teridentifikasi disetiap subkonsep pada konsep Archaebacteria dan Eubacteria. Urutan subkonsep yang teridentifikasi miskonsepsi dari yang memiliki persentase tertinggi hingga terendah adalah sebagai berikut : peranan bakteri dalam kehidupan (31%), cara bakteri mendapatkan nutrisi (28%), archaebacteria (26%), eubacteria (25%), struktur tubuh bakteri (25%), bentuk-bentuk bakteri (24%), dan reproduksi bakteri (22%). Dari hasil penelitian ini dapat disimpulkan bahwa 26 % siswa mengalami miskonsepsi pada konsep Archaebacteria dan Eubacteria sedangkan sisa persentase kategori lainnya didominasi oleh kategori memahami, menebak dan tidak paham konsep.

The 21st-century biology learning requires technology-based learning which is potential in improving students’ learning outcomes. This research aimed to examine the validity and effectiveness of web-based learning media on Archaebacteria and Eubacteria on learning process. This web-based learning media was structured by using 4-D Thiagarajan model. The research was conducted at three SSHSs of Samarinda. The data were obtained from validators’ (media expert, material expert, and linguist), teachers’, and students’ opinion by using questionnaire; while the effectiveness was determined based on students’ test score and their response. The results showed that the web-based learning media was very valid with the average score from experts as high as 95.27%. Moreover, this media has been proven to improve the student’s cognitive learning outcomes (gain score 0.39) and well responded by both teachers and students.

Penelitian ini bertujuan untuk mengungkap miskonsespsi siswa mengidentifikasi miskonsepsi siswa pada konsep Archaebacteria dan Eubacteria menggunakan two-tier multiple choice. Metode penelitian yang digunakan dalam penelitian ini adalah deskriptif. Subjek dalam penelitian ini adalah peserta didik Sekolah Menengah Atas ddi Jakarta pada tahun ajaran 2013-2014 yang berjumlah 35 orang siswa. Data tes diagnostik dikumpulkan menggunakan two-tier multiple choice untuk mengidentifikasi pemahaman konsep siswa ke dalam paham konsep, miskonsepsi, tidak paham dan menebak (4 kategori). Hasil menunjukkan 31.12% peserta didik mengalami miskonsepsi pada konsep Archaebacteria dan Eubacteria sedangkan sisanya didominasi oleh kategori tidak memahami.

ABSTRACT Group B Streptococcus (GBS) is a commensal of the gastrointestinal and genitourinary tracts, while a prevailing cause of neonatal disease worldwide. Of the various clonal complexes (CCs), CC17 is overrepresented in GBS-infected newborns for reasons that are still largely unknown. Here, we report a comprehensive genomic analysis of 626 CC17 isolates collected worldwide, identifying the genetic traits behind their successful adaptation to humans and the underlying differences between carriage and clinical strains. Comparative analysis with 923 GBS genomes belonging to CC1, CC19, and CC23 revealed that the evolution of CC17 is distinct from that of other human-adapted lineages and recurrently targets functions related to nucleotide and amino acid metabolism, cell adhesion, regulation, and immune evasion. We show that the most distinctive features of disease-specific CC17 isolates were frequent mutations in the virulence-associated CovS and Stk1 kinases, underscoring the crucial role of the entire CovRS regulatory pathway in modulating the pathogenicity of GBS. Importantly, parallel and convergent evolution of major components of the bacterial cell envelope, such as the capsule biosynthesis operon, the pilus, and Rib, reflects adaptation to host immune pressures and should be taken into account in the ongoing development of a GBS vaccine. The presence of recurrent targets of evolution not previously implicated in virulence also opens the way for uncovering new functions involved in host colonization and GBS pathogenesis. IMPORTANCE The incidence of group B Streptococcus (GBS) neonatal disease continues to be a significant cause of concern worldwide. Strains belonging to clonal complex 17 (CC17) are the most frequently responsible for GBS infections in neonates, especially among late-onset disease cases. Therefore, we undertook the largest genomic study of GBS CC17 strains to date to decipher the genetic bases of their remarkable colonization and infection ability. We show that crucial functions involved in different steps of the colonization or infection process of GBS are distinctly mutated during the adaptation of CC17 to the human host. In particular, our results implicate the CovRS two-component regulator of virulence in the differentiation between carriage- and disease-associated isolates. Not only does this work raise important implications for the ongoing development of a vaccine against GBS but might also drive the discovery of key functions for GBS adaptation and pathogenesis that have been overlooked until now. Author Video: An author video summary of this article is available.

Circadian clocks in plants, animals, fungi, and in photosynthetic bacteria have been well-described. Observations of circadian rhythms in non-photosynthetic Eubacteria have been sporadic, and the molecular basis for these potential rhythms remains unclear. Here, we present the published experimental and bioinformatical evidence for circadian rhythms in these non-photosynthetic Eubacteria. From this, we suggest that the timekeeping functions of these organisms will be best observed and studied in their appropriate complex environments. Given the rich temporal changes that exist in these environments, it is proposed that microorganisms both adapt to and contribute to these daily dynamics through the process of temporal mutualism. Understanding the timekeeping and temporal interactions within these systems will enable a deeper understanding of circadian clocks and temporal programs and provide valuable insights for medicine and agriculture.

Diabetes in India has distinct genetic, nutritional, developmental and socio-economic aspects; owing to the fact that changes in gut microbiota are associated with diabetes, we employed semiconductor-based sequencing to characterize gut microbiota of diabetic subjects from this region. We suggest consolidated dysbiosis of eubacterial, archaeal and eukaryotic components in the gut microbiota of newly diagnosed (New-DMs) and long-standing diabetic subjects (Known-DMs) compared to healthy subjects (NGTs). Increased abundance of phylum Firmicutes (p = 0.010) and Operational Taxonomic Units (OTUs) of Lactobacillus (p < 0.01) were observed in Known-DMs subjects along with the concomitant graded decrease in butyrate-producing bacterial families like Ruminococcaceae and Lachnospiraceae. Eukaryotes and fungi were the least affected components in these subjects but archaea, except Methanobrevibacter were significantly decreased in them. The two dominant archaea viz. Methanobrevibacater and Methanosphaera followed opposite trends in abundance from NGTs to Known-DMs subjects. There was a substantial reduction in eubacteria, with a noticeable decrease in Bacteroidetes phylum (p = 0.098) and an increased abundance of fungi in New-DMs subjects. Likewise, opportunistic fungal pathogens such as Aspergillus, Candida were found to be enriched in New-DMs subjects. Analysis of eubacterial interaction network revealed disease-state specific patterns of ecological interactions, suggesting the distinct behavior of individual components of eubacteria in response to the disease. PERMANOVA test indicated that the eubacterial component was associated with diabetes-related risk factors like high triglyceride (p = 0.05), low HDL (p = 0.03), and waist-to-hip ratio (p = 0.02). Metagenomic imputation of eubacteria depict deficiencies of various essential functions such as carbohydrate metabolism, amino acid metabolism etc. in New-DMs subjects. Results presented here shows that in diabetes, microbial dysbiosis may not be just limited to eubacteria. Due to the inter-linkedmetabolic interactions among the eubacteria, archaea and eukarya in the gut, it may extend into other two domains leading to trans-domain dysbiosis in microbiota. Our results thus contribute to and expand the identification of biomarkers in diabetes. [ABSTRACT FROM AUTHOR]

Initiation of DNA replication is required for cell viability and passage of genetic information to the next generation. Studies in Escherichia coli and Bacillus subtilis have established A TPases a ssociated with diverse cellular a ctivities (AAA+) as essential proteins required for loading of the replicative helicase at replication origins. AAA+ ATPases DnaC in E. coli and DnaI in B. subtilis have long been considered the paradigm for helicase loading during replication in bacteria. Recently, it has become increasingly clear that most bacteria lack DnaC/DnaI homologs. Instead, most bacteria express a protein homologous to the newly described DciA ( d na C /dna I a ntecedent) protein. DciA is not an ATPase, and yet it serves as a helicase operator, providing a function analogous to that of DnaC and DnaI across diverse bacterial species. The recent discovery of DciA and of other alternative mechanisms of helicase loading in bacteria has changed our understanding of DNA replication initiation. In this review, we highlight recent discoveries, detailing what is currently known about the replicative helicase loading process across bacterial species, and we discuss the critical questions that remain to be investigated.

Fluorescence in situ hybridization (FISH) is a molecular technique widely used for the detection and characterization of microbial populations. FISH is affected by a wide variety of abiotic and biotic variables and the way they interact with each other. This is translated into a wide variability of FISH procedures found in the literature. The aim of this work is to systematically study the effects of pH, dextran sulfate and probe concentration in the FISH protocol, using a general peptide nucleic acid (PNA) probe for the Eubacteria domain. For this, response surface methodology was used to optimize these 3 PNA-FISH parameters for Gram-negative ( Escherichia coli and Pseudomonas fluorescens ) and Gram-positive species ( Listeria innocua , Staphylococcus epidermidis and Bacillus cereus ). The obtained results show that a probe concentration higher than 300 nM is favorable for both groups. Interestingly, a clear distinction between the two groups regarding the optimal pH and dextran sulfate concentration was found: a high pH (approx. 10), combined with lower dextran sulfate concentration (approx. 2% [w/v]) for Gram-negative species and near-neutral pH (approx. 8), together with higher dextran sulfate concentrations (approx. 10% [w/v]) for Gram-positive species. This behavior seems to result from an interplay between pH and dextran sulfate and their ability to influence probe concentration and diffusion towards the rRNA target. This study shows that, for an optimum hybridization protocol, dextran sulfate and pH should be adjusted according to the target bacteria. [ABSTRACT FROM AUTHOR]

Elucidating the genomic features influencing prokaryotic growth rates has always been a study of interest. Previously, it was observed that overlapping genes (OGs) play a crucial role in the prokaryotic genome size reduction. This study is focused to explore whether OGs act as a potential correlate of prokaryotic growth rates. For this purpose, we compiled a dataset of 25 archaeal and 117 eubacterial genomes and analyzed the inter-correlation between the proportion of overlapping regions in these genomes with their growth rates. Here, we observed that the proportion of overlapping region holds a significant negative correlation with generation time in archaeal domain, whereas no correlation was observed in the eubacterial domain. However, after masking the effect of tRNA, rRNA multiplicity and environmental diversity, OGs show an independent effect over growth rates in the eubacterial domain as well as in the archaeal domain. Moreover, the influence of OGs on prokaryotic growth rates provides different delineations in archaeal and eubacterial domains. In archaea, both long overlap frequency (LOF) and short overlap frequency (SOF) influence the growth rates by increasing the degree of operonization. On the contrary, in the case of bacteria, neither SOF nor LOF plays any significant role in achieving faster growth rates. [ABSTRACT FROM AUTHOR]

We investigated microorganisms dwelling on rocks, walls and paintings in two votive chapels built in grottoes in the Region of Campania, Italy. One grotto was near the coast in an area with a Mediterranean climate, and the other grotto was inland on a mountain in an area with a cold continental climate. Color and distribution of biofilms in various areas of the grottoes were examined. Microbial components of biofilms were identified by light and electron microscopy and by molecular techniques (DNA analyses and Automatic rRNA Intergenic Spacer Analysis). Biofilms were also analyzed by X-ray diffraction to detect inorganic constituents deriving from rocks in the grottoes and walls of the churches and by X-ray fluorescence to detect the elements that made up the pigments of the mural paintings; optical cross sections were used to observe their relationships with substrata. Species of eubacteria, cyanobacteria and green algae were identified. Some of these species occurred in both grottoes, while others were exclusive to only one of the grottoes. The diversity of species, their common or exclusive occurrence in the grottoes, the relationships among microbial communities and the differences in color and distribution of biofilms were discussed on the basis of the different climatic factors affecting the two grottoes and the different inorganic components of substrata. [ABSTRACT FROM AUTHOR]

Bacterial evolution is characterized by strong purifying selection as well as rapid adaptive evolution in changing environments. In this context, the genomic GC content (genomic GC) varies greatly but presents some level of phylogenetic stability, making it challenging to explain based on current hypotheses. To illuminate the evolutionary mechanisms of the genomic GC, we analyzed the base composition and functional inventory of 11,083 representative genomes. A phylogenetically constrained bimodal distribution of the genomic GC, which mainly originated from parallel divergences in the early evolution, was demonstrated. Such variation of the genomic GC can be well explained by DNA replication and repair (DRR), in which multiple pathways correlate with the genomic GC. Furthermore, the biased conservation of various stress-related genes, especially the DRR-related ones, implies distinct adaptive processes in the ancestral lineages of high- or low-GC clades which are likely induced by major environmental changes. Our findings support that the mutational biases resulting from these legacies of ancient adaptation have changed the course of adaptive evolution and generated great variation in the genomic GC. This highlights the importance of indirect effects of natural selection, which indicates a new model for bacterial evolution. IMPORTANCE GC content has been shown to be an important factor in microbial ecology and evolution, and the genomic GC of bacteria can be characterized by great intergenomic heterogeneity, high intragenomic homogeneity, and strong phylogenetic inertia, as well as being associated with the environment. Current hypotheses concerning direct selection or mutational biases cannot well explain these features simultaneously. Our findings of the genomic GC showing that ancient adaptations have transformed the DRR system and that the resulting mutational biases further contributed to a bimodal distribution of it offer a more reasonable scenario for the mechanism. This would imply that, when thinking about the evolution of life, diverse processes of adaptation exist, and combined effects of natural selection should be considered.

BACKGROUND The utilization of different kinds of ion exchange membrane is a common practice in bioelectrochemical systems such as two-chambered microbial fuel cells ( MFCs). However, little is known about the effect of membrane materials on the anodic microbial community diversity. RESULTS The effect of two cationic and one anionic exchange membranes (Nafion N-117, Ultrex CMI-7000, and Ultrex AMI-7000) on the microbial community dynamics of Eubacteria and Archaea has been assessed in two-chambered MFCs. The experimental results indicated that the eubacterial community in the anodic chamber was not affected by the membrane materials, being predominantly populations of Bacteroidetes ( Porphyromonadaceae) and β-proteobacteria ( Alcaligenaceae and Comamonadaceae). On the other hand, the archaeal counterpart appears to be highly dependent on the type of membrane used, as was evidenced by the selective enrichment of Methanosarcina sp. in the MFC equipped with the membrane Nafion N-117 which was the MFC that showed the highest current production. CONCLUSIONS The results obtained in the present study suggest that membrane materials affect archaeal diversity whereas both anodofilic eubacteria and methanogenic archaea populations could play an important role in the overall MFC process performance. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Background: Only a fraction of the microbial species used for anaerobic digestion in biogas production plants are methanogenic archaea. We have analyzed the taxonomic profiles of eubacteria and archaea, a set of chemical key parameters, and biogas production in samples from nine production plants in seven facilities in Thuringia, Germany, including co-digesters, leach-bed, and sewage sludge treatment plants. Reactors were sampled twice, at a 1-week interval, and three biological replicates were taken in each case. Results: A complex taxonomic composition was found for both eubacteria and archaea, both of which strongly correlated with digester type. Plant-degrading Firmicutes as well as Bacteroidetes dominated eubacteria profiles in high biogas-producing co-digesters; whereas Bacteroidetes and Spirochaetes were the major phyla in leach-bed and sewage sludge digesters. Methanoculleus was the dominant archaea genus in co-digesters, whereas Methanosarcina and Methanosaeta were the most abundant methanogens in leachate from leach-bed and sewage sludge digesters, respectively. Conclusions: This is one of the most comprehensive characterizations of the microbial communities of biogas-producing facilities. Bacterial profiles exhibited very low variation within replicates, including those of semi-solid samples; and, in general, low variation in time. However, facility type correlated closely with the bacterial profile: each of the three reactor types exhibited a characteristic eubacteria and archaea profile. Digesters operated with solid feedstock, and high biogas production correlated with abundance of plant degraders (Firmicutes) and biofilm-forming methanogens (Methanoculleus spp.). By contrast, low biogas-producing sewage sludge treatment digesters correlated with high titers of volatile fatty acid-adapted Methanosaeta spp. [ABSTRACT FROM AUTHOR]

Five rumen-cannulated Polish Merino rams were used in a 5 × 5 Latin square design to investigate the effect of sucrose and polymers of fructose on the activity and characteristics of rumen protozoa and bacteria. The animals were fed concentrate and timothy hay containing either 33 g or 163 g fructan/kg. The low-fructan hay was supplied in the control diet and in the diets supplemented with sucrose and inulins. The high-fructan hay was the component of diet rich in timothy grass fructan. The control diet supplied 39.6 g fructan/day. The remaining diet offered 195.6 g of fructose-containing carbohydrates/day. The concentration of total SCFA was 7.2–9.5 mM/100 ml rumen fluid independently of diet and sampling time (P>0.05). The increase in sucrose or fructan contents lowered the proportions of acetate from 73.1 to 64.2 mM/100 mM (P≤0.002) and increased those of propionate and butyrate from 16.4 to 21.5 (P=0.003), and from 9.8 to 15.6 mM/100 mM (P≤0.004), respectively. The sampling time affected the proportions of propionate (P=0.01). The diets did not affect pH of rumen fluid except in the ration with timothy grass fructan (P=0.029). Irrespective of diet and sampling time (P≥0.05), the digestion velocities of timothy grass fructan, inulin, sucrose, starch, pectins, carboxymethylcellulose, and xylan were 1.3–2.9; 0.7–2.1; 4.8–9.6; 1.9–2.4, 0.3–0.5, 0.7–1.2, and 0.7–1.4 mM reducing sugars/g dry matter (DM) of rumen fluid per hour, respectively. A positive correlation was found between the degradation rate of timothy grass fructan and inulin or sucrose (P≤0.045). The total number of ciliates was 261–475×10 3 /ml, that of Entodinium , 231–439×10 3 /ml. The concentrations of Epidinium , Isotricha and Dasytricha were 14–24×10 3 /ml, 2.6–7.0×10 3 /ml, and 3.2–8.1×10 3 /ml, respectively (P≥0.05). The Diplodinium counts varied from 1.4 to 3.1×10 3 /ml in control animals to 4.0–7.3×10 3 /ml in sheep fed the diet rich in timothy grass fructan and Beneo P95 (P≤0.008). A postprandial decrease in the concentration of Diplodiniuum was found in animals fed the control ration and the rations rich in timothy grass fructan and Beneo HPX (P=0.017). It was stated that the sampling time influenced the composition of bacterial flora. The experiment showed that the applied doses of the examined carbohydrates did not affect the majority of the quantified parameters. The studies shall be continued using enlarged doses of fructose-containing carbohydrates. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate and compare the microbial community structures of eubacteria and archaea in the pit mud of Chinese Luzhou- flavor liquor from the wall (C) and bottom (C) of cellar through nested PCR-denaturing gradient gel electrophoresis (DGGE). The Shannon-Wiener index (H) calculated from the DGGE profiles showed that the community diversities of eubacteria and archaea in samples from C were almost higher than that from C. In addition, cluster analysis of the DGGE profiles revealed that some differences were found in the microbial community structure in samples from different locations. The closely relative microorganisms of all eubacterial 16S rRNA gene sequences fell into four phyla ( Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria), including 12 genera and 2 uncultured eubacteria. Moreover, 37.1 % eubacteria were affiliated with Clostridium. Particularly, genus Acinetobacter was absent in all samples from C but present in all samples from C. The closely relative microorganisms of all archaeal 16S rRNA gene sequences fell into four genera, which included Methanobrevibacter, Methanoculleus, Methanobacterium and Methanosaeta, while the dominant archaea in samples from C and C were similar. Results presented in this study provide further understanding of the spatial differences in microbial community structure in the pit mud, and is of great importance for the production and quality improvement of Luzhou- flavor liquor. [ABSTRACT FROM AUTHOR]

Modulation of the intracellular cyclic di-GMP (c-di-GMP) pool is central to the formation of structured bacterial communities. Genome annotations predict the presence of dozens of conserved c-di-GMP catalytic enzymes in many bacterial species, but the functionality and regulatory control of the vast majority remain underexplored. Here, we begin to fill this gap by utilizing an experimental evolution system in Pseudomonas fluorescens Pf0-1, which repeatedly produces a unique social behavior through bidirectional transitions between two distinct phenotypes converging on c-di-GMP modulation. Parallel evolution of 33 lineages captured 147 unique mutations among 191 evolved isolates in genes that are empirically demonstrated, bioinformatically predicted, or previously unknown to impact the intracellular pool of c-di-GMP. Quantitative chemistry confirmed that each mutation causing the phenotypic shift either amplifies or reduces c-di-GMP production. We identify missense or in-frame deletion mutations in numerous diguanylate cyclase genes that largely fall outside the conserved catalytic domain. We also describe a novel relationship between a regulatory component of branched-chain amino acid biosynthesis and c-di-GMP production, and predict functions of several other unexpected proteins that clearly impact c-di-GMP production. Sequential mutations that continuously disrupt or recover c-di-GMP production across discrete functional elements suggest a complex and underappreciated interconnectivity within the c-di-GMP regulome of P. fluorescens. IMPORTANCE Microbial communities comprise densely packed cells where competition for space and resources is fierce. Aging colonies of Pseudomonas fluorescens are known to repeatedly produce mutants with two distinct phenotypes that physically work together to spread away from the overcrowded population. We demonstrate that the mutants with one phenotype produce high levels of cyclic di-GMP (c-di-GMP) and those with the second phenotype produce low levels. C-di-GMP is an intracellular signaling molecule which regulates many bacterial traits that cause tremendous clinical and environmental problems. Here, we analyze 147 experimentally selected mutations, which manifest either of the two phenotypes, to identify key residues in diverse proteins that force or shut down c-di-GMP production. Our data indicate that the intracellular pool of c-di-GMP is modulated through the catalytic activities of many independent c-di-GMP enzymes, which appear to be in tune with several proteins with no known links to c-di-GMP modulation.

Among isolates of the eubacteria Brevundimonas bullata, Sphingomonas rhizogenes, Agrobacterium tumefaciens, Bacillus simplex, Acinetobacter johnsonii, Methylobacterium adhaesivum, and Deinococcus aquaticus isolated from the bottom layer on the medium, where the only source of organic matter was hydrolysate from diatoms, the activity of hydrolytic enzymes, such as proteases, β-xylosidase, β-glucosidase, β-galactosidase, chitobiase, and amylase, is revealed. An algicidal effect and degradation of siliceous diatom frustules are recorded in algal-bacterial cultures. These data show that the bottom water layer of Lake Baikal is inhabited by potential participants of silicon cycle, which degrade siliceous valves of diatoms. [ABSTRACT FROM AUTHOR]

Background: The gut microbiota has been shown to affect both fat storage and energy harvesting, suggesting that it plays a direct role in the development of obesity. The aim of this study was to investigate whether intestinal colonization by particular species/groups of the intestinal microbiota is related to body weight values in Estonian preschool children born in different years during the entire 1990s. Methods: Body weight, height, body mass index (BMI), and quantitative composition of cultivable gut microbiota (staphylococci, enterococci, streptococci, enterobacteria, lactobacilli, anaerobic gram-positive cocci, bifidobacteria, eubacteria, bacteroides, clostridia, and candida) were studied in 51 healthy 5-year-old children (40 were born between 1993 and 94 and 11 were born between 1996 and 97). Results: At the age of 5 years, median weight was 19.5 kg and median BMI was 15.3 kg/m2. Significantly higher BMI (p=0.006) was found in 5-year-old children born in late versus early 1990s during the development of socioeconomic situation of Estonia (2% rise in gross domestic product). The counts of the different gut bacteria did not show any association with weight and BMI in the 5-year-old children. However, the BMI values were in positive correlation with a relative share of anaerobic gram-positive bacteria, for example, bifidobacteria when adjusted for sex and year of birth (adj R2=0.459, p=0.026) and eubacteria (adj R2=0.484, p=0.014) in the community of cultured intestinal microbiota. The relative share of bacteroides showed a negative correlation with the childrens' weight (adj R2=-0.481, p=0.015). Conclusion: The body weight indices of preschool children of the general population are associated with the proportion of anaerobic intestinal microbiota and can be predicted by sex and particular socioeconomic situation from birth to 5 years of age [ABSTRACT FROM AUTHOR]

Abstract: On-deck CO2-Fe-manipulated incubation experiments were conducted using surface seawater collected from the Western Subarctic Gyre of the NW Pacific in the summer of 2008 to elucidate the impacts of ocean acidification and Fe enrichment on the abundance and community composition of phytoplankton and eubacteria in the study area. During the incubation, excluding the initial period, the mean partial pressures of CO2 in non-Fe-added bottles were 230, 419, 843, and 1124μatm, whereas those in Fe-added treatments were 152, 394, 791, and 1008μatm. Changes in the abundance and community composition of phytoplankton were estimated using HPLC pigment signatures with the program CHEMTAX and flow cytometry. A DGGE fingerprint technique targeting 16S rRNA gene fragments was also used to estimate changes in eubacterial phylotypes during incubation. The Fe addition induced diatom blooms, and subsequently stimulated the growth of heterotrophic bacteria such as Roseobacter, Phaeobacter, and Alteromonas in the post-bloom phase. In both the Fe-limited and Fe-replete treatments, concentrations of 19′-hexanoyloxyfucoxanthin, a haptophyte marker, and the cell abundance of coccolithophores decreased at higher CO2 levels (750 and 1000ppm), whereas diatoms exhibited little response to the changes in CO2 availability. The abundances of Synechococcus and small eukaryotic phytoplankton (<10μm) increased at the higher CO2 levels. DGGE band positions revealed that Methylobacterium of Alphaproteobacteria occurred solely at lower CO2 levels (180 and 380ppm) during the post-bloom phase. These results suggest that increases in CO2 level could affect not only the community composition of phytoplankton but also that of eubacteria. As these microorganisms play critical roles in the biological carbon pump and microbial loop, our results indicate that the progression of ocean acidification can alter the biogeochemical processes in the study area. [Copyright &y& Elsevier]

Phages and prophages are one of the principal modulators of microbial populations. However, much of their diversity is still poorly understood. Here, we extracted 33,624 prophages from 13,713 complete prokaryotic genomes to explore the prophage diversity and their relationships with their host. Our results reveal that prophages were present in 75% of the genomes studied. In addition, Enterobacterales were significantly enriched in prophages. We also found that pathogens are a significant reservoir of prophages. Finally, we determined that the prophage relatedness and the range of genomic hosts were delimited by the evolutionary relationships of their hosts. On a broader level, we got insights into the prophage population, identified in thousands of publicly available prokaryotic genomes, by comparing the prophage distribution and relatedness between them and their hosts. IMPORTANCE Phages and prophages play an essential role in controlling their host populations either by modulating the host abundance or providing them with genes that benefit the host. The constant growth in next-generation sequencing technology has caused the development of powerful computational tools to identify phages and prophages with high precision. Making it possible to explore the prophage populations integrated into host genomes on a large scale. However, it is still a new and under-explored area, and efforts are still required to identify prophage populations to understand their dynamics with their hosts.

A fundamental requirement for life is the replication of an organism's DNA. Studies in Escherichia coli and Bacillus subtilis have set the paradigm for DNA replication in bacteria. During replication initiation in E. coli and B. subtilis, the replicative helicase is loaded onto the DNA at the origin of replication by an ATPase helicase loader. However, most bacteria do not encode homologs to the helicase loaders in E. coli and B. subtilis. Recent work has identified the DciA protein as a predicted helicase operator that may perform a function analogous to the helicase loaders in E. coli and B. subtilis. DciA proteins, which are defined by the presence of a DUF721 domain (termed the DciA domain herein), are conserved in most bacteria but have only been studied in mycobacteria and gammaproteobacteria (Pseudomonas aeruginosa and Vibrio cholerae). Sequences outside the DciA domain in Mycobacterium tuberculosis DciA are essential for protein function but are not conserved in the P. aeruginosa and V. cholerae homologs, raising questions regarding the conservation and evolution of DciA proteins across bacterial phyla. To comprehensively define the DciA protein family, we took a computational evolutionary approach and analyzed the domain architectures and sequence properties of DciA domain-containing proteins across the tree of life. These analyses identified lineage-specific domain architectures among DciA homologs, as well as broadly conserved sequence-structural motifs. The diversity of DciA proteins represents the evolution of helicase operation in bacterial DNA replication and highlights the need for phylum-specific analyses of this fundamental biological process. IMPORTANCE Despite the fundamental importance of DNA replication for life, this process remains understudied in bacteria outside Escherichia coli and Bacillus subtilis. In particular, most bacteria do not encode the helicase-loading proteins that are essential in E. coli and B. subtilis for DNA replication. Instead, most bacteria encode a DciA homolog that likely constitutes the predominant mechanism of helicase operation in bacteria. However, it is still unknown how DciA structure and function compare across diverse phyla that encode DciA proteins. In this study, we performed computational evolutionary analyses to uncover tremendous diversity among DciA homologs. These studies provide a significant advance in our understanding of an essential component of the bacterial DNA replication machinery.

Abstract: A process combining an Anaerobic Baffled Reactor (ABR) and a constructed wetland was employed to treat nitrobenzene wastewater. The overall performance was examined throughout long-term operation with a hydraulic retention time (HRT) of 24h at 30±1°C. The effluent nitrobenzene concentration of the ABR and constructed wetland was less than 4.81 and 1.94mg/L, respectively, with an initial nitrobenzene concentration of 80mg/L at the steady-state periods. The corresponding removal efficiencies were 97.02% and 73.93%, respectively. Moreover, 97.29% of aniline produced in the ABR could be removed in the subsequent wetland. The number of sequenced clones from each library was sufficient to cover archaea and eubacteria diversity at the species level and to obtain a representation of the total microbial diversity in the ABR. The predominant microbial populations in the ABR were identified as Pseudomonas putida, Methanosaeta concilii and Methanothrix soehngenii. [Copyright &y& Elsevier]

Pseudomonas aeruginosa SB1 was isolated from conventional well water. It produced an extracellular fluorescent bright green pigment which diffused into the growth medium. The purified pigment showed absorption maxima of 690, 370, 312 nm and was identified as pyocyanine, a known inhibitor of eubacterial and eukaryotic growth. The effect of this pigment when tested on orange-red pigmented halophilic archaeal isolates showed remarkable inhibitory activity and resulted in total growth inhibition at 4.67 mg/ml pyocyanine in 24 h. The present work is the first report on eubacterial Pseudomonas aeruginosa SB1 showing anti-Haloarchaeal activity. [ABSTRACT FROM AUTHOR]

Abstract: Graphical techniques have become powerful tools for the visualization and analysis of complicated biological systems. However, we cannot give such a graphical representation in a 2D/3D space when the dimensions of the represented data are more than three dimensions. The proposed method, a combination dimensionality reduction approach (CDR), consists of two parts: (i) principal component analysis (PCA) with a newly defined parameter ρ and (ii) locally linear embedding (LLE) with a proposed graphical selection for its optional parameter k. The CDR approach with ρ and k not only avoids loss of principal information, but also sufficiently well preserves the global high-dimensional structures in low-dimensional space such as 2D or 3D. The applications of the CDR on characteristic analysis at different codon positions in genome show that the method is a useful tool by which biologists could find useful biological knowledge. [Copyright &y& Elsevier]

Abstract: The role of anaerobic digestion (AD) on the decontamination and biomethanization of a PCB-spiked sludge obtained from a Membrane Biological Reactor (MBR) pilot plant was investigated throughout a 10-month batch experiment. The study was carried out under mesophilic (35°C) and thermophilic (55°C) conditions and was monitored by means of an integrated chemical, microbiological and molecular biology strategy. Remarkable PCB depletions (higher than 50% of the overall spiked PCBs) and dechlorinations were achieved under methanogenic conditions. The process was not affected by yeast extract addition. Both acetoclastic and hydrogenotrophic methanogens, together with some fermentative eubacteria, were found to persist in all PCB biodegrading microcosms. This finding, together with those obtained from parallel microcosms where specific populations were selectively inhibited, suggested that native methanogens played a key role in the biodegradation and dechlorination of the spiked PCBs. Taken together, the results of this study indicate that AD is a feasible option for the decontamination and the efficient disposal (with the production of a CH4-rich biogas) of contaminated MBR sludge, which can be then employed as a fertilizer for agricultural purposes. [Copyright &y& Elsevier]