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1.
mSystems ; 9(3): e0121923, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38329353

RESUMO

Bumblebees are among the most abundant and important pollinators for sub-alpine and alpine flowering plant species in the Northern Hemisphere, but little is known about their adaptations to high elevations. In this article, we focused on two bumblebee species, Bombus friseanus and Bombus prshewalskyi, and their respective gut microbiota. The two species, distributed through the Hengduan Mountains of southwestern China, show species replacement at different elevations. We performed genome sequencing based on 20 worker bee samples of each species. Applying evolutionary population genetics and metagenomic approaches, we detected genes under selection and analyzed functional pathways between bumblebees and their gut microbes. We found clear genetic differentiation between the two host species and significant differences in their microbiota. Species replacement occurred in both hosts and their bacteria (Snodgrassella) with an increase in elevation. These extremely high-elevation bumblebees show evidence of positive selection related to diverse biological processes. Positively selected genes involved in host immune systems probably contributed to gut microbiota changes, while the butyrate generated by gut microbiota may influence both host energy metabolism and immune systems. This suggests a close association between the genomes of the host species and their microbiomes based on some degree of natural selection.IMPORTANCETwo closely related and dominant bumblebee species, distributed at different elevations through the Hengduan Mountains of southwestern China, showed a clear genomic signature of adaptation to elevation at the molecular level and significant differences in their respective microbiota. Species replacement occurred in both hosts and their bacteria (Snodgrassella) with an increase in elevation. Bumblebees' adaptations to higher elevations are closely associated with their gut microbiota through two biological processes: energy metabolism and immune response. Information allowing us to understand the adaptive mechanisms of species to extreme conditions is implicit if we are to conserve them as their environments change.


Assuntos
Microbioma Gastrointestinal , Neisseriaceae , Abelhas/genética , Animais , Microbioma Gastrointestinal/genética , Bactérias/genética , Neisseriaceae/genética , Metagenoma , Evolução Biológica
2.
Microbiol Spectr ; 11(6): e0312323, 2023 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-37882538

RESUMO

IMPORTANCE: Understanding the evolutionary relationships between the species in the Neisseriaceae family has been a persistent challenge in bacterial systematics due to high recombination rates in these species. Previous studies of this family have focused on Neisseria meningitidis and N. gonorrhoeae. However, previously understudied Neisseriaceae species are gaining new attention, with Kingella kingae now recognized as a common human pathogen and with Alysiella and Simonsiella being unique in the bacterial world as multicellular organisms. A better understanding of the genomic evolution of the Neisseriaceae can lead to the identification of specific genes and traits that underlie the remarkable diversity of this family.


Assuntos
Neisseriaceae , Humanos , Neisseriaceae/genética , Kingella , Filogenia , Genômica , Fenótipo , Neisseria gonorrhoeae
3.
Nat Commun ; 14(1): 2098, 2023 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-37055390

RESUMO

Much remains to be explored regarding the diversity of uncultured, host-associated microbes. Here, we describe rectangular bacterial structures (RBSs) in the mouths of bottlenose dolphins. DNA staining revealed multiple paired bands within RBSs, suggesting the presence of cells dividing along the longitudinal axis. Cryogenic transmission electron microscopy and tomography showed parallel membrane-bound segments that are likely cells, encapsulated by an S-layer-like periodic surface covering. RBSs displayed unusual pilus-like appendages with bundles of threads splayed at the tips. We present multiple lines of evidence, including genomic DNA sequencing of micromanipulated RBSs, 16S rRNA gene sequencing, and fluorescence in situ hybridization, suggesting that RBSs are bacterial and distinct from the genera Simonsiella and Conchiformibius (family Neisseriaceae), with which they share similar morphology and division patterning. Our findings highlight the diversity of novel microbial forms and lifestyles that await characterization using tools complementary to genomics such as microscopy.


Assuntos
Golfinho Nariz-de-Garrafa , Neisseriaceae , Animais , RNA Ribossômico 16S/genética , Hibridização in Situ Fluorescente , Neisseriaceae/genética , Boca , Estruturas Bacterianas
4.
mSystems ; 7(3): e0150021, 2022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35604118

RESUMO

Snodgrassella is a genus of Betaproteobacteria that lives in the gut of honeybees (Apis spp.) and bumblebees (Bombus spp). It is part of a conserved microbiome that is composed of a few core phylotypes and is essential for bee health and metabolism. Phylogenomic analyses using whole-genome sequences of 75 Snodgrassella strains from 4 species of honeybees and 14 species of bumblebees showed that these strains formed a monophyletic lineage within the Neisseriaceae family, that Snodgrassella isolates from Asian honeybees diverged early from the other species in their evolution, that isolates from honeybees and bumblebees were well separated, and that this genus consists of at least seven species. We propose to formally name two new Snodgrassella species that were isolated from bumblebees: i.e., Snodgrassella gandavensis sp. nov. and Snodgrassella communis sp. nov. Possible evolutionary scenarios for 107 species- or group-specific genes revealed very limited evidence for horizontal gene transfer. Functional analyses revealed the importance of small proteins, defense mechanisms, amino acid transport and metabolism, inorganic ion transport and metabolism and carbohydrate transport and metabolism among these 107 specific genes. IMPORTANCE The microbiome of honeybees (Apis spp.) and bumblebees (Bombus spp.) is highly conserved and represented by few phylotypes. This simplicity in taxon composition makes the bee's microbiome an emergent model organism for the study of gut microbial communities. Since the description of the Snodgrassella genus, which was isolated from the gut of honeybees and bumblebees in 2013, a single species (i.e., Snodgrassella alvi), has been named. Here, we demonstrate that this genus is actually composed of at least seven species, two of which (Snodgrassella gandavensis sp. nov. and Snodgrassella communis sp. nov.) are formally described and named in the present publication. We also report the presence of 107 genes specific to Snodgrassella species, showing notably the importance of small proteins and defense mechanisms in this genus.


Assuntos
Microbiota , Neisseriaceae , Animais , Abelhas , Filogenia , Neisseriaceae/genética
5.
Genomics ; 113(4): 2800-2811, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34157405

RESUMO

Acquiring new DNA allows the emergence of drug resistance in bacteria. Some Pasteurellaceae and Neisseriaceae species preferentially take up specific sequence tags. The study of such sequences is therefore relevant. They are over-represented in the genomes of the corresponding species. I found similar sequences to be present only in, but not in all, the genomes of the Pasteurellaceae and Neisseriaceae families. The genomic densities of these sequences are different both between species and between families. Interestingly, the family whose genomes harbor more of such sequences also shows more sequence types. A phylogenetic analysis allowed inferring the possible ancestral Neisseriacean sequence and a nucleotide-by-nucleotide analysis allowed inferring the potential ancestral Pasteurellacean sequence based on its genomic footprint. The method used for this work could be applied to other sequences, including transcription factor binding and repeated DNAs.


Assuntos
Neisseriaceae , Pasteurellaceae , Bactérias/genética , DNA/metabolismo , Neisseriaceae/genética , Neisseriaceae/metabolismo , Pasteurellaceae/genética , Pasteurellaceae/metabolismo , Filogenia
6.
Microb Genom ; 6(5)2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32375974

RESUMO

Mobile genetic elements (MGEs) are key factors responsible for dissemination of virulence determinants and antimicrobial-resistance genes amongst pathogenic bacteria. Conjugative MGEs are notable for their high gene loads donated per transfer event, broad host ranges and phylogenetic ubiquity amongst prokaryotes, with the subclass of chromosomally inserted integrative and conjugative elements (ICEs) being particularly abundant. The focus on a small number of model systems has biased the study of ICEs towards those conferring readily selectable phenotypes to host cells, whereas the identification and characterization of integrated cryptic elements remains challenging. Even though antimicrobial resistance and horizontally acquired virulence genes are major factors aggravating neisserial infection, conjugative MGEs of Neisseria gonorrhoeae and Neisseria meningitidis remain poorly characterized. Using a phenotype-independent approach based on atypical distributions of DNA uptake sequences (DUSs) in MGEs relative to the chromosomal background, we have identified two groups of chromosomally integrated conjugative elements in Neisseria: one found almost exclusively in pathogenic species possibly deriving from the genus Kingella, the other belonging to a group of Neisseria mucosa-like commensals. The former element appears to enable transfer of traditionally gonococcal-specific loci such as the virulence-associated toxin-antitoxin system fitAB to N. meningitidis chromosomes, whilst the circular form of the latter possesses a unique attachment site (attP) sequence seemingly adapted to exploit DUS motifs as chromosomal integration sites. In addition to validating the use of DUS distributions in Neisseriaceae MGE identification, the >170 identified ICE sequences provide a valuable resource for future studies of ICE evolution and host adaptation.


Assuntos
Cromossomos Bacterianos/genética , Elementos de DNA Transponíveis , Neisseriaceae/classificação , Plasmídeos/genética , Análise de Sequência de DNA/métodos , Conjugação Genética , Farmacorresistência Bacteriana Múltipla , Evolução Molecular , Transferência Genética Horizontal , Sequenciamento de Nucleotídeos em Larga Escala , Neisseriaceae/genética , Neisseriaceae/isolamento & purificação , Neisseriaceae/patogenicidade , Fenótipo , Filogenia , Simbiose , Fatores de Virulência/genética , Vocabulário
7.
Science ; 367(6477): 573-576, 2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-32001655

RESUMO

Honey bees are essential pollinators threatened by colony losses linked to the spread of parasites and pathogens. Here, we report a new approach for manipulating bee gene expression and protecting bee health. We engineered a symbiotic bee gut bacterium, Snodgrassella alvi, to induce eukaryotic RNA interference (RNAi) immune responses. We show that engineered S. alvi can stably recolonize bees and produce double-stranded RNA to activate RNAi and repress host gene expression, thereby altering bee physiology, behavior, and growth. We used this approach to improve bee survival after a viral challenge, and we show that engineered S. alvi can kill parasitic Varroa mites by triggering the mite RNAi response. This symbiont-mediated RNAi approach is a tool for studying bee functional genomics and potentially for safeguarding bee health.


Assuntos
Abelhas/imunologia , Abelhas/microbiologia , Microbioma Gastrointestinal/genética , Neisseriaceae/genética , Neisseriaceae/fisiologia , Varroidae/microbiologia , Animais , Engenharia Genética , Interferência de RNA , Simbiose
8.
Nat Commun ; 10(1): 5809, 2019 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-31863068

RESUMO

Transmissible plasmids spread genes encoding antibiotic resistance and other traits to new bacterial species. Here we report that laboratory populations of Escherichia coli with a newly acquired IncQ plasmid often evolve 'satellite plasmids' with deletions of accessory genes and genes required for plasmid replication. Satellite plasmids are molecular parasites: their presence reduces the copy number of the full-length plasmid on which they rely for their continued replication. Cells with satellite plasmids gain an immediate fitness advantage from reducing burdensome expression of accessory genes. Yet, they maintain copies of these genes and the complete plasmid, which potentially enables them to benefit from and transmit the traits they encode in the future. Evolution of satellite plasmids is transient. Cells that entirely lose accessory gene function or plasmid mobility dominate in the long run. Satellite plasmids also evolve in Snodgrassella alvi colonizing the honey bee gut, suggesting that this mechanism may broadly contribute to the importance of IncQ plasmids as agents of bacterial gene transfer in nature.


Assuntos
Escherichia coli/genética , Evolução Molecular , Transferência Genética Horizontal , Neisseriaceae/genética , Plasmídeos/genética , Animais , Abelhas/microbiologia , Replicação do DNA , Microbioma Gastrointestinal/genética
9.
Proc Natl Acad Sci U S A ; 116(51): 25909-25916, 2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31776248

RESUMO

Bees acquire carbohydrates from nectar and lipids; and amino acids from pollen, which also contains polysaccharides including cellulose, hemicellulose, and pectin. These potential energy sources could be degraded and fermented through microbial enzymatic activity, resulting in short chain fatty acids available to hosts. However, the contributions of individual microbiota members to polysaccharide digestion have remained unclear. Through analysis of bacterial isolate genomes and a metagenome of the honey bee gut microbiota, we identify that Bifidobacterium and Gilliamella are the principal degraders of hemicellulose and pectin. Both Bifidobacterium and Gilliamella show extensive strain-level diversity in gene repertoires linked to polysaccharide digestion. Strains from honey bees possess more such genes than strains from bumble bees. In Bifidobacterium, genes encoding carbohydrate-active enzymes are colocated within loci devoted to polysaccharide utilization, as in Bacteroides from the human gut. Carbohydrate-active enzyme-encoding gene expressions are up-regulated in response to particular hemicelluloses both in vitro and in vivo. Metabolomic analyses document that bees experimentally colonized by different strains generate distinctive gut metabolomic profiles, with enrichment for specific monosaccharides, corresponding to predictions from genomic data. The other 3 core gut species clusters (Snodgrassella and 2 Lactobacillus clusters) possess few or no genes for polysaccharide digestion. Together, these findings indicate that strain composition within individual hosts determines the metabolic capabilities and potentially affects host nutrition. Furthermore, the niche specialization revealed by our study may promote overall community stability in the gut microbiomes of bees.


Assuntos
Abelhas/microbiologia , Abelhas/fisiologia , Digestão , Microbioma Gastrointestinal/fisiologia , Plantas/química , Polissacarídeos/metabolismo , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Bifidobacterium/genética , Bifidobacterium/metabolismo , Gammaproteobacteria/genética , Gammaproteobacteria/metabolismo , Microbioma Gastrointestinal/genética , Trato Gastrointestinal/microbiologia , Regulação da Expressão Gênica , Genoma Bacteriano , Lactobacillus/genética , Metagenoma , Microbiota , Neisseriaceae/genética , Pólen/química
10.
Evolution ; 73(11): 2333-2342, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31584186

RESUMO

There has been a proliferation of studies demonstrating an organism's health is influenced by its microbiota. However, factors influencing beneficial microbe colonization and the evolution of these relationships remain understudied relative to host-pathogen interactions. Vertically transmitted beneficial microbes are predicted to show high levels of specificity in colonization, including genotype matching, which may transpire through coevolution. We investigate how host and bacterial genotypes influence colonization of a core coevolved microbiota member in bumble bees. The hindgut colonizing Snodgrassella alvi confers direct benefits, but, as an early colonizer, also facilitates the further development of a healthy microbiota. Due to predominantly vertical transmission promoting tight evolution between colonization factors of bacteria and host lineages, we predict that genotype-by-genotype interactions will determine successful colonization. Germ-free adult bees from seven bumble bee colonies (host genotypic units) were inoculated with one of six genetically distinct strains of S. alvi. Subsequent colonization within host and microbe genotypes combinations ranged from 0 to 100%, and an interaction between host and microbe genotypes determined colonization success. This novel finding of a genotype-by-genotype interaction determining colonization in an animal host-beneficial microbe system has implications for the ecological and evolutionary dynamics of host and microbe, including associated host-fitness benefits.


Assuntos
Abelhas/genética , Interações Hospedeiro-Patógeno/genética , Neisseriaceae/genética , Animais , Abelhas/microbiologia , Coevolução Biológica , Microbioma Gastrointestinal , Genótipo , Neisseriaceae/patogenicidade
11.
Microb Ecol ; 76(3): 588-591, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29520453

RESUMO

Use of antibiotics in medicine and farming contributes to increasing numbers of antibiotic-resistant bacteria in diverse environments. The ability of antibiotic resistance genes (ARG) to transfer between bacteria genera contributes to this spread. It is difficult to directly link antibiotic exposure to the spread of ARG in a natural environment where environmental settings and study populations cannot be fully controlled. We used managed honeybees in environments with contrasting streptomycin exposure (USA: high exposure, Norway: low exposure) and mapped the prevalence and spread of transferrable streptomycin resistance genes. We found a high prevalence of strA-strB genes in the USA compared to Norway with 17/90 and 1/90 positive samples, respectively (p < 0.00007). We identified strA-strB genes on a transferrable transposon Tn5393 in the honeybee gut symbiont Snodgrassella alvi. Such transfer of resistance genes increases the risk of the spread to new environments as honeybees are moved to new pollination sites.


Assuntos
Abelhas/microbiologia , Farmacorresistência Bacteriana , Neisseriaceae/efeitos dos fármacos , Neisseriaceae/isolamento & purificação , Estreptomicina/farmacologia , Animais , Abelhas/fisiologia , Elementos de DNA Transponíveis , Feminino , Trato Gastrointestinal/microbiologia , Masculino , Neisseriaceae/genética , Neisseriaceae/fisiologia , Simbiose
12.
FEMS Microbiol Ecol ; 94(2)2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29267902

RESUMO

Groundwater reservoirs constitute important freshwater resources. However, these ecosystems are highly vulnerable to contamination and have to rely on the resident microbiota to attenuate the impact of this contamination. Nitrate is one of the main contaminants found in groundwater, and denitrification is the main process that removes the compound. In this study, the response to nutrient load on indigenous microbial communities in groundwater from a low impacted aquifer in Uruguay was evaluated. Denitrification rates were measured in groundwater samples from three different sites with nitrate, acetate and pyrite amendments. Results showed that denitrification is feasible under in situ nitrate and electron donor concentrations, although the lack of readily available organic energy source would limit the attenuation of higher nitrate concentrations. DNA-stable isotope probing, combined with amplicon sequencing of 16S rRNA, nirS and nirK genes, was used to identify the active denitrifiers. Members of the phylum Betaproteobacteria were the dominant denitrifiers in two of three sites, with different families being observed; members of the genus Vogesella (Neisseriaceae) were key denitrifiers at one site, while the genera Dechloromonas (Rhodocyclaceae) and Comamonas (Comamonadaceae) were the main denitrifiers detected at the other sites.


Assuntos
Comamonadaceae/metabolismo , Desnitrificação/fisiologia , Água Subterrânea/química , Água Subterrânea/microbiologia , Neisseriaceae/metabolismo , Nitratos/análise , Nitratos/metabolismo , Rhodocyclaceae/metabolismo , Acetatos/metabolismo , Comamonadaceae/classificação , Comamonadaceae/genética , DNA , Sondas de DNA , Ferro/metabolismo , Marcação por Isótopo , Isótopos , Neisseriaceae/classificação , Neisseriaceae/genética , RNA Ribossômico 16S/genética , Rhodocyclaceae/classificação , Rhodocyclaceae/genética , Sulfetos/metabolismo , Uruguai
13.
Mol Ecol ; 27(8): 2057-2066, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29164717

RESUMO

The gut microbiome plays a key role in animal health, and perturbing it can have detrimental effects. One major source of perturbation to microbiomes, in humans and human-associated animals, is exposure to antibiotics. Most studies of how antibiotics affect the microbiome have used amplicon sequencing of highly conserved 16S rRNA sequences, as in a recent study showing that antibiotic treatment severely alters the species-level composition of the honeybee gut microbiome. But because the standard 16S rRNA-based methods cannot resolve closely related strains, strain-level changes could not be evaluated. To address this gap, we used amplicon sequencing of protein-coding genes to assess effects of antibiotics on fine-scale genetic diversity of the honeybee gut microbiota. We followed the population dynamics of alleles within two dominant core species of the bee gut community, Gilliamella apicola and Snodgrassella alvi, following antibiotic perturbation. Whereas we observed a large reduction in genetic diversity in G. apicola, S. alvi diversity was mostly unaffected. The reduction in G. apicola diversity accompanied an increase in the frequency of several alleles, suggesting resistance to antibiotic treatment. We find that antibiotic perturbation can cause major shifts in diversity and that the extent of these shifts can vary substantially across species. Thus, antibiotics impact not only species composition, but also allelic diversity within species, potentially affecting hosts if variants with particular functions are reduced or eliminated. Overall, we show that amplicon sequencing of protein-coding genes, without clustering into operational taxonomic units, provides an accurate picture of the fine-scale dynamics of microbial communities over time.


Assuntos
Abelhas/microbiologia , Microbioma Gastrointestinal/genética , Neisseriaceae/genética , Simbiose/genética , Animais , Antibacterianos/farmacologia , Abelhas/efeitos dos fármacos , Abelhas/genética , Microbioma Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/microbiologia , Variação Genética/genética , Interações entre Hospedeiro e Microrganismos/efeitos dos fármacos , Interações entre Hospedeiro e Microrganismos/genética , Neisseriaceae/efeitos dos fármacos , Filogenia , RNA Ribossômico 16S/genética
14.
J Antimicrob Chemother ; 73(3): 643-647, 2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-29244123

RESUMO

Background: Laribacter hongkongensis is a facultative anaerobic, non-fermentative, Gram-negative bacillus associated with community-acquired gastroenteritis and traveller's diarrhoea. No clinical MDR L. hongkongensis isolate has been reported yet. Methods: We performed WGS (PacBio and Illumina) on a clinical L. hongkongensis strain HLGZ1 with an MDR phenotype. Results: HLGZ1 was resistant to eight classes of commonly used antibiotics. Its complete genome was a single circular chromosome of 3 424 272 bp with a G + C content of 62.29%. In comparison with the reference strain HLHK9, HLGZ1 had a higher abundance of genes associated with DNA metabolism and recombination. Several inserts including two acquired resistance gene clusters (RC1 and RC2) were also identified. RC1 carried two resistance gene cassette arrays, aac(6')-Ib-cr-aadA2-Δqac-Δsul1-floR-tetR-tetG and arr-3-dfrA32-ereA2-Δqac-sul1, which shared significant nucleotide sequence identities with the MDR region of Salmonella Genomic Island 1 from Salmonella enterica serovar Typhimurium DT104. There was also an integron-like structure, intl1-arr3-dfrA27-Δqac-sul1-aph(3')-Ic, and a tetR-tetA operon located on RC2. MLST analysis identified HLGZ1 as ST167, a novel ST clustered with two strains previously isolated from frogs. Conclusions: This study provides insight into the genomic characteristics of MDR L. hongkongensis and highlights the possibilities of horizontal resistance gene transfer in this bacterium with other pathogens.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Genoma Bacteriano , Neisseriaceae/genética , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , China/epidemiologia , DNA Bacteriano/genética , Fezes/microbiologia , Gastroenterite/microbiologia , Transferência Genética Horizontal , Ilhas Genômicas , Genômica , Infecções por Bactérias Gram-Negativas/epidemiologia , Humanos , Integrons , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Neisseriaceae/efeitos dos fármacos , Neisseriaceae/isolamento & purificação , Salmonella enterica/genética , Sequenciamento Completo do Genoma
15.
mBio ; 8(6)2017 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-29233893

RESUMO

Microbial communities are shaped by interactions among their constituent members. Some Gram-negative bacteria employ type VI secretion systems (T6SSs) to inject protein toxins into neighboring cells. These interactions have been theorized to affect the composition of host-associated microbiomes, but the role of T6SSs in the evolution of gut communities is not well understood. We report the discovery of two T6SSs and numerous T6SS-associated Rhs toxins within the gut bacteria of honey bees and bumble bees. We sequenced the genomes of 28 strains of Snodgrassella alvi, a characteristic bee gut microbe, and found tremendous variability in their Rhs toxin complements: altogether, these strains appear to encode hundreds of unique toxins. Some toxins are shared with Gilliamella apicola, a coresident gut symbiont, implicating horizontal gene transfer as a source of toxin diversity in the bee gut. We use data from a transposon mutagenesis screen to identify toxins with antibacterial function in the bee gut and validate the function and specificity of a subset of these toxin and immunity genes in Escherichia coli Using transcriptome sequencing, we demonstrate that S. alvi T6SSs and associated toxins are upregulated in the gut environment. We find that S. alvi Rhs loci have a conserved architecture, consistent with the C-terminal displacement model of toxin diversification, with Rhs toxins, toxin fragments, and cognate immunity genes that are expressed and confer strong fitness effects in vivo Our findings of T6SS activity and Rhs toxin diversity suggest that T6SS-mediated competition may be an important driver of coevolution within the bee gut microbiota.IMPORTANCE The structure and composition of host-associated bacterial communities are of broad interest, because these communities affect host health. Bees have a simple, conserved gut microbiota, which provides an opportunity to explore interactions between species that have coevolved within their host over millions of years. This study examined the role of type VI secretion systems (T6SSs)-protein complexes used to deliver toxic proteins into bacterial competitors-within the bee gut microbiota. We identified two T6SSs and diverse T6SS-associated toxins in bacterial strains from bees. Expression of these genes is increased in bacteria in the bee gut, and toxin and immunity genes demonstrate antibacterial and protective functions, respectively, when expressed in Escherichia coli Our results suggest that coevolution among bacterial species in the bee gut has favored toxin diversification and maintenance of T6SS machinery, and demonstrate the importance of antagonistic interactions within host-associated microbial communities.


Assuntos
Antibiose , Toxinas Bacterianas/genética , Abelhas/microbiologia , Microbioma Gastrointestinal/fisiologia , Sistemas de Secreção Tipo VI/genética , Animais , Toxinas Bacterianas/isolamento & purificação , Toxinas Bacterianas/farmacologia , Escherichia coli/genética , Evolução Molecular , Gammaproteobacteria/classificação , Gammaproteobacteria/genética , Gammaproteobacteria/isolamento & purificação , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/genética , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Família Multigênica , Mutagênese , Neisseriaceae/classificação , Neisseriaceae/genética , Neisseriaceae/fisiologia , Simbiose , Sistemas de Secreção Tipo VI/classificação , Sistemas de Secreção Tipo VI/metabolismo , Sistemas de Secreção Tipo VI/toxicidade , Regulação para Cima
16.
Mol Ecol ; 26(23): 6590-6607, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29087008

RESUMO

How long-term antibiotic treatment affects host bacterial associations is still largely unknown. The honeybee-gut microbiota has a simple composition, so we used this gut community to investigate how long-term antibiotic treatment affects host-associated microbiota. We investigated the phylogenetic relatedness, genomic content (GC percentage, genome size, number of genes and CRISPR) and antibiotic-resistant genes (ARG) for strains from two abundant members of the honeybee core gut microbiota (Gilliamella apicola and Snodgrassella alvi). Domesticated honeybees are subjected to geographically different management policies, so we used two research apiaries, representing different antibiotic treatment regimens in their apiculture: low antibiotic usage (Norway) and high antibiotic usage (Arizona, USA). We applied whole-genome shotgun sequencing on 48 G. apicola and 22 S. alvi. We identified three predominating subgroups of G. apicola in honeybees from both Norway and Arizona. For G. apicola, genetic content substantially varied between subgroups and distance similarity calculations showed similarity discrepancy between subgroups. Functional differences between subgroups, such as pectin-degrading enzymes (G. apicola), were also identified. In addition, we identified horizontal gene transfer (HGT) of transposon (Tn10)-associated tetracycline resistance (Tet B) across the G. apicola subgroups in the Arizonan honeybees, using interspace polymorphisms in the Tet B determinant. Our results support that honeybee-gut symbiont subgroups can resist long-term antibiotic treatment and maintain functionality through acquisition of geographically distinct antibiotic-resistant genes by HGT.


Assuntos
Abelhas/microbiologia , Farmacorresistência Bacteriana/genética , Gammaproteobacteria/genética , Neisseriaceae/genética , Animais , Arizona , Composição de Bases , DNA Bacteriano/genética , Trato Gastrointestinal/microbiologia , Transferência Genética Horizontal , Genes Bacterianos , Tamanho do Genoma , Geografia , Microbiota , Noruega , Filogenia , Simbiose
17.
Int J Syst Evol Microbiol ; 67(7): 2274-2278, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28671534

RESUMO

Two chitin-degrading halophilic archaeal strains, MC-74T and MC-23, were isolated from commercial salt samples. Cells were motile, rod-shaped and stained Gram-negative. Colonies were vermillion-pigmented. Strains MC-74T and MC-23 were able to grow with 1.5-5.1 M NaCl (optimum, 2.6-3.1 M) at pH 6.0-10.0 (optimum, pH 7.0) and at 20-50 °C (optimum, 40 °C). The orthologous 16S rRNA gene sequence similarity between the two strains was 99.8 %, and the closest phylogenetic relative was Salinarchaeum laminariae JCM 17267T with 99.3-99.5 % similarity. The level of DNA-DNA relatedness between the two strains was 93 and 94 % (reciprocally), and those between the two strains and Salinarchaeumlaminariae JCM 17267T were 35-36 % and 38-39 % (reciprocally). The polar lipids of both strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate. Glycolipids were not detected. Based on the phenotypic and phylogenetic analyses, the strains represent a novel species of the genus Salinarchaeum, for which the name Salinarchaeum chitinilyticum sp. nov. is proposed. The type strain is MC-74T (=JCM 19597T=KCTC 4262T), isolated from solar salt produced in France. Strain MC-23, isolated from a commercial solar salt sample produced in China, is an additional strain of the species.


Assuntos
Halobacteriaceae/classificação , Filogenia , Cloreto de Sódio/análise , China , Quitina/metabolismo , DNA Arqueal/genética , França , Glicolipídeos/química , Halobacteriaceae/genética , Halobacteriaceae/isolamento & purificação , Neisseriaceae/genética , Hibridização de Ácido Nucleico , Fosfatidilgliceróis/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
18.
PLoS One ; 12(4): e0174506, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28406911

RESUMO

Two Gram-stain negative aerobic bacterial strains were isolated from the bark tissue of Populus × euramericana. The novel isolates were investigated using a polyphasic approach including 16S rRNA gene sequencing, genome sequencing, average nucleotide identity (ANI) and both phenotypic and chemotaxonomic assays. The genome core gene sequence and 16S rRNA gene phylogenies suggest that the novel isolates are different from the genera Snodgrassella and Stenoxybacter. Additionally, the ANI, G+C content, main fatty acids and phospholipid profile data supported the distinctiveness of the novel strain from genus Snodgrassella. Therefore, based on the data presented, the strains constitute a novel species of a novel genus within the family Neisseriaceae, for which the name Populibacter corticis gen. nov., sp. nov. is proposed. The type strain is 15-3-5T (= CFCC 13594T = KCTC 42251T).


Assuntos
Genoma Bacteriano/fisiologia , Neisseriaceae/genética , Filogenia , Casca de Planta/microbiologia , Populus/microbiologia , Neisseriaceae/isolamento & purificação
19.
PLoS One ; 12(1): e0169998, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28085929

RESUMO

Bacterial adaptation to different hosts requires transcriptomic alteration in response to the environmental conditions. Laribacter hongkongensis is a gram-negative, facultative anaerobic, urease-positive bacillus caused infections in liver cirrhosis patients and community-acquired gastroenteritis. It was also found in intestine from commonly consumed freshwater fishes and drinking water reservoirs. Since L. hongkongensis could survive as either fish or human pathogens, their survival mechanisms in two different habitats should be temperature-regulated and highly complex. Therefore, we performed transcriptomic analysis of L. hongkongensis at body temperatures of fish and human in order to elucidate the versatile adaptation mechanisms coupled with the temperatures. We identified numerous novel temperature-induced pathways involved in host pathogenesis, in addition to the shift of metabolic equilibriums and overexpression of stress-related proteins. Moreover, these pathways form a network that can be activated at a particular temperature, and change the physiology of the bacteria to adapt to the environments. In summary, the dynamic of transcriptomes in L. hongkongensis provides versatile strategies for the bacterial survival at different habitats and this alteration prepares the bacterium for the challenge of host immunity.


Assuntos
Adaptação Fisiológica/genética , Biomarcadores/metabolismo , Gastroenterite/genética , Genoma Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Neisseriaceae/genética , Gastroenterite/microbiologia , Humanos , Neisseriaceae/isolamento & purificação , RNA Bacteriano/genética , Estresse Fisiológico , Temperatura
20.
Antonie Van Leeuwenhoek ; 110(3): 415-428, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27915412

RESUMO

We investigated previously under explored cold aquatic environments of Andean Patagonia, Argentina. Oily sheens similar to an oil spill are frequently observed at the surface of water in creeks and small ponds in these places. Chemical analysis of a water sample revealed the occurrence of high concentrations of iron and the presence of a free insoluble indigoidine-derived pigment. A blue pigment-producing bacterium (strain EB) was isolated from the water sample and identified as Vogesella sp. by molecular analysis. The isolate was able to produce indigoidine and another derived-pigment (here called cryoindigoidine) with strong antifreeze properties. The production of the pigments depended on the cell growth at cold temperatures (below 15 °C), as well as on the attachment of cells to solid surfaces, and iron limitation in the media. The pigments produced by strain EB showed an inhibitory effect on the growth of diverse microorganisms such as Candida albicans, Escherichia coli and Staphylococcus aureus. In addition, pigmented cells were more tolerant to freezing than non-pigmented cells, suggesting a role of cryoindigoidine/indigoidine as a cold-protectant molecule. The possible roles of the pigments in strain EB physiology and its interactions with the iron-rich environment from which the isolate was obtained are discussed. Results of this study suggested an active role of strain EB in the investigated iron-oxidizing ecosystem.


Assuntos
Ferro/metabolismo , Neisseriaceae/fisiologia , Pigmentos Biológicos/metabolismo , Piperidonas/metabolismo , Aclimatação , Argentina , Candida albicans/efeitos dos fármacos , Temperatura Baixa , Microbiologia Ambiental , Escherichia coli/efeitos dos fármacos , Congelamento , Água Doce/química , Água Doce/microbiologia , Neisseriaceae/genética , Neisseriaceae/isolamento & purificação , Neisseriaceae/metabolismo , Filogenia , Pigmentos Biológicos/química , Piperidonas/química , Lagoas/química , Lagoas/microbiologia , RNA Ribossômico 16S/genética , Staphylococcus aureus/efeitos dos fármacos , Microbiologia da Água
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