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1.
Nucleic Acids Res ; 50(D1): D273-D278, 2022 01 07.
Article in English | MEDLINE | ID: mdl-34850116

ABSTRACT

Plasmids are known to contain genes encoding for virulence factors and antibiotic resistance mechanisms. Their relevance in metagenomic data processing is steadily growing. However, with the increasing popularity and scale of metagenomics experiments, the number of reported plasmids is rapidly growing as well, amassing a considerable number of false positives due to undetected misassembles. Here, our previously published database PLSDB provides a reliable resource for researchers to quickly compare their sequences against selected and annotated previous findings. Within two years, the size of this resource has more than doubled from the initial 13,789 to now 34,513 entries over the course of eight regular data updates. For this update, we aggregated community feedback for major changes to the database featuring new analysis functionality as well as performance, quality, and accessibility improvements. New filtering steps, annotations, and preprocessing of existing records improve the quality of the provided data. Additionally, new features implemented in the web-server ease user interaction and allow for a deeper understanding of custom uploaded sequences, by visualizing similarity information. Lastly, an application programming interface was implemented along with a python library, to allow remote database queries in automated workflows. The latest release of PLSDB is freely accessible under https://www.ccb.uni-saarland.de/plsdb.


Subject(s)
Bacteria/genetics , Databases, Genetic , Plasmids/chemistry , User-Computer Interface , Actinobacteria/genetics , Actinobacteria/pathogenicity , Bacteria/classification , Bacteria/pathogenicity , Bacteroidetes/genetics , Bacteroidetes/pathogenicity , Drug Resistance, Microbial/genetics , Firmicutes/genetics , Firmicutes/pathogenicity , Internet , Metagenomics/methods , Molecular Sequence Annotation , Plasmids/classification , Plasmids/metabolism , Proteobacteria/genetics , Proteobacteria/pathogenicity , Spirochaetales/genetics , Spirochaetales/pathogenicity , Tenericutes/genetics , Tenericutes/pathogenicity , Virulence/genetics
2.
Cell Rep ; 36(13): 109765, 2021 09 28.
Article in English | MEDLINE | ID: mdl-34592155

ABSTRACT

Gut microbial diversity decreases with aging, but existing studies have used stool samples, which do not represent the entire gut. We analyzed the duodenal microbiome in 251 subjects aged 18-35 (n = 32), 36-50 (n = 41), 51-65 (n = 96), and 66-80 (n = 82). Decreased duodenal microbial diversity in older subjects is associated with combinations of chronological age, number of concomitant diseases, and number of medications used, and also correlated with increasing coliform numbers (p < 0.0001). Relative abundance (RA) of phylum Proteobacteria increases in older subjects, with increased RA of family Enterobacteriaceae and coliform genera Escherichia and Klebsiella, and is associated with alterations in the RA of other duodenal microbial taxa and decreased microbial diversity. Increased RA of specific genera are associated with chronological age only (Escherichia, Lactobacillus, and Enterococcus), number of medications only (Klebsiella), or number of concomitant diseases only (Clostridium and Bilophila). These findings indicate the small intestinal microbiome changes significantly with age and the aging process.


Subject(s)
Aging/physiology , Gastrointestinal Microbiome/physiology , Intestine, Small/microbiology , Lactobacillus/pathogenicity , Duodenum/microbiology , Feces/microbiology , Humans , Proteobacteria/pathogenicity
3.
Sci Rep ; 11(1): 18364, 2021 09 15.
Article in English | MEDLINE | ID: mdl-34526632

ABSTRACT

Soda lakes are saline and alkaline ecosystems that are considered to have existed since the first geological records of the world. These lakes support the growth of ecologically and economically important microorganisms due to their unique geochemistry. Microbiota members of lakes are valuable models to study the link between community structure and abiotic parameters such as pH and salinity. Lake Van is the largest endroheic lake and in this study, bacterial diversity of lake water, sediment, and pearl mullet (inci kefali; Alburnus tarichi), an endemic species of fish which are collected from different points of the lake, are studied directly and investigated meticulously using a metabarcoding approach after pre-enrichment. Bacterial community structures were identified using Next Generation Sequencing of the 16S rRNA gene. The analysis revealed that the samples of Lake Van contain high level of bacterial diversity. Direct water samples were dominated by Proteobacteria, Cyanobacteria, and Bacteroidota, on the other hand, pre-enriched water samples were dominated by Proteobacteria and Firmicutes at phylum-level. In direct sediment samples Proteobacteria, whereas in pre-enriched sediment samples Firmicutes and Proteobacteria were determined at highest level. Pre-enriched fish samples were dominated by Proteobacteria and Firmicutes at phylum-level. In this study, microbiota members of Lake Van were identified by taxonomic analysis.


Subject(s)
Lakes/microbiology , Microbiota , Animals , Firmicutes/genetics , Firmicutes/isolation & purification , Firmicutes/pathogenicity , Fishes/microbiology , Geologic Sediments/microbiology , Proteobacteria/genetics , Proteobacteria/isolation & purification , Proteobacteria/pathogenicity , RNA, Ribosomal, 16S/genetics
4.
J Mol Neurosci ; 71(10): 2095-2106, 2021 Oct.
Article in English | MEDLINE | ID: mdl-33954858

ABSTRACT

Gut microbiome and plasma metabolome serve a role in the pathogenesis of ischemic stroke (IS). However, the relationship between the microbiota and metabolites remains unclear. This study aimed to reveal the specific asso-ciation between the microbiota and the metabolites in IS using integrated 16S rRNA gene sequencing and liquid chromatography-mass spectrometry (LC-MS) analysis. Male Sprague Dawley (SD) rats were divided into three groups: normal group (n = 8, Normal), model group (n = 9, IS), and sham-operated group (n = 8, Sham). Rats in the IS group were induced by middle cerebral artery occlusion (MCAO), and rats in the Sham group received an initial anesthesia and neck incision only. A neurological function test and 2,3,5-triphenyltetrazolium chloride (TTC) staining were used to assess the IS rat model. Then, the plasma samples were analyzed using untargeted LC-MS. The cecum samples were collected and analyzed using 16S rRNA sequencing. Pearson correlation analysis was performed to explore the association between the gut microbiota and the plasma metabolites. The 16S rRNA sequencing showed that the composition and diversity of the microbiota in the IS and control rats were significantly different. Compared with the Sham group, the abundance of the Firmicutes phylum was decreased, whereas Proteobacteria and Deferribacteres were increased in the IS group. Ruminococcus_sp_15975 and Lachnospiraceae_UCG_001 might be considered as biomarkers for the IS and Sham groups, respectively. LC-MS analysis revealed that many metabolites, such as L-leucine, L-valine, and L-phenylalanine, displayed different patterns between the IS and Sham groups. Pathway analysis indicated that these metabolites were mainly involved in mineral absorption and cholinergic synapse. Furthermore, integrated analysis correlated IS-related microbes with metabolites. For example, Proteobacteria were positively correlated with L-phenylalanine, while they were negatively correlated with eicosapentaenoic acid (EPA). Our results provided evidence of the relationship between the gut microbiome and plasma metabolome in IS, suggesting that these microflora-related metabolites might serve as potential diagnostic and therapeutic markers.


Subject(s)
Gastrointestinal Microbiome , Infarction, Middle Cerebral Artery/metabolism , Metabolome , Animals , Infarction, Middle Cerebral Artery/blood , Infarction, Middle Cerebral Artery/microbiology , Male , Proteobacteria/genetics , Proteobacteria/isolation & purification , Proteobacteria/pathogenicity , RNA, Ribosomal, 16S/genetics , Rats , Rats, Sprague-Dawley
5.
Sci Rep ; 10(1): 6068, 2020 04 08.
Article in English | MEDLINE | ID: mdl-32269258

ABSTRACT

Plant microbiota colonize all organs of a plant and play crucial roles including supplying nutrients to plants, stimulating seed germination, promoting plant growth, and defending plants against biotic and abiotic stress. Because of the economic importance, interactions between citrus and microbes have been studied relatively extensively, especially citrus-pathogen interactions. However, the spatial distribution of microbial taxa in citrus trees remains under-studied. In this study, Citrus reticulata cv. Chachiensis was examined for the spatial distribution of microbes by sequencing 16S rRNA genes. More than 2.5 million sequences were obtained from 60 samples collected from soil, roots, leaves, and phloem. The dominant microbial phyla from all samples were Proteobacteria, Actinobacteria and Acidobacteria. The composition and structure of microbial communities in different samples were analyzed by PCoA, CAP, Anosim and MRPP methods. Variation in microbial species between samples were analyzed and the indicator microbes of each sample group were identified. Our results suggested that the microbial communities from different tissues varied significantly and the microenvironments of tree tissues could affect the composition of its microbial community.


Subject(s)
Citrus/microbiology , Microbiota , Acidobacteria/genetics , Acidobacteria/pathogenicity , Actinobacteria/genetics , Actinobacteria/pathogenicity , Phloem/microbiology , Plant Leaves/microbiology , Plant Roots/microbiology , Proteobacteria/genetics , Proteobacteria/pathogenicity , RNA, Ribosomal, 16S/genetics , Soil Microbiology
6.
Microbiome ; 8(1): 25, 2020 02 24.
Article in English | MEDLINE | ID: mdl-32093774

ABSTRACT

BACKGROUND: The holistic view of bacterial symbiosis, incorporating both host and microbial environment, constitutes a major conceptual shift in studies deciphering host-microbe interactions. Interactions between Steinernema entomopathogenic nematodes and their bacterial symbionts, Xenorhabdus, have long been considered monoxenic two partner associations responsible for the killing of the insects and therefore widely used in insect pest biocontrol. We investigated this "monoxenic paradigm" by profiling the microbiota of infective juveniles (IJs), the soil-dwelling form responsible for transmitting Steinernema-Xenorhabdus between insect hosts in the parasitic lifecycle. RESULTS: Multigenic metabarcoding (16S and rpoB markers) showed that the bacterial community associated with laboratory-reared IJs from Steinernema carpocapsae, S. feltiae, S. glaseri and S. weiseri species consisted of several Proteobacteria. The association with Xenorhabdus was never monoxenic. We showed that the laboratory-reared IJs of S. carpocapsae bore a bacterial community composed of the core symbiont (Xenorhabdus nematophila) together with a frequently associated microbiota (FAM) consisting of about a dozen of Proteobacteria (Pseudomonas, Stenotrophomonas, Alcaligenes, Achromobacter, Pseudochrobactrum, Ochrobactrum, Brevundimonas, Deftia, etc.). We validated this set of bacteria by metabarcoding analysis on freshly sampled IJs from natural conditions. We isolated diverse bacterial taxa, validating the profile of the Steinernema FAM. We explored the functions of the FAM members potentially involved in the parasitic lifecycle of Steinernema. Two species, Pseudomonas protegens and P. chlororaphis, displayed entomopathogenic properties suggestive of a role in Steinernema virulence and membership of the Steinernema pathobiome. CONCLUSIONS: Our study validates a shift from monoxenic paradigm to pathobiome view in the case of the Steinernema ecology. The microbial communities of low complexity associated with EPNs will permit future microbiota manipulation experiments to decipher overall microbiota functioning in the infectious process triggered by EPN in insects and, more generally, in EPN ecology.


Subject(s)
Host Microbial Interactions , Microbiota , Proteobacteria/classification , Proteobacteria/pathogenicity , Rhabditida/microbiology , Symbiosis , Animals , Biological Control Agents , DNA Barcoding, Taxonomic , Larva/parasitology , Life Cycle Stages , Moths/parasitology , Rhabditida/physiology , Rhabditida Infections/parasitology , Virulence
7.
Nutrients ; 11(12)2019 Dec 07.
Article in English | MEDLINE | ID: mdl-31817895

ABSTRACT

Critically ill patients have an alteration in the microbiome in which it becomes a disease-promoting pathobiome. It is characterized by lower bacterial diversity, loss of commensal phyla, like Firmicutes and Bacteroidetes, and a domination of pathogens belonging to the Proteobacteria phylum. Although these alterations are multicausal, many of the treatments administered to these patients, like antibiotics, play a significant role. Critically ill patients also have a hyperpermeable gut barrier and dysregulation of the inflammatory response that favor the development of the pathobiome, translocation of pathogens, and facilitate the emergence of sepsis. In order to restore the homeostasis of the microbiome, several nutritional strategies have been evaluated with the aim to improve the management of critically ill patients. Importantly, enteral nutrition has proven to be more efficient in promoting the homeostasis of the gut microbiome compared to parenteral nutrition. Several nutritional therapies, including prebiotics, probiotics, synbiotics, and fecal microbiota transplantation, are currently being used, showing variable results, possibly due to the unevenness of clinical trial conditions and the fact that the beneficial effects of probiotics are specific to particular species or even strains. Thus, it is of great importance to better understand the mechanisms by which nutrition and supplement therapies can heal the microbiome in critically ill patients in order to finally implement them in clinical practice with optimal safety and efficacy.


Subject(s)
Critical Care/methods , Critical Illness/therapy , Gastrointestinal Microbiome , Nutritional Support , Bacteroidetes/physiology , Dysbiosis/microbiology , Fecal Microbiota Transplantation , Firmicutes/physiology , Homeostasis , Host Microbial Interactions , Humans , Prebiotics/administration & dosage , Probiotics/administration & dosage , Proteobacteria/pathogenicity , Synbiotics/administration & dosage
8.
Sci Rep ; 9(1): 14883, 2019 10 16.
Article in English | MEDLINE | ID: mdl-31619759

ABSTRACT

Control of common scab disease can be reached by resistant cultivars or suppressive soils. Both mechanisms are likely to translate into particular potato microbiome profiles, but the relative importance of each is not known. Here, microbiomes of bulk and tuberosphere soil and of potato periderm were studied in one resistant and one susceptible cultivar grown in a conducive and a suppressive field. Disease severity was suppressed similarly by both means yet, the copy numbers of txtB gene (coding for a pathogenicity determinant) were similar in both soils but higher in periderms of the susceptible cultivar from conducive soil. Illumina sequencing of 16S rRNA genes for bacteria (completed by 16S rRNA microarray approach) and archaea, and of 18S rRNA genes for micro-eukarytes showed that in bacteria, the more important was the effect of cultivar and diversity decreased from resistant cultivar to bulk soil to susceptible cultivar. The major changes occurred in proportions of Actinobacteria, Chloroflexi, and Proteobacteria. In archaea and micro-eukaryotes, differences were primarily due to the suppressive and conducive soil. The effect of soil suppressiveness × cultivar resistance depended on the microbial community considered, but differed also with respect to soil and plant nutrient contents particularly in N, S and Fe.


Subject(s)
Actinobacteria/growth & development , Archaea/growth & development , Disease Susceptibility/immunology , Plant Diseases/microbiology , Soil Microbiology , Solanum tuberosum/microbiology , Actinobacteria/classification , Actinobacteria/genetics , Actinobacteria/pathogenicity , Archaea/classification , Archaea/genetics , Archaea/pathogenicity , Chloroflexi/classification , Chloroflexi/genetics , Chloroflexi/growth & development , Chloroflexi/pathogenicity , Crops, Agricultural , Disease Resistance/drug effects , Eukaryotic Cells/metabolism , Genotyping Techniques , Iron/metabolism , Iron/pharmacology , Microbiota/genetics , Nitrogen/metabolism , Nitrogen/pharmacology , Plant Diseases/immunology , Proteobacteria/classification , Proteobacteria/genetics , Proteobacteria/growth & development , Proteobacteria/pathogenicity , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 18S/genetics , Solanum tuberosum/drug effects , Solanum tuberosum/immunology , Sulfur/metabolism , Sulfur/pharmacology , Virulence Factors/genetics , Virulence Factors/metabolism
9.
Med Sci Monit ; 25: 7312-7320, 2019 Sep 29.
Article in English | MEDLINE | ID: mdl-31563920

ABSTRACT

BACKGROUND Laparoscopic cholecystectomy (LC) is regarded as the criterion standard for gallstone therapy, but post-cholecystectomy syndrome (PCS) is a common complication. This study aimed to analyze and identify differences in gut microbiome in PCS patients. MATERIAL AND METHODS This study involved 8 PCS patients (RS1), 8 asymptomatic PCS patients (RS2), and 8 healthy individuals (RS3). Genomic DNA of gut microbiome was extracted and amplified with CTAB method. PCR products were sequenced with Illumina High-Through Sequencing. Sequencing data were analyzed with QIIME software. Effective sequence of bacterial 16S-rRNA gene was clustered into OTUs using UPARSE software. Species annotations were evaluated using Mothur software. QIIME software was used to conduct complexity analysis and calculate UniFrac distances. R software was used to generate PCoA plots. RESULTS Bacterial 16S-rDNA gene sequences showed that the effective species annotative data were more than 97%. According to Ternary plot, Firmicutes and Bacteroidetes had similar abundance and contents among the 3 groups. Contents of Proteobacteria in RS1 were higher compared to RS2 and RS3. Bacterial genomic DNAs samples were clustered together in the same group; however, distances were relative far between different groups. RS1 illustrated significantly higher abundance of Proteobacteria colonies compared to healthy people (p<0.05), and illustrated higher abundance of Verrucomicrobia and lower abundance of Bacteroidetes and Firmicutes, but without significant differences (p>0.05). CONCLUSIONS Gut microbiome of PCS patients was dominated by Proteobacteria in feces and contained little Firmicutes and Bacteroidetes. The enhanced abundance of Proteobacteria might be the highly pathogenic risk factor for chronic abdominal pain and diarrhea in PCS patients.


Subject(s)
Gastrointestinal Microbiome/genetics , Postcholecystectomy Syndrome/microbiology , Proteobacteria/pathogenicity , Abdominal Pain/etiology , Adult , Aged , Bacteria/genetics , China , DNA, Bacterial/genetics , Diarrhea/etiology , Feces/microbiology , Female , Gastrointestinal Microbiome/physiology , High-Throughput Nucleotide Sequencing , Humans , Male , Metabolomics/methods , Middle Aged , Postcholecystectomy Syndrome/complications , RNA, Ribosomal, 16S/genetics , Risk Factors , Sequence Analysis, DNA
10.
IUBMB Life ; 71(2): 152-165, 2019 02.
Article in English | MEDLINE | ID: mdl-30466159

ABSTRACT

It is now well appreciated that the human microbiome plays a significant role in a number of processes in the body, significantly affecting its metabolic, inflammatory, and immune homeostasis. Recent research has revealed that almost every mucosal surface in the human body is associated with a resident commensal microbiome of its own. While the gut microbiome and its role in regulation of host metabolism along with its alteration in a disease state has been well studied, there is a lacuna in understanding the resident microbiota of other mucosal surfaces. Among these, the scientific information on the role of lung microbiota in pulmonary diseases is currently severely limited. Historically, lungs have been considered to be sterile and lung diseases have only been studied in the context of bacterial pathogenesis. Recently however, studies have revealed a resilient microbiome in the upper and lower respiratory tracts and there is increased evidence on its central role in respiratory diseases. Knowledge of lung microbiome and its metabolic fallout (local and systemic) is still in its nascent stages and attracting immense interest in recent times. In this review, we will provide a perspective on lung-associated metabolic disorders defined for lung diseases (e.g., chronic obstructive pulmonary disease, asthma, and respiratory depression due to infection) and correlate it with lung microbial perturbation. Such perturbations may be due to altered biochemical or metabolic stress as well. Finally, we will draw evidence from microbiome and classical microbiology literature to demonstrate how specific lung morbidities associate with specific metabolic characteristics of the disease, and with the role of microbiome in this context. © 2018 IUBMB Life, 71(1):152-165, 2019.


Subject(s)
Asthma/metabolism , Cystic Fibrosis/metabolism , Lung Neoplasms/metabolism , Pneumonia, Bacterial/metabolism , Pulmonary Disease, Chronic Obstructive/metabolism , Actinobacteria/immunology , Actinobacteria/metabolism , Actinobacteria/pathogenicity , Asthma/immunology , Asthma/microbiology , Asthma/pathology , Cystic Fibrosis/immunology , Cystic Fibrosis/microbiology , Cystic Fibrosis/pathology , Firmicutes/immunology , Firmicutes/metabolism , Firmicutes/pathogenicity , Homeostasis/immunology , Humans , Lung/immunology , Lung/metabolism , Lung/microbiology , Lung/pathology , Lung Neoplasms/immunology , Lung Neoplasms/microbiology , Lung Neoplasms/pathology , Microbiota/immunology , Pneumonia, Bacterial/immunology , Pneumonia, Bacterial/microbiology , Pneumonia, Bacterial/pathology , Proteobacteria/immunology , Proteobacteria/metabolism , Proteobacteria/pathogenicity , Pulmonary Disease, Chronic Obstructive/immunology , Pulmonary Disease, Chronic Obstructive/microbiology , Pulmonary Disease, Chronic Obstructive/pathology , Respiratory Mucosa/immunology , Respiratory Mucosa/metabolism , Respiratory Mucosa/microbiology , Respiratory Mucosa/pathology
11.
J Invertebr Pathol ; 157: 74-79, 2018 09.
Article in English | MEDLINE | ID: mdl-30099012

ABSTRACT

Insect pests in the rice agroecosystem, particularly the leaf folder, Cnaphalocrosis medinalis (Guenee) and stem borer, Sesamia inferens (Walker), cause significant yield losses. These pests are generally managed by farmers by application of insecticides and a few biocontrol agents. As a component of integrated pest management, biocontrol agents play a dynamic role in pest control. Although diverse microbial communities are available in the rice ecosystem, bacterial genera such as Bacillus and Pseudomonas spp. are broadly used as biocontrol agents. Therefore, an attempt was made to identify other effective entomopathogenic bacteria to manage the above mentioned pests. In this study, the two entomopathogenic bacteria isolated from diseased pink stem borer (S. inferens Walker) larvae collected from rice fields were identified as Skermanella sp. (KX611462) and Serratia sp. (KX761232). The larvicidal activity of these two bacteria was evaluated against third instar larvae of C. medinalis and S. inferens in in vitro assays and on potted rice plants (Oryza sativa var. TN1). The results of this study demonstrated 50% (LC50) larval mortality of C. medinalis at 2.95 × 103 and 5.88 × 103 colony forming units (CFU) ml-1 for Skermanella sp. and Serratia sp., respectively, under in vitro conditions, 2.57 × 104 and 3.38 × 104 CFU ml-1, respectively, in whole plant assays. Similarly, the LC50 value for Skermanella sp. was 3.80 × 104 CFU ml-1 and Serratia sp. was 2.29 × 105 CFU ml-1 for S. inferens larvae. Our study reports the larvicidal activity of Skermanella sp. against C. medinalis and S. inferens.


Subject(s)
Larva/parasitology , Moths/parasitology , Pest Control, Biological/methods , Proteobacteria/pathogenicity , Serratia/pathogenicity , Animals
12.
Pediatr Pulmonol ; 53(10): 1340-1345, 2018 10.
Article in English | MEDLINE | ID: mdl-29943915

ABSTRACT

Asthma is the most common chronic disease in childhood. The pathogenesis of asthma is multifactorial and is thought to include environmental factors interacting with genetics during pregnancy and in the first years of life. In the last decades, a possible role of gut microbiota in allergic disease pathogenesis has been demonstrated. Next generation sequencing techniques have allowed the identification of a distinct microbiome in the healthy lungs. The lung microbiome is characterized by the prevalence of bacteria belonging to the phylum Bacteroidetes (mostly Prevotella and Veilonella spp) in healthy subjects and to the phylum Proteobacteria in asthmatics (mostly Haemophilus, Moraxella, and Neisseria spp). In asthma, as well as in other diseases, the lung microbiome composition changes due to a disruption of the delicate balance between immigration and elimination of bacteria. The lung microbiome can interact with the immune system, thus influencing inflammation. Early infections with viruses, such as respiratory syncytial virus, may alter lung microbiome composition favoring the emergence of Proteobacteria, a phylum which is also linked to severity of asthma and bronchial hyperreactivity. Lastly, antibiotics may alter the gut and lung microbiota and potentially disturb the relationship between microbiota and host. Therefore, antibiotics should be prescribed with increasing awareness of their potential harmful effect on the microbiota in young children with and without asthma. The potential effects of probiotics and prebiotics on lung microbiome are unknown.


Subject(s)
Asthma/microbiology , Lung/microbiology , Microbiota , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/therapeutic use , Asthma/immunology , Bacteria/drug effects , Bronchial Hyperreactivity/microbiology , Child , Humans , Inflammation/immunology , Inflammation/microbiology , Lung/immunology , Microbiota/drug effects , Proteobacteria/pathogenicity , Respiratory Syncytial Virus Infections/complications , Severity of Illness Index
13.
Cell Microbiol ; 20(6): e12850, 2018 06.
Article in English | MEDLINE | ID: mdl-29624823

ABSTRACT

Cyclic ß-1,2-D-glucans (CßG) are natural bionanopolymers present in the periplasmic space of many Proteobacteria. These molecules are sugar rings made of 17 to 25 D-glucose units linked exclusively by ß-1,2-glycosidic bonds. CßG are important for environmental sensing and osmoadaptation in bacteria, but most importantly, they play key roles in complex host-cell interactions such as symbiosis, pathogenesis, and immunomodulation. In the last years, the identification and characterisation of the enzymes involved in the synthesis of CßG allowed to know in detail the steps necessary for the formation of these sugar rings. Due to its peculiar structure, CßG can complex large hydrophobic molecules, a feature possibly related to its function in the interaction with the host. The capabilities of the CßG to function as molecular boxes and to solubilise hydrophobic compounds are attractive for application in the development of drugs, in food industry, nanotechnology, and chemistry. More importantly, its excellent immunomodulatory properties led to the proposal of CßG as a new class of adjuvants for vaccine development.


Subject(s)
Host-Pathogen Interactions , Proteobacteria/physiology , Proteobacteria/pathogenicity , Symbiosis , beta-Glucans/chemistry , beta-Glucans/metabolism , Biosynthetic Pathways , Hydrophobic and Hydrophilic Interactions
15.
Environ Microbiol ; 20(1): 1-15, 2018 01.
Article in English | MEDLINE | ID: mdl-29027348

ABSTRACT

The type VI secretion system (T6SS) is a bacterial nanomachine used to inject effectors into prokaryotic or eukaryotic cells and is thus involved in both host manipulation and interbacterial competition. The T6SS is widespread among Gram-negative bacteria, mostly within the Proteobacterium Phylum. This secretion system is commonly found in commensal and pathogenic plant-associated bacteria. Phylogenetic analysis of phytobacterial T6SS clusters shows that they are distributed in the five main clades previously described (group 1-5). The even distribution of the system among commensal and pathogenic phytobacteria suggests that the T6SS provides fitness and colonization advantages in planta and that the role of the T6SS is not restricted to virulence. This manuscript reviews the phylogeny and biological roles of the T6SS in plant-associated bacteria, highlighting a remarkable diversity both in terms of mechanism and function.


Subject(s)
Plants/microbiology , Proteobacteria/metabolism , Proteobacteria/pathogenicity , Type VI Secretion Systems/physiology , Bacterial Proteins/genetics , Phylogeny , Plant Diseases/microbiology , Proteobacteria/genetics , Type VI Secretion Systems/genetics , Virulence
16.
Microbiome ; 5(1): 159, 2017 12 11.
Article in English | MEDLINE | ID: mdl-29228991

ABSTRACT

BACKGROUND: The emergence and spread of antimicrobial resistance is the most urgent current threat to human and animal health. An improved understanding of the abundance of antimicrobial resistance genes and genes associated with microbial colonisation and pathogenicity in the animal gut will have a major role in reducing the contribution of animal production to this problem. Here, the influence of diet on the ruminal resistome and abundance of pathogenicity genes was assessed in ruminal digesta samples taken from 50 antibiotic-free beef cattle, comprising four cattle breeds receiving two diets containing different proportions of concentrate. RESULTS: Two hundred and four genes associated with antimicrobial resistance (AMR), colonisation, communication or pathogenicity functions were identified from 4966 metagenomic genes using KEGG identification. Both the diversity and abundance of these genes were higher in concentrate-fed animals. Chloramphenicol and microcin resistance genes were dominant in samples from forage-fed animals (P < 0.001), while aminoglycoside and streptomycin resistances were enriched in concentrate-fed animals. The concentrate-based diet also increased the relative abundance of Proteobacteria, which includes many animal and zoonotic pathogens. A high ratio of Proteobacteria to (Firmicutes + Bacteroidetes) was confirmed as a good indicator for rumen dysbiosis, with eight cases all from concentrate-fed animals. Finally, network analysis demonstrated that the resistance/pathogenicity genes are potentially useful as biomarkers for health risk assessment of the ruminal microbiome. CONCLUSIONS: Diet has important effects on the complement of AMR genes in the rumen microbial community, with potential implications for human and animal health.


Subject(s)
Animal Feed/analysis , Cattle/microbiology , Drug Resistance, Multiple, Bacterial/genetics , Genes, Bacterial , Microbiota , Rumen/microbiology , Animal Feed/adverse effects , Animals , Anti-Bacterial Agents/pharmacology , Bacteriocins/pharmacology , Bacteroidetes/drug effects , Bacteroidetes/genetics , Bacteroidetes/pathogenicity , Chloramphenicol/pharmacology , Firmicutes/drug effects , Firmicutes/genetics , Firmicutes/pathogenicity , Humans , Metagenomics/methods , Proteobacteria/drug effects , Proteobacteria/genetics , Proteobacteria/pathogenicity , Red Meat/analysis , Virulence
17.
Microbiome ; 5(1): 104, 2017 09 01.
Article in English | MEDLINE | ID: mdl-28859671

ABSTRACT

BACKGROUND: Although the plant microbiome is crucial for plant health, little is known about the significance of the seed microbiome. Here, we studied indigenous bacterial communities associated with the seeds in different cultivars of oilseed rape and their interactions with symbiotic and pathogenic microorganisms. RESULTS: We found a high bacterial diversity expressed by tight bacterial co-occurrence networks within the rape seed microbiome, as identified by llumina MiSeq amplicon sequencing. In total, 8362 operational taxonomic units (OTUs) of 40 bacterial phyla with a predominance of Proteobacteria (56%) were found. The three cultivars that were analyzed shared only one third of the OTUs. The shared core of OTUs consisted mainly of Alphaproteobacteria (33%). Each cultivar was characterized by having its own unique bacterial structure, diversity, and proportion of unique microorganisms (25%). The cultivar with the lowest bacterial abundance, diversity, and the highest predicted bacterial metabolic activity rate contained the highest abundance of potential pathogens within the seed. This data corresponded with the observation that seedlings belonging to this cultivar responded more strongly to the seed treatments with bacterial inoculants than other cultivars. Cultivars containing higher indigenous diversity were characterized as having a higher colonization resistance against beneficial and pathogenic microorganisms. Our results were confirmed by microscopic images of the seed microbiota. CONCLUSIONS: The structure of the seed microbiome is an important factor in the development of colonization resistance against pathogens. It also has a strong influence on the response of seedlings to biological seed treatments. These novel insights into seed microbiome structure will enable the development of next generation strategies combining both biocontrol and breeding approaches to address world agricultural challenges.


Subject(s)
Brassica napus/microbiology , Microbial Interactions , Microbiota/genetics , Proteobacteria/physiology , Seeds/microbiology , Symbiosis , Alphaproteobacteria/genetics , Alphaproteobacteria/isolation & purification , Alphaproteobacteria/metabolism , Bacteria/pathogenicity , Bacterial Physiological Phenomena , Genetic Variation , High-Throughput Nucleotide Sequencing , Microscopy, Confocal , Proteobacteria/genetics , Proteobacteria/isolation & purification , Proteobacteria/pathogenicity
18.
Nat Commun ; 8: 15784, 2017 06 07.
Article in English | MEDLINE | ID: mdl-28589945

ABSTRACT

It has been hypothesized that some antibiotic resistance genes (ARGs) found in pathogenic bacteria derive from antibiotic-producing actinobacteria. Here we provide bioinformatic and experimental evidence supporting this hypothesis. We identify genes in proteobacteria, including some pathogens, that appear to be closely related to actinobacterial ARGs known to confer resistance against clinically important antibiotics. Furthermore, we identify two potential examples of recent horizontal transfer of actinobacterial ARGs to proteobacterial pathogens. Based on this bioinformatic evidence, we propose and experimentally test a 'carry-back' mechanism for the transfer, involving conjugative transfer of a carrier sequence from proteobacteria to actinobacteria, recombination of the carrier sequence with the actinobacterial ARG, followed by natural transformation of proteobacteria with the carrier-sandwiched ARG. Our results support the existence of ancient and, possibly, recent transfers of ARGs from antibiotic-producing actinobacteria to proteobacteria, and provide evidence for a defined mechanism.


Subject(s)
Bacterial Proteins/genetics , Drug Resistance, Microbial/genetics , Proteobacteria/drug effects , Proteobacteria/genetics , Streptomyces/genetics , Acinetobacter/drug effects , Acinetobacter/genetics , Actinobacteria/drug effects , Actinobacteria/genetics , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , DNA Transposable Elements , Escherichia coli/genetics , Gene Transfer, Horizontal , Phylogeny , Proteobacteria/pathogenicity , Streptomyces/drug effects
19.
Microbiol Mol Biol Rev ; 80(3): 597-628, 2016 09.
Article in English | MEDLINE | ID: mdl-27307578

ABSTRACT

Bacterial sphingomyelinases and phospholipases are a heterogeneous group of esterases which are usually surface associated or secreted by a wide variety of Gram-positive and Gram-negative bacteria. These enzymes hydrolyze sphingomyelin and glycerophospholipids, respectively, generating products identical to the ones produced by eukaryotic enzymes which play crucial roles in distinct physiological processes, including membrane dynamics, cellular signaling, migration, growth, and death. Several bacterial sphingomyelinases and phospholipases are essential for virulence of extracellular, facultative, or obligate intracellular pathogens, as these enzymes contribute to phagosomal escape or phagosomal maturation avoidance, favoring tissue colonization, infection establishment and progression, or immune response evasion. This work presents a classification proposal for bacterial sphingomyelinases and phospholipases that considers not only their enzymatic activities but also their structural aspects. An overview of the main physiopathological activities is provided for each enzyme type, as are examples in which inactivation of a sphingomyelinase- or a phospholipase-encoding gene impairs the virulence of a pathogen. The identification of sphingomyelinases and phospholipases important for bacterial pathogenesis and the development of inhibitors for these enzymes could generate candidate vaccines and therapeutic agents, which will diminish the impacts of the associated human and animal diseases.


Subject(s)
Bacteria/pathogenicity , Fungi/pathogenicity , Phospholipase D/metabolism , Phosphoric Diester Hydrolases/metabolism , Sphingomyelin Phosphodiesterase/metabolism , Type C Phospholipases/metabolism , Bacteria/enzymology , Firmicutes/enzymology , Firmicutes/pathogenicity , Fungi/enzymology , Proteobacteria/enzymology , Proteobacteria/pathogenicity , Staphylococcus aureus/enzymology , Staphylococcus aureus/pathogenicity , Virulence Factors/metabolism
20.
PLoS Pathog ; 12(6): e1005629, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27322651

ABSTRACT

Heritable microbial symbionts have profound impacts upon the biology of their arthropod hosts. Whilst our current understanding of the dynamics of these symbionts is typically cast within a framework of vertical transmission only, horizontal transmission has been observed in a number of cases. For instance, several symbionts can transmit horizontally when their parasitoid hosts share oviposition patches with uninfected conspecifics, a phenomenon called superparasitism. Despite this, horizontal transmission, and the host contact structures that facilitates it, have not been considered in heritable symbiont epidemiology. Here, we tested for the importance of host contact, and resulting horizontal transmission, for the epidemiology of a male-killing heritable symbiont (Arsenophonus nasoniae) in parasitoid wasp hosts. We observed that host contact through superparasitism is necessary for this symbiont's spread in populations of its primary host Nasonia vitripennis, such that when superparasitism rates are high, A. nasoniae almost reaches fixation, causes highly female biased population sex ratios and consequently causes local host extinction. We further tested if natural interspecific variation in superparasitism behaviours predicted symbiont dynamics among parasitoid species. We found that A. nasoniae was maintained in laboratory populations of a closely related set of Nasonia species, but declined in other, more distantly related pteromalid hosts. The natural proclivity of a species to superparasitise was the primary factor determining symbiont persistence. Our results thus indicate that host contact behaviour is a key factor for heritable microbe dynamics when horizontal transmission is possible, and that 'reproductive parasite' phenotypes, such as male-killing, may be of secondary importance in the dynamics of such symbiont infections.


Subject(s)
Disease Transmission, Infectious/veterinary , Proteobacteria/pathogenicity , Symbiosis/physiology , Wasps/parasitology , Animals , Female , Male , Sex Ratio
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