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1.
Glycobiology ; 33(1): 57-74, 2023 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-36239418

RESUMO

Bacterial protein glycosylation is commonly mediated by oligosaccharyltransferases (OTases) that transfer oligosaccharides en bloc from preassembled lipid-linked precursors to acceptor proteins. Natively, O-linking OTases usually transfer a single repeat unit of the O-antigen or capsular polysaccharide to the side chains of serine or threonine on acceptor proteins. Three major families of bacterial O-linking OTases have been described: PglL, PglS, and TfpO. TfpO is limited to transferring short oligosaccharides both in its native context and when heterologously expressed in glycoengineered Escherichia coli. On the other hand, PglL and PglS can transfer long-chain polysaccharides when expressed in glycoengineered E. coli. Herein, we describe the discovery and functional characterization of a novel family of bacterial O-linking OTases termed TfpM from Moraxellaceae bacteria. TfpM proteins are similar in size and sequence to TfpO enzymes but can transfer long-chain polysaccharides to acceptor proteins. Phylogenetic analyses demonstrate that TfpM proteins cluster in distinct clades from known bacterial OTases. Using a representative TfpM enzyme from Moraxella osloensis, we determined that TfpM glycosylates a C-terminal threonine of its cognate pilin-like protein and identified the minimal sequon required for glycosylation. We further demonstrated that TfpM has broad substrate tolerance and can transfer diverse glycans including those with glucose, galactose, or 2-N-acetyl sugars at the reducing end. Last, we find that a TfpM-derived bioconjugate is immunogenic and elicits serotype-specific polysaccharide IgG responses in mice. The glycan substrate promiscuity of TfpM and identification of the minimal TfpM sequon renders this enzyme a valuable additional tool for expanding the glycoengineering toolbox.


Assuntos
Hexosiltransferases , Moraxellaceae , Animais , Camundongos , Moraxellaceae/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Filogenia , Hexosiltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Fímbrias , Polissacarídeos/metabolismo , Bactérias/metabolismo
2.
Food Res Int ; 158: 111579, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35840264

RESUMO

In the present context of growing antimicrobial resistance (AMR) concern, understanding the distribution of AMR determinants in food matrices such as milk is crucial to protect consumers and maintain high food safety standards. Herein, the resistome of different dairy farms was investigated through a shotgun metagenomic sequencing approach, taking advantage of in-line milk filters as promising tools. The application of both the reads-based and the assembly-based approaches has allowed the identification of numerous AMR determinants, enabling a comprehensive resolution of the resistome. Notably most of the species harboring AMR genes were predicted to be Gram-negative genera, namely Enterobacter, Acinetobacter, Escherichia, and Pseudomonas, pointing out the role of these bacteria as reservoirs of AMR determinants. In this context, the use of de novo assembly has allowed a more holistic AMR detection strategy, while the reads-based approach has enabled the detection of AMR genes from low abundance bacteria, usually undetectable by assembly-based methods. The application of both reads-based and assembly-based approaches, despite being computationally demanding, has facilitated the comprehensive characterization of a food chain resistome, while also allowing the construction of complete metagenome assembled genomes and the investigation of mobile genetic elements. Our findings suggest that milk filters can successfully be used to investigate the resistome of bulk tank milk through the application of the shotgun metagenomic sequencing. In accordance with our results, raw milk can be considered a source of AMR bacteria and genes; this points out the importance of properly informing food business operators about the risk associated with poor hygiene practices in the dairy production environment and consumers of the potential microbial food safety risks derived from raw milk products consumption. Translating these findings as risk assessment outputs heralds the next generation of food safety controls.


Assuntos
Anti-Infecciosos , Enterobacteriaceae , Animais , Antibacterianos/farmacologia , Bactérias , Farmacorresistência Bacteriana/genética , Metagenoma , Leite/microbiologia , Moraxellaceae/genética
3.
Curr Microbiol ; 78(1): 378-382, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33179156

RESUMO

The taxonomic position of strain H1T isolated from crude oil contaminated desert sands was determined. Strain H1T was Gram-stain-negative and cocci to short rod-shaped bacterium. It grew at 15-42ºC (optimum, 30-35ºC) and pH 6.5-8.8 (optimum, 7.0-7.5). No added NaCl was required for the growth. The isolate showed 98% 16S rRNA gene sequence similarity with the Alkanindiges illinoisensis GTI MVAB Hex1T, 95.5% with Alkanindiges hongkongensis HKU9T and < 95.2% with other members of the family Moraxellaceae of the phylum Proteobacteria. C10:0, C10:0 -2OH, C12:0 -3OH, C16:0, C16:0 N alcohol and C16:1ω6c/C16:1ω7c were present as major (5%) fatty acids with minor (< 5%) amounts of C12:0, C14:0, C14:1ω5c and C18:1ω9c in strain H1T. It contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and unidentified two unidentified lipids. Distinct morphological, physiological, phylogenetic, and genomic differences from the previously described taxa support the classification of strain H1T as a representative of a novel species in the genus Alkanindiges for which the name Alkanindiges hydrocarboniclasticus sp. nov. is proposed. The type strain is H1T (= JCM 31550T = KEMB 2255-480T). Emended description of the genus Alkanindiges is also proposed based on additional characteristics.


Assuntos
Moraxellaceae/classificação , Poluição por Petróleo , Filogenia , Areia/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Moraxellaceae/isolamento & purificação , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
4.
Anim Sci J ; 91(1): e13441, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32885570

RESUMO

Microbiota of individual cow milk, bulk tank milk, and feces of Jersey cows were examined. Samples were collected from two farms (F1 and F2) in cool (November, Nov) and hot (July, Jul) seasons. Milk yield and milk composition were similar between the two farms and between the two seasons. Prevalent taxa of the fecal microbiota, i.e. Ruminococcaceae, Bacteroidaceae, Lachnospiraceae, Rikenellaceae, and Clostridiaceae, were unaffected by the farm and season. Relative abundance of milk microbiota for Pseudomonadaceae, Enterobacteriaceae, and Streptococcaceae (F1 > F2) and Lactobacillaceae, Bifidobacteriaceae, and Cellulomonadaceae (F1 < F2) were different between the two farms, and those for Staphylococcaceae, Bacillaceae, Ruminococcaceae, and Veillonellaceae (Nov < Jul) and Methylobacteriaceae and Moraxellaceae (Nov > Jul) were different between the two seasons. The microbiota of bulk tank milk was numerically different from that of individual cow milk. Principal coordinate analysis indicated that the milk microbiota was unrelated to the fecal microbiota. The finding that relative abundance of Pseudomonadaceae and Moraxellaceae appeared greater than those reported for Holstein milk suggested that higher protein and fat content may result in a greater abundance of proteolytic and lipolytic taxa in Jersey cow milk.


Assuntos
Bovinos/metabolismo , Bovinos/microbiologia , Temperatura Baixa , Indústria de Laticínios , Fazendas , Gorduras/metabolismo , Fezes/microbiologia , Temperatura Alta , Proteínas do Leite/metabolismo , Leite/metabolismo , Leite/microbiologia , Estações do Ano , Animais , Feminino , Moraxellaceae , Proteólise , Pseudomonadaceae
5.
Int J Syst Evol Microbiol ; 70(10): 5479-5487, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32886597

RESUMO

Two novel bacteria, designated HYN0043T and HYN0046T, were isolated from a freshwater lake in Korea. 16S rRNA gene sequence phylogeny indicated that strain HYN0043T belongs to the genus Mucilaginibacter of the family Sphingobacteriaceae because it showed highest sequence similarity to Mucilaginibacter oryzae (98.2 %). The average nucleotide identity between strain HYN0043T and M. oryzae was 83.5 %, which is clearly below the suggested threshold for species demarcation. Strain HYN0046T was found to belong to the family Moraxellaceae and shared highest sequence similarity with Agitococcus lubricus (93.8 %). The average amino acid identity values between strain HYN0046T and representative type strains of closely related genera (Alkanindiges, Agitococcus and Acinetobacter) were 53.1-60.7 %, implying the novelty of the isolate at the genus level. Phenotypic characteristics (physiological, biochemical and chemotaxonomic) also supported the taxonomic novelty of the two isolates. Thus, we suggest the following names to accommodate strains HYN0043T and HYN0046T: Mucilaginibacter celer sp. nov. (type strain HYN0043T=KACC 19184T=NBRC 112738T) in the family Spingobacteriaceae and phylum Bacteroidetes and Aquirhabdus parva gen. nov., sp. nov. (type strain HYN0046T=KACC 19178T=NBRC 112739T) in the family Moraxellaceae and phylum Proteobacteria.


Assuntos
Bacteroidetes/classificação , Lagos/microbiologia , Moraxellaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Bacteroidetes/isolamento & purificação , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Moraxellaceae/isolamento & purificação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
6.
J Microbiol ; 58(7): 588-597, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32424577

RESUMO

Our study demonstrated that sleep deprivation resulted in homeostasis disorder of colon. Our study goes deeper into the positive effects of melatonin on small intestinal microbiota disorder caused by sleep deprivation. We successfully established a multiplatform 72 h sleep deprivation mouse model with or without melatonin supplementation, and analyzed the change of small intestinal microbiota using high-throughput sequencing of the 16S rRNA. We found melatonin supplementation suppressed the decrease of plasma melatonin level in sleep deprivation mice. Meanwhile, melatonin supplementation improved significantly the reduction in OTU numbers and the diversity and richness of jejunal microbiota and the abundance of Bacteroidaeae and Prevotellaceae, as well as an increase in the Firmicutes-to-Bacteroidetes ratio and the content of Moraxellaceae and Aeromonadaceae in the jejunum of sleep deprived-mice. Moreover, melatonin supplementation reversed the change of metabolic pathway in sleep deprived-mice, including metabolism, signal transduction mechanisms and transcription etc, which were related to intestinal health. Furthermore, melatonin supplementation inverted the sleep deprivation-induced a decline of anti-inflammatory cytokines (IL-22) and an increase of the ROS and proinflammatory cytokines (IL-17) in jejunum. These findings suggested that melatonin, similar to a probiotics agent, can reverse sleep deprivation-induced small intestinal microbiota disorder by suppressing oxidative stress and inflammation response.


Assuntos
Antioxidantes/farmacologia , Disbiose/tratamento farmacológico , Microbioma Gastrointestinal/efeitos dos fármacos , Jejuno/microbiologia , Melatonina/farmacologia , Privação do Sono/microbiologia , Aeromonadaceae/efeitos dos fármacos , Aeromonadaceae/isolamento & purificação , Animais , Bacteroidaceae/efeitos dos fármacos , Bacteroidaceae/isolamento & purificação , Firmicutes/efeitos dos fármacos , Firmicutes/isolamento & purificação , Inflamação , Interleucina-17/análise , Interleucinas/análise , Masculino , Camundongos , Camundongos Endogâmicos ICR , Moraxellaceae/efeitos dos fármacos , Moraxellaceae/isolamento & purificação , Estresse Oxidativo/efeitos dos fármacos , Prevotella/efeitos dos fármacos , Prevotella/isolamento & purificação , RNA Ribossômico 16S/genética , Transdução de Sinais/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacos , Transcrição Gênica/genética
7.
mSphere ; 4(3)2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-31043518

RESUMO

Laryngotracheal stenosis is an obstructive respiratory disease that leads to voicing difficulties and dyspnea with potential life-threatening consequences. The majority of incidences are due to iatrogenic etiology from endotracheal tube intubation; however, airway scarring also has idiopathic causes. While recent evidence suggests a microbial contribution to mucosal inflammation, the microbiota associated with different types of stenosis has not been characterized. High-throughput sequencing of the V4 region of the16S rRNA gene was performed to characterize the microbial communities of 61 swab samples from 17 iatrogenic and 10 adult idiopathic stenosis patients. Nonscar swabs from stenosis patients were internal controls, and eight swabs from four patients without stenosis represented external controls. Significant differences in diversity were observed between scar and nonscar samples and among sample sites, with decreased diversity detected in scar samples and the glottis region. Permutational analysis of variance (PERMANOVA) results revealed significant differences in community composition for scar versus nonscar samples, etiology type, sample site, groups (iatrogenic, idiopathic, and internal and external controls), and individual patients. Pairwise Spearman's correlation revealed a strong inverse correlation between Prevotella and Streptococcus among all samples. Finally, bacteria in the family Moraxellaceae were found to be distinctly associated with idiopathic stenosis samples in comparison with external controls. Our findings suggest that specific microbiota and community shifts are present with laryngotracheal stenosis in adults, with members of the family Moraxellaceae, including the known pathogens Moraxella and Acinetobacter, identified in idiopathic scar. Further work is warranted to elucidate the contributing role of bacteria on the pathogenesis of laryngotracheal stenosis.IMPORTANCE The laryngotracheal region resides at the intersection between the heavily studied nasal cavity and lungs; however, examination of the microbiome in chronic inflammatory conditions of the subglottis and trachea remains scarce. To date, studies have focused on the microbiota of the vocal folds, or the glottis, for laryngeal carcinoma, as well as healthy larynges, benign vocal fold lesions, and larynges exposed to smoking and refluxate. In this study, we seek to examine the structure and composition of the microbial community in adult laryngotracheal stenosis of various etiologies. Due to the heterogeneity among the underlying pathogenesis mechanisms and clinical outcomes seen in laryngotracheal stenosis disease, we hypothesized that different microbial profiles will be detected among various stenosis etiology types. Understanding differences in the microbiota for subglottic stenosis subtypes may shed light upon etiology-specific biomarker identification and offer novel insights into management approaches for this debilitating disease.


Assuntos
Bactérias/classificação , Laringoestenose/microbiologia , Microbiota , Traqueia/microbiologia , Estenose Traqueal/microbiologia , Acinetobacter/genética , Acinetobacter/isolamento & purificação , Adolescente , Adulto , Idoso , Bactérias/isolamento & purificação , Cicatriz/microbiologia , Constrição Patológica , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Laringoestenose/patologia , Masculino , Pessoa de Meia-Idade , Moraxellaceae/genética , Moraxellaceae/isolamento & purificação , Traqueia/patologia , Estenose Traqueal/patologia
8.
Curr Microbiol ; 76(4): 478-484, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30783797

RESUMO

A gram-stain-negative and strictly aerobic bacterium, designated strain HR-ET, was isolated from a water sample of the Han River in South Korea. Cells were catalase-negative and oxidase-positive motile rods with a flagellum. The strain grew at 10-37 °C and pH 7-8 and in the presence of 0-2% (w/v) NaCl. Ubiquinone-8 and summed features 3 (comprising C16:1ω7c and/or C16:1ω6c) and 8 (comprising C18:1ω7c and/or C18:1ω6c), C16:0, iso-C10:0 and C12:0 3-OH were identified as the major respiratory quinone and fatty acids (>5%), respectively. The 16S rRNA gene sequence of strain HR-ET shared the highest similarities with Perlucidibaca piscinae IMCC 1704T (98.1%), Perlucidibaca aquatica BK296T (96.8%), Paraperlucidibaca baekdonensis RL-2T (95.8%) and Paraperlucidibaca wandonensis WT-RY4T (95.7%). However, strain HR-ET formed a phylogenetic lineage distinct from members of the family Moraxellaceae, and a taxonomic analysis by RDP Naïve Bayesian rRNA Classifier classified strain HR-ET as a new genus of the family Moraxellaceae. In addition, analyses based on rpoD, secA and gyrB gene sequences also showed that strain HR-ET formed a lineage distinct from those of the genera Perlucidibaca and Paraperlucidibaca. Average nucleotide identity and in silico DNA-DNA hybridization values between strain HR-ET and the type strains of P. piscinae and P. aquatica were very low with 80.1 and 23.6% and 75.7 and 21.2%, respectively. The DNA G+C content was 67.0 mol%. Based on the phenotypic, genotypic and chemotaxonomic analyses, strain HR-ET represents a novel genus of the family Moraxellaceae, for which the name Amnimonas aquatica gen. nov., sp. nov. is proposed. The type strain of the type species is HR-ET (=KACC 19408T=JCM 32266T).


Assuntos
Moraxellaceae/classificação , Moraxellaceae/fisiologia , Filogenia , Rios/microbiologia , Microbiologia da Água , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos/genética , Moraxellaceae/química , Moraxellaceae/genética , Hibridização de Ácido Nucleico , Fenótipo , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Especificidade da Espécie , Ubiquinona/química
9.
mSphere ; 4(1)2019 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-30787115

RESUMO

Insertion sequences (IS) are fundamental mediators of genome plasticity with the potential to generate phenotypic variation with significant evolutionary outcomes. Here, a recently active miniature inverted-repeat transposon element (MITE) was identified in a derivative of Acinetobacter baumannii ATCC 17978 after being subjected to stress conditions. Transposition of the novel element led to the disruption of the hns gene, resulting in a characteristic hypermotile phenotype. DNA identity shared between the terminal inverted repeats of this MITE and coresident ISAba12 elements, together with the generation of 9-bp target site duplications, provides strong evidence that ISAba12 elements were responsible for mobilization of the MITE (designated MITE Aba12 ) within this strain. A wider genome-level survey identified MITE Aba12 in 30 additional Acinetobacter genomes at various frequencies and one Moraxella osloensis genome. Ninety MITE Aba12 copies could be identified, of which 40% had target site duplications, indicating recent transposition events. Elements ranged between 111 and 114 bp; 90% were 113 bp in length. Using the MITE Aba12 consensus sequence, putative outward-facing Escherichia coli σ70 promoter sequences in both orientations were identified. The identification of transcripts originating from the promoter in one direction supports the proposal that the element can influence neighboring host gene transcription. The location of MITE Aba12 varied significantly between and within genomes, preferentially integrating into AT-rich regions. Additionally, a copy of MITE Aba12 was identified in a novel 8.5-kb composite transposon, Tn6645, in the M. osloensis CCUG 350 chromosome. Overall, this study shows that MITE Aba12 is the most abundant nonautonomous element currently found in AcinetobacterIMPORTANCE One of the most important weapons in the armory of Acinetobacter is its impressive genetic plasticity, facilitating rapid genetic mutations and rearrangements as well as integration of foreign determinants carried by mobile genetic elements. Of these, IS are considered one of the key forces shaping bacterial genomes and ultimately evolution. We report the identification of a novel nonautonomous IS-derived element present in multiple bacterial species from the Moraxellaceae family and its recent translocation into the hns locus in the A. baumannii ATCC 17978 genome. The latter finding adds new knowledge to only a limited number of documented examples of MITEs in the literature and underscores the plastic nature of the hns locus in A. baumannii MITE Aba12 , and its predicted parent(s), may be a source of substantial adaptive evolution within environmental and clinically relevant bacterial pathogens and, thus, have broad implications for niche-specific adaptation.


Assuntos
Acinetobacter baumannii/genética , Elementos de DNA Transponíveis , Genoma Bacteriano , Sequências Repetidas Invertidas , Moraxellaceae/genética , Evolução Molecular
10.
Talanta ; 182: 536-543, 2018 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-29501189

RESUMO

The identification of microorganisms is very important in different fields and alternative methods are necessary for a rapid and simple identification. The use of fatty acids for bacterial identification is gaining attention as phenotypic characteristics are reflective of the genotype and are more easily analyzed. In this work, gas chromatography-vacuum ultraviolet spectroscopy (GC-VUV) was used to determine bacteria fatty acid methyl esters (FAMEs), to identify and discriminate different environmental bacteria based on their fatty acid profile. Microorganisms were grown in agar and their fatty acids extracted, saponified, and esterified before analysis. Unique FAME profiles were obtained for each microorganism mainly composed of branched, cyclopropane, hydroxy, saturated, and unsaturated fatty acid methyl esters. S. maltophilia showed a higher diversity of fatty acids while Bacillus species showed higher complexity in terms of branched-chain FAMEs, with several iso and anteiso forms. 12 different bacteria genera and 15 species were successfully differentiated based on their fatty acid profiles after performing PCA and cluster analysis. Some difficult to differentiate species, such as Bacillus sp., which are genetically very similar, were differentiated with the developed method.


Assuntos
Bactérias/isolamento & purificação , Cromatografia Gasosa/métodos , Ácidos Graxos/isolamento & purificação , Água Subterrânea/microbiologia , Espectroscopia Fotoeletrônica/métodos , Aeromonadaceae/classificação , Aeromonadaceae/isolamento & purificação , Aeromonadaceae/metabolismo , Alcaligenaceae/classificação , Alcaligenaceae/isolamento & purificação , Alcaligenaceae/metabolismo , Bacillaceae/classificação , Bacillaceae/isolamento & purificação , Bacillaceae/metabolismo , Bactérias/classificação , Bactérias/metabolismo , Análise por Conglomerados , Comamonadaceae/classificação , Comamonadaceae/isolamento & purificação , Comamonadaceae/metabolismo , Enterobacteriaceae/classificação , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/metabolismo , Ésteres , Ácidos Graxos/química , Ácidos Graxos/classificação , Moraxellaceae/classificação , Moraxellaceae/isolamento & purificação , Moraxellaceae/metabolismo , Análise de Componente Principal , Pseudomonadaceae/classificação , Pseudomonadaceae/isolamento & purificação , Pseudomonadaceae/metabolismo , Vácuo , Microbiologia da Água , Xanthomonadaceae/classificação , Xanthomonadaceae/isolamento & purificação , Xanthomonadaceae/metabolismo
11.
Eur J Mass Spectrom (Chichester) ; 24(3): 261-268, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29392979

RESUMO

The detection of post-translational modifications of proteins is an important comprehensive research area. Over the years, proteomic studies involving protein acetylation have attracted a great deal of attention. In the present study, we have focussed on the acetylation of histidine and the intrinsic stability of b1-ion of oxazolone ring and/or with side chain imidazole bicyclic product. The formation of oxazolone structure may occur when an amino moiety undergoes acetylation reaction and when it is present in the vicinity of the side chain imidazole moiety. Tryptic peptides generated from the proteins of Acenitobacter radioresistens MMC5-containing N-terminal histidine were explored in a standard proteomic workflow. Formation of [Formula: see text] ion with an oxazolone ring in these peptides has been supported by a tandem mass spectrometric study of a synthetic peptide and density functional theory calculations. The results obtained from this study have implications in understanding the fragmentation of the peptides generated in the proteomic workflows.


Assuntos
Proteínas de Bactérias/química , Histidina/química , Imidazóis/química , Moraxellaceae/química , Oxazolona/química , Acetilação , Íons/química , Estrutura Molecular , Peptídeos/química , Processamento de Proteína Pós-Traducional , Proteômica , Espectrometria de Massas em Tandem
12.
Medisan ; 22(1)ene. 2018. tab, ilus
Artigo em Espanhol | LILACS | ID: biblio-894670

RESUMO

Se efectuó un estudio con el empleo de la metodología de Gupta, en el Departamento de Biología y Geografía de la Universidad de Oriente en Santiago de Cuba, para determinar marcadores moleculares de tipo inserción en secuencias de las proteínas ADN polimerasa I y ADN polimerasa III (subunidad alfa), obtenidas de bases de datos internacionales y posteriormente alineadas con el programa ClustalX2. Las familias Moraxellaceae y Helicobacteraceae han sido ampliamente estudiadas, porque comprenden agentes patógenos causantes de numerosas enfermedades en humanos, pero pocas investigaciones han estado dirigidas a la identificación de las características moleculares que puedan distinguir a sus miembros de otros grupos de bacterias; de manera que los presentes resultados constituyen un aporte al conocimiento de la genética y la bioquímica de estas familias y proveen herramientas moleculares para la clasificación taxonómica y el diagnóstico de especies patógenas


A study with the use of Gupta methodology was carried out in the Biology and Geography Department of Oriente University in Santiago de Cuba, to determine molecular markers of insertion type in sequences of the DNA polimerase I proteins and DNA polimerase III (alpha subunity), obtained from international databases and later on aligned with the ClustalX2 program. The Moraxellaceae and Helicobacteraceae families have been broadly studied, because they comprise pathogen agents that cause numerous diseases in humans, but few investigations have been directed to the identification of the molecular characteristics that can distinguish their members from other groups of bacterias; so these results constitute a contribution to the knowledge of genetics and biochemistry of these families and provide molecular tools for the taxonomic classification and the diagnosis of pathogen species


Assuntos
Humanos , Helicobacter/citologia , Moraxellaceae/citologia , DNA Polimerase I , DNA Polimerase III , Biomarcadores , Marcadores Genéticos , Estudos Epidemiológicos
13.
Medisan ; 22(1)ene. 2018. ilus
Artigo em Espanhol | CUMED | ID: cum-70188

RESUMO

Se efectuó un estudio con el empleo de la metodología de Gupta, en el Departamento de Biología y Geografía de la Universidad de Oriente en Santiago de Cuba, para determinar marcadores moleculares de tipo inserción en secuencias de las proteínas ADN polimerasa I y ADN polimerasa III (subunidad alfa), obtenidas de bases de datos internacionales y posteriormente alineadas con el programa ClustalX2. Las familias Moraxellaceae y Helicobacteraceae han sido ampliamente estudiadas, porque comprenden agentes patógenos causantes de numerosas enfermedades en humanos, pero pocas investigaciones han estado dirigidas a la identificación de las características moleculares que puedan distinguir a sus miembros de otros grupos de bacterias; de manera que los presentes resultados constituyen un aporte al conocimiento de la genética y la bioquímica de estas familias y proveen herramientas moleculares para la clasificación taxonómica y el diagnóstico de especies patógenas(AU)


A study with the use of Gupta methodology was carried out in the Biology and Geography Department of Oriente University in Santiago de Cuba, to determine molecular markers of insertion type in sequences of the DNA polimerase I proteins and DNA polimerase III (alpha subunity), obtained from international databases and later on aligned with the ClustalX2 program. The Moraxellaceae and Helicobacteraceae families have been broadly studied, because they comprise pathogen agents that cause numerous diseases in humans, but few investigations have been directed to the identification of the molecular characteristics that can distinguish their members from other groups of bacterias; so these results constitute a contribution to the knowledge of genetics and biochemistry of these families and provide molecular tools for the taxonomic classification and the diagnosis of pathogen species(AU)


Assuntos
Humanos , Helicobacter/citologia , Moraxellaceae/citologia , Estruturas Celulares , Biologia Celular
14.
PLoS Negl Trop Dis ; 11(10): e0005975, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28968382

RESUMO

A longitudinal study was undertaken in infants living in the Maela refugee camp on the Thailand-Myanmar border between 2007 and 2010. Nasopharyngeal swabs were collected monthly, from birth to 24 months of age, with additional swabs taken if the infant was diagnosed with pneumonia according to WHO clinical criteria. At the time of collection, swabs were cultured for Streptococcus pneumoniae and multiple serotype carriage was assessed. The bacterial 16S rRNA gene profiles of 544 swabs from 21 infants were analysed to see how the microbiota changes with age, respiratory infection, antibiotic consumption and pneumococcal acquisition. The nasopharyngeal microbiota is a somewhat homogenous community compared to that of other body sites. In this cohort it is dominated by five taxa: Moraxella, Streptococcus, Haemophilus, Corynebacterium and an uncharacterized Flavobacteriaceae taxon of 93% nucleotide similarity to Ornithobacterium. Infant age correlates with certain changes in the microbiota across the cohort: Staphylococcus and Corynebacterium are associated with the first few months of life while Moraxella and the uncharacterised Flavobacteriaceae increase in proportional abundance with age. Respiratory illness and antibiotic use often coincide with an unpredictable perturbation of the microbiota that differs from infant to infant and in different illness episodes. The previously described interaction between Dolosigranulum and Streptococcus was observed in these data. Monthly sampling demonstrates that the nasopharyngeal microbiota is in flux throughout the first two years of life, and that in this refugee camp population the pool of potential bacterial colonisers may be limited.


Assuntos
Antibacterianos/uso terapêutico , Bactérias/isolamento & purificação , Microbiota , Nasofaringe/microbiologia , Infecções Respiratórias/epidemiologia , Fatores Etários , Antibacterianos/efeitos adversos , Bactérias/classificação , Bactérias/genética , Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Estudos de Coortes , Feminino , Humanos , Lactente , Recém-Nascido , Estudos Longitudinais , Masculino , Moraxellaceae/genética , Moraxellaceae/isolamento & purificação , Pneumonia/epidemiologia , RNA Ribossômico 16S/genética , Refugiados , Infecções Respiratórias/microbiologia , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificação
15.
Int J Syst Evol Microbiol ; 67(7): 2296-2300, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28741994

RESUMO

A Gram-staining-negative, non-motile, non-pigmented, strictly aerobic and rod-shape bacterium, designated BK296T, was isolated from stream water originating from a limestone cave in Samcheok, Korea. Optimal growth of strain BK296T was observed at 30 °C, pH 7.0-8.0 and without NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BK296T belonged to the genus Perlucidibaca, forming a robust clade with a member of the genus, and was most closely related to Perlucidibaca piscinae (97.8 %). The average nucleotide identity value between strain BK296T and Perlucidibacapiscinae IMCC1704T was 79.8 %, and the genome-to-genome distance was 17.5 % on mean. The G+C content of the DNA of strain BK296T was 55.7 mol%. The major fatty acids were summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0, C12 : 0 3-OH and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The major isoprenoid quinone was ubiquinone Q-8. On the basis of phenotypic, genotypic and phylogenetic analyses, strain BK296T (=KCTC 52162T=JCM 31377T) represents a novel species of the genus Perlucidibaca, for which the name Perlucidibaca aquatica sp. nov. is proposed.


Assuntos
Água Doce/microbiologia , Moraxellaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , Cavernas , DNA Bacteriano/genética , Ácidos Graxos/química , Moraxellaceae/genética , Moraxellaceae/isolamento & purificação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/química
16.
Res Microbiol ; 167(9-10): 731-744, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27475037

RESUMO

Uncultivable microorganisms account for over 99% of all species on the planet, but their functions are yet not well characterized. Though many cultivable degraders for n-alkanes have been intensively investigated, the roles of functional n-alkane degraders remain hidden in the natural environment. This study introduces the novel magnetic nanoparticle-mediated isolation (MMI) technology in Nigerian soils and successfully separates functional microbes belonging to the families Oxalobacteraceae and Moraxellaceae, which are dominant and responsible for alkane metabolism in situ. The alkR-type n-alkane monooxygenase genes, instead of alkA- or alkP-type, were the key functional genes involved in the n-alkane degradation process. Further physiological investigation via a BIOLOG PM plate revealed some carbon (Tween 20, Tween 40 and Tween 80) and nitrogen (tyramine, l-glutamine and d-aspartic acid) sources promoting microbial respiration and n-alkane degradation. With further addition of promoter carbon or nitrogen sources, the separated functional alkane degraders significantly improved n-alkane biodegradation rates. This suggests that MMI is a promising technology for separating functional microbes from complex microbiota, with deeper insight into their ecological functions and influencing factors. The technique also broadens the application of the BIOLOG PM plate for physiological research on functional yet uncultivable microorganisms.


Assuntos
Alcanos/metabolismo , Técnicas Bacteriológicas/métodos , Moraxellaceae/isolamento & purificação , Oxalobacteraceae/isolamento & purificação , Petróleo/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Biotransformação , Carbono/metabolismo , Magnetismo , Metaboloma , Análise em Microsséries , Nanopartículas , Nitrogênio/metabolismo , Fenótipo
18.
Carbohydr Res ; 422: 5-12, 2016 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-26795079

RESUMO

The heptose-deficient inner core of the lipopolysaccharide of several pathogenic strains of the Moraxellaceae family (Moraxella, Acinetobacter) and of Bartonella henselae, respectively, comprises an α-D-glucopyranose attached to position 5 of Kdo. In continuation of the synthesis of fragments of Acinetobacter haemolyticus LPS, the branched α-Glcp-(1 → 5)[α-Kdo-(2 → 4)]-α-Kdo trisaccharide motif was elaborated. The glycosylation of a suitably protected, α-(2 → 4)-interlinked Kdo-disaccharide was achieved in high yield and fair anomeric selectivity using a 4,6-O-benzylidene N-phenyltrifluoroacetimidate glucosyl donor. Subsequent regioselective reductive benzylidene opening afforded a trisaccharide acceptor, which was extended with ß-D-glucopyranosyl and isomaltosyl residues. Global deprotection provided tri- to pentasaccharide structures corresponding to the inner core region of A. haemolyticus lipopolysaccharide.


Assuntos
Dissacarídeos/química , Dissacarídeos/síntese química , Glucose/química , Lipopolissacarídeos/química , Moraxellaceae/química , Técnicas de Química Sintética , Estereoisomerismo , Trissacarídeos/química
19.
Int J Syst Evol Microbiol ; 66(1): 201-205, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26498187

RESUMO

A Gram-stain-negative, non-spore-forming and coccus-shaped bacterial strain, designated 4DR5T, was isolated from freshwater and its taxonomic position was investigated using a polyphasic approach. Growth occurred at 10-40 °C (optimum 30 °C), at pH 6-9 (optimum pH 7) and in the presence of 0-0.4 % (w/v) NaCl (optimum 0 %) on R2A agar. On the basis of 16S rRNA gene sequence similarity, strain 4DR5T was assigned to the family Moraxellaceae of the class Gammaproteobacteria, and its closest related taxa were species of the genera Perlucidibaca (93.67 % sequence similarity), Agitococcus (93.07 %), Paraperlucidibaca (92.31-92.38 %), Alkanindiges (91.79 %) and Acinetobacter (90.24-91.23 %). The predominant isoprenoid quinone detected in strain 4DR5T was Q-10. The major cellular fatty acids were a summed feature consisting of C16 : 1ω7c and/or C16 : 1ω6c, one consisting of C18 : 1ω7c and/or C18 : 1ω6c, and C16 : 0. The major polar lipid was phosphatidylethanolamine. The genomic DNA G+C content of the strain was 61.2 mol%. The phylogenetic, chemotaxonomic and biochemical data not only supported the affiliation of strain 4DR5T to the family Moraxellaceae, but also separated it from other established genera within the family. Therefore, the novel isolate evidently represents a novel species of a new genus of Moraxellaceae, for which the name Fluviicoccus keumensis gen. nov., sp. nov. is proposed. The type strain of Fluviicoccus keumensis is 4DR5T ( = KCTC 32475T = JCM 19370T).


Assuntos
Água Doce/microbiologia , Moraxellaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Dados de Sequência Molecular , Moraxellaceae/genética , Moraxellaceae/isolamento & purificação , Fosfatidiletanolaminas/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química
20.
Int J Syst Evol Microbiol ; 65(Pt 1): 11-14, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25267870

RESUMO

An aerobic, Gram-stain-negative, non-motile coccus, designated strain GVCNT2(T), was isolated from the tonsils of a healthy adult female. Cells were oxidase- and catalase-positive, positive for the production of esterase (C4), esterase lipase (C8) and leucine arylamidase, and weakly positive for naphthol-AS-BI-phosphohydrolase and alkaline phosphatase. Cells were also capable of hydrolysing DNA. Growth was observed at 20-37 °C and in the presence of up to 1.5% NaCl. Phylogenetic analysis of near full-length 16S rRNA gene sequences indicated that the strain exhibited closest sequence similarity to Moraxella boevrei ATCC 700022(T) (94.68%) and an uncultured, unspeciated bacterial clone (strain S12-08; 99%). The major fatty acids were C18:1ω9c, C18 : 0, C16:0 and C16:1ω6c/C16:1ω7c. The DNA G+C content of strain GVCNT2(T) was 40.7 mol%. The major respiratory quinone identified was Q-8. Strain GVCNT2(T) exhibited a comparable phenotypic profile to other members of the genus Moraxella but could be distinguished based on its ability to produce acid (weakly) from d-glucose, melibiose, l-arabinose and rhamnose and on its ability to hydrolyse DNA. On the basis of phenotypic and phylogenetic differences from other members of the family Moraxellaceae, strain GVCNT2(T) is considered to represent a novel species of a new genus, for which the name Faucicola mancuniensis gen. nov., sp. nov. is proposed. The type strain of Faucicola mancuniensis is GVCNT2(T) ( =DSM 28411(T) =NCIMB 14946(T)).


Assuntos
Moraxellaceae/classificação , Orofaringe/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Feminino , Humanos , Dados de Sequência Molecular , Moraxellaceae/genética , Moraxellaceae/isolamento & purificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
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