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1.
Microb Genom ; 6(12)2020 12.
Article in English | MEDLINE | ID: mdl-33206037

ABSTRACT

Sellimonas intestinalis is a Gram-positive and anaerobic bacterial species previously considered as uncultivable. Although little is known about this Lachnospiraceae family member, its increased abundance has been reported in patients who have recovered from intestinal homeostasis after dysbiosis events. In this context, the aim of the present study was to take advantage of a massive in vitro culture protocol that allowed the recovery of extremely oxygen-sensitive species from faecal samples, which led to isolation of S. intestinalis. Whole genome analyses of 11 S. intestinalis genomes revealed that this species has a highly conserved genome with 99.7 % 16S rRNA gene sequence similarity, average nucleotide polymorphism results >95, and 50.1 % of its coding potential being part of the core genome. Despite this, the variable portion of its genome was informative enough to reveal the existence of three lineages (lineage-I including isolates from Chile and France, lineage-II from South Korea and Finland, and lineage-III from China and one isolate from the USA) and evidence of some recombination signals. The identification of a cluster of orthologous groups revealed a high number of genes involved in metabolism, including amino acid and carbohydrate transport as well as energy production and conversion, which matches with the metabolic profile previously reported for microbiota from healthy individuals. Additionally, virulence factors and antimicrobial resistance genes were found (mainly in lineage-III), which could favour their survival during antibiotic-induced dysbiosis. These findings provide the basis of knowledge about the potential of S. intestinalis as a bioindicator of intestinal homeostasis recovery and contribute to advancing the characterization of gut microbiota members with beneficial potential.


Subject(s)
Clostridiales/classification , Drug Resistance, Bacterial , Whole Genome Sequencing/methods , Bacterial Proteins/genetics , Clostridiales/genetics , Clostridiales/isolation & purification , Feces/microbiology , Gene Regulatory Networks , Healthy Volunteers , High-Throughput Nucleotide Sequencing , Humans , Phylogeny , RNA, Ribosomal, 16S/genetics
2.
Curr Microbiol ; 76(6): 713-722, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30968206

ABSTRACT

Despite the broad assessment of sponge bacterial diversity through cultivation-independent and dependent strategies, the knowledge focusing on cultivable anaerobes from this holobiont is still incipient. Plakina is a genus with the highest number of described species from the smallest of poriferan classes, Homoscleromorpha. The Brazilian Atlantic coast has been presenting itself as a hotspot for the discovery of new plakinidae species, with initial surveys just now concerning to characterize their microbiome. The current study aimed to isolate and identify strict anaerobes from recently described species of Plakina collected at the coast of Cabo Frio, RJ. Samples of four sympatric morphotypes of Plakina cyanorosea and Plakina cabofriense were collected on the coast of Cabo Frio, RJ. Using five different culture media, a total of 93 bacterial isolates were recovered, among which 60 were strict anaerobes and, ultimately, 34 remaining viable. A total of 76.5% from these strains were mostly identified as Clostridium bifermentans by mass spectrometry and 82.4% identified by 16S rRNA sequencing, almost all of them affiliated to the genus Paraclostridium, and with one isolate identified as Clostridium butyricum by both techniques. None of the anaerobic bacteria exhibited antimicrobial activity by the adopted screening test. The present work highlights not only the need for cultivation and characterization of the anaerobic microbiota from marine sponges but also adds the existing scarce knowledge of culturable bacterial communities from Homoscleromorph sponges from Brazilian coast.


Subject(s)
Bacteria, Anaerobic/classification , Bacteria, Anaerobic/isolation & purification , Clostridiales/classification , Clostridiales/isolation & purification , Porifera/microbiology , Aerobiosis , Anaerobiosis , Animals , Anti-Infective Agents/metabolism , Aquatic Organisms/microbiology , Atlantic Ocean , Bacteria, Anaerobic/chemistry , Bacteria, Anaerobic/genetics , Bacteriological Techniques , Brazil , Clostridiales/chemistry , Clostridiales/genetics , Clostridium bifermentans , Clostridium butyricum , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Mass Spectrometry , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
3.
Int J Syst Evol Microbiol ; 69(4): 1142-1148, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30767850

ABSTRACT

A novel Gram-stain-positive, non-motile, non-spore-forming coccus-shaped obligately anaerobic bacterium was recovered from a fecal sample obtained from an individual from a traditional community located on the southern coast of Peru. The results of analysis based on 16S rRNA gene sequencing indicated the novel bacterium to be phylogenetically distinct from other genera of members of the Peptoniphilaceae family, sharing a loose affinity with the genera Ezakiella, Finegoldia, Gallicola and Parvimonas. The major cellular fatty acids of the novel isolate were determined to be C16:0, C17:1ω8c, and C18:1ω9c. The DNA G+C content was 29.9 mol%. End products of metabolism from peptone yeast glucose broth (PYG) were determined to be acetate and methyl succinate. The diagnostic diamino acid present in the cell wall was lysine. On the basis of the phenotypic, chemotaxonomic and phylogenetic results the organism is a member of a novel genus belonging to the family Peptoniphilaceae for which the name Citroniella saccharovorans gen nov. sp. nov., is proposed. The type strain is M6.X9T (DSM 29873T=CCUG 66799T).


Subject(s)
Clostridiales/classification , Feces/microbiology , Phylogeny , Bacterial Typing Techniques , Base Composition , Clostridiales/isolation & purification , DNA, Bacterial/genetics , Fatty Acids/chemistry , Humans , Peru , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
4.
J Biotechnol ; 257: 178-186, 2017 Sep 10.
Article in English | MEDLINE | ID: mdl-28595834

ABSTRACT

The strictly anaerobic Peptoniphilaceae bacterium str. ING2-D1G (=DSM 28672=LMG 28300) was isolated from a mesophilic laboratory-scale completely stirred tank biogas reactor (CSTR) continuously co-digesting maize silage, pig and cattle manure. Based on 16S rRNA gene sequence comparison, the closest described relative to this strain is Peptoniphilus obesi ph1 showing 91.2% gene sequence identity. The most closely related species with a validly published name is Peptoniphilus indolicus DSM 20464T whose 16S rRNA gene sequence is 90.6% similar to the one of strain ING2-D1G. The genome of the novel strain was completely sequenced and manually annotated to reconstruct its metabolic potential regarding anaerobic digestion of biomass. The strain harbors a circular chromosome with a size of 1.6 Mb that contains 1466 coding sequences, 53 tRNA genes and 4 ribosomal RNA (rrn) operons. The genome carries a 28,261bp prophage insertion comprising 47 phage-related coding sequences. Reconstruction of fermentation pathways revealed that strain ING2-D1G encodes all enzymes for hydrogen, lactate and acetate production, corroborating that it is involved in the acido- and acetogenic phase of the biogas process. Comparative genome analyses of Peptoniphilaceae bacterium str. ING2-D1G and its closest relative Peptoniphilus obesi ph1 uncovered rearrangements, deletions and insertions within the chromosomes of both strains substantiating a divergent evolution. In addition to genomic analyses, a physiological and phenotypic characterization of the novel isolate was performed. Grown in Brain Heart Infusion Broth with added yeast extract, cells were spherical to ovoid, catalase- and oxidase-negative and stained Gram-positive. Optimal growth occurred between 35 and 37°C and at a pH value of 7.6. Fermentation products were acetate, butanoate and carbon dioxide.


Subject(s)
Biofuels/microbiology , Bioreactors/microbiology , Clostridiales/classification , Clostridiales/genetics , Clostridiales/isolation & purification , Genome, Bacterial , Phylogeny , Animals , Bacterial Typing Techniques , Base Sequence , Cattle , Clostridiales/physiology , DNA, Bacterial , Fatty Acids/metabolism , Fermentation , Genes, Bacterial/genetics , Manure/microbiology , Metabolic Networks and Pathways , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Silage/microbiology , Swine , Zea mays
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