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1.
Curr Res Struct Biol ; 6: 100112, 2023.
Article in English | MEDLINE | ID: mdl-38046895

ABSTRACT

Reducing inflammation by diet is a major goal for prevention or lowering symptoms of a variety of diseases, such as auto-immune reactions and cancers. Natural polysaccharides are increasingly gaining attention due to their potential immunomodulating capacity. Structures of those molecules are highly important for their effects on the innate immune system, cytokine production and secretion, and enzymes in immune cells. Such polysaccharides include ß-glucans, pectins, fucoidans, and fructans. To better understand the potential of these immunomodulatory molecules, it is crucial to enhance dedicated research in the area. A bibliometric analysis was performed to set a starting observation point. Major pillars of inflammation, such as pattern recognition receptors (PRRs), enzymatic production of inflammatory molecules, and involvement in specific pathways such as Nuclear-factor kappa-B (NF-kB), involved in cell transcription, survival, and cytokine production, and mitogen-activated protein kinase (MAPK), a regulator of genetic expression, mitosis, and cell differentiation. Therefore, the outcomes from polysaccharide applications in those scenarios are discussed.

2.
Plants (Basel) ; 12(14)2023 Jul 24.
Article in English | MEDLINE | ID: mdl-37514364

ABSTRACT

Pectin, a plant-derived polysaccharide, possesses immense technological and biological application value. Several variables influence pectin's physicochemical aspects, resulting in different fermentations, interactions with receptors, and other functional properties. Some of those variables are molecular weight, degree of methylation and blockiness, and monosaccharide composition. Cancer cell cytotoxicity, important fermentation-related byproducts, immunomodulation, and technological application were found in cell culture, animal models, and preclinical and clinical assessments. One of the greater extents of recent pectin technological usage involves nanoencapsulation methods for many different compounds, ranging from chemotherapy and immunotherapy to natural extracts from fruits and other sources. Structural modification (modified pectin) is also utilized to enhance the use of dietary fiber. Although pectin is already recognized as a component of significant importance, there is still a need for a comprehensive review that delves into its intricate relationships with biological effects, which depend on the source and structure of pectin. This review covers all levels of clinical research, including cell culture, animal studies, and clinical trials, to understand how the plant source and pectin structures influence the biological effects in humans and some technological applications of pectin regarding human health.

3.
Int J Mol Sci ; 23(19)2022 Sep 30.
Article in English | MEDLINE | ID: mdl-36232855

ABSTRACT

Beyond the problem in public health that protist-generated diseases represent, understanding the variety of mechanisms used by these parasites to interact with the human immune system is of biological and medical relevance. Giardia lamblia is an early divergent eukaryotic microorganism showing remarkable pathogenic strategies for evading the immune system of vertebrates. Among various multifunctional proteins in Giardia, arginine deiminase is considered an enzyme that plays multiple regulatory roles during the life cycle of this parasite. One of its most important roles is the crosstalk between the parasite and host. Such a molecular "chat" is mediated in human cells by membrane receptors called Toll-like receptors (TLRs). Here, we studied the importance of the 3D structure of giardial arginine deiminase (GlADI) to immunomodulate the human immune response through TLRs. We demonstrated the direct effect of GlADI on human TLR signaling. We predicted its mode of interaction with TLRs two and four by using the AlphaFold-predicted structure of GlADI and molecular docking. Furthermore, we showed that the immunomodulatory capacity of this virulent factor of Giardia depends on the maintenance of its 3D structure. Finally, we also showed the influence of this enzyme to exert specific responses on infant-like dendritic cells.


Subject(s)
Giardia , Giardiasis , Animals , Humans , Hydrolases , Immunity , Immunomodulation , Molecular Docking Simulation , Toll-Like Receptors
4.
Sci Rep ; 12(1): 4028, 2022 03 07.
Article in English | MEDLINE | ID: mdl-35256749

ABSTRACT

Human triosephosphate isomerase (HsTIM) is a central glycolytic enzyme and is overexpressed in cancer cells with accelerated glycolysis. Triple-negative breast cancer is highly dependent on glycolysis and is typically treated with a combination of surgery, radiation therapy, and chemotherapy. Deamidated HsTIM was recently proposed as a druggable target. Although thiol-reactive drugs affect cell growth in deamidated HsTIM-complemented cells, the role of this protein as a selective target has not been demonstrated. To delve into the usefulness of deamidated HsTIM as a selective target, we assessed its natural accumulation in breast cancer cells. We found that deamidated HsTIM accumulates in breast cancer cells but not in noncancerous cells. The cancer cells are selectively programmed to undergo cell death with thiol-reactive drugs that induced the production of methylglyoxal (MGO) and advanced glycation-end products (AGEs). In vivo, a thiol-reactive drug effectively inhibits the growth of xenograft tumors with an underlying mechanism involving deamidated HsTIM. Our findings demonstrate the usefulness of deamidated HsTIM as target to develop new therapeutic strategies for the treatment of cancers and other pathologies in which this post translationally modified protein accumulates.


Subject(s)
Breast Neoplasms , Triose-Phosphate Isomerase , Female , Glycolysis , Humans , Proteins/metabolism , Pyruvaldehyde/metabolism , Sulfhydryl Compounds , Triose-Phosphate Isomerase/metabolism
5.
Int J Mol Sci ; 22(17)2021 Aug 31.
Article in English | MEDLINE | ID: mdl-34502400

ABSTRACT

Giardiasis represents a latent problem in public health due to the exceptionally pathogenic strategies of the parasite Giardia lamblia for evading the human immune system. Strains resistant to first-line drugs are also a challenge. Therefore, new antigiardial therapies are urgently needed. Here, we tested giardial arginine deiminase (GlADI) as a target against giardiasis. GlADI belongs to an essential pathway in Giardia for the synthesis of ATP, which is absent in humans. In silico docking with six thiol-reactive compounds was performed; four of which are approved drugs for humans. Recombinant GlADI was used in enzyme inhibition assays, and computational in silico predictions and spectroscopic studies were applied to follow the enzyme's structural disturbance and identify possible effective drugs. Inhibition by modification of cysteines was corroborated using Ellman's method. The efficacy of these drugs on parasite viability was assayed on Giardia trophozoites, along with the inhibition of the endogenous GlADI. The most potent drug against GlADI was assayed on Giardia encystment. The tested drugs inhibited the recombinant GlADI by modifying its cysteines and, potentially, by altering its 3D structure. Only rabeprazole and omeprazole decreased trophozoite survival by inhibiting endogenous GlADI, while rabeprazole also decreased the Giardia encystment rate. These findings demonstrate the potential of GlADI as a target against giardiasis.


Subject(s)
Giardia lamblia/drug effects , Giardiasis/drug therapy , Hydrolases/metabolism , Animals , Antiprotozoal Agents/pharmacology , Computer Simulation , Cysteine/chemistry , Drug Evaluation, Preclinical/methods , Drug Repositioning/methods , Giardia lamblia/pathogenicity , Giardiasis/immunology , Gold Sodium Thiomalate/pharmacology , Humans , Hydrolases/drug effects , Hydrolases/ultrastructure , Omeprazole/pharmacology , Proton Pump Inhibitors/pharmacology , Rabeprazole , Thiamine/analogs & derivatives , Thiamine/pharmacology , Trophozoites/drug effects
6.
Front Immunol ; 12: 658354, 2021.
Article in English | MEDLINE | ID: mdl-34122415

ABSTRACT

The diverse and dynamic microbial community of the human gastrointestinal tract plays a vital role in health, with gut microbiota supporting the development and function of the gut immune barrier. Crosstalk between microbiota-gut epithelium and the gut immune system determine the individual health status, and any crosstalk disturbance may lead to chronic intestinal conditions, such as inflammatory bowel diseases (IBD) and celiac disease. Microbiota-derived metabolites are crucial mediators of host-microbial interactions. Some beneficially affect host physiology such as short-chain fatty acids (SCFAs) and secondary bile acids. Also, tryptophan catabolites determine immune responses, such as through binding to the aryl hydrocarbon receptor (AhR). AhR is abundantly present at mucosal surfaces and when activated enhances intestinal epithelial barrier function as well as regulatory immune responses. Exogenous diet-derived indoles (tryptophan) are a major source of endogenous AhR ligand precursors and together with SCFAs and secondary bile acids regulate inflammation by lowering stress in epithelium and gut immunity, and in IBD, AhR expression is downregulated together with tryptophan metabolites. Here, we present an overview of host microbiota-epithelium- gut immunity crosstalk and review how microbial-derived metabolites contribute to host immune homeostasis. Also, we discuss the therapeutic potential of bacterial catabolites for IBD and celiac disease and how essential dietary components such as dietary fibers and bacterial tryptophan catabolites may contribute to intestinal and systemic homeostasis.


Subject(s)
Bacteria/metabolism , Gastrointestinal Microbiome , Host Microbial Interactions , Intestinal Mucosa/metabolism , Intestinal Mucosa/microbiology , Animals , Bile Acids and Salts/metabolism , Dietary Fiber , Disease Susceptibility , Gastrointestinal Microbiome/immunology , Homeostasis , Host Microbial Interactions/immunology , Humans , Inflammatory Bowel Diseases/etiology , Inflammatory Bowel Diseases/metabolism , Ligands , Receptors, Aryl Hydrocarbon/metabolism , Tryptophan/metabolism
7.
J Biomater Appl ; 36(4): 638-647, 2021 10.
Article in English | MEDLINE | ID: mdl-33541171

ABSTRACT

Cell-encapsulation is used for preventing therapeutic cells from being rejected by the host. The technology to encapsulate cells in immunoprotective biomaterials, such as alginate, commonly involves application of an electrostatic droplet generator for reproducible manufacturing droplets of similar size and with similar surface properties. As many factors influencing droplet formation are still unknown, we investigated the impact of several parameters and fitted them to equations to make procedures more reproducible and allow optimal control of capsule size and properties. We demonstrate that droplet size is dependent on an interplay between the critical electric potential (Uc,), the needle size, and the distance between the needle and the gelation bath, and that it can be predicted with the equations proposed. The droplet formation was meticulously studied and followed by a high-speed camera. The X-ray photoelectron analysis demonstrated optimal gelation and substitution of sodium with calcium on alginate surfaces while the atomic force microscopy analysis demonstrated a low but considerable variation in surface roughness and low surface stiffness. Our study shows the importance of documenting critical parameters to guarantee reproducible manufacturing of beads with constant and adequate size and preventing batch-to-batch variations.


Subject(s)
Biocompatible Materials/chemistry , Capsules/chemistry , Static Electricity , Alginates/chemistry , Gels/chemistry , Particle Size , Photoelectron Spectroscopy/methods , Surface Properties , Surface Tension , Viscosity
8.
Sci Rep ; 10(1): 1690, 2020 02 03.
Article in English | MEDLINE | ID: mdl-32015377

ABSTRACT

Dietary fibers have been shown to exert immune effects via interaction with pattern recognition receptors (PRR) such as toll-like receptors (TLR) and nucleotide-binding oligomerization domain (NOD)-like receptors. Pectin is a dietary fiber that interacts with PRR depending on its chemical structure. Papaya pectin retains different chemical structures at different ripening stages. How this influence PRR signaling is unknown. The aim of this work was to determine how ripening influences pectin structures and their ability to interact with TLR2, 3, 4, 5 and 9, and NOD1 and 2. It was evaluated the interaction of the water-soluble fractions rich in pectin extracted from unripe to ripe papayas. The pectin extracted from ripe papayas activated all the TLR and, to a lesser extent, the NOD receptors. The pectin extracted from unripe papayas also activated TLR2, 4 and 5 but inhibited the activation of TLR3 and 9. The differences in pectin structures are the higher methyl esterification and smaller galacturonan chains of pectin from ripe papayas. Our finding might lead to selection of ripening stages for tailored modulation of PRR to support or attenuate immunity.


Subject(s)
Carica/metabolism , Pectins/metabolism , Receptors, Immunologic/metabolism , CARD Signaling Adaptor Proteins/metabolism , Dietary Fiber/metabolism , Receptors, Pattern Recognition/metabolism , Signal Transduction/physiology , Toll-Like Receptors/metabolism
9.
PeerJ ; 2: e427, 2014.
Article in English | MEDLINE | ID: mdl-25024905

ABSTRACT

Five novel strains of Photobacterium (A-394T, A-373, A-379, A-397 and A-398) were isolated from bleached coral Madracis decactis (scleractinian) in the remote St Peter & St Archipelago (SPSPA), Mid-Atlantic Ridge, Brazil. Healthy M. decactis specimens were also surveyed, but no strains were related to them. The novel isolates formed a distinct lineage based on the 16S rRNA, recA, and rpoA gene sequences analysis. Their closest phylogenetic neighbours were Photobacterium rosenbergii, P. gaetbulicola, and P. lutimaris, sharing 96.6 to 95.8% 16S rRNA gene sequence similarity. The novel species can be differentiated from the closest neighbours by several phenotypic and chemotaxonomic markers. It grows at pH 11, produces tryptophane deaminase, presents the fatty acid C18:0, but lacks C16:0 iso. The whole cell protein profile, based in MALDI-TOF MS, distinguished the strains of the novel species among each other and from the closest neighbors. In addition, we are releasing the whole genome sequence of the type strain. The name Photobacterium sanctipauli sp. nov. is proposed for this taxon. The G + C content of the type strain A-394(T) (= LMG27910(T) = CAIM1892(T)) is 48.2 mol%.

10.
Curr Microbiol ; 69(4): 405-11, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24824949

ABSTRACT

Three novel isolates (A-354(T), A-328, and A-384) were retrieved from apparently healthy scleractinian Madracis decactis in the remote St Peter & St Paul Archipelago, Mid-Atlantic Ridge, Brazil. The novel isolates formed a distinct lineage based on the phylogenetic reconstruction using the 16S rRNA and pyrH gene sequences. They fell into the Mediterranei clade and their closest phylogenetic neighbour was V. mediterranei species, sharing upto 98.1 % 16S rRNA gene sequence similarity. Genomic analysis including in silico DDH, MLSA, AAI and genomic signature distinguished A-354(T) from V. mediterranei LMG 19703 (=AK1) with values of 33.3, 94.2, 92 %, and 11.3, respectively. Phenotypically, the novel isolates can be differentiated from V. mediterranei based on the four following features. They do not grow at 8 % NaCl; use D-gluconic acid but not L-galactonic acid lactone as carbon source; and do not have the fatty acid C18:0. Differentiation from both the other Mediterranei clade species (V. maritimus and V. variabilis) is supported by fifteen features. The novel species show lysine decarboxylase and tryptophan deaminase, but not gelatinase and arginine dihydrolase activity; produce acetoin; use α-D-lactose, N-acetyl-D-galactosamine, myo-Inositol, D-gluconic acid, and ß-hydroxy-D,L-butyric acid; and present the fatty acids C14:0 iso, C15:0 anteiso, C16:0 iso, C17:0 anteiso, and C17:1x8c . Whole-cell protein profiles, based on MALDI-TOF, showed that the isolates are not clonal and also distinguished them from the closes phylogenetic neighbors. The name Vibrio madracius sp. nov. is proposed to encompass these novel isolates. The G+C content of the type strain A-354(T) (=LMG 28124(T)=CBAS 482(T)) is 44.5 mol%.


Subject(s)
Anthozoa/microbiology , Vibrio/classification , Vibrio/isolation & purification , Animals , Bacterial Typing Techniques , Brazil , DNA, Bacterial/genetics , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Vibrio/genetics
11.
Syst Appl Microbiol ; 36(2): 116-27, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23333025

ABSTRACT

The Central Andean Highlands are the center of origin of the potato plant (Solanum tuberosum). Ages of mutualism between potato plants and soil bacteria in this region support the hypothesis that Andean soils harbor interesting plant growth-promoting (PGP) bacteria. Therefore, the aim of this study was to isolate rhizobacteria from Andean ecosystems, and to identify those with PGP properties. A total of 585 bacterial isolates were obtained from eight potato fields in the Andes and they were screened for suppression of Phytophthora infestans and Rhizoctonia solani. Antagonistic mechanisms were determined and antagonistic isolates were further tested for phosphate solubilization, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and production of NH3- and indole-3-acetic acid (IAA). PGP was studied in healthy and R. solani diseased plantlets under growth room conditions. Performance was compared to the commercial strain B. subtilis FZB24(®) WG. Isolates were dereplicated with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), and identified with 16S rRNA gene sequencing and multi locus sequence analysis (MLSA). A total of 10% of the isolates were effective antagonists, of which many were able to solubilize phosphate, and produce IAA, ACC deaminase, NH3 and hydrogen cyanide (HCN). During growth room experiments, 23 antagonistic isolates were associated with plant growth-promotion and/or disease suppression. Ten isolates had a statistically significant impact on test parameters compared to the uninoculated control. Three isolates significantly promoted plant growth in healthy plantlets compared to the commercial strain, and seven isolates outperformed the commercial strain in in vitro R. solani diseased plantlets.


Subject(s)
Antibiosis , Bacteria/isolation & purification , Bacteria/metabolism , Plant Growth Regulators/metabolism , Soil Microbiology , Solanum tuberosum/growth & development , Solanum tuberosum/microbiology , Bacteria/chemistry , Bacteria/classification , Bolivia , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Peru , Phytophthora infestans/growth & development , RNA, Ribosomal, 16S/genetics , Rhizoctonia/growth & development , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
12.
Int J Syst Evol Microbiol ; 63(Pt 3): 1119-1125, 2013 Mar.
Article in English | MEDLINE | ID: mdl-22729025

ABSTRACT

The phylogenetic position of a cellulose-producing acetic acid bacterium, strain ID13488, isolated from commercially available Colombian homemade fruit vinegar, was investigated. Analyses using nearly complete 16S rRNA gene sequences, nearly complete 16S-23S rRNA gene internal transcribed spacer (ITS) sequences, as well as concatenated partial sequences of the housekeeping genes dnaK, groEL and rpoB, allocated the micro-organism to the genus Gluconacetobacter, and more precisely to the Gluconacetobacter xylinus group. Moreover, the data suggested that the micro-organism belongs to a novel species in this genus, together with LMG 1693(T), a non-cellulose-producing strain isolated from vinegar by Kondo and previously classified as a strain of Gluconacetobacter xylinus. DNA-DNA hybridizations confirmed this finding, revealing a DNA-DNA relatedness value of 81 % between strains ID13488 and LMG 1693(T), and values <70 % between strain LMG 1693(T) and the type strains of the closest phylogenetic neighbours. Additionally, the classification of strains ID13488 and LMG 1693(T) into a single novel species was supported by amplified fragment length polymorphism (AFLP) and (GTG)5-PCR DNA fingerprinting data, as well as by phenotypic data. Strains ID13488 and LMG 1693(T) could be differentiated from closely related species of the genus Gluconacetobacter by their ability to produce 2- and 5-keto-d-gluconic acid from d-glucose, their ability to produce acid from sucrose, but not from 1-propanol, and their ability to grow on 3 % ethanol in the absence of acetic acid and on ethanol, d-ribose, d-xylose, sucrose, sorbitol, d-mannitol and d-gluconate as carbon sources. The DNA G+C content of strains ID13488 and LMG 1693(T) was 58.0 and 60.7 mol%, respectively. The major ubiquinone of LMG 1693(T) was Q-10. Taken together these data indicate that strains ID13488 and LMG 1693(T) represent a novel species of the genus Gluconacetobacter for which the name Gluconacetobacter medellinensis sp. nov. is proposed. The type strain is LMG 1693(T) ( = NBRC 3288(T) = Kondo 51(T)).


Subject(s)
Acetic Acid , Cellulose/biosynthesis , Gluconacetobacter/classification , Phylogeny , Amplified Fragment Length Polymorphism Analysis , Bacterial Typing Techniques , Base Composition , Colombia , DNA, Bacterial/genetics , Fatty Acids/analysis , Genes, Bacterial , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
13.
Int J Syst Evol Microbiol ; 62(Pt 7): 1457-1464, 2012 Jul.
Article in English | MEDLINE | ID: mdl-21841003

ABSTRACT

Several Gram-negative-staining, facultatively anaerobic bacterial isolates were obtained from Eucalyptus seedlings showing symptoms of bacterial blight and dieback in Colombia, Rwanda and South Africa. Partial 16S rRNA gene sequencing, together with partial gyrB sequencing, placed the isolates in the genus Pantoea and indicated that they constituted three novel species. Multilocus sequence analysis (MLSA) based on partial sequences of gyrB, rpoB, infB and atpD revealed Pantoea dispersa, Pantoea eucrina and Pantoea cypripedii as their closest phylogenetic relatives. DNA-DNA hybridization studies confirmed the classification of the new isolates as three novel species and phenotypic tests allowed them to be differentiated from their closest phylogenetic neighbours. The names Pantoea rodasii sp. nov. [type strain LMG 26273(T)=BD 943(T) (deposited with the Plant Pathogenic and Plant Protecting Bacteria Collection, South Africa)=BCC 581(T) (deposited with the Bacterial Culture Collection, Forestry and Agricultural Institute, South Africa)], Pantoea rwandensis sp. nov. (type strain LMG 26275(T)=BD 944(T)=BCC 571(T)) and Pantoea wallisii sp. nov. (type strain LMG 26277(T)=BD 946(T)=BCC 682(T)) are proposed.


Subject(s)
Eucalyptus/microbiology , Pantoea/classification , Pantoea/isolation & purification , Anaerobiosis , Bacterial Typing Techniques , Cluster Analysis , Colombia , DNA Gyrase/genetics , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Multilocus Sequence Typing , Nucleic Acid Hybridization , Pantoea/genetics , Pantoea/physiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Rwanda , Sequence Analysis, DNA , South Africa
14.
Mem Inst Oswaldo Cruz ; 106(4): 394-9, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21739025

ABSTRACT

An online scheme to assign Stenotrophomonas isolates to genomic groups was developed using the multilocus sequence analysis (MLSA), which is based on the DNA sequencing of selected fragments of the housekeeping genes ATP synthase alpha subunit (atpA), the recombination repair protein (recA), the RNA polymerase alpha subunit (rpoA) and the excision repair beta subunit (uvrB). This MLSA-based scheme was validated using eight of the 10 Stenotrophomonas species that have been previously described. The environmental and nosocomial Stenotrophomonas strains were characterised using MLSA, 16S rRNA sequencing and DNA-DNA hybridisation (DDH) analyses. Strains of the same species were found to have greater than 95% concatenated sequence similarity and specific strains formed cohesive readily recognisable phylogenetic groups. Therefore, MLSA appeared to be an effective alternative methodology to amplified fragment length polymorphism fingerprint and DDH techniques. Strains of Stenotrophomonas can be readily assigned through the open database resource that was developed in the current study (www.steno.lncc.br/).


Subject(s)
DNA, Bacterial/genetics , Multilocus Sequence Typing/methods , RNA, Ribosomal, 16S/genetics , Stenotrophomonas/genetics , Bacterial Typing Techniques , DNA-Directed RNA Polymerases/genetics , Humans , Phylogeny
15.
Mem. Inst. Oswaldo Cruz ; 106(4): 394-399, June 2011. ilus, tab
Article in English | LILACS | ID: lil-592180

ABSTRACT

An online scheme to assign Stenotrophomonas isolates to genomic groups was developed using the multilocus sequence analysis (MLSA), which is based on the DNA sequencing of selected fragments of the housekeeping genes ATP synthase alpha subunit (atpA), the recombination repair protein (recA), the RNA polymerase alpha subunit (rpoA) and the excision repair beta subunit (uvrB). This MLSA-based scheme was validated using eight of the 10 Stenotrophomonas species that have been previously described. The environmental and nosocomial Stenotrophomonas strains were characterised using MLSA, 16S rRNA sequencing and DNA-DNA hybridisation (DDH) analyses. Strains of the same species were found to have greater than 95 percent concatenated sequence similarity and specific strains formed cohesive readily recognisable phylogenetic groups. Therefore, MLSA appeared to be an effective alternative methodology to amplified fragment length polymorphism fingerprint and DDH techniques. Strains of Stenotrophomonas can be readily assigned through the open database resource that was developed in the current study (www.steno.lncc.br/).


Subject(s)
Humans , DNA, Bacterial , Multilocus Sequence Typing/methods , Stenotrophomonas , Bacterial Typing Techniques , DNA-Directed RNA Polymerases , Phylogeny
16.
Int J Syst Evol Microbiol ; 61(Pt 2): 362-368, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20305064

ABSTRACT

Eight Vibrio isolates originating from the marine corals Mussismilia hispida and Phyllogorgia dilatata and the zoanthids Palythoa caribaeorum and Palythoa variabilis in Brazil and the Pacific white shrimp (Litopenaeus vannamei) in Ecuador were studied by means of a polyphasic approach. The novel isolates formed a tight monophyletic group in the genus Vibrio and were closely related to species of the Vibrio harveyi group, to which they showed more than 99 % 16S rRNA gene sequence similarity. Analysis based on concatenated sequences of the following seven genes, 16S rRNA, gyrB, recA, rpoA, topA, pyrH and mreB (5633 bp in length), showed clear separation between the isolates and species of the V. harveyi group. Amplified fragment length polymorphism (AFLP) analysis, performed previously, revealed that a representative isolate of this group, LMG 20370, was clearly separate from known Vibrio species (it belonged to the so-called AFLP cluster A31). DNA-DNA hybridization (DDH) experiments with representative isolates and type strains of the V. harveyi species group revealed high DDH between the novel isolates (more than 74 %) and less than 70 % DDH towards type strains of related Vibrio species, proving the novel species status of the isolates. Phenotypically, the novel species belongs to the arginine dihydrolase (A)-negative, lysine decarboxylase (L)-positive and ornithine decarboxylase (O)-positive (A-/L+/O+) cluster reported previously. Most species of the V. harveyi group (i.e. Vibrio rotiferianus, V. harveyi, V. parahaemolyticus and V. alginolyticus) also belong to this A-/L+/O+ cluster. However, several phenotypic features can be used for the identification of the novel species. In contrast to its closest phylogenetic neighbours, the novel species exhibits esterase (C4) and N-acetyl-ß-glucosaminidase activities, but it does not produce acetoin, does not use citrate, α-ketoglutaric acid or propionic acid and does not ferment melibiose. The novel species can also be differentiated on the basis of the presence of the fatty acids C(17 : 0,) C(17 : 1)ω8c, iso-C(17 : 0) and iso-C(13 : 0) and the absence of the fatty acid C(18 : 0). The name Vibrio communis sp. nov. is proposed for this taxon. Strain R-40496(T) (=LMG 25430(T) =CAIM 1816(T)) is the type strain.


Subject(s)
Anthozoa/microbiology , Penaeidae/microbiology , Phylogeny , Vibrio/classification , Amplified Fragment Length Polymorphism Analysis , Animals , Bacterial Typing Techniques , Brazil , DNA, Bacterial/genetics , Ecuador , Fatty Acids/analysis , Molecular Sequence Data , Multilocus Sequence Typing , Phenotype , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Vibrio/genetics , Vibrio/isolation & purification
17.
Int J Syst Evol Microbiol ; 61(Pt 1): 60-64, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20154332

ABSTRACT

A Gram-negative, aerobic bacterium, designated R-40509(T), was isolated from mucus of the reef builder coral (Mussismilia hispida) located in the São Sebastião Channel, São Paulo, Brazil. The strain was oxidase-positive and catalase-negative, and required Na(+) for growth. Its phylogenetic position was in the genus Marinobacterium and the closest related species were Marinobacterium sediminicola, Marinobacterium maritimum and Marinobacterium stanieri; the isolate exhibited 16S rRNA gene sequence similarities of 97.5-98.0 % with the type strains of these species. 16S rRNA gene sequence similarities with other type strains of the genus Marinobacterium were below 96 %. DNA-DNA hybridizations between strain R-40509(T) and the type strains of the phylogenetically closest species of the genus Marinobacterium revealed less than 70 % DNA-DNA relatedness, supporting the novel species status of the strain. Phenotypic characterization revealed that the strain was able to grow at 15-42 °C and in medium containing up to 9 % NaCl. The isolate could be differentiated from phenotypically related species by several features, including its ability to utilize d-alanine, l-alanine, bromosuccinic acid, ß-hydroxybutyric acid and α-ketovaleric acid, but not acetate or l-arabinose. It produced acetoin (Voges-Proskauer), but did not have esterase lipase (C8) or catalase activities. It possessed C(18 : 1)ω7c (35 %), summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c; 25 %) and C(16 : 0) (22 %) as major cellular fatty acids. The DNA G+C content was 58.5 mol%. The name Marinobacterium coralli sp. nov. is proposed to accommodate this novel isolate; the type strain is R-40509(T) (=LMG 25435(T) =CAIM 1449(T)).


Subject(s)
Alteromonadaceae/classification , Alteromonadaceae/isolation & purification , Anthozoa/microbiology , Alteromonadaceae/genetics , Alteromonadaceae/physiology , Animals , Bacterial Typing Techniques , Base Composition , Brazil , Catalase/metabolism , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fatty Acids/analysis , Molecular Sequence Data , Nucleic Acid Hybridization , Oxidoreductases/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sodium Chloride/metabolism , Temperature
18.
Int J Syst Evol Microbiol ; 61(Pt 4): 926-931, 2011 Apr.
Article in English | MEDLINE | ID: mdl-20495025

ABSTRACT

A Gram-negative, rod-shaped, non-spore-forming and nitrogen-fixing bacterium, designated ICB 89(T), was isolated from stems of a Brazilian sugar cane variety widely used in organic farming. 16S rRNA gene sequence analysis revealed that strain ICB 89(T) belonged to the genus Stenotrophomonas and was most closely related to Stenotrophomonas maltophilia LMG 958(T), Stenotrophomonas rhizophila LMG 22075(T), Stenotrophomonas nitritireducens L2(T), [Pseudomonas] geniculata ATCC 19374(T), [Pseudomonas] hibiscicola ATCC 19867(T) and [Pseudomonas] beteli ATCC 19861(T). DNA-DNA hybridization together with chemotaxonomic data and biochemical characteristics allowed the differentiation of strain ICB 89(T) from its nearest phylogenetic neighbours. Therefore, strain ICB 89(T) represents a novel species, for which the name Stenotrophomonas pavanii sp. nov. is proposed. The type strain is ICB 89(T) ( = CBMAI 564(T)  = LMG 25348(T)).


Subject(s)
Nitrogen Fixation , Saccharum/microbiology , Stenotrophomonas/classification , Stenotrophomonas/isolation & purification , Brazil , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Nucleic Acid Hybridization , Organic Agriculture , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Stenotrophomonas/genetics , Stenotrophomonas/physiology
19.
Int J Syst Evol Microbiol ; 61(Pt 5): 1170-1175, 2011 May.
Article in English | MEDLINE | ID: mdl-20562247

ABSTRACT

A Gram-negative, aerobic bacterium, designated strain R-40503(T), was isolated from mucus of the reef-builder coral Mussismilia hispida, located in the São Sebastião Channel, São Paulo, Brazil. Phylogenetic analyses revealed that strain R-40503(T) belongs to the genus Marinomonas. The 16S rRNA gene sequence similarity of R-40503(T) was above 97 % with the type strains of Marinomonas vaga, M. basaltis, M. communis and M. pontica, and below 97 % with type strains of the other Marinomonas species. Strain R-40503(T) showed less than 35 % DNA-DNA hybridization (DDH) with the type strains of the phylogenetically closest Marinomonas species, demonstrating that it should be classified into a novel species. Amplified fragment length polymorphism (AFLP), chemotaxonomic and phenotypic analyses provided further evidence for the proposal of a novel species. Concurrently, a close genomic relationship between M. basaltis and M. communis was observed. The type strains of these two species showed 78 % DDH and 63 % AFLP pattern similarity. Their phenotypic features were very similar, and their DNA G+C contents were identical (46.3 mol%). Collectively, these data demonstrate unambiguously that Marinomonas basaltis is a later heterotypic synonym of Marinomonas communis. Several phenotypic features can be used to discriminate between Marinomonas species. The novel strain R-40503(T) is clearly distinguishable from its neighbours. For instance, it shows oxidase and urease activity, utilizes l-asparagine and has the fatty acid C(12 : 1) 3-OH but lacks C(10 : 0) and C(12 : 0). The name Marinomonas brasilensis sp. nov. is proposed, with the type strain R-40503(T) ( = R-278(T)  = LMG 25434(T)  = CAIM 1459(T)). The DNA G+C content of strain R-40503(T) is 46.5 mol%.


Subject(s)
Anthozoa/microbiology , Marinomonas/classification , Marinomonas/isolation & purification , Amplified Fragment Length Polymorphism Analysis , Animals , Base Composition , Brazil , DNA, Bacterial/genetics , Fatty Acids/metabolism , Marinomonas/genetics , Marinomonas/metabolism , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Seawater/microbiology
20.
Int J Syst Evol Microbiol ; 60(Pt 12): 2843-2848, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20081016

ABSTRACT

Four novel isolates (R-40508(T), R-40507, R-40903 and R-21419) were obtained from different cnidarian species (Phyllogorgia dilatata, Merulina ampliata and Palythoa caribaeorum) from different places in Brazil and Australia. The novel isolates formed a tight phylogenetic group based on 16S rRNA, recA, topA, ftsZ, mreB and rpoA gene sequences. Their closest phylogenetic neighbours were the type strains of Photobacterium leiognathi, P. rosenbergii and P. halotolerans, sharing 97.1-97.5 % 16S rRNA gene sequence similarity. DNA-DNA hybridization between a representative strain (R-40508(T)) and the type strains of these Photobacterium species revealed less than 20 % relatedness, showing that the new isolates belong to a novel species. Several phenotypic features allow the differentiation of the novel species from its closest phylogenetic neighbours. It has gelatinase and lipase activity and can utilize melibiose, but it cannot grow on 6 % NaCl. In addition, the novel species has the fatty acid iso-C(16 : 0), but lacks the fatty acids C(17 : 0), C(17 : 0) cyclo, iso-C(17 : 0), C(17 : 1)ω8c and iso-C(17 : 1)ω9c. The name Photobacterium jeanii sp. nov. is proposed for this species, with the type strain R-40508(T) (=LMG 25436(T) =CAIM 1817(T)). The G+C content of the type strain is 45.5mol%.


Subject(s)
Anthozoa/microbiology , Photobacterium/classification , Phylogeny , Animals , Australia , Bacterial Typing Techniques , Base Composition , Brazil , DNA, Bacterial/genetics , Fatty Acids/chemistry , Genes, Bacterial , Molecular Sequence Data , Nucleic Acid Hybridization , Photobacterium/genetics , Photobacterium/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
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