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PLoS Neglected Tropical Diseases Apr 2021Naja atra is a major venomous snake found in Taiwan. The bite of this snake causes extensive wound necrosis or necrotizing soft tissue infection. Conventional microbial...
An investigation of conventional microbial culture for the Naja atra bite wound, and the comparison between culture-based 16S Sanger sequencing and 16S metagenomics of the snake oropharyngeal bacterial microbiota.
Naja atra is a major venomous snake found in Taiwan. The bite of this snake causes extensive wound necrosis or necrotizing soft tissue infection. Conventional microbial culture-based techniques may fail to identify potential human pathogens and render antibiotics ineffective in the management of wound infection. Therefore, we evaluated 16S Sanger sequencing and next-generation sequencing (NGS) to identify bacterial species in the oropharynx of N. atra. Using conventional microbial culture methods and the VITEK 2 system, we isolated nine species from snakebite wounds. On the basis of the 16S Sanger sequencing of bacterial clones from agar plates, we identified 18 bacterial species in the oropharynx of N. atra, including Morganella morganii, Proteus vulgaris, and Proteus mirabilis, which were also present in the infected bite wound. Using NGS of 16S metagenomics, we uncovered more than 286 bacterial species in the oropharynx of N. atra. In addition, the bacterial species identified using 16S Sanger sequencing accounted for only 2% of those identified through NGS of 16S metagenomics. The bacterial microbiota of the oropharynx of N. atra were modeled better using NGS of 16S metagenomics compared to microbial culture-based techniques. Stenotrophomonas maltophilia, Acinetobacter baumannii, and Proteus penneri were also identified in the NGS of 16S metagenomics. Understanding the bacterial microbiota that are native to the oropharynx of N. atra, in addition to the bite wound, may have additional therapeutic implications regarding empiric antibiotic selection for managing N. atra bites.
Topics: Adult; Aged; Animals; Anti-Bacterial Agents; Bacteria; Female; High-Throughput Nucleotide Sequencing; Humans; Male; Metagenomics; Middle Aged; Naja naja; Oropharynx; RNA, Ribosomal, 16S; Snake Bites; Taiwan; Wound Infection
PubMed: 33857127
DOI: 10.1371/journal.pntd.0009331 -
European Journal of Biochemistry Aug 1996An acidic O-specific polysaccharide isolated from Proteus penneri 52 contains D-galacturonic acid, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2-deoxy-D-galactose in...
An acidic O-specific polysaccharide isolated from Proteus penneri 52 contains D-galacturonic acid, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2-deoxy-D-galactose in the ratio 1:2:2. On the basis of sugar analysis and NMR spectroscopy, which included one-dimensional TOCSY, two-dimensional COSY, heteronuclear 13C, 1H-COSY, and rotating-frame NOE spectroscopy, the following structure of the pentasaccharide repeating unit of the O-specific polysaccharide was established: [sequence: see text] Cross-reactivity of the anti-P-penneri 52 O-serum with other strains of P. penneri isolated in Germany. Poland, USA, and Canada is discussed and a new Proteus serogroup is proposed.
Topics: Carbohydrate Conformation; Carbohydrate Sequence; Cross Reactions; Immune Sera; Magnetic Resonance Spectroscopy; Molecular Sequence Data; O Antigens; Oligosaccharides; Proteus
PubMed: 8797860
DOI: 10.1111/j.1432-1033.1996.0245h.x -
European Journal of Biochemistry Aug 2001The acidic O-specific polysaccharide chain (O-antigen) of the lipopolysaccharide (LPS) of Proteus mirabilis strain D52 was studied using chemical analyses along with...
The acidic O-specific polysaccharide chain (O-antigen) of the lipopolysaccharide (LPS) of Proteus mirabilis strain D52 was studied using chemical analyses along with 1H-NMR and 13C-NMR spectroscopy, including 2D COSY, TOCSY, ROESY, H-detected 1H,13C and 1H,31P HMQC experiments. The polysaccharide was found to contain D-ribitol 5-phosphate (D-Rib-ol-5-P) and ethanolamine phosphate (Etn-P) and has the following structure: D-Rib-ol-5-P (3) approximately 75% EtnP(6)-->2)-beta-D-Galp-(1-->3)-alpha-D-GlcpNAc-(1-->3)-beta-D-Glcp-(1-->3)-beta-D-GlcpNAc-(1-->). This structure is identical with that of the O-polysaccharide of P. mirabilis O33 strain 59/57, and, hence, P. mirabilis D52 belongs to the same Proteus serogroup O33. Serological studies with O-antiserum against P. mirabilis D52 confirmed this but showed that the LPS species of P. mirabilis 59/57 and D52 are not identical, having different epitopes in the core region. A serological cross-reactivity of P. mirabilis D52 O-antiserum was observed with LPS of two other Proteus strains, P. mirabilis O16 and P. penneri 103, which have structurally different O-polysaccharides. The role of charged groups, Rib-ol-5-P and Etn-P in the immunospecificity is discussed.
Topics: Animals; Blotting, Western; Carbohydrate Sequence; Carbohydrates; Electrophoresis, Polyacrylamide Gel; Ethanolamines; Hemolysis; Immunoenzyme Techniques; Lipopolysaccharides; Magnetic Resonance Spectroscopy; Models, Chemical; Molecular Sequence Data; Polysaccharides; Proteus mirabilis; Rabbits; Ribitol
PubMed: 11488930
DOI: 10.1046/j.1432-1327.2001.02356.x -
Microbiological Research Nov 2019Endophytic bacteria isolated from cactus were characterized and assessed for their capability to induce drought tolerance and growth promotion in tomato. A total of...
Endophytic bacteria isolated from cactus were characterized and assessed for their capability to induce drought tolerance and growth promotion in tomato. A total of 191-bacteria representing 13-genera and 18-species were isolated from wild cactus, Euphorbia trigonas. Bacillus (58), Lysinibacillus (36), Enterobacter (29), Stenotrophomonas (18), Lelliottia (12) and Pseudomonas (12) were the most represented genera. 16S rDNA sequence (>1400-bp) comparison placed the bacterial isolates with Bacillus xiamenensis; Bacillus megaterium; Bacillus cereus; Bacillus amyloliquefaciens; Bacillus velezensis; Brevibacillus brevis; Lysinibacillus fusiformis; Enterobacter cloacae; Lelliottia nimipressuralis; Proteus penneri; Sphingobacterium multivorum; Klebsiella pneumoniae; Pseudomonas putida; Pseudomonas aeruginosa; Stenotrophomonas maltophilia; Citrobacter freundii; Chryseobacterium indologenes and Paracoccus sp. Bacillus xiamenensis was identified for the first time as plant endophyte. Upon bacterization, the endophytes triggered germination and growth promotion in tomato as indicated by 118 % and 52 % more root-biomass under drought-free and drought-induced conditions, respectively. Bacillus amyloliquefaciens CBa_RA37 and B. megaterium RR10 displayed broad spectrum endophytism in tomato. Bacterization of tomato with cactus endophyte showed altered oxidative status, stomatal and photosystem II functioning, internal leaf temperature and relative water content suggestive of physiological de-stressing from moisture stress. Activity of oxidative stress enzymes such as guaiacol peroxidase and catalase was also indicative of endophyte assisted de-stressing of tomato. Re-irrigation on 20-days of drought infliction showed 86.9% recovery of B. amyloliquefaciens CBa_RA37 primed tomato when non-primed plantlets succumbed. The cactus endophytic bacterial strain B. amyloliquefaciens CBa_RA37 showed promise for low-cost, efficient and environmentally friendly bio-inoculant technology to mitigate drought in arid zones of Asian and African continents.
Topics: Acclimatization; Bacillus; Biomass; Cactaceae; Cameroon; DNA, Ribosomal; Desert Climate; Droughts; Endophytes; Solanum lycopersicum; Phylogeny; Plant Development; Plant Leaves; Plant Roots; RNA, Ribosomal, 16S; Rifamycins; Sequence Analysis; Soil Microbiology; Stress, Physiological
PubMed: 31442862
DOI: 10.1016/j.micres.2019.126302 -
European Journal of Biochemistry Apr 1991O-Specific polysaccharide was obtained by mild acid degradation of Proteus penneri strain 16 lipopolysaccharide and found to contain D-glucose, D-glucuronic acid,...
O-Specific polysaccharide was obtained by mild acid degradation of Proteus penneri strain 16 lipopolysaccharide and found to contain D-glucose, D-glucuronic acid, 2-acetamido-2-deoxy-D-glucose, and 3,6-dideoxy-3-[(R)-3-hydroxybutyramido]- D-galactose in the ratio of 2:1:1:1 as well as a small proportion of O-acetyl groups. On the basis of one-dimensional 1H-NMR13C-NMR and NOE spectroscopy, two-dimensional homonuclear-shift-correlated spectroscopy with one-step and two-step relayed coherence transfer and heteronuclear 1H/13C NMR shift-correlated spectroscopy, it was concluded that the O-specific polysaccharide of P. penneri strain 16 has the following structure: (formula; see text) This structure was confirmed by methylation analysis and structural analysis of a linear tetrasaccharide fragment prepared by cleavage of the polysaccharide with anhydrous hydrogen fluoride followed by conversion of the alpha-tetrosyl fluoride obtained in to the corresponding free oligosaccharide and alditol. O-Acetyl groups were tentatively located at position 3 of the glucuronic acid residue and at position 4 of the 6-substituted glucose residue, the degree of acetylation being less than 20% of the total. Cross-reactions of P. penneri strain 16 anti-(O-specific polysaccharide) antiserum with lipopolysaccharides from several other Proteus strains and the role of 3,6-dideoxy-3-(R)-3-hydroxybutyramido-D-galactose in the serological specificity of P. penneri strain 16 are discussed.
Topics: Carbohydrate Conformation; Carbohydrate Sequence; Electrophoresis, Polyacrylamide Gel; Immune Sera; Immunoblotting; Indicators and Reagents; Lipopolysaccharides; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Oligosaccharides; Polysaccharides, Bacterial; Proteus vulgaris
PubMed: 2015828
DOI: 10.1111/j.1432-1033.1991.tb15886.x -
Antimicrobial Agents and Chemotherapy May 2018The complete nucleotide sequences of six IMP-4-encoding plasmids recovered from isolates of wildlife origin were characterized. Sequencing data showed that plasmids of...
Characterization of the Complete Nucleotide Sequences of IMP-4-Encoding Plasmids, Belonging to Diverse Inc Families, Recovered from Enterobacteriaceae Isolates of Wildlife Origin.
The complete nucleotide sequences of six IMP-4-encoding plasmids recovered from isolates of wildlife origin were characterized. Sequencing data showed that plasmids of different incompatibility groups (IncM, IncI1, IncF, and nontypeable [including an IncX5_2 and two pPrY2001-like]) carried the -carrying integrons In809 or In1460. Most of the plasmids carried an (A) region, and -like, , and genes. Finally, plasmid analysis revealed the involvement of two different IS- and Tn-associated mechanisms in the mobilization of IMP-4-encoding integrons.
Topics: Anti-Bacterial Agents; Enterobacteriaceae; Enterobacteriaceae Infections; Gene Transfer, Horizontal; Plasmids
PubMed: 29483121
DOI: 10.1128/AAC.02434-17 -
Antimicrobial Agents and Chemotherapy Jan 2002We report on a case of a postneurosurgical meningitis due to ceftriaxone-susceptible Proteus penneri, with selection of a ceftriaxone-resistant isolate following...
Postneurosurgical meningitis due to Proteus penneri with selection of a ceftriaxone-resistant isolate: analysis of chromosomal class A beta-lactamase HugA and its LysR-type regulatory protein HugR.
We report on a case of a postneurosurgical meningitis due to ceftriaxone-susceptible Proteus penneri, with selection of a ceftriaxone-resistant isolate following treatment with ceftriaxone. The isolates presented identical patterns by pulsed-field gel electrophoresis and produced a single beta-lactamase named HugA with an isoelectric point of 6.7. The ceftriaxone-resistant isolate hyperproduced the beta-lactamase (increase in the level of production, about 90-fold). The sequences of the hugA beta-lactamase gene and its regulator, hugR, were identical in both P. penneri strains and had 85.96% homology with those of Proteus vulgaris. The HugA beta-lactamase belongs to molecular class A, and the transcriptional regulator HugR belongs to the LysR family.
Topics: Adult; Amino Acid Sequence; Base Sequence; Ceftriaxone; Chromosomes, Bacterial; DNA, Bacterial; Drug Resistance; Humans; Meningitis, Bacterial; Molecular Sequence Data; Postoperative Complications; Proteus; beta-Lactamases
PubMed: 11751137
DOI: 10.1128/AAC.46.1.216-219.2002 -
Indian Journal of Anaesthesia Jul 2011
PubMed: 22013265
DOI: 10.4103/0019-5049.84842 -
European Journal of Biochemistry Jan 2000Lipopolysaccharide of Proteus penneri strain 63 was degraded by mild acid to give a high molecular mass O-specific polysaccharide that was isolated by gel-permeation...
Lipopolysaccharide of Proteus penneri strain 63 was degraded by mild acid to give a high molecular mass O-specific polysaccharide that was isolated by gel-permeation chromatography. Sugar and methylation analyses and NMR spectroscopic studies, including two-dimensional 1H, 1H COSY, TOCSY rotating-frame NOE spectroscopy, H-detected 1H,13C and 1H,31P heteronuclear multiple-quantum coherence (HMQC), and 1H, 13C HMQC-TOCSY experiments, demonstrated the following structure of the polysaccharide: where FucNAc is 2-acetamido-2,6-dideoxygalactose and PEtn is 2-aminoethyl phosphate. The polysaccharide studied shares some structural features, such as the presence of D-GlcNAc6PEtn and an alpha-L-FucNAc-(1-->3)-D-GlcNAc disaccharide, with other Proteus O-specific polysaccharides. A marked cross-reactivity of P. penneri 63 O-antiserum with P. vulgaris O12 was observed and substantiated by a structural similarity of the O-specific polysaccharides of the two strains. In spite of this, the polysaccharide of P. penneri 63 has the unique structure among Proteus O-antigens, and therefore a new, separate serogroup, O68, is proposed for this strain.
Topics: Carbohydrate Conformation; Carbohydrate Sequence; Cross Reactions; Magnetic Resonance Spectroscopy; Molecular Sequence Data; O Antigens; Polysaccharides, Bacterial; Proteus; Serotyping
PubMed: 10632731
DOI: 10.1046/j.1432-1327.2000.01041.x -
Antimicrobial Agents and Chemotherapy Sep 1984Patterns of susceptibility of 45 Proteus penneri clinical isolates to 14 antimicrobial agents were evaluated by a macrobroth dilution method. All strains were highly...
Patterns of susceptibility of 45 Proteus penneri clinical isolates to 14 antimicrobial agents were evaluated by a macrobroth dilution method. All strains were highly susceptible to ceftizoxime, ceftazidime, moxalactam, cefoxitin, gentamicin, tobramycin, netilmicin, and, with few exceptions, to amikacin, piperacillin, and cefoperazone. Most strains were susceptible to cefotaxime and ceftriaxone. All strains were resistant to cefazolin and cefsulodin.
Topics: Anti-Bacterial Agents; Microbial Sensitivity Tests; Proteus
PubMed: 6508270
DOI: 10.1128/AAC.26.3.419