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Multilamellar and Multivesicular Outer Membrane Vesicles Produced by a Buttiauxella agrestis Mutant.Applied and Environmental Microbiology Oct 2020Outer membrane vesicles (OMVs) are naturally released from Gram-negative bacteria and play important roles in various biological functions. Released vesicles are not...
Outer membrane vesicles (OMVs) are naturally released from Gram-negative bacteria and play important roles in various biological functions. Released vesicles are not uniform in shape, size, or characteristics, and little is known about this diversity of OMVs. Here, we show that deletion of , which encodes a part of the Tol-Pal system, leads to the production of multiple types of vesicles and increases overall vesicle production in the high-vesicle-forming type strain JCM 1090. The Δ mutant produced small OMVs and multilamellar/multivesicular OMVs (M-OMVs) as well as vesicles with a striking similarity to the wild type. M-OMVs, previously undescribed, contained triple-lamellar membrane vesicles and multiple vesicle-incorporating vesicles. Ultracentrifugation enabled the separation and purification of each type of OMV released from the Δ mutant, and visualization by quick-freeze deep-etch and replica electron microscopy indicated that M-OMVs are composed of several lamellar membranes. Visualization of intracellular compartments of Δ mutant cells showed that vesicles were accumulated in the broad periplasm, which is probably due to the low linkage between the outer and inner membranes attributed to the Tol-Pal defect. The outer membrane was invaginating inward by wrapping a vesicle, and the precursor of M-OMVs existed in the cell. Thus, we demonstrated a novel type of bacterial OMV and showed that unconventional processes enable the Δ mutant to form unique vesicles. Membrane vesicle (MV) formation has been recognized as a common mechanism in prokaryotes, and MVs play critical roles in intercellular interaction. However, a broad range of MV types and their multiple production processes make it difficult to gain a comprehensive understanding of MVs. In this work, using vesicle separation and electron microscopic analyses, we demonstrated that diverse types of outer membrane vesicles (OMVs) were released from an engineered strain, JCM 1090 Δ mutant. We also discovered a previously undiscovered type of vesicle, multilamellar/multivesicular outer membrane vesicles (M-OMVs), which were released by this mutant using unconventional processes. These findings have facilitated considerable progress in understanding MV diversity and expanding the utility of MVs in biotechnological applications.
Topics: Bacterial Proteins; Enterobacteriaceae; Mutation; Periplasmic Proteins
PubMed: 32801184
DOI: 10.1128/AEM.01131-20 -
Microbiological Research Jul 2021The association of different species of endophytic bacteria with the rhizosphere of the host plants can stimulate growth, development and acclimatization, offering a...
The association of different species of endophytic bacteria with the rhizosphere of the host plants can stimulate growth, development and acclimatization, offering a greater quantity of seedlings, in addition to reducing the cycle, providing economic return to the producer. The objective of this study was to evaluate the effect of introduction four bacterial isolates through inoculation into the root system in three banana cultivars (Prata Anã, Grande Naine and BRS Princesa) in the acclimatization phase. The evaluated treatments were: control (nutrient broth without bacteria); Bacillus cereus strain 1 (BC1); Bacillus cereus strain 2 (BC2); Bacillus thuringiensis (BT); Buttiauxella agrestis (BA). The morphological characteristics related to the development of the plants (total height and pseudostem diameter) were evaluated throughout the acclimatization period. After 90 days of transplanting and acclimatization, root length, leaf number, dry root weight, pseudostem and leaf, leaf area, internal carbon concentration, stomatal conductance, photosynthesis rate, transpiration rate, leaf temperature and chlorophyll were evaluated. The bacteria showed different results in relation to the studied cultivars. Considering the morphological and physiological characteristics observed in this study, B. thuringiensis for the cultivars Prata Anã and Grande Naine and the B. agrestis for the cultivar BRS Princesa are recommended for the process of acclimatization of banana seedlings, as they stimulated growth of the plant, increasing the dry mass, besides promoting the growth of roots. In this way, they improved the physiological aspects of the plants and reduced the period of acclimatization of the banana.
Topics: Adaptation, Physiological; Agricultural Inoculants; Bacillus; Chlorophyll; Endophytes; Enterobacteriaceae; Musa; Photosynthesis; Plant Roots; Seedlings
PubMed: 33765636
DOI: 10.1016/j.micres.2021.126750 -
Microbiology Resource Announcements May 2021We report here the complete genome sequence of DSM 9389, which harbors eight 16S rRNA genes classified into three types. The genome sequence of this strain showed a...
We report here the complete genome sequence of DSM 9389, which harbors eight 16S rRNA genes classified into three types. The genome sequence of this strain showed a high average nucleotide identity (97.3%) with that of the highly membrane vesicle-producing strain ATCC 33320.
PubMed: 33986092
DOI: 10.1128/MRA.00301-21 -
Genome Announcements Oct 2014MI agar is routinely used for quantifying Escherichia coli in drinking water. A suspect E. coli colony isolated from a water sample was identified as Buttiauxella...
MI agar is routinely used for quantifying Escherichia coli in drinking water. A suspect E. coli colony isolated from a water sample was identified as Buttiauxella agrestis. The whole genome sequence of B. agrestis was determined to understand the genetic basis for its phenotypic resemblance to E. coli on MI agar.
PubMed: 25323724
DOI: 10.1128/genomeA.01060-14 -
Frontiers in Microbiology 2017Membrane vesicles (MVs) are secreted from a wide range of microbial species and transfer their content to other cells. Although MVs play critical roles in bacterial...
Membrane vesicles (MVs) are secreted from a wide range of microbial species and transfer their content to other cells. Although MVs play critical roles in bacterial communication, whether MVs selectively interact with bacterial cells in microbial communities is unclear. In this study, we investigated the specificity of the MV-cell interactions and evaluated the potential of MVs to target bacterial cells for delivery. MV association with bacterial cells was examined using a fluorescent membrane dye to label MVs. MVs derived from the enterobacterium specifically interacted with cells of the parent strain but interacted less specifically with those of other genera tested in this study. Electron microscopic analyses showed that MVs were not only attached on cells but also fused to them. The interaction energy, which was characterized by hydrodynamic diameter and zeta potential based on the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, was significant low between MVs and cells in , compared to those between MVs and cells of other genera. Similar specific interaction was also occurred between MVs and cells of six other species belonging to spp. harboring plasmid pBBR1MCS-1 secreted plasmid-containing MVs (p-MVs), and plasmid DNA in p-MVs was transferred to the same species. Moreover, antibiotic-associated MVs enabled effective killing of target species; the survival rate of was lower than those of and in the presence of gentamicin-associated MVs derived from . Altogether, we provide the evidence that MVs selectively interact with target bacterial cells and offer a new avenue for controlling specific bacterial species using bacterial MVs in microbial communities.
PubMed: 28439261
DOI: 10.3389/fmicb.2017.00571 -
International Journal of Infectious... May 2014Surgical site infections (SSI) are postoperative complications that constitute a major public health problem. We present a rare case report of infection by Buttiauxella...
Surgical site infections (SSI) are postoperative complications that constitute a major public health problem. We present a rare case report of infection by Buttiauxella agrestis, a member of the Enterobacteriaceae family, occurring after a cesarean delivery in a young woman with no comorbidities. The authors further discuss the origin of this infection.
Topics: Adolescent; Anti-Bacterial Agents; Cesarean Section; Enterobacteriaceae; Enterobacteriaceae Infections; Female; Humans; Postoperative Complications; Pregnancy
PubMed: 24641982
DOI: 10.1016/j.ijid.2014.01.025 -
Molecules (Basel, Switzerland) May 2022Acetyl xylan esterases (AXEs) are enzymes capable of hydrolysing the acetyl bonds in acetylated xylan, allowing for enhanced activity of backbone-depolymerizing enzymes....
Acetyl xylan esterases (AXEs) are enzymes capable of hydrolysing the acetyl bonds in acetylated xylan, allowing for enhanced activity of backbone-depolymerizing enzymes. Bioprospecting novel AXE is essential in designing enzyme cocktails with desired characteristics targeting the complete breakdown of lignocellulose. In this article, we report the characterisation of a novel AXE identified as Gene_id_40363 in the metagenomic library analysed from the gut microbiota of the common black slug. The conserved domain description was identified with an NCBI BLASTp search using the translated nucleotide sequence as a query. The activity of the recombinant enzyme was tested on various synthetic substrates and acetylated substrates. The protein sequence matched the conserved domain described as putative hydrolase and aligned closely to an uncharacterized esterase from , hence the designation as BaAXE. BaAXE showed low sequence similarity among characterized CE family proteins with an available 3D structure. BaAXE was active on 4-nitrophenyl acetate, reporting a specific activity of 78.12 U/mg and a Km value of 0.43 mM. The enzyme showed optimal activity at 40 °C and pH 8 and showed high thermal stability, retaining over 40% activity after 2 h of incubation from 40 °C to 100 °C. BaAXE hydrolysed acetyl bonds, releasing acetic acid from acetylated xylan and β-D-glucose pentaacetate. BaAXE has great potential for biotechnological applications harnessing its unique characteristics. In addition, this proves the possibility of bioprospecting novel enzymes from understudied environments.
Topics: Acetylesterase; Animals; Gastrointestinal Microbiome; Gastropoda; Substrate Specificity; Xylans
PubMed: 35566348
DOI: 10.3390/molecules27092999 -
Antimicrobial Agents and Chemotherapy Oct 2002Extensive biochemical testing and 16S rRNA and rpoB sequence analysis revealed that clinical strain CF01Ent1, initially identified as Buttiauxella agrestis by the use of...
Extensive biochemical testing and 16S rRNA and rpoB sequence analysis revealed that clinical strain CF01Ent1, initially identified as Buttiauxella agrestis by the use of Api 32 biochemical strips, is a new organism in the Enterobacteriaceae family. It produced an inducible AmpC-type beta-lactamase whose sequence shares 69 to 72% identity with those of the other AmpC-type beta-lactamases of ENTEROBACTERIACEAE: This enzyme exhibits an atypical high affinity for all beta-lactams tested.
Topics: Amino Acid Sequence; Anti-Bacterial Agents; Bacterial Proteins; Base Sequence; DNA-Directed RNA Polymerases; Enterobacteriaceae; Enzyme Induction; Microbial Sensitivity Tests; Molecular Sequence Data; RNA, Ribosomal, 16S; Sequence Analysis, DNA; beta-Lactam Resistance; beta-Lactamases; beta-Lactams
PubMed: 12234870
DOI: 10.1128/AAC.46.10.3316-3319.2002 -
Antimicrobial Agents and Chemotherapy Jun 1988One hundred and sixty-nine strains of new species of the family Enterobacteriaceae, isolated mainly from the environment, were tested to determine their susceptibilities...
One hundred and sixty-nine strains of new species of the family Enterobacteriaceae, isolated mainly from the environment, were tested to determine their susceptibilities to 13 antibiotics and 4 antiseptics or disinfectants. All the species were susceptible to aminoglycosides, doxycycline, and trimethoprim but were resistant to chloramphenicol. Susceptibility to beta-lactams varied more among the strains. However, all the strains were cefotaxime susceptible, apart from some Buttiauxella agrestis strains for which MICs were greater than 256 micrograms/ml. The antiseptic MBCs were similar to those published elsewhere for species of the Enterobacteriaceae of clinical origin. No resistance to chlorhexidine was observed. On the other hand, the environmental strains presented a greater resistance to active chlorine than did the reference strains.
Topics: Anti-Bacterial Agents; Anti-Infective Agents, Local; Disinfectants; Enterobacteriaceae; Microbial Sensitivity Tests
PubMed: 3415208
DOI: 10.1128/AAC.32.6.873 -
Irish Veterinary Journal 2015Antimicrobial resistance (AMR) is the most significant threat to global public health and ascertaining the role wild birds play in the epidemiology of resistance is...
BACKGROUND
Antimicrobial resistance (AMR) is the most significant threat to global public health and ascertaining the role wild birds play in the epidemiology of resistance is critically important. This study investigated the prevalence of AMR Gram-negative bacteria among long-distance migratory East Canadian High Arctic (ECHA) light-bellied Brent geese found wintering on the east coast of Ireland.
FINDINGS
In this study a number of bacterial species were isolated from cloacal swabs taken from ECHA light-bellied Brent geese. Nucleotide sequence analysis identified five species of Gram-negative bacteria; the dominant isolated species were Pantoea spp. (n = 5) followed by Buttiauxella agrestis (n = 2). Antimicrobial susceptibility disk diffusion results identified four of the Pantoea spp. strains, and one of the Buttiauxella agrestis strains resistant to amoxicillin-clavulanic acid.
CONCLUSION
To our knowledge this is the first record of AMR bacteria isolated from long distance migratory ECHA light-bellied Brent geese. This indicates that this species may act as reservoirs and potential disseminators of resistance genes into remote natural ecosystems across their migratory range. This population of geese frequently forage (and defecate) on public amenity areas during the winter months presenting a potential human health risk.
PubMed: 27651892
DOI: 10.1186/s13620-016-0072-7