Biodegradation of ochratoxin A by Brevundimonas diminuta HAU429: Characterized performance, toxicity evaluation and functional enzymes.

Ochratoxin A (OTA) is a notorious mycotoxin commonly contaminating food products worldwide. In this study, an OTA-degrading strain Brevundimonas diminuta HAU429 was isolated by using hippuryl-L-phenylalanine as the sole carbon source. The biodegradation of OTA by strain HAU429 was a synergistic effect of intracellular and extracellular enzymes, which transformed OTA into ochratoxin α (OTα) through peptide bond cleavage. Cytotoxicity tests and cell metabolomics confirmed that the transformation of OTA into OTα resulted in the detoxification of its hepatotoxicity since OTA but not OTα disturbed redox homeostasis and induced oxidative damage to hepatocytes. Genome mining identified nine OTA hydrolase candidates in strain HAU429. They were heterologously expressed in Escherichia coli, and three novel amidohydrolase BT6, BT7 and BT9 were found to display OTA-hydrolyzing activity. BT6, BT7 and BT9 showed less than 45 % sequence identity with previously identified OTA-degrading amidohydrolases. BT6 and BT7 shared 60.9 % amino acid sequence identity, and exhibited much higher activity towards OTA than BT9. BT6 and BT7 could completely degrade 1 µg mL-1 of OTA within 1 h and 50 min, while BT9 hydrolyzed 100 % of OTA in the reaction mixture by 12 h. BT6 was the most thermostable retaining 38 % of activity after incubation at 70 °C for 10 min, while BT7 displayed the highest tolerance to ethanal remaining 76 % of activity in the presence of 6 % ethanol. This study could provide new insights towards microbial OTA degradation and promote the development of enzyme-catalyzed OTA detoxification during food processing.

Peiella sedimenti gen. nov., sp. nov., a novel taxon within the family Caulobacteraceae isolated from sediment of a river.

A Gram-stain-negative bacterium, designated XZ-24T, was isolated from sediment of a river in Mianyang city, Sichuan province, PR China. Cells (1.0-2.0 µm long and 0.4-0.5 µm in width) were strictly aerobic, non-spore-forming, rod shaped, prosthecate and motile by means of a polar flagellum. Growth occurred at 10-37 °C (optimum, 30 °C), at pH 5.0-9.0 (optimum pH 7.0) and with 0-3.0 % (w/v) NaCl (optimum 1.0 % NaCl). The results of phylogenetic analysis based on genomes and 16S rRNA gene sequences indicated that XZ-24T formed a distinct phyletic branch within the family Caulobacteraceae and was most closely related to members of the genera Brevundimonas, Caulobacter and Phenylobacterium with 95.3-96.5 % 16S rRNA gene sequence similarities. The average amino acid identities (AAI) between XZ-24T and species of the family Caulobacteraceae were 47.0-64.5 %, which were below the genus boundary (70 %). The predominant cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0, C18 : 1ω7c 11-methyl and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), the isoprenoid quinone was Q-10, and the major polar lipids were 1,2-di-O-acyl-3-O-α-d-glucopyranuronosyl glycerol; 1,2-di-O-acyl-3-O-[d-glucopyranosyl-(1→4)-α-d glucopyranuronosyl] glycerol and phosphatidylglycerol. The genome size of XZ-24T was 2.64 Mb with a DNA G+C content of 68.9 %. On the basis of the evidence presented in this study, strain XZ-24T represents a novel species of a novel genus in the family Caulobacteraceae, for which the name Peiella sedimenti gen. nov., sp. nov. (Type strain XZ-24T=CCTCC AB 20 23 094T=KCTC 8038T) is proposed.

Seasonal changes of prokaryotic microbial community structure in Zhangjiayan Reservoir and its response to environmental factors.

As a typical sub-deep reservoir in the upper reaches of the Yangtze River in the southwest region, Zhangjiayan Reservoir is also an important source of drinking water. Exploring the role of microorganisms in the material cycle of water bodies is of great significance for preventing the exacerbation of eutrophication in the reservoir. In this study, water samples from the overlying water of five points in the reservoir were collected four times in spring (April), summer (July), autumn (November), and winter (January) of 2022-2023 using a gas-tight water sampler. Physicochemical factors were measured, and the microbial community structure was analyzed by high-throughput MiSeq sequencing of the V3-V4 hypervariable region of 16S rRNA gene in order to explore the relationship between physicochemical factors and microbial community structure and the dominant microbial populations that affect eutrophication of the reservoir. The following results were obtained through analysis. Among the 20 overlying water samples from Zhangjiayan Reservoir, a total of 66 phyla, 202 classes, 499 orders, 835 families, 1716 genera, and 27,904 ASVs of the bacterial domain were detected. The phyla Proteobacteria and Actinobacteria were dominant in the microbial community of the overlying water in Zhangjiayan Reservoir. At the genus level, hgcI_clade and Actinobacteria had the highest abundance and was the dominant population. The microbial community in the water of Zhangjiayan Reservoir has a high level of diversity. The diversity index ranked by numerical order was winter > autumn > summer > spring. Significant differences were found in the composition and structure of the microbial community between the spring/summer and autumn/winter seasons (p < 0.05). Total phosphorus, dissolved total phosphorus, soluble reactive phosphorus, and dissolved oxygen have a significant impact on the composition and structure of the microbial community (p < 0.01). The bacterial community in the overlying water of Zhangjiayan Reservoir showed a mainly positive correlation. Sphingomonas, Brevundimonas, and Blastomonas were the central populations of the bacterial community in the overlying water of Zhangjiayan Reservoir. This study indicates that environmental factors, such as phosphorus and other nutrients, have a significant impact on the formation of the microbial community structure in different seasons. Sphingomonas, Brevundimonas, and Blastomonas are key populations that may have a significant impact on eutrophication in Zhangjiayan Reservoir.

Honokiol affects the composition of gut microbiota and the metabolism of lipid and bile acid in methionine-choline deficiency diet-induced NASH mice.

Honokiol (HNK), one of the main active components of Magnolia officinalis, has a positive effect on non-alcoholic steatohepatitis (NASH). However, the effects of HNK on the composition of serum lipids and bile acids (BAs) and gut microbiota (GM) of NASH mice are still unknown.C57BL/6 mice were fed with methionine-choline deficiency (MCD) diet and gavaged with HNK (20 mg/kg/d) for 8 weeks, then the serum lipids and BAs were detected by LC-MS, the composition of ileum microflora and the mRNA expression of hepatic BAs homeostasis related genes were analyzed by 16S rDNA sequencing and RT-qPCR, respectively. HNK treatment decreased the degree of hepatic lipid drops, inflammatory cell infiltration and fibrosis. Meantime, the serum levels of 34 lipids and 4 BAs in MCD mice were significantly altered by HNK treatment, as well as the increased abundance of Ruminococcaceae, Caulobacteraceae and Brevundimonas, and the decreased abundance of Firmicutes and Dubosiella. Besides, HNK treatment increased the hepatic mRNA expression of Oatp1b2 in MCD mice. The ameliorating effect of HNK on NASH may be partly related to its correction on the disorders of GM, serum lipids and BAs of MCD mice.

Purification and characterization of the enzymes from Brevundimonas naejangsanensis that degrade ochratoxin A and B.

Ochratoxin A (OTA) and Ochratoxin B (OTB) co-contaminate many types of agricultural products. Screening enzymes that degrade both OTA and OTB has significance in food safety. In this study, four novel OTA and OTB degrading enzymes, namely BnOTase1, BnOTase2, BnOTase3, and BnOTase4, were purified from the metabolites of the Brevundimonas naejangsanensis ML17 strain. These four enzymes hydrolyzed OTA into OTα and hydrolyzed OTB into OTß. BnOTase1, BnOTase2, BnOTase3, and BnOTase4 have the apparent Km values for hydrolyzing OTA of 19.38, 0.92, 12.11, 1.09 µmol/L and for hydrolyzing OTB of 0.76, 2.43, 0.60, 0.64 µmol/L respectively. OTα and OTß showed no significant cytotoxicity to HEK293 cells, suggesting that these enzymes mitigate the toxicity of OTA and OTB. The discovery of the novel OTA and OTB degrading enzymes enriches the research on ochratoxin control and provides objects for protein rational design.

Biodegradation of textile dye Rhodamine-B by Brevundimonas diminuta and screening of their breakdown metabolites.

The carcinogenic Rhodamine-B dye is recalcitrant which could cause serious hazards to human beings. Degradation with the application of unique bacterial strain is a sustainable technique. The bioremediation technique showed great potential to degrade a variety of recalcitrant pollutants like dyes. In this study, Brevundimonas diminuta, was selected for the breakdown of toxic textile dye Rhodamine-B. This bacterium showed 90-95% of degradation at the optimum conditions like 10 mg L-1 of concentration of dye, pH 7 and temperature of 30 °C. Further UV-Visible spectrophotometry, FT-IR spectral scan, GC-MS analysis depicted the breakdown products like Methyl 18-fluoro-octadec-9-enoate, Methyl 18-fluoro-octadec-9-enoate and d-Homo-24-nor-17-oxachola-20,22-diene-3,16-dione,7-(acetyloxy)-1, 23 tri-epoxy-4,4,8-trimethyl. The degradation was confirmed by the changes in the functional groups, change in molecular weight and charge to-mass ratio. These results suggested that this strain is a deserving organism for the degradation of dye compounds.

An EVOH nanofibrous sterile membrane with a robust and antifouling surface for high-performance sterile filtration via glutaraldehyde crosslinking and a plasma-assisted process.

Constructing a sterile membrane with a robust and antifouling surface is a powerful means to improve the sterile filtration efficiency of sterile membranes. In this work, a robust EVOH nanofibrous sterile membrane was facilely fabricated by the method of in situ crosslinking with glutaraldehyde and surface plasma treatment. The resultant EVOH nanofibrous sterile membrane possessed a carboxylated-crosslinked surface, with high hydrophilicity, which generated high chemical stability, high-temperature steam resistance, and an ultrahigh antifouling performance against bovine serum albumin, ribonucleic acid and nanoparticle pollutants. Moreover, the membrane also exhibited a reasonably high primary water permeance (4522.2 LMH bar-1 at 0.2 MPa), as well as an absolute interception rate (100%) of Escherichia coli, Staphylococcus aureus cells and Brevundimonas diminuta superior to the state-of-the-art sterile membrane. Moreover, the modified membrane packed syringe-driven filter presented 100% interception (LRV ≥ 7) to Brevundimonas diminuta and high permeation flux (from 10.8 to 41.8 L·h-1) in a wide operating pressure range of 0.1 MPa to 0.6 MPa, indicating its potential in real bio-separation applications. This work provides a facile strategy for the preparation of a high-performance sterile membrane for biological drug product sterilization.

Characterization of Two Highly Arsenic-Resistant Caulobacteraceae Strains of Brevundimonas nasdae: Discovery of a New Arsenic Resistance Determinant.

Arsenic (As), distributed widely in the natural environment, is a toxic substance which can severely impair the normal functions in living cells. Research on the genetic determinants conferring functions in arsenic resistance and metabolism is of great importance for remediating arsenic-contaminated environments. Many organisms, including bacteria, have developed various strategies to tolerate arsenic, by either detoxifying this harmful element or utilizing it for energy generation. More and more new arsenic resistance (ars) determinants have been identified to be conferring resistance to diverse arsenic compounds and encoded in ars operons. There is a hazard in mobilizing arsenic during gold-mining activities due to gold- and arsenic-bearing minerals coexisting. In this study, we isolated 8 gold enrichment strains from the Zijin gold and copper mine (Longyan, Fujian Province, China) wastewater treatment site soil, at an altitude of 192 m. We identified two Brevundimonas nasdae strains, Au-Bre29 and Au-Bre30, among these eight strains, having a high minimum inhibitory concentration (MIC) for As(III). These two strains contained the same ars operons but displayed differences regarding secretion of extra-polymeric substances (EPS) upon arsenite (As(III)) stress. B. nasdae Au-Bre29 contained one extra plasmid but without harboring any additional ars genes compared to B. nasdae Au-Bre30. We optimized the growth conditions for strains Au-Bre29 and Au-Bre30. Au-Bre30 was able to tolerate both a lower pH and slightly higher concentrations of NaCl. We also identified folE, a folate synthesis gene, in the ars operon of these two strains. In most organisms, folate synthesis begins with a FolE (GTP-Cyclohydrolase I)-type enzyme, and the corresponding gene is typically designated folE (in bacteria) or gch1 (in mammals). Heterologous expression of folE, cloned from B. nasdae Au-Bre30, in the arsenic-hypersensitive strain Escherichia coli AW3110, conferred resistance to As(III), arsenate (As(V)), trivalent roxarsone (Rox(III)), pentavalent roxarsone (Rox(V)), trivalent antimonite (Sb(III)), and pentavalent antimonate (Sb(V)), indicating that folate biosynthesis is a target of arsenite toxicity and increased production of folate confers increased resistance to oxyanions. Genes encoding Acr3 and ArsH were shown to confer resistance to As(III), Rox(III), Sb(III), and Sb(V), and ArsH also conferred resistance to As(V). Acr3 did not confer resistance to As(V) and Rox(V), while ArsH did not confer resistance to Rox(V).

First report of Brevundimonas aurantiaca human infection: infective endocarditis on aortic bioprostheses and supracoronary aortic graft acquired by water dispenser of domestic refrigerator.

Infective endocarditis (IE) is a feared life-threatening complication that requires a multidisciplinary approach. Although a variety of microorganisms have caused IE, Brevundimonas aurantiaca human infection has never been reported previously. To our knowledge, this is the first reported case of endocarditis and human infection due to B. aurantiaca.

Comparative Genomic Analysis Reveals Potential Pathogenicity and Slow-Growth Characteristics of Genus Brevundimonas and Description of Brevundimonas pishanensis sp. nov.

The genus Brevundimonas consists of Gram-negative bacteria widely distributed in environment and can cause human infections. However, the genomic characteristics and pathogenicity of Brevundimonas remain poorly studied. Here, the whole-genome features of 24 Brevundimonas type strains were described. Brevundimonas spp. had relatively small genomes (3.13 ± 0.29 Mb) within the family Caulobacteraceae but high G+C contents (67.01 ± 2.19 mol%). Two-dimensional hierarchical clustering divided those genomes into 5 major clades, in which clades II and V contained nine and five species, respectively. Interestingly, phylogenetic analysis showed a one-to-one match between core and accessory genomes, which suggested coevolution of species within the genus Brevundimonas. The unique genes were annotated to biological functions like catalytic activity, signaling and cellular processes, multisubstance metabolism, etc. The majority of Brevundimonas spp. harbored virulence-associated genes icl, tufA, kdsA, htpB, and acpXL, which encoded isocitrate lyase, elongation factor, 2-dehydro-3-deoxyphosphooctonate aldolase, heat shock protein, and acyl carrier protein, respectively. In addition, genomic islands (GIs) and phages/prophages were identified within the Brevundimonas genus. Importantly, a novel Brevundimonas species was identified from the feces of a patient (suffering from diarrhea) by the analyses of biochemical characteristics, phylogenetic tree of 16S rRNA gene, multilocus sequence analysis (MLSA) sequences, and genomic data. The name Brevundimonas pishanensis sp. nov. was proposed, with type strain CHPC 1.3453 (= GDMCC 1.2503T = KCTC 82824T). Brevundimonas spp. also showed obvious slow growth compared with that of Escherichia coli. Our study reveals insights into genomic characteristics and potential virulence-associated genes of Brevundimonas spp., and provides a basis for further intensive study of the pathogenicity of Brevundimonas. IMPORTANCEBrevundimonas spp., a group of bacteria from the family Caulobacteraceae, is associated with nosocomial infections, deserve widespread attention. Our study elucidated genes potentially associated with the pathogenicity of the Brevundimonas genus. We also described some new characteristics of Brevundimonas spp., such as small chromosome size, high G+C content, and slow-growth phenotypes, which made the Brevundimonas genus a good model organism for in-depth studies of growth rate traits. Apart from the comparative analysis of the genomic features of the Brevundimonas genus, we also reported a novel Brevundimonas species, Brevundimonas pishanensis, from the feces of a patient with diarrhea. Our study promotes the understanding of the pathogenicity characteristics of Brevundimonas species bacteria.

Pseudaquidulcibacter saccharophilus gen. nov., sp. nov., a novel member of family Caulobacteraceae, isolated from a water purification facility with supplement of starch as a carbon source.

During a survey of microbial communities in the influent (ambient water) and effluent of a water purification facility with aeration and supplement of starch as carbon source, a novel bacterial strain, designated SZ9T, was isolated from the effluent sample. Colonies of strain SZ9T were small (approximately 0.5-1.0 mm in diameter), creamy-white, circular, smooth, translucent and convex. Cells were facultative anaerobic, motile by means of a single polar flagellum, rod-shaped, multiplied by binary fission, Gram-stain-negative, oxidase-positive and catalase-negative. Growth occurred at 10-40 °C (optimum, 28 °C) and pH 5.5-8.0 (optimum, pH 7.5). The range of NaCl concentration for growth was 0-1.0 % (w/v), with an optimum of 0-0.5 % (w/v). Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain SZ9T formed a lineage within the family Caulobacteraceae of the class Alphaproteobacteria and showed the highest 16S rRNA gene sequence similarities to Aquidulcibacter paucihalophilus TH1-2T (92.44%), followed by Vitreimonas flagellata SYSU XM001T (89.61 %), Asprobacter aquaticus DRW22-8T (89.49 %) and Hyphobacterium vulgare WM6T (89.49%). The predominant fatty acids (>10 % of the total fatty acids) of strain SZ9T was summed feature 3 (comprising C16 : 1 ω6c and/or C16 : 1 ω7c), summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c) and C16 : 0. The sole respiratory quinone was ubiquinone-10, and the major polar lipids were phosphatidylcholine and two unidentified glycolipids. The whole genome of strain SZ9T was 2 842 140 bp in size, including 2769 protein-coding genes, 37 tRNA genes and two rRNA genes, and the genomic G+C content was 41.4 mol%. The orthologous average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between strain SZ9T and other genera within the family Caulobacteraceae were 64.50-66.62 %, 46.96-54.17 % and 27.70-31.70 %, respectively. Therefore, based on the results of phenotypic, chemotaxonomic and phylogenetic analyses, the isolated strain SZ9T could be distinguished from other genera, suggesting that it represents a novel species of a novel genus in the family Caulobacteraceae, for which the name Pseudaquidulcibacter saccharophilus gen. nov., sp. nov is proposed. The type strain is SZ9T (=CCTCC AB2021029T=KCTC 82788T).

Post-VX exposure treatment of rats with engineered phosphotriesterases.

The biologically stable and highly toxic organophosphorus nerve agent (OP) VX poses a major health threat. Standard medical therapy, consisting of reactivators and competitive muscarinic receptor antagonists, is insufficient. Recently, two engineered mutants of the Brevundimonas diminuta phosphotriesterase (PTE) with enhanced catalytic efficiency (kcat/KM = 21 to 38 × 106 M-1 min-1) towards VX and a preferential hydrolysis of the more toxic P(-) enantiomer were described: PTE-C23(R152E)-PAS(100)-10-2-C3(I106A/C59V/C227V/E71K)-PAS(200) (PTE-2), a single-chain bispecific enzyme with a PAS linker and tag having enlarged substrate spectrum, and 10-2-C3(C59V/C227V)-PAS(200) (PTE-3), a stabilized homodimeric enzyme with a double PASylation tag (PAS-tag) to reduce plasma clearance. To assess in vivo efficacy, these engineered enzymes were tested in an anesthetized rat model post-VX exposure (~ 2LD50) in comparison with the recombinant wild-type PTE (PTE-1), dosed at 1.0 mg kg-1 i.v.: PTE-2 dosed at 1.3 mg kg-1 i.v. (PTE-2.1) and 2.6 mg kg-1 i.v. (PTE-2.2) and PTE-3 at 1.4 mg kg-1 i.v. Injection of the mutants PTE-2.2 and PTE-3, 5 min after s.c. VX exposure, ensured survival and prevented severe signs of a cholinergic crisis. Inhibition of erythrocyte acetylcholinesterase (AChE) could not be prevented. However, medulla oblongata and diaphragm AChE activity was partially preserved. All animals treated with the wild-type enzyme, PTE-1, showed severe cholinergic signs and died during the observation period of 180 min. PTE-2.1 resulted in the survival of all animals, yet accompanied by severe signs of OP poisoning. This study demonstrates for the first time efficient detoxification in vivo achieved with low doses of heterodimeric PTE-2 as well as PTE-3 and indicates the suitability of these engineered enzymes for the development of highly effective catalytic scavengers directed against VX.

Brevundimonas diminuta isolated from mines polluted soil immobilized cadmium (Cd2+) and zinc (Zn2+) through calcium carbonate precipitation: Microscopic and spectroscopic investigations.

The toxic metal(loid)s TMs resistant bacterium Brevundimonas diminuta was isolated for the first time from mines polluted soil in Fengxian, China, and assessed for its potential for Cd and Zn precipitation in Cd and Zn co-contaminated aqueous solution at various Cd and Zn levels (20, 40, 80, 160, and 200 mg L-1), pH values (5, 6, 7, 8, and 9), and temperatures (20, 25, 30, and 35 °C). B. diminuta showed a high resistance to both Cd and Zn and was able to precipitate up to 99.2 and 99.7% of dissolved Cd and Zn respectively, at a pH of 7 and temperature of 30 °C. B. diminuta reduced the dissolved concentrations of Cd and Zn below the threshold levels in water. The 3D-EEM analysis revealed the presence of extracellular polymeric substances (EPS) such as tryptophan indicating bacterial growth under Cd/Zn stress. FTIR showed polysaccharides, CO32-, CaCO3, PO43-, and proteins, which may enhance bacterial growth and metal precipitation. SEM-EDS confirmed the leaf-like and granular shape of the biological precipitation and reduction in the percent weight of TMs, which promoted the adhesion/adsorption of Cd2+, Zn2+, and Ca2+. Moreover, XRD analysis confirmed the precipitation of Cd, Zn, and Ca in the form of CdCO3/Cd3(PO4)2, ZnCO3/ZnHPO4/Zn2(OH)PO4/Zn3(PO4)2, and CaCO3/Ca5(PO3)4OH, respectively. These findings indicate that Brevundimonas diminuta can be used for the bioremediation of TMs-contaminated aquatic environments.

Rice straw particles covered with Brevundimonas naejangsanensis DD1 cells can synergistically remove doxycycline from water using adsorption and biotransformation.

Doxycycline (DC) is a second generation tetracycline antibiotic and its occurrence in the aquatic environment due to the discharge of municipal and agricultural wastes has called for technologies to effectively remove DC from water. The objective of the study was to characterize the synergistic benefits of adsorption and biotransformation in removing DC from water using rice straw particles (RSPs) covered with DC degrading bacteria, Brevundimonas naejangsanensis strain DD1. First, optimal experimental conditions were identified for individual processes, i.e., hydrolysis, adsorption, and biotransformation, in terms of their performance of removing DC from water. Then, synergistic effects between adsorption and biotransformation were demonstrated by adding DD1-covered RSPs (DD1-RSPs) to DC-containing solution. Results suggest that DC was quickly adsorbed onto RSPs and the adsorbed DC was subsequently biotransformed by the DD1 cells on RSPs. The adsorption of DC to DD1-RSPs can be well described using the pseudo-second-order kinetics and the Langmuir isotherm. The DD1 cells on RSPs converted DC to several biotransformation products through a series of demethylation, dehydration, decarbonylation, and deamination. This study demonstrated that adsorption and biotransformation could work synergistically to remove DC from water.

Molecular mechanisms of the CdnG-Cap5 antiphage defense system employing 3',2'-cGAMP as the second messenger.

Cyclic-oligonucleotide-based antiphage signaling systems (CBASS) are diverse and abundant in bacteria. Here, we present the biochemical and structural characterization of two CBASS systems, composed of CdnG and Cap5, from Asticcacaulis sp. and Lactococcus lactis. We show that CdnG from Asticcacaulis sp. synthesizes 3',2'-cGAMP in vitro, and 3',2'-cGAMP is the biological signaling molecule that activates Cap5 for DNA degradation. Crystal structures of Cap5, together with the SAVED domain in complex with 3',2'-cGAMP, provide insight into the architecture of Cap5 as well as molecular recognition of 3',2'-cGAMP by the SAVED domain of Cap5. Amino acid conservation of the SAVED domain of Cap5, together with mutational studies, led us to propose a mechanism of Back-to-Front stacking of two SAVED domains, mediated by 3',2'-cGAMP, to activate HNH nuclease domain for DNA degradation. This study of the most abundant CBASS system provides insights into the mechanisms employed by bacteria in their conflicts against phage.

First Isolation and Clinical Case of Brevundimonasdiminuta in a Newborn with Low Birth Weight, in Democratic Republic of Congo: A Case Report.

Brevundimonas diminuta is rarely described in clinical specimens, never at the umbilical stump. Most of the reported cases are in patients with underlying pathologies. We must integrate this microorganism in the etiological agents of nosocomial infections, but much remains to be understood about its virulence. We present a case of umbilical stump infection (omphalitis) caused by B. diminuta, in a preterm and hypotrophic new-born and discuss the diagnosis of this bacterium and its role as responsible of nosocomial neonatal infections.

High arsenic tolerance in Brevundimonas aurantiaca PFAB1 from an arsenic-rich Indian hot spring

BACKGROUND Arsenic contamination in the ground water of rural India is a recurrent problem and decon tamination is mostly based on the chemical or physical treatments until now. Microbial bioremediation is eco-friendly, cheap, time-efficient and does not produce any toxic by-products. RESULT In the present study, a high arsenic tolerant bacteria Brevundimonas aurantiaca PFAB1 was iso lated from Panifala hot spring located in West Bengal, India. Previously Panifala was also reported to be an arsenic-rich hot spring. B. aurantiaca PFAB1 exhibited both positive arsenic reductase and arsenite oxidase activity. It was tolerant to arsenite up to 90 mM and arsenate up to 310 mM. Electron microscopy has proved significant changes in cellular micromorphology and stalk appearance under the presence of arsenic in growth medium. Bioaccumulation of arsenic in As (III) treated cells were 0.01% of the total cell weight, while 0.43% in case of As (V) treatment. CONCLUSIONS All experimental lines of evidence prove the uptake/accumulation of arsenic within the bac terial cell. All these features will help in the exploitation of B. aurantiaca PFAB1 as a potent biological weapon to fight arsenic toxicity in the near future

Genome-Based Taxonomy of Brevundimonas with Reporting Brevundimonas huaxiensis sp. nov.

Brevundimonas is a genus of Gram-negative bacteria widely distributed in nature and is also an opportunistic pathogen causing health care-associated infections. Brevundimonas strain 090558T was recovered from a blood culture of a cancer patient and was subjected to genome sequencing and analysis. The average nucleotide identity and in silico DNA-DNA hybridization values between 090558T and type strains of Brevundimonas species were 78.76% to 93.94% and 19.8% to 53.9%, respectively, below the cutoff to define bacterial species. Detailed phenotypic tests were performed, suggesting that 090558T can be differentiated from other Brevundimonas species by its ability to assimilate sodium acetate but not to utilize glucose, trypsin, or ß-glucosidase. Strain 090558T (GDMCC 1.1871T or KCTC 82165T) therefore represents a novel Brevundimonas species, for which the name Brevundimonas huaxiensis sp. nov. is proposed. All Brevundimonas genomes available in GenBank (accessed on 25 January 2021) were retrieved, discarding those labeled "excluded from RefSeq" by GenBank, and included 82 genomes for precise species curation. In addition to the 21 Brevundimonas species with genomes of type strains available, we identified 29 Brevundimonas taxa that either belong to the 12 Brevundimonas species without available genomes of type strains or represent novel species. We found that more than half (57.3%) of the 82 Brevundimonas genomes need to be corrected for species assignation, including species mislabeling of a type strain. Our analysis highlights the complexity of Brevundimonas taxonomy. We also found that only some Brevundimonas species are associated with human infections, and more studies are warranted to understand their pathogenicity and epidemiology. IMPORTANCEBrevundimonas is a genus of the family Caulobacteraceae and comprises 33 species. Brevundimonas can cause various infections but remains poorly studied. In this study, we reported a novel Brevundimonas species, Brevundimonas huaxiensis, based on genome and phenotype studies of strain 090558T recovered from human blood. We then examined the species assignations of all Brevundimonas genomes (n = 82) in GenBank and found that in addition to the known Brevundimonas species with genome sequences of type strains available, there are 29 Brevundimonas taxa based on genome analysis, which need to be further studied using phenotype-based methods to establish their species status. Our study significantly updates the taxonomy of Brevundimonas and enhances our understanding of this genus of clinical relevance. The findings also encourage future studies on the characterization of novel Brevundimonas species.

Urinary tract infection by atypical uropathogens in dogs.

Bacterial urinary tract infection (UTI) is a common condition affecting dogs. Urine culture and antimicrobial susceptibility test, associated with the identification of underlying cause, are of primary importance in order to select a correct treatment, especially in presence of comorbidities. Two cases of immunecompromised dogs affected by urinary tract infection (UTI) have been described: the first, probably immunosuppressed due to old age, was in poor body condition, with severe odontolithiasis and periodontitis; the second was affected by chronic kidney disease in advanced stage. Urine cultures isolated two rare and atypical pathogens, Moellerella wisconsensis and Brevundimonas vesicularis, both showing sensitivity versus floroquinolones which were selected for the treatment. After a 4 weeks treatment, a second culture demonstrated the resolution of infection in both cases, in absence of clinical signs.To date neither of the two bacteria have been reported as cause of UTI in dog.

Inhibition of an organophosphate-detoxifying bacterial phosphotriesterase by albumin and plasma thiol components.

The phosphotriesterase of the bacterium Brevundimonas diminuta (BdPTE) is a naturally occurring enzyme that catalyzes the hydrolysis of organophosphate (OP) nerve agents as well as pesticides and offers a potential treatment of corresponding intoxications. While BdPTE mutants with improved catalytic efficiencies against several OPs have been described, unexpectedly, less efficient breakdown of an OP was observed upon application in an animal model compared with in vitro measurements. Here, we describe detailed inhibition studies with the high-activity BdPTE mutant 10-2C3(C59M/C227A) by human plasma components, indicating that this enzyme is inhibited by serum albumin. The inhibitory activity is mediated by depletion of crucial zinc ions from the BdPTE active site, either via the known high-affinity zinc binding site of albumin or via chemical complex formation with its free thiol side chain at position Cys34. Albumin pre-charged with zinc ions or carrying a chemically blocked Cys34 side chain showed significantly reduced inhibitory activity; in fact, the combination of both measures completely abolished BdPTE inhibition. Consequently, the available zinc ion concentration in blood plays an important role for BdPTE activity in vivo and should be taken into account for therapeutic development and application of a catalytic OP scavenger.