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1.
Emerg Infect Dis ; 30(5): 1060-1062, 2024 May.
Article in English | MEDLINE | ID: mdl-38666619

ABSTRACT

We report a case of Sphingobium yanoikuyae bacteremia in an 89-year-old patient in Japan. No standard antimicrobial regimen has been established for S. yanoikuyae infections. However, ceftriaxone and ceftazidime treatments were effective in this case. Increased antimicrobial susceptibility data are needed to establish appropriate treatments for S. yanoikuyae.


Subject(s)
Anti-Bacterial Agents , Bacteremia , Sphingomonadaceae , Aged, 80 and over , Humans , Male , Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/microbiology , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/microbiology , Japan , Microbial Sensitivity Tests , Sphingomonadaceae/genetics , Sphingomonadaceae/isolation & purification , Sphingomonadaceae/drug effects
2.
Biochem J ; 477(15): 2821-2840, 2020 08 14.
Article in English | MEDLINE | ID: mdl-32686828

ABSTRACT

Our previous studies have shown the existence of organophosphate hydrolase (OPH) as a part of the inner membrane associated Ton complex (ExbB/ExbD and TonB) of Sphingobium fuliginis. We now show its involvement in iron uptake by establishing direct interactions with ferric-enterobactin. The interactions between OPH and ferric-enterobactin were not affected even when the active site architecture is altered by substituting active site aspartate with either alanine or asparagine. Protein docking studies further substantiated these findings and predicted the existence of ferric-enterobactin binding site that is different from the catalytic site of OPH. A lysine residue (82K) found at the predicted ferric-enterobactin binding site facilitated interactions between OPH and ferric-enterobactin. Substitution of lysine with alanine did not affect triesterase activity, but it abrogated OPH ability to interact with both ferric-enterobactin and ExbD, strengthening further the fact that the catalytic site is not the site for binding of these ligands. In the absence of interactions between OPHK82A and ExbD, OPHK82A failed to target membrane in E. coli cells. The Sphingobium fuliginis TonB-dependent transport (SfTonBDT) system was reconstituted in E. coli GS027 cells generated by deleting the exbD and tonB genes. The E. coli GS030 cells having SfTonBDT system with OPH showed increased iron uptake. Such an increase was not seen in E. coli GS029, cells having SfTonBDT system generated either by omitting OPH or by including its variants, OPHD301A, OPHD301N suggesting a role for OPH in enhanced iron uptake.


Subject(s)
Bacterial Proteins/metabolism , Iron/pharmacokinetics , Membrane Proteins/metabolism , Phosphoric Monoester Hydrolases/metabolism , Sphingomonadaceae/metabolism , Bacterial Proteins/genetics , Binding Sites , Biological Transport , Catalytic Domain , Circular Dichroism , Enterobactin/metabolism , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/metabolism , Genetic Complementation Test , Iron/metabolism , Lysine/metabolism , Membrane Proteins/genetics , Mutation , Phosphoric Monoester Hydrolases/genetics , Sphingomonadaceae/drug effects , Sphingomonadaceae/genetics
3.
Int J Med Microbiol ; 310(2): 151396, 2020 Feb.
Article in English | MEDLINE | ID: mdl-32005588

ABSTRACT

The occurrence of antibiotic resistance bacteria has become a major threat to public health. We have recently discovered a transcriptional activator that belongs to MarR family, EstR, and an esterase B (EstB) with a newly proposed de-arenethiolase activity from Sphingobium sp. SM42. De-arenethiolase activity involves the removal of the small aromatic side chain of cephalosporin antibiotics as an excellent leaving group by the enzymatic CS bond cleavage. Here, we report the regulation of estB through EstR as an activator in response to a third generation cephalosporin, cefoperazone, antibiotic. Cefoperazone induced the expression of estB in wild type Sphingobium sp., but not in the estR knockout strain, and the induction was restored in the complemented strain. Moreover, we revealed the importance of EstB localization in periplasm. Since EsB has the ability to inactivate selected ß-lactam antibiotics in vitro, it is possible that the enzyme works at the periplasmic space of Gram negative bacteria similar to ß-lactamases. EstB was genetically engineered by incorporating NlpA binding motif, or OmpA signal sequence, or SpyTag-SpyCatcher to the estB gene to mobilize it to different compartments of periplasm; inner membrane, outer membrane, and periplasmic space, respectively. Surprisingly, we found that Sphingobium sp. SM42 and E. coli expressing EstB at the periplasm were more sensitive to cefoperazone. The possible drug enhancement mechanism by enzyme was proposed. This work might lead to a novel strategy to tackle antibiotic resistance problem.


Subject(s)
Cefoperazone/pharmacology , Cephalosporins/pharmacology , Periplasm/enzymology , Serine Endopeptidases/genetics , Sphingomonadaceae/drug effects , Transcription Factors/genetics , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Periplasm/drug effects , Protein Sorting Signals , Sphingomonadaceae/enzymology , Sphingomonadaceae/genetics
4.
Appl Microbiol Biotechnol ; 102(6): 2803-2815, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29417199

ABSTRACT

Sphingomonads are isolated at exceptionally high frequency from organic polluted environments and assumed to be more hydrophobic than other Gram-negative bacteria. However, the potential roles of cell-surface hydrophobicity (CSH) in the cell survival in polluted environment, as well as the mechanisms underlying the CSH of sphingomonads, remain unclear. Sphingobium hydrophobicum C1T is a highly hydrophobic sphingomonad isolated from electronic-waste contaminated sediment. In this study, we found that exposure to the typical pollutants in electronic-waste contaminated sediment, such as the heavy metal ion Pb and the organic compound deca-brominated diphenyl ether (deca-BDE), resulted in the development of even higher CSH of the hydrophobic strain C1T; but no significant change was observed in the low CSH of its hydrophilic variant C2. The hydrophobic strain C1T achieved higher biomass yield in standing conditions and adsorbed more amounts of hydrophobic deca-BDE than its hydrophilic variant C2, suggesting that the high CSH potentially enhanced the adaptation of hydrophobic strain to colonize in sediment and adsorb hydrophobic nutrients. The identification of the bacterial cell-surface constituents showed that the high CSH of S. hydrophobicum was contributed greatly by outer-membrane proteins, particularly membrane transporters functioning as enhancers for nutrient uptake and stress sustainment. This study will enhance our understanding of the adaptive strategies of sphingomonads in contaminated environments. It will be of great importance to enhance the CSH of sphingomonads and utilize them in cleaning up the environment from organic pollution.


Subject(s)
Adaptation, Biological , Electronic Waste , Hydrophobic and Hydrophilic Interactions , Sphingomonadaceae/physiology , Surface Properties , Adsorption , Bacterial Outer Membrane Proteins/metabolism , Environmental Microbiology , Environmental Pollutants/metabolism , Halogenated Diphenyl Ethers/metabolism , Lead/metabolism , Membrane Transport Proteins/metabolism , Sphingomonadaceae/drug effects , Sphingomonadaceae/isolation & purification
5.
Wei Sheng Wu Xue Bao ; 57(3): 399-410, 2017 Mar 04.
Article in Zh | MEDLINE | ID: mdl-29756438

ABSTRACT

Objective: The present study aims to analyze the chemotaxis genes and proteins of several PAH-degrading Novosphingobium strains, and the chemotaxis of these strains toward aromatic compounds and intermediates. Methods: Based on genome comparative analysis, we identified the chemotaxis genes organization and proteins distribution. We used drop and swarm plate assays to detect the chemotaxis of these strains toward aromatic compounds and intermediates of TCA cycle. Results: We found that all these Novosphingobium strains showed chemotaxis, but the chemotatic ability varied. The completed genome sequenced strains N. pentaromativorans F2, N. pentaromativorans US6-1, N. pentaromativorans PP1Y, Novosphingobium sp. AP12, Novosphingobium sp. Rr 2-17, and Novosphingobium nitrogenifigens DSM 19370 contained MCP, CheW, CheA, CheB, CheR and CheY. Strain F2, US6-1 and PP1Y, shared a consistent order of chemotaxis genes in "che" cluster. The chemotatic system of these Novosphingobium strains belonged to the Fla chemotactic system. Conclusion: These strains all contained a complete chmotaxis pathway. Their chemotactic ability toward aromatic compounds and intermediates varied, and the chemotaxis of US6-1 was obvious.


Subject(s)
Chemotaxis , Sphingomonadaceae/physiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Chemotaxis/drug effects , Hydrocarbons, Aromatic/pharmacology , Sphingomonadaceae/drug effects , Sphingomonadaceae/genetics , Trichloroacetic Acid/pharmacology
6.
Bioorg Med Chem Lett ; 24(13): 2863-6, 2014 Jul 01.
Article in English | MEDLINE | ID: mdl-24825299

ABSTRACT

The biochemical mechanisms that marine sponges have developed as a chemical defense to protect themselves against micro and subsequent macrobiofouling process might comprise a potential alternative for the preventing attack of biofilm forming bacteria. The present study investigated the antimicrobial activity of a series of major secondary metabolites isolated from the sponges Fasciospongia cavernosa and Axinella donnani against fouling bacteria. Secomanoalide (1), dehydromanoalide (2) and cavernosine (3) have been isolated from F. cavernosa. Their structures were determined by MS, (1)H NMR spectra analyses and by comparison with those reported in the literature. The most promising activity was exhibited by the metabolites from A. donnani, that is, cerebroside (5) against three strains Aeromonas hydrophila subsp. salmonicida A449 and Erythrobacter litoralis. Our investigation revealed that combined metabolites 1, 2 and 3 retained strong activity but individual metabolite had moderate activity indicating that activity probably results from synergistic interactions between multiple compounds. The antibacterial screening of compounds 3, 5 and synergistic effect of 1-3 against fouling bacteria has been studied for the first time. Further, isolation of manoalide related compounds and their synergistic screening can be accelerated for the development of new biofilm inhibitors.


Subject(s)
Aeromonas hydrophila/drug effects , Aeromonas salmonicida/drug effects , Anti-Bacterial Agents/pharmacology , Porifera/chemistry , Sphingomonadaceae/drug effects , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Dose-Response Relationship, Drug , Microbial Sensitivity Tests , Molecular Structure , Structure-Activity Relationship
7.
Plant Physiol Biochem ; 214: 108958, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39053315

ABSTRACT

Uranium (U) contamination of rice is an urgent ecological and agricultural problem whose effective alleviation is in great demand. Sphingopyxis genus has been shown to remediate heavy metal-contaminated soils. Rare research delves into the mitigation of uranium (U) toxicity to rice by Sphingopyxis genus. In this study, we exposed rice seedlings for 7 days at U concentrations of 0, 10, 20, 40, and 80 mg L-1 with or without the Sphingopyxis sp. YF1 in the rice nutrient solution. Here, we firstly found YF1 colonized on the root of rice seedlings, significantly mitigated the growth inhibition, and counteracted the chlorophyll content reduction in leaves induced by U. When treated with 1.1 × 107 CFU mL-1 YF1 with the amendment of 10 mg L-1 U, the decrease of U accumulation in rice seedling roots and shoots was the largest among all treatments; reduced by 39.3% and 32.1%, respectively. This was associated with the redistribution of the U proportions in different organelle parts, leading to the alleviation of the U damage to the morphology and structure of rice root. Interestingly, we found YF1 significantly weakens the expression of antioxidant enzymes genes (CuZnSOD,CATA,POD), promotes the up-regulation of metal-transporters genes (OsHMA3 and OsHMA2), and reduces the lipid peroxidation damage induced by U in rice seedlings. In summary, YF1 is a plant-probiotic with potential applications for U-contaminated rice, benefiting producers and consumers.


Subject(s)
Oryza , Plant Roots , Uranium , Oryza/drug effects , Oryza/metabolism , Oryza/growth & development , Oryza/genetics , Uranium/toxicity , Plant Roots/metabolism , Plant Roots/drug effects , Plant Roots/growth & development , Sphingomonadaceae/metabolism , Sphingomonadaceae/genetics , Sphingomonadaceae/drug effects , Seedlings/drug effects , Seedlings/metabolism , Seedlings/growth & development , Plant Leaves/metabolism , Plant Leaves/drug effects , Gene Expression Regulation, Plant/drug effects , Plant Proteins/metabolism , Plant Proteins/genetics , Chlorophyll/metabolism
8.
Appl Environ Microbiol ; 78(14): 4802-15, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22582058

ABSTRACT

Novosphingobium nitrogenifigens Y88(T) (Y88) is a free-living, diazotrophic Alphaproteobacterium, capable of producing 80% of its biomass as the biopolymer polyhydroxybutyrate (PHB). We explored the potential utility of this species as a polyhydroxybutyrate production strain, correlating the effects of glucose, nitrogen availability, dissolved oxygen concentration, and extracellular pH with polyhydroxybutyrate production and changes in the Y88 proteomic profile. Using two-dimensional differential in-gel electrophoresis and tandem mass spectrometry, we identified 217 unique proteins from six growth conditions. We observed reproducible, characteristic proteomic signatures for each of the physiological states we examined. We identified proteins that changed in abundance in correlation with either nitrogen fixation, dissolved oxygen concentration, or acidification of the growth medium. The proteins that correlated with nitrogen fixation were identified either as known nitrogen fixation proteins or as novel proteins that we predict play roles in aspects of nitrogen fixation based on their proteomic profiles. In contrast, the proteins involved in central carbon and polyhydroxybutyrate metabolism were constitutively abundant, consistent with the constitutive polyhydroxybutyrate production that we observed in this species. Three proteins with roles in detoxification of reactive oxygen species were identified in this obligate aerobe. The most abundant protein in all experiments was a polyhydroxyalkanoate granule-associated protein, phasin. The full-length isoform of this protein has a long, intrinsically disordered Ala/Pro/Lys-rich N-terminal segment, a feature that appears to be unique to sphingomonad phasins. The data suggest that Y88 has potential as a PHB production strain due to its aerobic tolerance and metabolic orientation toward polyhydroxybutyrate accumulation, even in low-nitrogen growth medium.


Subject(s)
Bacterial Proteins/metabolism , DNA-Binding Proteins/metabolism , Nitrogen Fixation/physiology , Polyhydroxyalkanoates/biosynthesis , Proteomics/methods , Reactive Oxygen Species/pharmacology , Sphingomonadaceae/drug effects , Bacterial Proteins/genetics , DNA-Binding Proteins/genetics , Drug Resistance, Bacterial , Phenotype , Sphingomonadaceae/classification , Sphingomonadaceae/growth & development , Sphingomonadaceae/metabolism , Tandem Mass Spectrometry
9.
Appl Environ Microbiol ; 77(16): 5697-706, 2011 Aug 15.
Article in English | MEDLINE | ID: mdl-21705522

ABSTRACT

Sphingomonadaceae (n = 86) were isolated from a drinking water treatment plant (n = 6), tap water (n = 55), cup fillers for dental chairs (n = 21), and a water demineralization filter (n = 4). The bacterial isolates were identified based on analysis of the 16S rRNA gene sequence, and intraspecies variation was assessed on the basis of atpD gene sequence analysis. The isolates were identified as members of the genera Sphingomonas (n = 27), Sphingobium (n = 28), Novosphingobium (n = 12), Sphingopyxis (n = 7), and Blastomonas (n = 12). The patterns of susceptibility to five classes of antibiotics were analyzed and compared for the different sites of isolation and taxonomic groups. Colistin resistance was observed to be intrinsic (92%). The highest antibiotic resistance prevalence values were observed in members of the genera Sphingomonas and Sphingobium and for beta-lactams, ciprofloxacin, and cotrimoxazole. In tap water and in water from dental chairs, antibiotic resistance was more prevalent than in the other samples, mainly due to the predominance of isolates of the genera Sphingomonas and Sphingobium. These two genera presented distinct patterns of association with antibiotic resistance, suggesting different paths of resistance development. Antibiotic resistance patterns were often related to the species rather than to the site or strain, suggesting the importance of vertical resistance transmission in these bacteria. This is the first study demonstrating that members of the family Sphingomonadaceae are potential reservoirs of antibiotic resistance in drinking water.


Subject(s)
Drinking Water/microbiology , Drug Resistance, Multiple, Bacterial , Genetic Variation , Sphingomonadaceae/drug effects , Bacterial Typing Techniques , Base Sequence , Biofilms , Cluster Analysis , Culture Media/chemistry , Dental Equipment/microbiology , Disease Reservoirs , Genes, Bacterial , Genes, rRNA , Molecular Sequence Data , Phenotype , RNA, Ribosomal, 16S/genetics , Sphingomonadaceae/classification , Sphingomonadaceae/genetics , Sphingomonadaceae/isolation & purification , Water Microbiology
10.
mSphere ; 6(4): e0041221, 2021 08 25.
Article in English | MEDLINE | ID: mdl-34378981

ABSTRACT

Rhizoremediation is based on the ability of microorganisms to metabolize nutrients from plant root exudates and, thereby, to cometabolize or even mineralize toxic environmental contaminants. Novosphingobium sp. HR1a is a bacterial strain able to degrade a wide variety of polycyclic aromatic hydrocarbons (PAHs). Here, we have demonstrated that the number of CFU in microcosms vegetated with clover was almost 2 orders of magnitude higher than that in nonvegetated microcosms or microcosms vegetated with rye-grass or grass. Strain HR1a was able to eliminate 92% of the phenanthrene in the microcosms with clover after 9 days. We have studied the molecular basis of the interaction between strain HR1a and clover by phenomic, metabolomic, and transcriptomic analyses. By measuring the relative concentrations of several metabolites exudated by clover both in the presence and in the absence of the bacteria, we identified some compounds that were probably consumed in the rhizosphere; the transcriptomic analyses confirmed the expression of genes involved in the catabolism of these compounds. By using a transcriptional fusion of the green fluorescent protein (GFP) to the promoter of the gene encoding the dioxygenase involved in the degradation of PAHs, we have demonstrated that this gene is induced at higher levels in clover microcosms than in nonvegetated microcosms. Therefore, the positive interaction between clover and Novosphingobium sp. HR1a during rhizoremediation is a result of the bacterial utilization of different carbon and nitrogen sources released during seedling development and the capacity of clover exudates to induce the PAH degradation pathway. IMPORTANCE The success of an eco-friendly and cost-effective strategy for soil decontamination is conditioned by the understanding of the ecology of plant-microorganism interactions. Although many studies have been published about the bacterial metabolic capacities in the rhizosphere and about rhizoremediation of contaminants, there are fewer studies dealing with the integration of bacterial metabolic capacities in the rhizosphere during PAH bioremediation, and some aspects still remain controversial. Some authors have postulated that the presence of easily metabolizable carbon sources in root exudates might repress the expression of genes required for contaminant degradation, while others found that specific rhizosphere compounds can induce such genes. Novosphingobium sp. HR1a, which is our model organism, has two characteristics desirable in bacteria for use in remediation: its ubiquity and the capacity to degrade a wide variety of contaminants. We have demonstrated that this bacterium consumes several rhizospheric compounds without repression of the genes required for the mineralization of PAHs. In fact, some compounds even induced their expression.


Subject(s)
Medicago/chemistry , Phenanthrenes/metabolism , Plant Roots/chemistry , Sphingomonadaceae/physiology , Biodegradation, Environmental , Medicago/drug effects , Phenanthrenes/analysis , Phenanthrenes/pharmacology , Plant Extracts/pharmacology , Soil Microbiology , Sphingomonadaceae/drug effects
11.
Appl Environ Microbiol ; 76(7): 2075-85, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20118361

ABSTRACT

The effects of the chiral pharmaceuticals atenolol and propranolol on Pseudomonas putida, Pseudomonas aeruginosa, Micrococcus luteus, and Blastomonas natatoria were investigated. The growth dynamics of exposed cultures were monitored using a Bioscreen instrument. In addition, Fourier-transform infrared (FT-IR) spectroscopy with appropriate chemometrics and high-performance liquid chromatography (HPLC) were employed in order to investigate the phenotypic changes and possible degradation of the drugs in exposed cultures. For the majority of the bacteria studied there was not a statistically significant difference in the organism's phenotype when it was exposed to the different enantiomers or mixtures of enantiomers. In contrast, the pseudomonads appeared to respond differently to propranolol, and the two enantiomers had different effects on the cellular phenotype. This implies that there were different metabolic responses in the organisms when they were exposed to the different enantiomers. We suggest that our findings may indicate that there are widespread effects on aquatic communities in which active pharmaceutical ingredients are present.


Subject(s)
Antimetabolites/pharmacology , Atenolol/pharmacology , Metabolome/drug effects , Micrococcus luteus/drug effects , Propranolol/pharmacology , Pseudomonas/drug effects , Sphingomonadaceae/drug effects , Antimetabolites/metabolism , Atenolol/metabolism , Chromatography, High Pressure Liquid , Micrococcus luteus/chemistry , Micrococcus luteus/growth & development , Propranolol/metabolism , Pseudomonas/chemistry , Pseudomonas/growth & development , Spectroscopy, Fourier Transform Infrared , Sphingomonadaceae/chemistry , Sphingomonadaceae/growth & development
12.
Sci Rep ; 10(1): 12177, 2020 07 22.
Article in English | MEDLINE | ID: mdl-32699224

ABSTRACT

Iron, an essential element for all organisms, acts as a cofactor of enzymes in bacterial degradation of recalcitrant aromatic compounds. The bacterial family, Sphingomonadaceae comprises various degraders of recalcitrant aromatic compounds; however, little is known about their iron acquisition system. Here, we investigated the iron acquisition system in a model bacterium capable of degrading lignin-derived aromatics, Sphingobium sp. strain SYK-6. Analyses of SYK-6 mutants revealed that FiuA (SLG_34550), a TonB-dependent receptor (TBDR), was the major outer membrane iron transporter. Three other TBDRs encoded by SLG_04340, SLG_04380, and SLG_10860 also participated in iron uptake, and tonB2 (SLG_34540), one of the six tonB comprising the Ton complex which enables TBDR-mediated transport was critical for iron uptake. The ferrous iron transporter FeoB (SLG_36840) played an important role in iron uptake across the inner membrane. The promoter activities of most of the iron uptake genes were induced under iron-limited conditions, and their regulation is controlled by SLG_29410 encoding the ferric uptake regulator, Fur. Although feoB, among all the iron uptake genes identified is highly conserved in Sphingomonad strains, the outer membrane transporters seem to be diversified. Elucidation of the iron acquisition system promises better understanding of the bacterial degradation mechanisms of aromatic compounds.


Subject(s)
Bacterial Proteins/metabolism , Benzene Derivatives/chemistry , Iron/metabolism , Lignin/chemistry , Sphingomonadaceae/metabolism , Bacterial Outer Membrane/metabolism , Bacterial Proteins/genetics , Cation Transport Proteins/genetics , Cation Transport Proteins/metabolism , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Hydroxybenzoates/chemistry , Hydroxybenzoates/pharmacology , Lignin/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mutagenesis , Promoter Regions, Genetic , Sphingomonadaceae/drug effects , Sphingomonadaceae/genetics , Vanillic Acid/chemistry , Vanillic Acid/pharmacology
13.
J Dent ; 35(5): 438-51, 2007 May.
Article in English | MEDLINE | ID: mdl-17303302

ABSTRACT

UNLABELLED: Although many studies have highlighted the problem of biofilm growth in dental chair unit waterlines (DUWs), no long-term studies on the efficacy of DUW disinfection using a large number of dental chair units (DCUs) have been reported. OBJECTIVES: To investigate the long-term (21 months) efficacy of the Planmeca Waterline Cleaning System (WCS) to maintain the quality of DUW output water below the American Dental Association (ADA) recommended standard of < or =200cfu/mL of aerobic heterotrophic bacteria using once weekly disinfection with the hydrogen peroxide-and silver ion-containing disinfectant Planosil. METHODS: Microbiological quality of DUW output water was monitored by culture on R2A agar for 10 DCUs fitted with the WCS. The presence of biofilm in DUWs was examined by electron microscopy. RESULTS: During the first 9 months a high prevalence (28/300 disinfection cycles; 9.3%) of intermittent DUW disinfection failure occurred in 8/10 DCUs due to operator omission to disinfect all DUWs (10/28 failed cycles), incorrect compressed air pressure failing to distribute the disinfectant properly (4/28 failed cycles) and physical blockage of disinfectant intake valves due to corrosion effects of Planosil (14/28 failed cycles). On rectification of these faults through engineering redesign and procedural changes, no further cases of intermittent DUW disinfection failure were observed. Independently of these factors, a rapid and consistent decline in efficacy of DUW disinfection occurred in 4/10 DCUs following the initial 9 months of once weekly disinfection. There was a highly significant difference (P<0.0001) in the prevalence of strongly catalase-positive Novosphingobium and Sphingomonas bacterial species (mean average prevalence of 37.1%) in DUW output water from these 4 DCUs compared to the other 6 DCUs and DCU supply water (prevalence <1%), which correlated with biofilm presence in the DUWs and indicated selective pressure for maintenance of these species by prolonged disinfectant usage. Planosil was reformulated to a more concentrated form (Planosil Forte) and when used once weekly was found to maintain bacterial density in output water below the ADA standard for all 10 DCUs. CONCLUSIONS: A variety of factors can contribute to failure of DUW disinfection in the long-term, including human error, disinfectant corrosion of equipment and natural selection of naturally disinfectant-tolerant bacterial species.


Subject(s)
Dental Disinfectants , Dental Equipment , Disinfection/methods , Water Microbiology , Analysis of Variance , Bacterial Typing Techniques , Corrosion , DNA, Bacterial/analysis , Dental Disinfectants/chemistry , Dental Disinfectants/pharmacology , Drug Resistance, Bacterial , Equipment Failure , Hydrogen Peroxide , Prospective Studies , Silver/chemistry , Silver/pharmacology , Sphingomonadaceae/drug effects
14.
Sci Rep ; 7(1): 8262, 2017 08 15.
Article in English | MEDLINE | ID: mdl-28811613

ABSTRACT

Nickel acts as cofactor for a number of enzymes of many bacteria species. Its homeostasis is ensured by proteins working as ion efflux or accumulation systems. These mechanisms are also generally adopted to counteract life-threatening high extra-cellular Ni2+ concentrations. Little is known regarding nickel tolerance in the genus Sphingobium. We studied the response of the novel Sphingobium sp. ba1 strain, able to adapt to high Ni2+ concentrations. Differential gene expression in cells cultured in 10 mM Ni2+, investigated by RNA-seq analysis, identified 118 differentially expressed genes. Among the 90 up-regulated genes, a cluster including genes coding for nickel and other metal ion efflux systems (similar to either cnrCBA, nccCBA or cznABC) and for a NreB-like permease was found. Comparative analyses among thirty genomes of Sphingobium species show that this cluster is conserved only in two cases, while in the other genomes it is partially present or even absent. The differential expression of genes encoding proteins which could also work as Ni2+-accumulators (HupE/UreJ-like protein, NreA and components of TonB-associated transport and copper-homeostasis systems) was also detected. The identification of Sphingobium sp. ba1 strain adaptive mechanisms to nickel ions, can foster its possible use for biodegradation of poly-aromatic compounds in metal-rich environments.


Subject(s)
Nickel/adverse effects , Sphingomonadaceae/drug effects , Sphingomonadaceae/genetics , Biodegradation, Environmental , Gene Expression Profiling , Gene Expression Regulation, Bacterial/drug effects , Genomics/methods , High-Throughput Nucleotide Sequencing , Sequence Analysis, RNA , Sphingomonadaceae/growth & development , Sphingomonadaceae/metabolism , Transcriptome
15.
FEMS Microbiol Ecol ; 58(3): 364-72, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17117981

ABSTRACT

The survival and effectiveness of a bioaugmentation strain in its target environment depend not only on physicochemical parameters in the soil but also on the physiological state of the inoculated organism. This study examined the effect of variations in inoculum pretreatment on the survival, metabolic activity (measured as rRNA content) and polycyclic aromatic hydrocarbon (PAH)-catabolic gene expression of Sphingobium yanoikuyae B1 in an aged PAH-contaminated soil. RNA denaturing gradient gel electrophoresis analysis showed stable colonization of PAH-contaminated soil by S. yanoikuyae B1 after four pretreatments (growth in complex or minimal medium, starvation, or acclimation to phenanthrene). By contrast, extractable CFUs decreased with time for all four treatments, and significantly faster for Luria Bertani-grown inocula, suggesting that these cells adhered strongly to soil particles while remaining metabolically active. Pretreatment of the inoculum had a dramatic effect on the expression of genes specific to the PAH-degradation pathway. The highest levels of bphC and xylE expression were seen for inocula that had been precultivated on complex medium, and degradation of PAHs was significantly enhanced in soils treated with these inocula. The results suggest that using complex media instead of minimal media for cultivating bioaugmentation inocula may improve the subsequent efficiency of contaminant biodegradation in the soil.


Subject(s)
Gene Expression Regulation, Bacterial/drug effects , Polycyclic Aromatic Hydrocarbons/pharmacology , Soil Microbiology , Soil Pollutants/pharmacology , Sphingomonadaceae/drug effects , Sphingomonadaceae/metabolism , Biodegradation, Environmental , Molecular Structure , Polycyclic Aromatic Hydrocarbons/metabolism , RNA, Bacterial/biosynthesis , RNA, Bacterial/drug effects , Reverse Transcriptase Polymerase Chain Reaction
17.
J Gen Appl Microbiol ; 62(1): 14-7, 2016.
Article in English | MEDLINE | ID: mdl-26923126

ABSTRACT

In the present study, spheroplasts from the aerobic anoxygenic photosynthetic marine bacterium Erythrobacter litoralis were generated and cultivated. In the presence of penicillin, the spheroplasts grew and enlarged in marine broth without undergoing cell division. However, continuous light inhibited their enlargement, and they were therefore cultivated in the dark. Cellular DNA was quantified at various time points (0, 24, and 48 h) and temperatures (20°C, 25°C, and 30°C) using real-time quantitative PCR. The DNA content was highest at 30°C in the absence of penicillin, whereas there was no observable change with exposure to penicillin at all evaluated temperatures. During growth, larger spheroplasts were more frequently observed at 25°C in the presence of penicillin. These results demonstrate that the optimal culture conditions for the enlargement of spheroplasts in E. litoralis differ from those required for cell division.


Subject(s)
Darkness , Spheroplasts/growth & development , Sphingomonadaceae/growth & development , Anti-Bacterial Agents/metabolism , DNA, Bacterial/analysis , Penicillins/metabolism , Real-Time Polymerase Chain Reaction , Spheroplasts/cytology , Spheroplasts/drug effects , Sphingomonadaceae/cytology , Sphingomonadaceae/drug effects , Temperature
18.
Chemosphere ; 144: 330-7, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26378869

ABSTRACT

A mercury resistant bacterial strain, SA2, was isolated from soil contaminated with mercury. The 16S rRNA gene sequence of this isolate showed 99% sequence similarity to the genera Sphingobium and Sphingomonas of α-proteobacteria group. However, the isolate formed a distinct phyletic line with the genus Sphingobium suggesting the strain belongs to Sphingobium sp. Toxicity studies indicated resistance to high levels of mercury with estimated EC50 values 4.5 mg L(-1) and 44.15 mg L(-1) and MIC values 5.1 mg L(-1) and 48.48 mg L(-1) in minimal and rich media, respectively. The strain SA2 was able to volatilize mercury by producing mercuric reductase enzyme which makes it potential candidate for remediating mercury. ICP-QQQ-MS analysis of Hg supplemented culture solutions confirmed that almost 79% mercury in the culture suspension was volatilized in 6 h. A very small amount of mercury was observed to accumulate in cell pellets which was also evident according to ESEM-EDX analysis. The mercuric reductase gene merA was amplified and sequenced. The deduced amino acid sequence demonstrated sequence homology with α-proteobacteria and Ascomycota group.


Subject(s)
Bacterial Proteins/metabolism , Mercury/metabolism , Oxidoreductases/metabolism , Soil Pollutants/metabolism , Sphingomonadaceae/metabolism , Amino Acid Sequence , Bacterial Proteins/genetics , Base Sequence , Biodegradation, Environmental , Mercury/toxicity , Molecular Sequence Data , Oxidoreductases/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Soil Pollutants/toxicity , Sphingomonadaceae/drug effects , Sphingomonadaceae/genetics , Sphingomonadaceae/growth & development
19.
J Hazard Mater ; 300: 121-128, 2015 Dec 30.
Article in English | MEDLINE | ID: mdl-26164069

ABSTRACT

Soils are exposed to various types of chemical contaminants due to anthropogenic activities; however, research on persistent organic pollutants and the existence of antibiotic resistance genes (ARGs) is limited. To our knowledge, the present work for the first time focused on the bioremediation of soil co-contaminated with pyrene and tetracycline/sulfonamide-resistance genes. After 90 days of incubation, the pyrene concentration and the abundance of the four ARGs (tetW, tetM, sulI, and sulII) significantly decreased in different treatment conditions (p<0.05). The greatest pyrene removal (47.8%) and greatest decrease in ARG abundance (from 10(-7) to 10(-8) ARG copies per 16S rRNA copy) were observed in microcosms with a combination of bacterial and sophorolipid treatment. Throughout the incubation, pyrene bioaccessibility constantly declined in the microcosm inoculated with bacteria. However, an increased pyrene bioaccessibility and ARG abundance at day 40 were observed in soil treated with sophorolipid alone. Tenax extraction methods and linear correlation analysis indicated a strong positive relationship between the rapidly desorbing fraction (Fr) of pyrene and ARG abundance. Therefore, we conclude that bioaccessible pyrene rather than total pyrene plays a major role in the maintenance and fluctuation of ARG abundance in the soil.


Subject(s)
Drug Resistance, Microbial/genetics , Genes, Bacterial , Pyrenes/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Sphingomonadaceae/genetics , Biodegradation, Environmental , Lipids/pharmacology , Pyrenes/analysis , RNA, Ribosomal, 16S/genetics , Soil Pollutants/analysis , Sphingomonadaceae/drug effects , Sphingomonadaceae/metabolism , Surface-Active Agents/pharmacology
20.
FEMS Microbiol Lett ; 361(1): 8-9, 2014 12.
Article in English | MEDLINE | ID: mdl-25288103

ABSTRACT

The genome sequence of a Sphingobium strain capable of tolerating high concentrations of Ni ions, and exhibiting natural kanamycin resistance, is presented. The presence of a transposon derived kanamycin resistance gene and several genes for efflux-mediated metal resistance may explain the observed characteristics of the new Sphingobium isolate.


Subject(s)
Drug Resistance, Bacterial/genetics , Genome, Bacterial/genetics , Kanamycin/pharmacology , Nickel/toxicity , Sphingomonadaceae/genetics , Sphingomonadaceae/drug effects , Sphingomonadaceae/isolation & purification
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