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 effectsABSTRACT
Recently, the coupling of biofuel fermentation broths and pervaporation has been receiving increasing attention. Some challenges, such as the destructive effects of constituents of the real fermentation broth on the membrane performances, the lethal effects of the membrane surface chemical modifiers on the microorganisms, and being expensive, are against this concept. For the first time, a continuous study on the one-step and low-cost preparation of superhydrophobic membranes for bioethanol separation is made to address these challenges. In our previous work, spraying as a fast, scalable, and low-cost procedure was applied to fabricate the one-layered active-layer hydrophobic (OALH) silicalite-1/polydimethylsiloxane (PDMS) membrane on the low-cost mullite support. In this work, the spraying method was adopted to fabricate a two-layered active-layer superhydrophobic (TALS) silicalite-1/PDMS membrane, where the novel active layer consisted of two layers with different hydrophobicities and densities. Contact-angle measurements, surface charge determination, scanning electron microscopy, atomic force microscopy, and pervaporation separation using a 5 wt % ethanol solution were used to statically evaluate the fouling/biofouling resistance and pervaporation performances of OALH and TALS membranes in this study. The TALS membrane presented a better resistance and performance. For dynamic experiments, the Box-Behnken design was used to identify the effects of substrates, microorganisms, and nutrient contents as the leading indicators of fermentation broth on the TALS membrane performances for the long-term utilization. The maximum performances of 1.88 kg/m2·h, 32.34, and 59.04 kg/m2·h concerning the permeation flux, separation factor, and pervaporation separation index were obtained, respectively. The dynamic fouling/biofouling resistance of the TALS membrane was also characterized using energy-dispersive X-ray spectroscopy of all the tested membranes. The TALS membrane demonstrated the synergistic resistance of membrane fouling and biofouling. Eventually, the novel TALS membrane was found to have potential for biofuel recovery, especially bioethanol.
Subject(s)
Biofouling/prevention & control , Biofuels , Dimethylpolysiloxanes/chemistry , Hydrogen-Ion Concentration , Hydrophobic and Hydrophilic Interactions , Membranes, Artificial , Sphingomonadaceae/physiologyABSTRACT
BACKGROUND: Numerous studies have reported the health-promoting effects of exopolysaccharides (EPSs) in in vitro models; however, a functional evaluation of EPSs will provide additional knowledge of EPS-microbe interactions by in vivo intestinal microbial model. In the present study, high-throughput amplicon sequencing, short-chain fatty acid (SCFAs) and intestinal inflammation evaluation were performed to explore the potential benefits of exopolysaccharides (EPSs) and EPS-producing Lactobacillus (HNUB20 group) using the healthy zebrafish (Danio rerio) model. RESULTS: The results based on microbial taxonomic analysis revealed that the abundance of four genera, Ochrobactrum, Sediminibacterium, Sphingomonas and Sphingobium, were increased in the control group in comparison to HNUB20 group. Pelomonas spp. levels were significantly higher and that of the genera Lactobacillus and Brachybacterium were significantly decreased in EPS group compared with control group. PICRUSt based functional prediction of gut microbiota metabolic pathways indicated that significantly lower abundance was found for transcription, and membrane transport, whereas folding, sorting and degradation and energy metabolism had significantly higher abundance after HNUB20 treatment. Two metabolic pathways, including metabolism and endocrine functions, were more abundant in the EPS group than control group. Similar to the HNUB20 group, transcription was also decreased in the EPS group compared with the control group. However, SCFAs and immune indexes indicated EPS and HNUB20 performed limited efficacy in the healthy zebrafish. CONCLUSIONS: The present intestinal microbial model-based study indicated that EPSs and high-yield EPS-producing Lactobacillus can shake the structure of intestinal microbiota, but cannot change SCFAs presence and intestinal inflammation.
Subject(s)
Gastrointestinal Microbiome/physiology , Intestines/microbiology , Lactobacillus/physiology , Polysaccharides, Bacterial/pharmacology , Zebrafish/microbiology , Actinobacteria/physiology , Animals , Bacteroidetes/physiology , Comamonadaceae/physiology , Fatty Acids, Volatile/metabolism , Female , Male , Metabolic Networks and Pathways/genetics , Ochrobactrum/physiology , Polysaccharides, Bacterial/biosynthesis , Sphingomonadaceae/physiology , Sphingomonas/physiology , Transcription, GeneticABSTRACT
A novel carbofuran-degrading strain CFD-1 was isolated and preliminarily identified as Sphingbium sp. This strain was able to utilize carbofuran as the sole carbon source for growth. The carbofuran hydrolase gene cehA was cloned from strain CFD-1 and expressed in Escherichia coli. CehA could hydrolyze carbamate pesticides including carbofuran and carbaryl efficiently, while it showed poor hydrolysis ability against isoprocarb, propoxur, oxamyl and aldicarb. CehA displayed maximal enzymatic activity at 40⯰C and pH 7.0. The apparent Km and Kcat values of CehA for carbofuran were 133.22⯱â¯5.70⯵M and 9.48⯱â¯0.89 s-1, respectively. The site-directed mutation experiment showed that His313, His315, His453 and His495 played important roles in the hydrolysis of carbofuran by CehA. Furthermore, the sequence of cehA is highly conserved among different carbofuran-degrading strains, and there are mobile elements around cehA, indicating that it may be transferred horizontally between different strains.
Subject(s)
Carbofuran/metabolism , Pesticides/metabolism , Sphingomonadaceae/physiology , Amino Acids/metabolism , Biodegradation, Environmental , Carbamates , Carbaryl/metabolism , Hydrolases/metabolism , HydrolysisABSTRACT
Vanillate and syringate are major intermediate metabolites generated during the microbial degradation of lignin. In Sphingobium sp. SYK-6, vanillate is O demethylated to protocatechuate by LigM; protocatechuate is then catabolized via the protocatechuate 4,5-cleavage pathway. Syringate is O demethylated to gallate by consecutive reactions catalyzed by DesA and LigM, and then gallate is subjected to ring cleavage by DesB. Here, we investigated the transcriptional regulation of desA, ligM, and desB involved in vanillate and syringate catabolism. Quantitative reverse transcription-PCR analyses indicated that the transcription of these genes was induced 5.8-37-fold in the presence of vanillate and syringate. A MarR-type transcriptional regulator, SLG_12870 (desR), was identified as the gene whose product bound to the desB promoter region. Analysis of a desR mutant indicated that the transcription of desB, ligM, and desR is negatively regulated by DesR. Purified DesR bound to the upstream regions of desB, ligM, and desR, and the inverted repeat sequences similar to each other in these regions were suggested to be essential for DNA binding of DesR. Vanillate and syringate inhibited DNA binding of DesR, indicating that these compounds are effector molecules of DesR. The transcription of desA was found to be regulated by an as-yet unidentified regulator.
Subject(s)
Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Lignin/metabolism , Repressor Proteins/metabolism , Sphingomonadaceae/physiology , Bacterial Proteins/genetics , Metabolic Networks and Pathways/genetics , Oxidoreductases, O-Demethylating/genetics , Oxidoreductases, O-Demethylating/metabolism , Promoter Regions, Genetic/genetics , Transcription, Genetic , Vanillic Acid/metabolismABSTRACT
Strain SYSU M10002T was isolated from a water sample collected from the coastal region of Pearl River estuary, Guangdong Province, southern China. The taxonomic position of the isolate was investigated by polyphasic taxonomic approaches. The isolate was found to be Gram-negative, non-motile, short rods and aerobic. The strain was able to grow at 14-37 °C, pH 6.0-10.0 and in the presence of up to 0.5% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SYSU M10002T is a member of the family Sphingomonadaceae, with high sequence similarity to Sphingorhabdus buctiana T5T (95.1%). Overall genomic related indices between the genome of strain SYSU M10002T and those of related strains were low to moderate (AAI values < 64.3%; POCP values < 58%), indicating that strain SYSU M10002T represents a novel lineage within the family Sphinogomonadaceae. Strain SYSU M10002T contained homospermidine as its polyamine. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, two unidentified phospholipids and an unidentified aminolipid. Ubiquinone Q-9 (44.9%) and Q-10 (43.2%) were the dominant respiratory quinones, along with a minor amount of Q-8 (11.9%). The predominant cellular fatty acids (> 10%) identified were summed feature 3 (C16:1ω7c and/or C16:1ω6c), summed feature 8 (C18:1ω7c) and C14:0 2-OH. The genomic DNA G+C content was 64.0%. Based on the analyses of the phenotypic, genotypic and phylogenetic characteristics, strain SYSU M10002T is determined to represent a novel species of a novel genus, for which the name Aestuariisphingobium litorale gen. nov., sp. nov. is proposed. The type strain of the species is SYSU M10002T (= KCTC 52944T = NBRC 112961T).
Subject(s)
Rivers/microbiology , Sphingomonadaceae/classification , Sphingomonadaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , China , Cluster Analysis , Cytosol/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Estuaries , Fatty Acids/analysis , Phospholipids/analysis , Phylogeny , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spermidine/analysis , Sphingomonadaceae/genetics , Sphingomonadaceae/physiologyABSTRACT
A novel Gram-staining negative, yellow-pigmented, non-motile, aerobic and rod-shaped bacterium, designated MAH-11T, was isolated from rhizosphere of Pinus koraiensis and was characterised by using a polyphasic taxonomic approach. The colonies were smooth, circular and 0.3-1.0 mm in diameter when grown on R2A agar for 3 days. The strain was positive for both catalase and oxidase tests. Optimum growth temperature and pH were 28-30 °C and 7.0, respectively. Cell growth occurs on R2A agar, nutrient agar, Luria-Bertani agar and tryptone soya agar but not on MacConkey agar. The novel strain was found to be able to hydrolyse esculin but not casein, gelatin, starch, L-tyrosine, DNA, L-arginine, urea, Tween 20 and Tween 80. On the basis of 16S rRNA gene sequence analysis, strain MAH-11T belongs to the genus Sphingobium and is closely related to Sphingobium quisquiliarum P25T (98.1%), Sphingobium vermicomposti VC-230T (97.8%), Sphingobium mellinum WI4T (97.5%), Sphingobium barthaii KK22T (97.2%) and Sphingobium fuliginis TKPT (97.2%). In DNA-DNA hybridization tests, the DNA relatedness values between strain MAH-11T and its close phylogenetic neighbors were below 45.0%. The DNA G+C content was 64.5 mol% and the predominant respiratory quinone was identified as ubiquinone-10. The major cellular fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c), summed feature 3 (C16:1ω7c and/or C16:1ω6c) and C16:0. The DNA-DNA hybridization results in combination with chemotaxonomic and physiological data demonstrated that strain MAH-11T represents a novel species within the genus Sphingobium, for which the name Sphingobium chungangianum is proposed. The type strain is MAH-11T (= KACC 19836T = CGMCC 1.13749T).
Subject(s)
Pinus/growth & development , Soil Microbiology , Sphingomonadaceae/classification , Sphingomonadaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , Cluster Analysis , Cytosol/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Nucleic Acid Hybridization , Phylogeny , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Rhizosphere , Sequence Analysis, DNA , Sphingomonadaceae/genetics , Sphingomonadaceae/physiologyABSTRACT
N-Acylhomoserine lactone (AHL)-degrading enzyme is identified from the various environments and applied for quorum-sensing inhibition. In this study, we isolated two AHL-degrading strains, Sphingopyxis sp. EG6 and FD7, from the industrial cooling water samples. When the eight Sphingopyxis type strains were checked for the AHL-degrading activity, two strains, Sphingopyxis alaskensis DSM 13593 and Sphingopyxis bauzanensis DSM 22271, showed high AHL-degrading activity. The complete genome sequences of EG6 and FD7 revealed the presence of gene homolog of qsdS, which encodes AHL-lactonase in Sphingomonas ursincola. The qsdS gene is seated between putative gene homologs involved in 3-isopropylmalate dehydratase large (leuC2) and small (leuD) subunits in the genome of EG6, FD7, DSM 13593, and DSM 22271, but completely disappeared between leuC2 and leuD in the genome sequences of Sphingopyxis type strains without AHL-degrading activity. Purified His-tagged QsdS showed high AHL-degrading activity and catalyzed AHL ring opening by hydrolyzing lactones. In addition, heterologous expression of qsdS in Pseudomonas aeruginosa resulted in reduction of biofilm formation. These results suggested that the AHL-degrading activity in Sphingopyxis is useful as an effective agent for biofilm inhibition.
Subject(s)
Carboxylic Ester Hydrolases , Lactones/metabolism , Sphingomonadaceae/enzymology , Sphingomonadaceae/genetics , Amino Acid Sequence , Base Sequence , Biofilms/growth & development , Carboxylic Ester Hydrolases/analysis , Carboxylic Ester Hydrolases/genetics , Carboxylic Ester Hydrolases/metabolism , Chromosome Mapping , Cloning, Molecular , Genome, Bacterial , Phylogeny , Pseudomonas aeruginosa/enzymology , Pseudomonas aeruginosa/genetics , Quorum Sensing/genetics , Sequence Analysis, DNA , Sphingomonadaceae/physiology , Sphingomonas/enzymology , Sphingomonas/geneticsABSTRACT
A pale yellow bacterial strain, designated JJ-A5T, was isolated form an agricultural soil from Jeju Island in Republic of Korea. Cells of the strain were Gram-stain-negative, motile, flagellated and rod-shaped. The strain grew at 15-30°C, pH 6.0-9.0, and in the presence of 0-1.5% (w/v) NaCl. Growth occurred on R2A, but not on Luria-Bertani agar, nutrient agar, trypticase soy agar and MacConkey agar. The strain utilized alachlor as a sole carbon source for growth. The strain JJ-A5T showed 16S rRNA gene sequence similarities lower than 95.4% with members of the family Sphingomonadaceae. Phylogenetic analysis showed that the strain belongs to the family Sphingomonadaceae and strain JJ-A5T was distinctly separated from established genera of this family. The strain contained Q-10 as dominant ubiquinone and spermidine as major polyamine. The predominant cellular fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c), summed feature 3 (C16:1ω7c and/or C16:1ω6c), 11-methyl C18:1ω7c, C16:0 and C14:0 2-OH. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, and phosphatidylcholine. The DNA G + C content of the strain was 62.7 mol%. On the basis of the phenotypic, genomic and chemotaxonomic characteristics, strain JJ-A5T is considered to represent a novel genus and species within the family Sphingomonadaceae, for which the name Tardibacter chloracetimidivorans gen. nov., sp. nov. is proposed. The type strain of Tardibacter chloracetimidivorans is JJ-A5T (= KACC 19450T = NBRC 113160T).
Subject(s)
Agriculture , Phylogeny , Soil Microbiology , Sphingomonadaceae/classification , Sphingomonadaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/analysis , Islands , Nucleic Acid Hybridization , Phospholipids/analysis , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA , Soil , Species Specificity , Spermidine/analysis , Sphingomonadaceae/genetics , Sphingomonadaceae/physiology , Ubiquinone/analysisABSTRACT
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 & purificationABSTRACT
AIM: To analyse the physiological response of Sphingobium sp. 22B to water stress. METHODS AND RESULTS: The strain was grown under excess of carbon source and then subjected to low (60RH) and high (18RH) water stress conditions for 96 h. Quantification of trehalose, glycogen, polyhydroxybutyrate (PHB) and transmission electron microscopy (TEM) was studied. Genes linked with desiccation were searched in Sphingobium sp. 22B and Sphingomonas 'sensu latu' genomes and their transcripts were quantified by real-time PCR. Results showed that, in the absence of water stress, strain 22B accumulated 4·76 ± 1·41% of glycogen, 0·84 ± 1·62% of trehalose and 44·9 ± 6·4% of PHB per cellular dry weight. Glycogen and trehalose were mobilized under water stressed conditions, this mobilization was significantly higher in 60RH in comparison to 18RH. Gene treY was upregulated sixfold in 60RH relative to control condition. TEM and quantification of PHB revealed that PHB was mobilized under 60RH condition accompanied by the downregulation of the phbB gene. TEM images showed an extracellular amorphous matrix in 18RH and 60RH. Major differences were found in the presence of aqpZ and trehalose genes between strain 22B and Sphingomonas genomes. CONCLUSION: Strain 22B showed a carbon conservative metabolism capable of accumulation of three types of endogenous carbon sources. The strain responds to water stress by changing the expression pattern of genes related to desiccation, formation of an extracellular amorphous matrix and mobilization of the carbon sources according to the degree of water stress. Trehalose, glycogen and PHB may have multiple functions in different degrees of desiccation. The robust endowment of molecular responses to desiccation shown in Sphingobium sp. 22B could explain its survival in semi-arid soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the physiology implicated in the toleration of the PAH-degrading strain Sphingobium sp 22B to environmental desiccation may improve the bioaugmentation technologies in semi-arid hydrocarbon-contaminated soils.
Subject(s)
Adaptation, Physiological/physiology , Microbial Viability , Sphingomonadaceae/physiology , Water/metabolism , Argentina , Chile , Glycogen/metabolism , Humidity , Soil Microbiology , Sphingomonadaceae/genetics , Sphingomonadaceae/metabolism , Trehalose/metabolismABSTRACT
As the development of marine economy, the submarine battery with the seawater electrolyte has obtained more and more attentions. Owing to the conventional electrochemical catalysts of the cathodes in seawater battery are expensive, it is to seek the new biological catalysts to improve the electrochemical performance of the cathode and reduce the cost of seawater battery. A novel marine bacterial strain (Strain SQ-32) phylogenetically related to the Erythrobactercitreus strain has been isolated from the sea-bed sludge in the Yellow Sea of China successfully. The electrochemical measurements, which include the cyclic voltammetry, potentiostatic polarization, and electrochemical impedance spectroscopy, have been conducted in synthetic seawater. The electrochemical testing results show that the Strain SQ-32 is a cold-tolerant bacterium, which may exhibit a catalytic activity for the ORR in synthetic seawater at a freezing temperature. The SEM photo demonstrates that the Strain SQ-32 displays a rod-shaped characteristic, which has a diameter of 0.4µm and a length of about 1-2.5µm. By the testing of Gram staining, the Strain SQ-32 has been identified as a Gram-negative bacterium. The chemical analytical result reveals that the bacterium cell of Strain SQ-32 contains 1.92mgg-1 (DCW) of coenzyme Q10, which is a possible impact factor on the electro-catalytic effect on the Strain SQ-32. The exploitation of Strain SQ-32 may boost the development of the biocathode of seawater battery at a low temperature.
Subject(s)
Biocatalysis , Cold Temperature , Oxygen/metabolism , Phylogeny , Sphingomonadaceae/classification , Sphingomonadaceae/metabolism , Carbon/chemistry , Electrochemistry , Electrodes , Glass/chemistry , Oxidation-Reduction , Sphingomonadaceae/physiology , Ubiquinone/analogs & derivatives , Ubiquinone/metabolismABSTRACT
Corals harbor a wide diversity of bacteria associated with their mucus. These bacteria can play an important role in nutrient cycling, degradation of xenobiotics and defense against pathogens by producing antimicrobial compounds. However, the diversity of the cultivable heterotrophic bacteria, especially in the Brazilian coral species, remains poorly understood. The present work compares the diversity of cultivable bacteria isolated from the mucus and surrounding environments of four coral species present along the Brazilian coast, and explores the antibacterial activity of these bacteria. Bacteria belonging to the phyla Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes were isolated. The mucus environment presented a significantly different bacteria composition, compared to the water and sediment environments, with high abundance of Alcanivorax, Acinetobacter, Aurantimonas and Erythrobacter. No difference in the inhibition activity was found between the isolates from mucus and from the surrounding environment. Eighty-three per cent of the bacteria isolated from the mucus presented antimicrobial activity against Serratia marcescens, an opportunistic coral pathogen, suggesting that they might play a role in maintaining the health of the host. Most of the bacteria isolates that presented positive antimicrobial activity belonged to the genus Bacillus.
Subject(s)
Anthozoa/microbiology , Antibiosis , Bacillus/physiology , Microbiota/physiology , Seawater/microbiology , Serratia marcescens/physiology , Acinetobacter/classification , Acinetobacter/genetics , Acinetobacter/isolation & purification , Acinetobacter/physiology , Actinobacteria/classification , Actinobacteria/genetics , Actinobacteria/isolation & purification , Actinobacteria/physiology , Alcanivoraceae/classification , Alcanivoraceae/genetics , Alcanivoraceae/isolation & purification , Alcanivoraceae/physiology , Animals , Bacillus/classification , Bacillus/genetics , Bacillus/isolation & purification , Brazil , Genetic Variation , Heterotrophic Processes , Phylogeny , RNA, Ribosomal, 16S/genetics , Seasons , Sphingomonadaceae/classification , Sphingomonadaceae/genetics , Sphingomonadaceae/isolation & purification , Sphingomonadaceae/physiologyABSTRACT
It is believed that sphingomonads are ubiquitously distributed in environments. However detailed information about their community structure and their co-relationship with environmental parameters remain unclear. In this study, novel sphingomonads-specific primers based on the 16S rRNA gene were designed to investigate the distribution of sphingomonads in 10 different niches. Both in silico and in-practice tests on pure cultures and environmental samples showed that Sph384f/Sph701r was an efficient primer set. Illumina MiSeq sequencing revealed that community structures of sphingomonads were significantly different among the 10 samples, although 12 sphingomonad genera were present in all samples. Based on RDA analysis and Monte Carlo permutation test, sphingomonad community structure was significantly correlated with limnetic and marine habitat types. Among these niches, the genus Sphingomicrobium showed strong positive correlation with marine habitats, whereas genera Sphingobium, Novosphingobium, Sphingopyxis, and Sphingorhabdus showed strong positive correlation with limnetic habitats. Our study provided direct evidence that sphingomonads are ubiquitously distributed in environments, and revealed for the first time that their community structure can be correlated with habitats.
Subject(s)
Ecosystem , Phylogeny , RNA, Ribosomal, 16S , Sphingomonadaceae/genetics , DNA, Bacterial , DNA, Ribosomal , Sphingomonadaceae/physiologyABSTRACT
Manganese (Mn) is an important metal in geochemical cycles. Some microorganisms can oxidize Mn(II) to Mn oxides, which can, in turn, affect the global cycles of other elements by strong sorption and oxidation effects. Microbe-microbe interactions have important roles in a number of biological processes. However, how microbial interactions affect Mn(II) oxidation still remains unknown. Here, we investigated the interactions between two bacteria (Arthrobacter sp. and Sphingopyxis sp.) in a co-culture, which exhibited Mn(II)-oxidizing activity, although neither were able to oxidize Mn(II) in isolation. We demonstrated that the Mn(II)-oxidizing activity in co-culture was most likely induced via contact-dependent interactions. The expressed Mn(II)-oxidizing protein in the co-culture was purified and identified as a bilirubin oxidase belonging to strain Arthrobacter. Full sequencing of the bilirubin oxidase-encoding gene (boxA) was performed. The Mn(II)-oxidizing protein and the transcripts of boxA were detected in the co-culture, but not in either of the isolated cultures. This indicate that boxA was silent in Arthrobacter monoculture, and was activated in response to presence of Sphingopyxis in the co-culture. Further, transcriptomic analysis by RNA-Seq, extracellular superoxide detection and cell density quantification by flow cytometry indicate induction of boxA gene expression in Arthrobacter was co-incident with a stress response triggered by co-cultivation with Sphingopyxis. Our findings suggest the potential roles of microbial physiological responses to stress induced by other microbes in Mn(II) oxidation and extracellular superoxide production.
Subject(s)
Arthrobacter/physiology , Bacterial Proteins/genetics , Gene Expression Regulation, Bacterial , Manganese/metabolism , Microbial Interactions , Sphingomonadaceae/physiology , Arthrobacter/genetics , Bacterial Proteins/metabolism , Oxidation-Reduction , Sphingomonadaceae/geneticsABSTRACT
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/pharmacologyABSTRACT
Although the source of drinking water (DW) used in hospitals is commonly disinfected, biofilms forming on water pipelines are a refuge for bacteria, including possible pathogens that survive different disinfection strategies. These biofilm communities are only beginning to be explored by culture-independent techniques that circumvent the limitations of conventional monitoring efforts. Hence, theories regarding the frequency of opportunistic pathogens in DW biofilms and how biofilm members withstand high doses of disinfectants and/or chlorine residuals in the water supply remain speculative. The aim of this study was to characterize the composition of microbial communities growing on five hospital shower hoses using both 16S rRNA gene sequencing of bacterial isolates and whole-genome shotgun metagenome sequencing. The resulting data revealed a Mycobacterium-like population, closely related to Mycobacterium rhodesiae and Mycobacterium tusciae, to be the predominant taxon in all five samples, and its nearly complete draft genome sequence was recovered. In contrast, the fraction recovered by culture was mostly affiliated with Proteobacteria, including members of the genera Sphingomonas, Blastomonas, and Porphyrobacter.The biofilm community harbored genes related to disinfectant tolerance (2.34% of the total annotated proteins) and a lower abundance of virulence determinants related to colonization and evasion of the host immune system. Additionally, genes potentially conferring resistance to ß-lactam, aminoglycoside, amphenicol, and quinolone antibiotics were detected. Collectively, our results underscore the need to understand the microbiome of DW biofilms using metagenomic approaches. This information might lead to more robust management practices that minimize the risks associated with exposure to opportunistic pathogens in hospitals.
Subject(s)
Bacterial Physiological Phenomena , Biofilms/growth & development , Cross Infection/genetics , Cross Infection/microbiology , Hospitals , Water Microbiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/pathogenicity , Chlorine , Culture Techniques , DNA, Bacterial/analysis , Disinfectants/pharmacology , Disinfection , Drug Resistance, Bacterial , Genome, Bacterial , Metagenome , Microbiota/genetics , Mycobacterium/physiology , Ohio , Phylogeny , Proteobacteria/physiology , RNA, Ribosomal, 16S/genetics , Sphingomonadaceae/physiology , Water SupplyABSTRACT
We have identified coproporphyrins including structurally new zincmethylphyrins I and III as growth factors A-F for the previously uncultured bacterial strain, Leucobacter sp. ASN212, from a supernatant of 210 l of Sphingopyxis sp. GF9 culture. Growth factors A-F induced significant growth of strain ASN212 at the concentrations of picomolar to nanomolar which would otherwise be unculturable in liquid medium or on agar plate. More interestingly, we found that the growth factors functioned as self-toxic compounds for the growth-factor producing strain GF9 at the picomolar to nanomolar levels. As a variety of bacteria could potentially produce coproporphyrins, our findings suggest that these compounds function as a novel class of signal molecules across a boundary at phylum level in the complex bacterial communities.
Subject(s)
Actinobacteria/drug effects , Actinobacteria/growth & development , Coproporphyrins/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Sphingomonadaceae/metabolism , Symbiosis , Actinobacteria/physiology , Coproporphyrins/isolation & purification , Culture Media/chemistry , Intercellular Signaling Peptides and Proteins/isolation & purification , Models, Molecular , Molecular Structure , Sphingomonadaceae/physiologyABSTRACT
A novel rod-shaped binary fission, and yellow-pigmented bacterial strain, JLT 2480(T), was isolated from surface seawater in the East Pacific Ocean. The strain is Gram negative and oxidase negative. Phylogenetic analyses based on 16S rRNA gene sequence indicate that strain JLT 2480(T) falls in the family Sphingomonadaceae, sharing highest similarity (95.6 %) with the species Blastomonas ursincola. The DNA G+C content of JLT 2480(T) is 65.5 mol%, and the sole respiratory quinone is coenzyme Q10. The predominant polar lipids are sphingoglycolipids (SGL1 and SGL2), phosphatidylglycerols, phosphatidylethanolamines, phospholipids, glycolipids, and phosphatidylcholines. The predominant cellular fatty acids are C16:0, C18:0, C18:1ω7c, C12:0, and C16:1ω7c. Strain JLT 2480(T) is distinct from the B. ursincola type strain DSM 9006(T) as reflected by major chemotaxonomic distinctions between the two. Furthermore, two notable characteristics of the genus Blastomonas, that is, the presence of bacteriochlorophyll a and the puf genes, are not detected in JLT 2480(T). On the basis of present evidence, we consider JLT 2480(T) to be a novel species in a new genus of the family Sphingomonadaceae, and propose the name Binariimonas pacifica gen. nov., sp. nov., with strain JLT 2480(T) (=CGMCC 1.12850(T) = DSM 28646(T)) to be the type strain for genus Binariimonas.
Subject(s)
Aquatic Organisms/classification , Aquatic Organisms/isolation & purification , Seawater/microbiology , Sphingomonadaceae/classification , Sphingomonadaceae/isolation & purification , Aquatic Organisms/genetics , Aquatic Organisms/physiology , Bacterial Typing Techniques , Base Composition , Cluster Analysis , Cytosol/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fatty Acids/analysis , Glycolipids/analysis , Microscopy, Electron, Transmission , Molecular Sequence Data , Pacific Ocean , Phospholipids/analysis , Phylogeny , Pigments, Biological/analysis , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sphingomonadaceae/genetics , Sphingomonadaceae/physiologyABSTRACT
Strain THG-SQA7(T), a Gram-negative, strictly aerobic, non-motile, rod-shaped bacterium was isolated from rhizosphere soil of a rose in PR China. Strain THG-SQA7(T) is closely related to the members of the genus Sphingobium, showing the highest 16S rRNA gene sequence similarities with Sphingobium lactosutens KACC 18100(T) (98.2%) and Sphingobium abikonense KCTC 2864(T) (98.1%). The DNA-DNA relatedness between strain THG-SQA7(T) and S. lactosutens KACC 18100(T) and S. abikonense KCTC 2864(T) was 26.2 ± 0.9 and 28.3 ± 1.2%, respectively. Chemotaxonomic data showed that strain THG-SQA7(T) possesses ubiquinone Q-10 as the predominant respiratory quinone, and C(18:1)ω7c, C(16:0), summed feature 3 (C(16:1)ω7c and/or C(16:1)ω6c) and C(14:0) 2OH as the major fatty acids. The major polar lipids were found to be phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, sphingoglycolipid, diphosphatidylglycerol and phosphatidyldimethylethanolamine. Based on these results, together with phenotypic characterization, a novel species, Sphingobium soli sp. nov. is proposed.with the type strain is THG-SQA7(T) (=CCTCC AB 2015125(T) = KCTC 42607(T)).