Biodegradation and metabolic pathway of sulfamethoxazole by Sphingobacterium mizutaii.

Sulfamethoxazole (SMX) is the most commonly used antibiotic in worldwide for inhibiting aquatic animal diseases. However, the residues of SMX are difficult to eliminate and may enter the food chain, leading to considerable threats on human health. The bacterial strain Sphingobacterium mizutaii LLE5 was isolated from activated sludge. This strain could utilize SMX as its sole carbon source and degrade it efficiently. Under optimal degradation conditions (30.8 °C, pH 7.2, and inoculum amount of 3.5 × 107 cfu/mL), S. mizutaii LLE5 could degrade 93.87% of 50 mg/L SMX within 7 days. Four intermediate products from the degradation of SMX were identified and a possible degradation pathway based on these findings was proposed. Furthermore, S. mizutaii LLE5 could also degrade other sulfonamides. This study is the first report on (1) degradation of SMX and other sulfonamides by S. mizutaii, (2) optimization of biodegradation conditions via response surface methodology, and (3) identification of sulfanilamide, 4-aminothiophenol, 5-amino-3-methylisoxazole, and aniline as metabolites in the degradation pathway of SMX in a microorganism. This strain might be useful for the bioremediation of SMX-contaminated environment.

Exemplifying endophytes of banana (Musa paradisiaca) for their potential role in growth stimulation and management of Fusarium oxysporum f. sp cubense causing panama wilt.

In the present study, potentiality of endophytic microorganisms such as Rigidiporus vinctus AAU EF, Trichoderma reesei UH EF, and Sphingobacterium tabacisoli UH EB in the management of panama wilt and growth promotion of banana was assessed through artificial inoculation. During the study, a total of 220 bacterial and 110 fungal endophytes were isolated from root, pseudostem, and leaf samples of banana, and they were evaluated against Fusarium oxysporum f. sp cubense causing panama wilt. Out of total 330 bacterial and fungal endophytes, only five endophytes exhibited antagonism against Fusarium oxysporum f. sp cubense, out of which only three isolates, namely Trichoderma reesei UH EF, Rigidiporus vinctus AAU EF, and Sphingobacterium tabacisoli UH EB, produced indole acetic acid, siderophore, and hydrogen cyanide, except one bacterial strain Sphingobacterium tabacisoli UH EB which does not produce hydrogen cyanide. Furthermore, these three endophytes were identified through cultural and morphological characteristics as well as by the sequencing internal transcribed spacer (ITS) and 16S rRNA gene sequences analysis for bacteria, respectively. The response of host plant to endophyte inoculation was assessed by measuring the change in four growth parameters; plant height, pseudo stem girth (diameter), number of roots, and total number of leaves. The application of endophytes, irrespective of isolate and treatment type promoted the overall growth of the plant growth when compared with diseased plants with significant higher values recorded for all parameters assessed. The endophytes reported as growth promoters were found to have significant inhibition effect on Foc which can evidenced with lowest AUDPC values and epidemic rate at 99.09 units2 and 0.02 unit/day, respectively.

Endophytic bacteria promote growth of the medicinal legume Clitoria ternatea L. by chemotactic activity.

Part of the native root nodule endophytic microflora referring to members of the genera Proteobacteria and Sphingobacteria were used to test their bioefficacy as seed biopriming. These were quantified for their plant growth promoting (PGP) attributes such as IAA production, P and K-solubilization and ACC deaminase production. Results showed that significantly highest IAA was produced by E. hormaechi RCT10. The highest P-solubilization was observed with S. maltophila RCT31 it was solubilizing all the substrate tri-calcium phosphate, di-calcium phosphate, and zinc phosphate. Significantly highest K-solubilization was observed with S. maltophila RCT31 followed by E. turicensis RCT5. However, the maximum zinc solubilization was reported with S. maltophila RCT31 followed by E. turicensis RCT5. The maximum ACC deaminase was quantified in the bacterium. Results revealed that the E. hormaechi RCT10 utilized seed leechates most effectively while root exudates were maximally utilized by S. maltophila RCT31. The pots experiment proves that S. maltophila RCT31 was the most effective bacterium and it was replication vis-à-vis field experiment. In particular, S. maltophila RCT31 holds strong potential to be possibly used as a bioformulation for the medicinal legume, as an economical and eco-friendly alternative to agrochemicals.

Complete Genome Sequence of Sphingobacterium psychroaquaticum Strain SJ-25, an Aerobic Bacterium Capable of Suppressing Fungal Pathogens.

Using antagonistic bacterium is an effective method to control plant disease by fungal pathogens. An aerobic bacterium designated SJ-25, capable of suppressing Fusarium graminearum, Exserohilum turcicum, Pythium aphanidermatum, and Cochliobolus sativus, was isolated from farmland soil. The phylogenetic analysis revealed that strain SJ-25 belongs to the species of Sphingobacterium psychroaquaticum. The genome of strain SJ-25 consists of a 4,396,535-bp chromosome with a G+C content of 41.7 mol%; including 3696 CDS, 64 tRNA genes and six rRNA operons. Genomic analysis revealed that its genome contains multiple genes responsible for biosynthesis of siderophore, methyl 4-hydroxybenzoate, chitinase, giving strain SJ-25 the antagonistic ability on fungi pathogen. Strain SJ-25 harbors sets genes responsible for production of 2, 3-butanediol and salicylic acid, which could elicit the induced systemic resistance of the host plant. This genome sequence could be used as a basis material for further exploration of antagonistic mechanisms on fungi, widening our understanding of the ecological role of the genus Sphingobacterium in farmland ecosystem.

Isolation and prospection of diazotrophic rhizobacteria associated with sugarcane under organic management.

Microorganisms associated with organic management are essential in nutrient transformation and release for plant use. The present study aimed to isolate, identify and characterize plant growth promoting diazotrophic rhizobacteria associated with sugarcane under organic management. Rhizospheres of organic sugarcane varieties IAC 911099 and CTC4 were sampled and inoculated onto nitrogen free NFb and Burk media. The isolated microorganisms were screened in vitro concerning their ability to produce plant growth promoting factors. Eighty-one bacteria were isolated; 45.6% were positive for the nifH gene and produced at least one of the evaluated plant growth promotion factors. The production of indole-3-acetic acid was observed in 46% of the isolates, while phosphate solubilization was observed in 86.5%. No isolates were hydrogen cyanide producers, while 81% were ammonia producers, 19% produced cellulases and 2.7%, chitinases. Microorganisms belonging to the Burkholderia genus were able to inhibit Fusarium moniliforme growth in vitro. Plant growth promoting microorganisms associated with organic sugarcane, especially belonging to Burkholderia, Sphingobium, Rhizobium and Enterobacter genera, can be environmentally friendly alternatives to improve sugarcane production.

The first Korean case of Sphingobacterium spiritivorum bacteremia in a patient with acute myeloid leukemia.

Sphingobacterium spiritivorum has been rarely isolated from clinical specimens of immunocompromised patients, and there have been no case reports of S. spiritivorum infection in Korea to our knowledge. We report a case of S. spiritivorum bacteremia in a 68-yr-old woman, who was diagnosed with acute myeloid leukemia and subsequently received chemotherapy. One day after chemotherapy ended, her body temperature increased to 38.3℃. A gram-negative bacillus was isolated in aerobic blood cultures and identified as S. spiritivorum by an automated biochemical system. A 16S rRNA sequencing analysis confirmed that the isolate was S. spiritivorum. The patient received antibiotic therapy for 11 days but died of septic shock. This is the first reported case of human S. spiritivorum infection in Korea. Although human infection is rare, S. spiritivorum can be a fatal opportunistic pathogen in immunocompromised patients.

Bacterial sphingophospholipids containing non-hydroxy fatty acid activate murine macrophages via Toll-like receptor 4 and stimulate bacterial clearance.

Sphingobacterium spiritivorum has five unusual sphingophospholipids (SPLs). Our previous study determined the complete chemical structures of these SPLs. The compositions of the long-chain bases/fatty acids in the ceramide portion, isoheptadecasphingosine/isopentadecanoate or isoheptadecasphingosine/2-hydroxy isopentadecanoate, are characteristic. The immune response against bacterial lipid components is considered to play important roles in microbial infections. It is reported that several bacterial sphingolipids composed of ceramide are recognized by CD1-restricted T and NKT cells and that a non-peptide antigen is recognized by γδ T cells. In this study, we demonstrated that these bacterial SPLs activated murine bone marrow macrophages (BMMs) via Toll-like receptor (TLR) 4 but not TLR2, although they slightly activated CD1d-restricted NKT and γδT cells. Interestingly, this TLR 4-recognition pathway of bacterial SPLs involves the fatty acid composition of ceramide in addition to the sugar moiety. A non-hydroxy fatty acid composed of ceramide was necessary to activate murine BMMs. The bacterial survival was significantly higher in TLR4-KO mice than in TLR2-KO and wild-type mice. The results indicate that activation of the TLR4-dependent pathway of BMMs by SPLs induced an innate immune response and contributed to bacterial clearance.

Hailstones: a window into the microbial and chemical inventory of a storm cloud.

Storm clouds frequently form in the summer period in temperate climate zones. Studies on these inaccessible and short-lived atmospheric habitats have been scarce. We report here on the first comprehensive biogeochemical investigation of a storm cloud using hailstones as a natural stochastic sampling tool. A detailed molecular analysis of the dissolved organic matter in individual hailstones via ultra-high resolution mass spectrometry revealed the molecular formulae of almost 3000 different compounds. Only a small fraction of these compounds were rapidly biodegradable carbohydrates and lipids, suitable for microbial consumption during the lifetime of cloud droplets. However, as the cloud environment was characterized by a low bacterial density (Me = 1973 cells/ml) as well as high concentrations of both dissolved organic carbon (Me = 179 µM) and total dissolved nitrogen (Me = 30 µM), already trace amounts of easily degradable organic compounds suffice to support bacterial growth. The molecular fingerprints revealed a mainly soil origin of dissolved organic matter and a minor contribution of plant-surface compounds. In contrast, both the total and the cultivable bacterial community were skewed by bacterial groups (γ-Proteobacteria, Sphingobacteriales and Methylobacterium) that indicated the dominance of plant-surface bacteria. The enrichment of plant-associated bacterial groups points at a selection process of microbial genera in the course of cloud formation, which could affect the long-distance transport and spatial distribution of bacteria on Earth. Based on our results we hypothesize that plant-associated bacteria were more likely than soil bacteria (i) to survive the airborne state due to adaptations to life in the phyllosphere, which in many respects matches the demands encountered in the atmosphere and (ii) to grow on the suitable fraction of dissolved organic matter in clouds due to their ecological strategy. We conclude that storm clouds are among the most extreme habitats on Earth, where microbial life exists.

Transient response of microbial communities in a water well field to application of an impressed current.

Deterioration of water wells due to clogging and corrosion over time is a common problem where solutions may be costly and ineffective. Pilot studies have suggested that impressed current or cathodic protection may be used to reduce microbially-induced declines in water well performance. Two water wells in an alluvial aquifer close to the North Saskatchewan River were selected to study the response of subsurface microbial communities to the application of an impressed current as an anti-fouling technology. The treated well was exposed to an impressed current while the untreated well was used as a reference site. Biofilms grown on in situ coupons under the influence of the impressed current were significantly (p < 0.05) thicker (mean thickness = 67.3 µm) when compared to the biofilms (mean thickness = 19.3 µm) grown outside the electric field. Quantitative PCR analyses showed significantly (p < 0.05) higher numbers of total bacteria, iron- and nitrate-reducers in the electrified zone. Molecular analysis revealed that the predominant bacteria present in biofilms grown under the influence of the impressed current belonged to Rhodobacter spp., Sediminibacterium spp. and Geobacter spp. In addition to favouring the growth of biofilms, direct microscopic and ICP-AES analyses revealed that the impressed current also caused the deposition of iron and manganese on, and in the vicinity of, the well screen. Together, these factors contributed to rapid clogging leading to reduced specific pumping capacities of the treated well. The study revealed that the impressed current system was not effective as an anti-fouling technology but actually promoted both microbial growth and physical clogging in this aquifer.

Biofilm, ice recrystallization inhibition and freeze-thaw protection in an epiphyte community.

Microbial communities found on the surface of overwintering plants may be exposed to low temperatures as well as multiple freeze-thaw events. To explore the adaptive mechanisms of these epiphytes, with the objective of identifying products for freeze-protection, enrichment libraries were made from frost-exposed leaves. Of 15 identified bacteria from 60 individual clones, approximately half had ice-association activities, with the great majority showing high freeze-thaw resistance. Isolates with ice nucleation activity and ice recrystallization inhibition activity were recovered. Of the latter, two (Erwinia billingiae J10, and Sphingobacterium kitahiroshimense Y2) showed culture and electron microscopic evidence of motility and/or biofilm production. Mass spectrometric characterization of the E. billingiae extracellular polymeric substance (EPS) identified the major proteins as 35 kDa outer membrane protein A and F, supporting its biofilm character. The addition of the EPS preparation increased the freeze-thaw survival of the more susceptible bacteria 1000-10000 times, and protection was at least partially dependent on the protein component.

Sphingobacterium shayense sp. nov., isolated from forest soil.

A Gram-staining-negative, yellow-coloured, strictly aerobic, non-spore-forming, rod-shaped bacterium, designated HS39(T), isolated from a soil sample collected from a natural Populus euphratica forest in Xinjiang, China, was characterized using a polyphasic approach. The isolate grew optimally at 30-37 °C, at pH 6.5-8.0 and with 0-3 % NaCl. Analysis of the 16S rRNA gene sequence of strain HS39(T) revealed that it is a member of the genus Sphingobacterium. Sphingobacterium mizutaii ATCC 33299(T) was the nearest relative (94.0 % 16S rRNA gene sequence similarity). The G+C content of the genomic DNA was 40.2 mol%. The major fatty acids were iso-C15:0, iso-C17:0 3-OH and summed feature 3 (comprising C16:1ω6c and/or C16:1ω7c). The predominant isoprenoid quinone was MK-7. On the basis of phenotypic properties and phylogenetic inference, strain HS39(T) represents a novel species of the genus Sphingobacterium, for which the name Sphingobacterium shayense sp. nov. is proposed. The type strain is HS39(T) (=CCTCC AB 209006(T) =NRRL B-59203(T)).

Dyadobacter psychrophilus sp. nov., a psychrophilic bacterium isolated from soil.

A Gram-negative, aerobic bacterium, designated strain BZ26(T), was isolated from hydrocarbon-contaminated soil. The strain was psychrophilic, showing good growth over a temperature range of 1-20 degrees C. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BZ26(T) was related to members of the genus Dyadobacter and had highest 16S rRNA gene sequence similarity to Dyadobacter alkalitolerans 12116(T) (98.1 %), Dyadobacter koreensis KCTC 12537(T) (97.5 %) and Dyadobacter ginsengisoli Gsoil 043(T) (97.2 %). Strain BZ26(T) had MK-7 as the major menaquinone and summed feature 3 (C(16 : 1)omega7c/iso-C(15 : 0) 2-OH), C(16 : 1)omega5c and iso-C(15 : 0) as major fatty acids. The genomic DNA G+C content of strain BZ26(T) was 48.9 mol%. On the basis of phenotypic characteristics and genotypic analysis, strain BZ26(T) is considered to represent a novel species of the genus Dyadobacter, for which the name Dyadobacter psychrophilus sp. nov. is proposed. The type strain is BZ26(T) (=DSM 22270(T) =CGMCC 1.8951(T)).

Molecular and biochemical characterization of a novel xylanase from the symbiotic Sphingobacterium sp. TN19.

A xylanase-encoding gene, designated xynA19, was cloned from Sphingobacterium sp. TN19--a symbiotic bacterium isolated from the gut of Batocera horsfieldi larvae--and expressed in Escherichia coli BL21 (DE3). The full-length xynA19 (1,155 bp in length) encodes a 384-residue polypeptide (XynA19) containing a predicted signal peptide of 24 residues and a catalytic domain belonging to glycosyl hydrolase family 10 (GH 10). The deduced amino acid sequence of XynA19 is most similar (53.1% identity) to an endo-1,4-beta-xylanase from Prevotella bryantii B(1)4. Phylogenetic analysis of GH 10 Bacteroidia xylanases indicated that GH 10 xylanases from Sphingobacteria were separated into two clusters, and XynA19 is more closely related to the xylanases of Bacteroidia from gut or rumen than to those of Flavobacteria and Sphingobacteria from other sources. Recombinant XynA19 (r-XynA19) showed apparent optimal activity at pH 6.5 and 45 degrees C. Compared with thermophilic and mesophilic counterparts, r-XynA19 was more active at low temperatures, retaining >65% of its maximum activity at 20-28 degrees C and approximately 40% even at 10 degrees C, and modeling indicated that XynA19 has fewer hydrogen bonds and salt bridges. These properties suggest that XynA19 has various potential applications, especially in aquaculture and the food industry.

Sphingobacterium bambusae sp. nov., isolated from soil of bamboo plantation.

A Gram-negative, non-motile, non-spore-forming bacterial strain designated IBFC2009(T) was isolated from soil of a bamboo plantation. The strain could grow at 11 degrees C approximately 39 degrees C, pH 6.0-9.0, and in the presence of 0 approximately 5% NaCl. Based on 16S rRNA gene sequence analysis, Strain IBFC2009(T) belonged to the genus Sphingobacterium and showed the highest sequence similarity of 94.6% (S. composti T5-12(T)) with the type strains within the genus. The major fatty acids were summed feature 3 (iso-C(15:0) 2-OH and/or C(16:1) omega7c, 34.4%), iso-C(15:0) (22.4%), C(16:0) 3-OH (15.2%), and iso-C(17:0) 3-OH (12.8%). The G+C content of the genomic DNA was 41.0 mol%. According to the phenotypic and genotypic characteristics, Strain IBFC2009(T) should represent a novel species of the genus Sphingobacterium, for which the name Sphingobacterium bambusae sp. nov. is proposed. The type strain is IBFC2009(T) (=CCTCC AB 209162(T) =KCTC 22814(T)).

Sphingobacterium anhuiense sp. nov., isolated from forest soil.

A Gram-negative bacterium, designated strain CW 186(T), was isolated from forest soil in Anhui province, China. Cells of strain CW 186(T) were strictly aerobic, non-motile and rod-shaped. The strain grew optimally at 25-30 degrees C and pH 6.0-8.0. The major cellular fatty acids of strain CW 186(T) were iso-C(15:0) (32.2%), iso-C(17:0) 3-OH (9.8%) and summed feature 3 (iso-C(15:0) 2-OH and/or C(16:1)omega7c; 33.7%). The predominant isoprenoid quinone was MK-7. The G+C content of the genomic DNA was 36.3 mol%. Phylogenetic analysis using 16S rRNA gene sequences showed that strain CW 186(T) formed a monophyletic cluster with Sphingobacterium daejeonense LMG 23402(T), Sphingobacterium composti LMG 23401(T), Sphingobacterium composti DSM 18850(T), Sphingobacterium mizutaii ATCC 33299(T) and Sphingobacterium spiritivorum ATCC 33861(T). Sequence similarities were less than 94% (the maximal similarity was about 93.9% to S. composti LMG 23401(T)) to Sphingobacterium species with validly published names. A polyphasic taxonomic study including chemotaxonomic and phylogenetic analyses demonstrated that strain CW 186(T) should be classified as representing a novel species of the genus Sphingobacterium, for which the name Sphingobacterium anhuiense sp. nov. is proposed. The type strain is CW 186(T) (=KCTC 22209(T)=CCTCC AB 207197(T)).

Sphingobacterium siyangense sp. nov., isolated from farm soil.

The taxonomic position of a novel Gram-negative strain, designated SY1(T), isolated from a farm-soil sample obtained from Jiangsu Province, PR China, was characterized by using a polyphasic approach. The cells were non-motile, non-spore-forming rods. The organism grew optimally at 30-37 degrees C and at pH 6.0-8.0. Based on 16S rRNA gene sequence analysis, strain SY1(T) is a member of the genus Sphingobacterium; Sphingobacterium multivorum JCM 21156(T) was the nearest relative (98.5 % sequence similarity). The predominant fatty acids of strain SY1(T) were iso-C(15 : 0) (32.9 %), C(16 : 0) (10.9 %) and summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c; 24.1 %). The DNA G+C content was 38.5 mol%. The low level of DNA-DNA relatedness (2.2 %) to S. multivorum JCM 21156(T) in combination with differential morphological and biochemical properties demonstrated that strain SY1(T) (=KCTC 22131(T)=CGMCC 1.6855(T)) should be classified as representing a novel species of the genus Sphingobacterium for which the name Sphingobacterium siyangense sp. nov. is proposed.

Sphingobacterium daejeonense sp. nov., isolated from a compost sample.

A Gram-negative, strictly aerobic, rod-shaped, non-motile, non-spore-forming bacterial strain, designated TR6-04(T), was isolated from compost and characterized taxonomically by using a polyphasic approach. The organism grew optimally at 30 degrees Celsius and at pH 6.5-7.0. The isolate was positive for catalase and oxidase tests but negative for gelatinase, indole and H(2)S production. Comparative 16S rRNA gene sequence analysis showed that strain TR6-04(T) fell within the radiation of the cluster comprising Sphingobacterium species and clustered with Sphingobacterium mizutaii ATCC 33299(T) (96.7 % sequence similarity); the similarity to sequences of other species within the family Sphingobacteriaceae was less than 92.0 %. The G+C content of the genomic DNA was 38.7 mol%. The predominant respiratory quinone was MK-7. The major fatty acids were iso-C(15 : 0), iso-C(17 : 0) 3-OH and summed feature 4 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c). These chemotaxonomic data supported the affiliation of strain TR6-04(T) to the genus Sphingobacterium. However, on the basis of its phenotypic properties and phylogenetic distinctiveness, strain TR6-04(T) (=KCTC 12579(T)=LMG 23402(T)=CCUG 52468(T)) should be classified as the type strain of a novel species, for which the name Sphingobacterium daejeonense sp. nov. is proposed.

Intraspecific comparative analysis of the species Salinibacter ruber.

Salinibacter ruber is the first extremely halophilic member of the Bacteria domain of proven environmental relevance in hypersaline brines at or approaching NaCl saturation, that has been brought to pure culture. A collection of 17 strains isolated from five different geographical locations (Mallorca, Alicante, Ebro Delta, Canary Islands, and Peruvian Andes) were studied following the currently accepted taxonomic approach. Additionally, random amplification of genomic DNA led to the phenetic analysis of the intraspecific diversity. Altogether the taxonomic study indicated that S. ruber remained highly homogeneous beyond any geographical barrier. However, genomic fingerprints indicated that populations from different isolation sites could still be discriminated.