A medicinal herb Cassia alata attenuates quorum sensing in Chromobacterium violaceum and Pseudomonas aeruginosa.

Quorum sensing (QS) has been shown to play a crucial role in the pathogenesis in many bacteria, and attenuation of QS is one of the targets of antimicrobial therapy with particular interest in combating drug resistance. This study reports the QS inhibitory activity of metabolites from Cassia alata L. (Ca. alata), an important medicinal herb widely used in the treatment of microbial infections. For investigating the QS inhibition (QSI), the potential of Ca. alata L., initially, metabolites of the leaves extracted using ethanol was tested against biosensor strain Chromobacterium violaceum CV026 and C. violaceum wild-type strains. Furthermore, a purified fraction rich in flavonoids (F-AF) was used for establishing QSI activity by studying the inhibition of violacein production in C. violaceum, and QS controlled virulence and biofilm formation in Pseudomonas aeruginosa PAO1. The study results showed 50% inhibition of violacein production in C. violaceum at 0·05 mg ml-1 concentration of F-AF. In P. aeruginosa PAO1, it inhibited the tested virulence factors and biofilm formation significantly. The F-AF contained major flavonoids namely, quercetin, quercetrin and kaempferol displaying QSI activity individually against the test organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: Present study demonstrates the quorum sensing inhibitory activity of metabolites from Cassia alata, an important medicinal herb which is commonly used worldwide in the treatment of infections caused by microorganisms. An extract prepared from the leaves of the plant showed activity against quorum sensing in Chromobacterium violaceum and was also effective against attenuating the quorum sensing controlled virulence factors in Pseudomonas aeruginosa. Activity is attributed to the rich flavonoid composition of the plant. Results of the present investigation throw an insight into the possibility of developing drug formulations using the isolated compounds against infections caused by quorum sensing-mediated pathogenicity of bacteria.

Anti-quorum sensing activity of flavonoid-rich fraction from Centella asiatica L. against Pseudomonas aeruginosa PAO1.

BACKGROUND/PURPOSE: Inhibition of quorum sensing (QS), a cell-density dependent regulation of gene expression in bacteria by autoinducers is an attractive strategy for the development of antipathogenic agents. METHODS: In this study, the anti-QS activity of the ethanolic extract of the traditional herb Centella asiatica was investigated by the biosensor bioassay using Chromobacterium violaceum CV026. The effect of ethyl acetate fraction (CEA) from the bioassay-guided fractionation of ethanol extract on QS-regulated violacein production in C. violaceum ATCC12472 and pyocyanin production, proteolytic and elastolytic activities, swarming motility, and biofilm formation in Pseudomonas aeruginosa PAO1 were evaluated. Possible mechanism of QS-inhibitory action on autoinducer activity was determined by measuring the acyl homoserine lactone using C. violaceum ATCC31532. Anti-QS compounds in the CEA fraction were identified using thin layer chromatography biosensor overlay assay. RESULTS: Ethanol extract of C. asiatica showed QS inhibition in C. violaceum CV026. Bioassay-guided fractionation of ethanol extract revealed that CEA was four times more active than the ethanol extract. CEA, at 400 µg/mL, completely inhibited violacein production in C. violaceum ATCC12472 without significantly affecting growth. CEA also showed inhibition of QS-regulated phenotypes, namely, pyocyanin production, elastolytic and proteolytic activities, swarming motility, and biofilm formation in P. aeruginosa PAO1 in a concentration-dependent manner. Thin layer chromatography of CEA with biosensor overlay showed anti-QS spot with an Rf value that corresponded with that of standard kaempferol. CONCLUSION: The anti-QS nature of C. asiatica herb can be further exploited for the formulation of drugs targeting bacterial infections where pathogenicity is mediated through QS.

Zeaxanthin production by novel marine isolates from coastal sand of India and its antioxidant properties.

Zeaxanthin carotenoids are class of commercially important natural products and diverse biomolecules produced by plants and many microorganisms. Bacteria often produce a cocktail of polar and nonpolar carotenoids limiting their industrial applications. Marine members of the family Flavobacteriaceae are known to produce potential carotenoids such as astaxanthin and zeaxanthin. A few bacterial species have been reported for the predominant production zeaxanthin. Here, we report the molecular identification of the zeaxanthin as a major carotenoid produced by two novel bacteria (YUAB-SO-11 and YUAB-SO-45) isolated from sandy beaches of South West Coast of India and the effect of carbon sources on the production of zeaxanthin. The strains were identified based on the 16S rRNA gene sequencing as a member of genus Muricauda. The closest relatives of YUAB-SO-11 and YUAB-SO-45 were Muricauda aquimarina (JCM 11811(T)) (98.9 %) and Muricauda olearia (JCM 15563(T)) (99.2 %), respectively, indicating that both of these strains might represent a novel species. The highest level of zeaxanthin production was achieved (YUAB-SO-11, 1.20 ± 0.11 mg g(-1)) and (YUAB-SO-45, 1.02 ± 0.13 mg g(-1)) when cultivated in marine broth supplemented with 2 % NaCl (pH 7) and incubated at 30 °C. Addition of 0.1 M glutamic acid, an intermediate of citric acid cycle, enhanced the zeaxanthin production as 18 and 14 % by the strains YUAB-SO-11 and YUAB-SO-45 respectively. The zeaxanthin showed in vitro nitric oxide scavenging, inhibition of lipid peroxidation, and 2,2-diphenyl-1-picryl hydrazyl scavenging activities higher than the commercial zeaxanthin. The results of this study suggest that two novel strains YUAB-SO-11 and YUAB-SO-45 belonging to genus Muricauda produce zeaxanthin as a predominant carotenoid, and higher production of zeaxanthin was achieved on glutamic acid supplementation. The pigment showed good in vitro antioxidant activity, which can be exploited further for commercial applications.

Hydrolysis of acid and alkali presoaked lignocellulosic biomass exposed to electron beam irradiation.

In this study, synergetic effect of mild acid and alkali with electron beam irradiation (EBI) on the enzymatic hydrolysis of a selected grass biomass was assessed. Biomass samples prepared by soaking with 1% H2SO4, or 1% NaOH, were exposed to 75 and 150 kGy of EBI. Water presoaked biomass was used as control. Hydrolysis of pretreated samples was carried out using cellulase (15 FPU/g biomass) for 120 h. Structural changes were studied by FTIR and XRD analyses. Reducing sugar and glucose yields from enzymatic hydrolysis were significantly higher in acid and alkali presoaked EBI exposed samples. Theoretical glucose yield showed 40% increase from control in alkali presoaked EBI exposed (150 kGy) samples. Removal of hemicellulose, decreased crystallinity and structural changes were major factors for the combined treatment effect favoring the hydrolysis.

Molecular detection and phylogenetic characterization of Gordonia species in heavily oil-contaminated soils.

This study was undertaken to assess genetic diversity among Gordonia species present in heavily oil-contaminated sites using both a culture-dependent and a culture-independent (PCR-denaturing gradient gel electrophoresis (DGGE)) approach. Soil samples for this purpose were collected from 8 different heavily (crude) oil-contaminated industrial park sites located around Kaohsiung County, Taiwan. Using Gordonia-specific PCR-DGGE, a significant increase in Gordonia species diversity was noted in 1% heavily oil-enriched soil. A total of 67 strains were scored and identified as Gordonia after genus-specific PCR amplification and sequencing. BOX-PCR fingerprinting of culturable Gordonia showed wide strain diversity. A total of 33 different strains were identified from most of the sampling sites. Based on gyrB gene sequence analysis, all Gordonia strains could be segregated into five major clusters. Gordonia amicalis was the predominant species in all oil-amended soil samples. Isolates sharing <98.5% gyrB gene sequence similarities with Gordonia type strains represent indigenous novel Gordonia species. Variations in phenotypic characteristics further confirm the presence of a wide range of species and strain diversity among Gordonia isolates. Based on the genotypic and phenotypic details obtained here, we conclude that heavily oil-contaminated soil supports diverse indigenous Gordonia strains.

Arenimonas malthae sp. nov., a gammaproteobacterium isolated from an oil-contaminated site.

A Gram-negative, rod-shaped bacterium (CC-JY-1(T)) was isolated on nutrient agar from a soil sample collected from an oil-contaminated site located in Chyai county, Taiwan. 16S rRNA gene sequence analysis demonstrated that this isolate is unique, showing 96.7 % sequence similarity to the type strain of Arenimonas donghaensis and similarities of 93.0-93.8 % to species of the genera Thermomonas, Lysobacter and Silanimonas. The presence of ubiquinone Q-8, a polar lipid profile consisting of the major compounds diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine and the fatty acid profile were in accordance with the phylogenetic affiliation of CC-JY-1(T). DNA-DNA reassociation experiments between CC-JY-1(T) and A. donghaensis KACC 11381(T) resulted in a mean relatedness value of 32 %, indicating that strain CC-JY1(T) represents a novel species in the genus Arenimonas, for which we propose the name Arenimonas malthae sp. nov. The type strain is CC-JY-1(T) (=CCUG 53596(T) =CIP 109310(T)).

Sphingobium olei sp. nov., isolated from oil-contaminated soil.

The taxonomic status of a yellow-coloured bacterial isolate from an oil-contaminated soil sample was determined using a polyphasic taxonomic approach. Comparative analysis of 16S rRNA gene sequences showed that the novel isolate formed a distinct phyletic line within the genus Sphingobium. The generic assignment was confirmed by chemotaxonomic data, which revealed: a fatty acid profile that is characteristic of the genus Sphingobium consisting of straight-chain saturated and unsaturated as well as 2-OH fatty acids; a ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone; a polar lipid pattern consisting of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine and sphingoglycolipid, and spermidine as the major polyamine component. Genotypic and phenotypic data show that the new isolate merits classification as a representative of a novel species of the genus Sphingobium, for which the name Sphingobium olei sp. nov. is proposed. The type strain is IMMIB HF-1T (=DSM 18999T=CCUG 54329T).

Brachybacterium phenoliresistens sp. nov., isolated from oil-contaminated coastal sand.

A coccoid- to ovoid-shaped, Gram-positive, non-motile bacterial strain, designated phenol-AT, was isolated from an oil-contaminated coastal sand sample collected from Pingtung County, southern Taiwan, and characterized by use of a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that the novel strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Brachybacterium in the family Dermabacteraceae, class Actinobacteria. The closest neighbours were Brachybacterium rhamnosum LMG 19848T (96.9% 16S rRNA gene sequence similarity), Brachybacterium nesterenkovii DSM 9573T (97.0%) and Brachybacterium muris C3H-21T (96.3%). The peptidoglycan type of strain phenol-AT was variation A4gamma with meso-diaminopimelic acid as the diagnostic cell-wall diamino acid. The isolate contained MK-7 as the major component of the quinone system. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, and unidentified phospholipids and glycolipids. The predominant fatty acid was anteiso-C15:0 (59.5%); significant amounts of iso-C16:0 (9.4%), iso-C14:0 (9.5%) and anteiso-C17:0 (10.8%) were also present. The isolate was also distinguished from recognized members of the genus Brachybacterium on the basis of several phenotypic and biochemical characteristics. It is evident from the genotypic, chemotaxonomic and phenotypic data that isolate phenol-AT represents a novel species of the genus Brachybacterium, for which the name Brachybacterium phenoliresistens sp. nov. is proposed. The type strain is phenol-AT (=LMG 23707T=BCRC 17589T).

Pseudoxanthomonas spadix sp. nov., isolated from oil-contaminated soil.

A bacterial isolate from a sample of oil-contaminated soil was characterized using a polyphasic taxonomic approach. Comparative analysis of the 16S rRNA gene sequence showed that this isolate constituted a distinct phyletic line within the genus Pseudoxanthomonas, displaying >3.7 % sequence divergence with respect to recognised Pseudoxanthomonas species. The genus assignment was confirmed by a chemotaxonomic analysis, which revealed the presence of a fatty acid profile characteristic of members of the genus Pseudoxanthomonas (straight-chain saturated, unsaturated and branched-chain fatty acids of the iso/anteiso type and 3-hydroxylated fatty acids) and the presence of a ubiquinone with eight isoprene units (Q-8) as the predominant respiratory quinone. The novel isolate was distinguishable from other members of the genus Pseudoxanthomonas on the basis of a combination of phenotypic properties. The genotypic and phenotypic data show that the strain represents a novel species of the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas spadix sp. nov. is proposed. The type strain is IMMIB AFH-5(T) (=DSM 18855(T)=CCUG 53828(T)).

Williamsia serinedens sp. nov., isolated from an oil-contaminated soil.

The taxonomic status of a bacterium designated strain IMMIB SR-4(T) isolated from an oil-contaminated soil sample was characterized by using a polyphasic approach. Chemotaxonomic investigations revealed the presence of cell-wall chemotype IV, short-chain mycolic acids that co-migrated with those extracted from members of the genus Williamsia and that on pyrolysis GC produce C(16 : 0) and C(18 : 0) fatty acids, and dihydrogenated menaquinone with nine isoprene units as the predominant menaquinone. The generic assignment was confirmed by 16S rRNA gene sequence analysis. Comparative analysis of the 16S rRNA gene sequence showed that strain IMMIB SR-4(T) formed a distinct phyletic line within the genus Williamsia, displaying sequence similarities of 95.5-98.1 % with the type strains of recognized Williamsia species. Strain IMMIB SR-4(T) was distinguished from the type strains of recognized species of the genus Williamsia based on a set of phenotypic features. The genotypic and phenotypic data indicated that strain IMMIB SR-4(T) represents a novel species of the genus Williamsia, for which the name Williamsia serinedens sp. nov. is proposed. The type strain is IMMIB SR-4(T) (=DSM 45037(T)=CCUG 53151(T)).