Uranium (U)-tolerant aerobic chemo-heterotrophic bacteria were isolated from the sub-surface soils of U-rich deposits in Domiasiat, North East India. The bacterial community explored at molecular level by amplified ribosomal DNA restriction analysis (ARDRA) resulted in 51 distinct phylotypes. Bacterial community assemblages at the U mining site with the concentration of U ranging from 20 to 100 ppm, were found to be most diverse. Representative bacteria analysed by 16S rRNA gene sequencing were affiliated to Firmicutes (51%), Gammaproteobacteria (26%), Actinobacteria (11%), Bacteroidetes (10%) and Betaproteobacteria (2%). Representative strains removed more than 90% and 53% of U from 100 μM and 2 mM uranyl nitrate solutions, respectively, at pH 3.5 within 10 min of exposure and the activity was retained until 24 h. Overall, 76% of characterized isolates possessed phosphatase enzyme and 53% had PIB-type ATPase genes. This study generated baseline information on the diverse indigenous U-tolerant bacteria which could serve as an indicator to estimate the environmental impact expected to be caused by mining in the future. Also, these natural isolates efficient in uranium binding and harbouring phosphatase enzyme and metal-transporting genes could possibly play a vital role in the bioremediation of metal-/radionuclide-contaminated environments.
Fifty three fungi isolated from soils of different microhabitats of eastern Himalayan range (3,400~3,600 msl) were screened for mycosynthesis of silver nanaoparticles (AgNPs) and their efficacy as antimicrobials were assessed in combination with commonly used antibiotics. Three isolates Aspergillus terreus SP5, Paecilomyces lilacinus SF1 and Fusarium sp. MP5 identified based on morphological and 18S rRNA gene sequences were found to synthesize AgNPs. These nanoparticles were characterized by visual observation followed by UV-visible spectrophotometric analysis. The AgNPs synthesized by Aspergillus terreus SP5, Paecilomyces lilacinus SF1 and Fusarium sp. MP5 showed absorbance maxima at 412, 419, and 421 nm respectively in the visible region. Transmission electron microscopy micrograph showed formation of spherical AgNPs of 5~50 nm size. The antimicrobial activity of the mycosynthesized nanoparticles were investigated alone and in combination with commonly used antibiotics for analysis of growth inhibition zone against test organisms, namely, Staphylococcus aureus MTCC96, Streptococcus pyogenes MTCC1925, Salmonella enterica MTCC735 and Enterococcus faecalis MTCC2729. The mycosynthesized nanoparticles showed potent antibacterial activity and interestingly their syngergistic effect with erythromycin, methicillin, chloramphenicol and ciprofloxacin was significantly higher as compared to inhibitions by AgNPs alone. The present study indicates that silver nanoparticles synthesized using soil borne indigenous fungus of high altitudes show considerable antimicrobial activity, deserving further investigation for potential applications.
Three endophytic fungi Aspergillus tamarii PFL2, Aspergillus niger PFR6 and Penicllium ochrochloron PFR8 isolated from an ethno-medicinal plant Potentilla fulgens L. were used for the biosynthesis of silver nanoparticles. Scanning and transmission electron microscopic analysis were performed to study the structural morphology of the biosynthesized silver nanoparticles. The electron microscopy study revealed the formation of spherical nanosized silver particles with different sizes. The nanoparticles synthesized using the fungus A. tamarii PFL2 was found to have the smallest average particle size (3.5 ±3 nm) as compared to the nanoparticles biosynthesized using other two fungi A. niger PFR6 and P. ochrochloron PFR8 which produced average particle sizes of 8.7 ±6 nm and 7.7 ±4.3 nm, respectively. The energy dispersive X-ray spectroscopy (EDS) technique in conjunction with scanning electron microscopy was used for the elemental analysis of the nanoparticles. The selected area diffraction pattern recorded from single particle in the aggregates of nanoparticles revealed that the silver particles are crystalline in nature.
Enrichment-based methods targeted at uranium-tolerant populations among the culturable, aerobic, chemo-heterotrophic bacteria from the subsurface soils of Domiasiat (India's largest sandstone-type uranium deposits, containing an average ore grade of 0.1 % U(3)O(8)), indicated a wide occurrence of Serratia marcescens. Five representative S. marcescens isolates were characterized by a polyphasic taxonomic approach. The phylogenetic analyses of 16S rRNA gene sequences showed their relatedness to S. marcescens ATCC 13880 (≥99.4% similarity). Biochemical characteristics and random amplified polymorphic DNA profiles revealed significant differences among the representative isolates and the type strain as well. The minimum inhibitory concentration for uranium U(VI) exhibited by these natural isolates was found to range from 3.5-4.0 mM. On evaluation for their uranyl adsorption properties, it was found that all these isolates were able to remove nearly 90-92% (21-22 mg/L) and 60-70% (285-335 mg/L) of U(VI) on being challenged with 100 μM (23.8 mg/L) and 2 mM (476 mg/L) uranyl nitrate solutions, respectively, at pH 3.5 within 10 min of exposure. his capacity was retained by the isolates even after 24 h of incubation. Viability tests confirmed the tolerance of these isolates to toxic concentrations of soluble uranium U(VI) at pH 3.5. This is among the first studies to report uranium-tolerant aerobic chemoheterotrophs obtained from the pristine uranium ore-bearing site of Domiasiat.
In the present study, an attempt to evaluate the antimicrobial and antioxidant activity of fungal endophytes inhabiting Emblica officinalis has been made keeping in view the medicinal importance of the selected host plant in Indian traditional practices. A total of four endophytic fungi belonging to Phylum Ascomycetes were isolated from different parts of the plant which were characterized morphologically and by using rDNA-internal transcribed spacer. The most frequently isolated endophyte was Phomopsis sp. The antioxidant activity by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assay, and total phenol were evaluated using ethanolic extract of endophytic fungi. DPPH activities in all the ethanolic extract increased with the increase in concentrations. Endophytes, Phomopsis sp. and Xylaria sp. showed highest antioxidant activity and also had the higher levels of phenolics. Antimicrobial activity of fungal extract were tested against four bacteria namely, Escherichia coli MTCC730, Enteroccocus faecalis MTCC2729, Salmonella enterica ser. paratyphi MTCC735 and Streptococcus pyogenes MTCC1925, and the fungus Candida albicans MTCC183. In general, the fungal extracts inhibited the growth of test organisms except E. coli.
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