Molecular identification and screening of mushrooms for antibacterial property against Pseudomonas aeruginosa and Staphylococcus aureus

In this study, 18 mushrooms were collected from Gandhi Krishi Vigyana Kendra (GKVK) campus, University of Agricultural Sciences, Bangalore (India) and identified by Internal Transcribed Spacer (ITS) region using National Centre for Biotechnology Information (NCBI) data base and screened for antibacterial property against two skin infection causing bacteria viz.,Pseudomonas aeruginosa and Staphylococcus aureus in dogs. Eleven mushrooms extract inhibited both the bacteria showing their antimicrobials as broad spectrum while inhibiting gram positive as well as gram negative bacteria.  The highest zone of inhibition was recorded from the extract of Lyophyllum sp. The extract of two mushrooms viz., Phlebopus portentosus and Termitomyces sp. inhibited only P. aeruginosa, a gram negative bacterium  and the extract of three mushrooms viz., Agaricus sp, Coprinellus disseminates and Agaricus blazei inhibited only S. aureus, a gram positive bacterium, suggesting spectral specificity of their antimicrobial compounds.  However,  the extracts of two mushrooms viz.,Macrocybe gigantea and Schizophyllum commune did not show inhibition of any of the pathogens.  Hence, this study suggests that the occurrence of potential pharmaceuticals in mushrooms can be exploited for control of multidrug resistant bacteria

Field evaluation of nursery bed inoculated arbuscular mycorrhiza and rootdip inoculated Azotobacter chroococcum and Aspergillus awamori on aerobic rice

Field evaluation of nursery bed inoculated with Glomus fasciculatum and root dip treatement with Azotobacter chroococcum and Aspergillus awamori was carried out on aerobic rice. All the inoculated treatments with G. fasciculatum either singly or incombination showed increased growth and yield of rice compared to control and dual and triple inoculation were performed better than single inoculation treatment. G. fasciculatum inoculated seedlings in nursery bed shown better root colonization in field after transplantation compared to un-inoculated plants. The root dip inoculation with A. chroococcum and A. awamori during transplantation also increased the population of N2 fixer and Phosphate solubilizers besides increasing the population of general microflora in the rhizosphere. The results revealed the possibility of nursery inoculation of arbuscular mycorrhiza and root dip inoculation of other biofertilizers for aerobic rice

Mushroom diversity in the Biligiri rangana hills of Karnataka (India)

Evergreen forests of Biligiri rangana hills (BR hills) spread over an area of 540 sq. KM over eastern most edge of Western Ghats in Karnataka. Climatic conditions are more favorable for establishment of mushrooms and complete their life cycle. In this study, fourteen mushroom species were collected from BR hills region during monsoon (June through September 2013) with the help of Solega tribe inhabited the region since many years. Of the fourteen mushrooms, two mushrooms were identified as Ganoderma lucidum and Polyporous flabelliformis based on their phenotypic characters. The other 12 mushrooms were identified by ITS (Internal Transcribed spacer)region sequence homology as Termitomyces sp. (BRM-3)., Auricularia delicate (BRM-4), Termitomyces microcarpus (BRM-5), Amanita sp. (BRM-6), Podoscypha petalodes (BRM-7), Agaricaceae sp. (BRM-8), Macrolepiota sp.(BRM-9), Calvatia holothurioides (BRM-10), Gymnopillus crociphyllus (BRM-11), Coprinus comatus (BRM-12), Gyrodontium sacchari (BRM-13) and Clitocybeafffellea (BRM-14). Among the fourteen mushrooms, three species viz., Termito-myces species, Auricularia delicate and Termitomyces microcarpus were edible. The others were non edible/poisonous species. This study reports the diverse mushroom species as addition to biodiversity at BR hills

Molecular characterization of Lepidopteran specific Bacillus thuringiensis strains isolated from Hilly Zone Soils of Karnataka, India

Bacillus thuringiensis (Bt) strains pathogenic to Lepidopteran insects and native to hilly zone soils of Karnataka (India) were explored. 19 strains were isolated from the soils and identified by morphological and microscopic characters. Toxicity level of the Bt isolates was tested by treating third Instar larvae of silkworm (Bombyx mori L.) and mulberry leaf roller (Diaphania pulverulentalis). Mortality rate of the insect larvae treated with Bt isolates was ranged from 20 to 80%. A few isolates namely, Bt2, Bt6, Bt8, and Bt14 strains were more virulent (caused >50% death) than others. Detection of cry genes using cry gene specific primers by polymerase chain reaction (PCR) revealed the presence of at least one of the cry genes. In one of the isolates (Bt9), the cry gene was not detected. The Cry1Ac gene was abundant and it was detected in 13 isolates. Cluster analysis for genetic diversity showed two major groups and four sub groups.Keywords: Molecular characterization, Bacillus thuringiensis, bioassay, genetic diversity, Cry genes, hilly zoneAfrican Journal of Biotechnology Vol. 12(20), pp. 2924-293

Response of Coleus aromaticus to Glomus fasciculatum and other beneficial soil microflora

Stem cuttings of Coleus aromaticus were treated with vesicular arbuscular mycorrhiza (Glomus fasciculatum) and beneficial microorganisms viz., Bacillus coagulans, Pseudomonas fluorescens, Azotobacter chroococcum and Trichoderma harzianum either singly or in combination of organisms. The number of leaves and branches; total biomass, nitrogen and phosphorus contents were higher in plants treated with G. fasciculatum alone. Synergistic effect was noticed especially in plants treated with G. fasciculatum + P. fluorescens. Mycorrhizal colonization in roots was also maximum when V AM was applied alone. &nbsp

Response of Coleus aromaticus to Glomus fasciculatum and other beneficial soil microflora

Stem cuttings of Coleus aromaticus were treated with vesicular arbuscular mycorrhiza (Glomus fasciculatum) and beneficial microorganisms viz., Bacillus coagulans, Pseudomonas fluorescens, Azotobacter chroococcum and Trichoderma harzianum either singly or in combination of organisms. The number of leaves and branches; total biomass, nitrogen and phosphorus contents were higher in plants treated with G. fasciculatum alone. Synergistic effect was noticed especially in plants treated with G. fasciculatum + P. fluorescens. Mycorrhizal colonization in roots was also maximum when V AM was applied alone. &nbsp

Bacillus cereus nositelj plazmida pXO1 s genom pag uzrokuje u goveda s oslabljenim imunosnim sustavom smrtonosnu septikemiju sličnu bedrenici - kratko priopćenje.

Bacillus cereus is ubiquitous in nature and while most isolates appear to be harmless, some are associated with food-borne illnesses, wound infections, endocarditis, osteomyelitis, endophthalmitis and urinary tract infections in humans. Recently, a few isolates have been identified as the causative agents of anthrax-like severe pneumonia in humans, and these isolates were found to harbor most of the B. anthracis virulence plasmid pXO1. Here we report the characterization of three clinical B. cereus isolates recovered from heart blood and spleen samples of cattle which had died with ‘anthrax like’ symptoms. Apart from the cultural characterizations, primers targeting the 16S rRNA gene of B. cereus were designed and used on these isolates. The isolates were found to harbor the pXO1 plasmid and lacked pXO2 plasmid. Further characterization of the pXO1 plasmid revealed that the isolates contained pag, lef and cya genes, which code for protective antigen, lethal factor and edema factor toxins responsible for eliciting an ‘anthrax like disease’ in cattle. The sequencing and phylogenetic analysis of partial pag gene sequences of B. cereus isolates were identical to pag gene sequences on the pXO1 of B. anthracis. In a pathogenicity test on mice, B. cereus isolates, when inoculated by the intra peritoneal route, caused mortality of the mice within 6 hours post inoculation.Bacillus cereus je posvudašnja bakterija. Većina izolata te bakterije je neškodljiva, a neki mogu uzrokovati bolesti koje se prenose hranom, infekcije rana, endokarditis, osteomijelitis, endoftalmitis i infekcije mokraćnog sustava u ljudi. Nedavno je identificirano nekoliko izolata koji uzrokuju tešku upalu pluća u čovjeka sličnu onoj kod bedrenice. Ti izolati većinom nose virulentni plazmid pXO1 vrste B. anthracis. U ovom radu određena su obilježja triju kliničkih izolata vrste B. cereus izdvojenih iz krvi sadržane u srcu i uzoraka slezene goveda uginulih pod znakovima sličnima bedrenici. Osim određivanja kulturalnih obilježja, pripremljene su i početnice za gen 16S rRNA vrste B. cereus koje su bile rabljene za identifikaciju izolata. Ustanovljeno je da izolati nose plazmid pXO1, a nedostaje im plazmid pXO2. Daljnja karakterizacija plazmida pXO1 pokazala je da izolati sadrže gene pag, lef i cya koji kodiraju za zaštitni antigen, letalni čimbenik i edemski čimbenik, toksine koji su odgovorni za pojavu bolesti u goveda slične bedrenici. Određivanje slijeda i filogenetska analiza dijela sekvencija gena pag izolata B. cereus pokazala je da su oni istovjetni sekvencijama gena pag na pXO1 bakterije B. anthracis. U testu patogenosti na miševima, izolati B. cereus prouzročili su njihovo uginuće šest sati nakon intraperitonejske inokulacije

Molecular identification of phosphate solubilizing bacterium (Alcaligenes faecalis) and its interaction effect with Bradyrhizobium japonicum on growth and yield of soybeanGlycine max L.)

A phosphate solubilizing bacterium was isolated from the rhizosphere soil of upland rice and identified by 16S rRNA gene sequencing. The gene sequence showed 99% homology with Alcaligenes faecalis. Based on the gene sequence homology, it was identified as A. faecalis. Interaction effect of this bacterium on growth and yield of soybean was studied under glass house conditions by inoculating the bacterium either alone or in combination with Bradyrhizobium japonicum and Bacillus megaterium or both. The inoculated plants showed significantly taller plant height, more number of leaves, higher numbers of pods, plant dry weight and grain yield compared to un-inoculated ones (control). The triple inoculation was found superior compared to single as well as dual inoculations. Nitrogen and phosphorus content of the plant tissue was also higher in triple inoculation compared to others, which indicates their synergistic interaction in the rhizosphere of soybean.Keywords: Alcaligenes faecalis, Glycine max, phosphate solubilization, 16S rRNA gene sequencing.African Journal of Biotechnology, Vol 13(31) 3450-345

Equilibrium unfolding studies of monellin: the double-chain variant appears to be more stable than the single-chain variant

To improve our understanding of the contributions of different stabilizing interactions to protein stability, including that of residual structure in the unfolded state, the small sweet protein monellin has been studied in both its two variant forms, the naturally occurring double-chain variant (dcMN) and the artificially created single-chain variant (scMN). Equilibrium guanidine hydrochloride-induced unfolding studies at pH 7 show that the standard free energy of unfolding, ΔG°<SUB>U</SUB>, of dcMN to unfolded chains A and B and its dependence on guanidine hydrochloride (GdnHCl) concentration are both independent of protein concentration, while the midpoint of unfolding has an exponential dependence on protein concentration. Hence, the unfolding of dcMN like that of scMN can be described as two-state unfolding. The free energy of dissociation, ΔG°<SUB>d</SUB>, of the two free chains, A and B, from dcMN, as measured by equilibrium binding studies, is significantly lower than Δ G°<SUB>U</SUB>, apparently because of the presence of residual structure in free chain B. The value of ΔG°<SUB>U</SUB>, at the standard concentration of 1 M, is found to be ~5.5 kcal mol<SUP>−1</SUP> higher for dcMN than for scMN in the range from pH 4 to 9, over which unfolding appears to be two-state. Hence, dcMN appears to be more stable than scMN. It seems that unfolded scMN is stabilized by residual structure that is absent in unfolded dcMN and/or that native scMN is destabilized by strain that is relieved in native dcMN. The value of ΔG°<SUB>U</SUB> for both protein variants decreases with an increase in pH from 4 to 9, apparently because of the thermodynamic coupling of unfolding to the protonation of a buried carboxylate side chain whose pK<SUB>a</SUB> shifts from 4.5 in the unfolded state to 9 in the native state. Finally, it is shown that although the thermodynamic stabilities of dcMN and scMN are very different, their kinetic stabilities with respect to unfolding in GdnHCl are very similar