Secondary metabolites and anti-microbial/anti-oxidant profiles in Ocimum spp.: Role of soil physico-chemical characteristics as eliciting factors.

Ocimum has long been used as a medicinal plant, although little information is available about its bioactive ingredients, and the influence of soil properties on modulation of secondary metabolites in Ocimum has yet to be ascertained. In this study, we present a thorough survey of all potential metabolic compounds in O. sanctum and O. basilicum. In both species, certain compounds (e.g., quercetin, kaempferol, catechin, and S-adenosyl homocysteine) were detected coincidently. In the case of O. basilicum, other vital phenolic acids (e.g., ursolic, vanilic, coumaric, and syringic acids) were identified. The aqueous extracts (AEs) of Ocimum recorded decrease of 6-94% in the proliferation of pathogenic bacteria (e.g., Listeria monocytogenes, Staphylococcus sp., Salmonella sp., and Bacillus sp.). The AEs also showed effective antioxidant activity by reducing free radicals by a factor of 1.04-1.13. Root-zone soil samples of both Ocimum spp. were collected from strategic locations with varying levels of key soil attributes (e.g., soil organic carbon (SOC), microbial biomass carbon (MBC), urease, and phosphatase). At high levels of SOC, MBC, and soil enzymes, the bioactivity of Ocimum spp. was observed to be promoted, especially with respect to secondary metabolite expression, anti-pathogenic activity, and anti-oxidant properties. As such, the findings of strong correlations between secondary metabolite concentrations and bioactivity attributes in Ocimum suggest the potent role of soil quality in eliciting the production of secondary metabolite in association with bioactivity in Ocimum spp.

Detoxification and eco-friendly recycling of brick kiln coal ash using Eisenia fetida: A clean approach through vermitechnology.

Brick kiln coal ashes (BKCAs) are one of the major toxic byproducts of the rapidly growing construction industry in developing countries. However, eco-friendly recycling avenues for BKCAs are yet to be explored. The major objectives of the present research were to evaluate the viability of vermitechnology in transforming BKCAs into valuable products, and to examine the metal detoxification potential of Eisenia fetida BKCA-based feedstocks. BKCAs were mixed in large scale with cow dung (CD) in 1:1 and 2:1 ratios, for vermicomposting and aerobic composting; performance was assessed in comparison with CD. Vermiconverted-BKCA was then used as organic fertilizer for rice grown in poorly fertile soil. Acidic nature of BKCA feedstocks was neutralized by 30-86% in the vermireactors. Total N and available P concentrations significantly increased in the vermireactors supplemented with considerable mineralization of total organic C. Exorbitantly high K and S contents were pacified to a normal range after vermicomposting. Greater improvement in microbial biomass, respiration, fungal and bacterial growth was observed under vermicomposting against aerobic composting. Consequently, urease and phosphatase activity increased by 1-4 folds in the BKCA based vermibeds. Bioavailability of toxic metals reduced by 41-74% in the vermicomposted BKCAs. High metal accumulation by the earthworms resulted in substantial reduction of pollution load in the finished product. The field experiment demonstrated that vermiconverted-BKCA could be utilized as potential organic fertilizer for rice production, soil fertility rejuvenation, and metal detoxification. Overall, the study reveals that E. fetida could be used as an efficient contender for sanitization of toxic BKCAs.

Appraisal of lignocellusoic biomass degrading potential of three earthworm species using vermireactor mediated with spent mushroom substrate: Compost quality, crystallinity, and microbial community structural analysis.

Spent mushroom substrate (SMS) is a recalcitrant lignocellulosic waste. Recycling of SMS through composting has been reported; however, the process is lengthy due to its complex biochemical composition. Although vermitechnology is known for its high efficiency, it has rarely been applied to recycle SMS. In this study, the qualitative value of vermicomposted SMS mediated by three earthworm species (i.e., Eisenia fetida, Eudrilus eugeniae, and Perionyx excavatus) was evaluated on the basis of nutrient availability, microbial activity, phospholipid fatty acid (PLFA) profiles, and seed germination assays. Degradation profiles of the lignocellulosic substrate in the vermireactors were assessed by monitoring the changes in crystallinity and distribution of functional groups using X-ray diffraction (XRD) and Fourier transform infrared spectroscopy, respectively. Total organic carbon decreased by 1.4-3.5 folds with approximately 2.1-2.4 folds increase in nitrogen and phosphorus availability in all vermibeds. Interestingly, pH declined in the Eisenia and Eudrilus systems but increased in the Perionyx-vermibeds. XRD-derived crystallinity index was reduced significantly by 1.37 folds in Perionyx-vermicompost with concurrent microbial enrichment. Further, profuse abundance of vital functional groups (CO, NH, and OH) was clearly observed in the vermicompost with Perionyx followed by that with Eisenia. Moreover, PLFA illustrated significant variations in fatty acid distributions and microbial communities of the three vermicomposting systems. The seed germination assay showed that the germination index and relative root-shoot vigor of Perionyx-vermicompost treated seeds were 1.05-1.30 times greater than those of Eisenia and Eudrilus vermicompost treated ones. The results suggest that SMS degradability was affected by the growth of a healthy microbial community through vermicomposting.

Phosphate deficiency induced biofilm formation of Burkholderia on insoluble phosphate granules plays a pivotal role for maximum release of soluble phosphate.

Involvement of biofilm formation process during phosphate (P) solubilization by rhizobacterial strains is not clearly understood. Scanning electron microscopic observations revealed prominent biofilm development on tricalcium phosphate as well as on four different rock phosphate granules by two P solubilizing rhizobacteria viz. Burkholderia tropica P4 and B. unamae P9. Variation in the biofilm developments were also observed depending on the total P content of insoluble P used. Biofilm quantification suggested a strong correlation between the amounts of available P and degrees of biofilm formation. Lower concentrations of soluble P directed both the organisms towards compact biofilm development with maximum substratum coverage. Variation in the production of extracellular polymeric substances (EPS) in the similar pattern also suggested its close relationship with biofilm formation by the isolates. Presence of BraI/R quorum sensing (QS) system in both the organisms were detected by PCR amplification and sequencing of two QS associated genes viz. braR and rsaL, which are probably responsible for biofilm formation during P solubilization process. Overall observations help to hypothesize for the first time that, biofilm on insoluble P granules creates a close environment for better functioning of organic acids secreted by Burkholderia strains for maximum P solubilization during P deficient conditions.

Longterm storage of post-packaged bread by controlling spoilage pathogens using Lactobacillus fermentum C14 isolated from homemade curd.

One potent lactic acid bacterial strain C14 with strong antifungal activity was isolated from homemade curd. Based on morphological as well as biochemical characters and 16S rDNA sequence homology the strain was identified as Lactobacillus fermentum. It displayed a wide antimicrobial spectrum against both Gram-positive and Gram-negative pathogenic bacteria, and also against number of food spoilage, plant and human pathogenic fungi. The cell free supernatant (CFS) of the strain C14 was also effective against the fungi tested. Inhibition of radial growth of Penicillium digitatum, Trichophyton rubrum and Mucor sp. was noticed in the presence of CFS of C14 even at low concentration (1%). More than 94.3 ± 1.6% and 91.5 ± 2.2% inhibition of conidial germination of P. digitatum and Mucor sp. were noticed in the presence of 10-fold-concentrated CFS of C14. Massive deformation of the fungal mycelia was observed by SEM studies, and losses of cellular proteins and DNA are also evident upon its treatment with C14. HPLC analysis revealed the presence of phenyl lactic acid, lactic acid along with some unidentified compounds in the antifungal extract. Challenge experiment showed immense potential of the strain C14 in preventing the spoilage of bread samples caused by Mucor sp. and Bacillus subtilis. The bread samples remained fresh upto 25 days even after inoculation with Mucor sp. (3.7 × 104 spores /ml) and B. subtilis (4.6 × 104 CFU /ml). Along with the antifungal properties, the isolated lactic acid bacterial strain also showed very good antioxidant activities. Unchanged level of liver enzymes serum glutamic pyruvic transaminase and serum glutamic oxaloacetic transaminase in albino mice upon feeding with C14 also suggested non-toxic nature of the bacterial isolate.

Silver Nanoparticles as Antibacterial and Anticancer Materials Against Human Breast, Cervical and Oral Cancer Cells.

Silver nanoparticles contribute a giant share to the realm of modern nanobiotechnology. Their utility as antimicrobial agents is also well documented. Green synthesis of nanoparticle has several advantages over its chemical synthesis. In the present study, Thuja occidentalis leaf extract mediated silver nanoparticles were prepared without using a stabilizing agent and tested for their anticancer and anti-microbial activity. Thuja occidentalis leaf extract mediated silver nanoparticles were prepared under ambient conditions which showed a narrow size distribution within the range of 10­15 nm, with average particle size of 12.7 nm. Interestingly, these nanoparticles exhibited anti-cancer activity against human breast (MCF 7, MDA MB 231) and cervical cancer (HeLa) as well as mouth epidermoid carcinoma (KB) cell lines at a concentration range of 6.25­50 µg/mL. Contrarily, they are compatible with human peripheral blood mononuclear cells and rat hepatocytes. Moreover, their efficient inhibitory effect was witnessed against Bacillus subtilis, Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium and Pseudomonas aeruginosa with inhibitory concentration at 5­10 µg/mL. The prepared nanoparticles were highly biocompatible and have strong potential in the development of non-toxic chemotherapy with antibacterial attributes.

Modeling of Cancer Classifier to Predict Site of Origin.

Cancer classification based on site of origin is very significant research issue for prediction and treatment of cancer. This paper is addressing the problem of cancer classification for Homo Sapiens genes composed of amino acid chain. Cancer gene network is realized by equivalent electrical circuits based on hydrophilic/ hydrophobic property of amino acid and a classifier is modeled to determine the cancer origin. The phase value, peak gain value and shape of Nyquist curve of network model are investigated to characterize different types of cancer gene origins. The model achieves 81.09% of classification accuracy and proves to be more sensitive and simple, since it shows 69% better performance compare to the existing nucleotide based method. The proposed classifier successfully predicts the site of origin of 93 cancer gene samples.

Application of Euclidean distance measurement and principal component analysis for gene identification.

Gene systems are extremely complex, heterogeneous, and noisy in nature. Many statistical tools which are used to extract relevant feature from genes provide fuzzy and ambiguous information. High-dimensional gene expression database available in public domain usually contains thousands of genes. Efficient prediction method is demanding nowadays for accurate identification of such database. Euclidean distance measurement and principal component analysis methods are applied on such databases to identify the genes. In both methods, prediction algorithm is based on homology search approach. Digital Signal Processing technique along with statistical method is used for analysis of genes in both cases. A two-level decision logic is used for gene classification as healthy or cancerous. This binary logic minimizes the prediction error and improves prediction accuracy. Superiority of the method is judged by receiver operating characteristic curve.

Role of phosphate solubilizing Burkholderia spp. for successful colonization and growth promotion of Lycopodium cernuum L. (Lycopodiaceae) in lateritic belt of Birbhum district of West Bengal, India.

Profuse growth of Lycpodium cernuum L. was found in phosphate deficient red lateritic soil of West Bengal, India. Interaction of vesicular-arbuscular mycorrhiza (VAM) with Lycopodium rhizoids were described earlier but association of PGPR with their rhizoids were not studied. Three potent phosphate solubilizing bacterial strains (P4, P9 and P10) associated with L. cernuum rhizoids were isolated and identified by 16S rDNA homologies on Ez-Taxon database as Burkholderia tropica, Burkholderia unamae and Burkholderia cepacia respectively. Day wise kinetics of phosphate solubilization against Ca3(PO4)2 suggested P4 (580.56±13.38 µg ml(-1)) as maximum mineral phosphate solubilizer followed by P9 (517.12±17.15 µg ml(-1)) and P10 (485.18±14.23 µg ml(-1)) at 28 °C. Release of bound phosphates by isolated strains from ferric phosphate (FePO4), aluminum phosphate (AlPO4) and four different complex rock phosphates indicated their very good phosphate solubilizng efficacy. Nitrogen independent solubilizition also supports their nitrogen fixing capabilities. Inhibition of P solubilization by calcium salts and induction by EDTA suggested pH dependent chelation of metal cations by all of the isolates. Rhizoidal colonization potentials of Burkholderia spp. were confirmed by in planta experiment and also using scanning electron microscope (SEM). Increases of total phosphate content in Lycopodium plants upon soil treatment with these isolates were also recorded. In addition siderophore production on CAS agar medium, tryptophan dependent IAA production and antifungal activities against pathogenic fungi by rhizospheric isolates deep-rooted that they have definite role in nutrient mobilization for successful colonization of L. cernuum in nutrient deficient lateritic soil.

Performance Analysis of Network Model to Identify Healthy and Cancerous Colon Genes.

Modeling of cancerous and healthy Homo Sapiens colon gene using electrical network is proposed to study their behavior. In this paper, the individual amino acid models are designed using hydropathy index of amino acid side chain. The phase and magnitude responses of genes are examined to screen out cancer from healthy genes. The performance of proposed modeling technique is judged using various performance measurement metrics such as accuracy, sensitivity, specificity, etc. The network model performance is increased with frequency, which is analyzed using the receiver operating characteristic curve. The accuracy of the model is tested on colon genes and achieved maximum 97% at 10-MHz frequency.

Improved gene prediction by principal component analysis based autoregressive Yule-Walker method.

Spectral analysis using Fourier techniques is popular with gene prediction because of its simplicity. Model-based autoregressive (AR) spectral estimation gives better resolution even for small DNA segments but selection of appropriate model order is a critical issue. In this article a technique has been proposed where Yule-Walker autoregressive (YW-AR) process is combined with principal component analysis (PCA) for reduction in dimensionality. The spectral peaks of DNA signal are used to detect protein-coding regions based on the 1/3 frequency component. Here optimal model order selection is no more critical as noise is removed by PCA prior to power spectral density (PSD) estimation. Eigenvalue-ratio is used to find the threshold between signal and noise subspaces for data reduction. Superiority of proposed method over fast Fourier Transform (FFT) method and autoregressive method combined with wavelet packet transform (WPT) is established with the help of receiver operating characteristics (ROC) and discrimination measure (DM) respectively.

A behavioral study of healthy and cancer genes by modeling electrical network.

In recent years, gene network modeling is gaining popularity in genomics to monitor the activity profile of genes. More specifically, the objective of the network modeling concept is to study the genetic behavior associated with disease. Previous researchers have designed network model at nucleotide level which produces more complexity for designing circuits mostly in case of gene expression studies. Whereas the authors have designed the present network model, based on amino acid level which is simpler as well as more appropriate for prediction of the genetic abnormality. In the present concept, SISO continuous and discrete system models of genes are realized using Foster network. The model is designed based on hydropathy index value of amino acids to study the biological system behavior. The time and phase response in continuous (s) domain and pole-zero distribution in discrete (z) domain are used as measurement metric in the present study. The simulated responses of the system show genetic instability for cancer genes which truly reflects the medical reports. The proposed modeling concept can be used, to accurately identify or separate out the diseased genes from healthy genes.

Anti-bacterial activity of Achatina CRP and its mechanism of action.

The physiological role of C-reactive protein (CRP), the classical acute-phase protein, is not well documented, despite many reports on biological effects of CRP in vitro and in model systems in vivo. It has been suggested that CRP protects mice against lethal toxicity of bacterial infections by implementing immunological responses. In Achatina fulica CRP is a constitutive multifunctional protein in haemolymph and considered responsible for their survival in the environment for millions of years. The efficacy of Achatina CRP (ACRP) was tested against both Salmonella typhimurium and Bacillus subtilis infections in mice where endogenous CRP level is negligible even after inflammatory stimulus. Further, growth curves of the bacteria revealed that ACRP (50 microg/mL) is bacteriostatic against gram negative salmonellae and bactericidal against gram positive bacilli. ACRP induced energy crises in bacterial cells, inhibited key carbohydrate metabolic enzymes such as phosphofructokinase in glycolysis, isocitrate dehydrogenase in TCA cycle, isocitrate lyase in glyoxylate cycle and fructose-1,6-bisphosphatase in gluconeogenesis. ACRP disturbed the homeostasis of cellular redox potential as well as reduced glutathione status, which is accompanied by an enhanced rate of lipid peroxidation. Annexin V-Cy3/CFDA dual staining clearly showed ACRP induced apoptosis-like death in bacterial cell population. Moreover, immunoblot analyses also indicated apoptosis-like death in ACRP treated bacterial cells, where activation of poly (ADP-ribose) polymerase-1 (PARP) and caspase-3 was noteworthy. It is concluded that metabolic impairment by ACRP in bacterial cells is primarily due to generation of reactive oxygen species and ACRP induced anti-bacterial effect is mediated by metabolic impairment leading to apoptosis-like death in bacterial cells.

Effective gene prediction by high resolution frequency estimator based on least-norm solution technique.

Linear algebraic concept of subspace plays a significant role in the recent techniques of spectrum estimation. In this article, the authors have utilized the noise subspace concept for finding hidden periodicities in DNA sequence. With the vast growth of genomic sequences, the demand to identify accurately the protein-coding regions in DNA is increasingly rising. Several techniques of DNA feature extraction which involves various cross fields have come up in the recent past, among which application of digital signal processing tools is of prime importance. It is known that coding segments have a 3-base periodicity, while non-coding regions do not have this unique feature. One of the most important spectrum analysis techniques based on the concept of subspace is the least-norm method. The least-norm estimator developed in this paper shows sharp period-3 peaks in coding regions completely eliminating background noise. Comparison of proposed method with existing sliding discrete Fourier transform (SDFT) method popularly known as modified periodogram method has been drawn on several genes from various organisms and the results show that the proposed method has better as well as an effective approach towards gene prediction. Resolution, quality factor, sensitivity, specificity, miss rate, and wrong rate are used to establish superiority of least-norm gene prediction method over existing method.

Sunlight-induced rapid and efficient biogenic synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum Linn. with enhanced antibacterial activity.

BACKGROUND: Nanotechnology is now regarded as a distinct field of research in modern science and technology with multifaceted areas including biomedical applications. Among the various approaches currently available for the generation of metallic nanoparticles, biogenic synthesis is of increasing demand for the purpose of green nanotechnology. Among various natural sources, plant materials are the most readily available template-directing matrix offering cost-effectiveness, eco-friendliness, and easy handling. Moreover, the inherent pharmacological potentials of these medicinal plant extracts offer added biomedical implementations of the synthesized metal nanoparticles. RESULTS: A robust practical method for eco-friendly synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum (Tulsi) as both reducing and capping agent, under the influence of direct sunlight has been developed without applying any other chemical additives. The nanoparticles were characterized with the help of UV-visible spectrophotometer and transmission electron microscopy (TEM). The prepared silver nanoparticles exhibited considerable antibacterial activity. The effects were more pronounced on non-endospore-forming Gram-positive bacteria viz., Staphylococcus aureus, Staphylococcus epidermidis, and Listeria monocytogenes than endospore-forming species Bacillus subtilis. The nanoparticles also showed prominent activity on Gram-negative human pathogenic Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and plant pathogenic Pantoea ananatis. A bactericidal mode of action was observed for both Gram-positive and Gram-negative bacteria by the nanoparticles. CONCLUSIONS: We have developed a very simple, efficient, and practical method for the synthesis of silver nanoparticles using aqueous leaf extract of O. sanctum under the influence of direct sunlight. The biosynthesis of silver nanoparticles making use of such a traditionally important medicinal plant without applying any other chemical additives, thus offers a cost-effective and environmentally benign route for their large-scale commercial production. The nanoparticles dispersed in the mother solution showed promising antibacterial efficacy. Graphical AbstractSunlight-induced rapid and efficient biogenic synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum Linn. with enhanced antibacterial activity.

Phosphate solubilizing ability of Emericella nidulans strain V1 isolated from vermicompost.

Phosphorus is one of the key factors that regulate soil fertility. Its deficiencies in soil are largely replenished by chemical fertilizers. The present study was aimed to isolate efficient phosphate solubilizing fungal strains from Eisenia fetida vermicompost. Out of total 30 fungal strains the most efficient phosphate solubilizing one was Emericella (Aspergillus) nidulans V1 (MTCC 11044), identified by custom sequencing of beta-tubulin gene and BLAST analysis. This strain solubilized 13 to 36% phosphate from four different rock phosphates. After three days of incubation of isolated culture with black Mussorie phosphate rock, the highest percentage of phosphate solubilization was 35.5 +/- 1.01 with a pH drop of 4.2 +/- 0.09. Kinetics of solubilization and acid production showed a linear relationship until day five of incubation. Interestingly, from zero to tenth day of incubation, solubility of soil phosphate increased gradually from 4.31 +/- 1.57 to 13.65 +/- 1.82 (mg kg(-1)) recording a maximum of 21.23 +/- 0.54 on day 45 in respect of the V1 isolate. Further, enhanced phosphorus uptake by Phaseolus plants with significant pod yield due to soil inoculation of Emericella nidulans V1 (MTCC 11044), demonstrated its prospect as an effective biofertilizer for plant growth.

A new pentacyclic triterpene with potent antibacterial activity from Limnophila indica Linn. (Druce).

A new pentacyclic triterpenoid constituent, characterized as 3-oxo-olean-12(13),18(19)-dien-29α-carboxylic acid (1) on the basis of detailed spectral studies, was isolated from the aerial parts and roots of Limnophila indica (Scrophulariaceae). Compound 1 exhibited considerable antibacterial activity against three Gram-positive bacteria viz. Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes (MICs within a range of 25-30 µg/ml) and moderate activity against four Gram-negative bacteria Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and Pantoea ananatis (MICs within a range of 30-100 µg/ml). The plant pathogenic bacterium P. ananatis and human pathogenic S. typhimurium responded at comparatively higher concentrations of the compound 1, which were 75 and 100 µg/ml respectively. The compound inhibited the growth of Gram-positive B. subtilis and Gram-negative P. aeruginosa completely with a clear bactericidal mode of action at their MIC values. The compound upon treatment on both B. subtilis and P. aeruginosa released substantial amount of nucleic acid in the external medium and also effected the change of morphology towards pleomorphicity, thereby indicating its probable action on cell membrane. Furthermore, the triterpenoid 1 was found not to inhibit a probiotic lactic acid bacterium Lactococcus lactis subsp. lactis LABW4 under in vitro condition and to possess no toxicity in Swiss albino mice.

A comparative antibacterial evaluation of raw and processed Guñja (Abrus precatorius Linn.) seeds.

BACKGROUND: Seed of Guñja (Abrus precatorius Linn.), a known poisonous drug, is used extensively in various ayurvedic formulations with great therapeutic significance. Ayurveda recommends the administration of Guñja in diseases like Indralupta (alopecia), Sotha (edema), Krmi (helminthes), Kustha (skin diseases), Kandu (itching), Prameha (urinary disorders) etc., after being treated with specific Sodhana (purification) procedures. OBJECTIVE: To assess the antimicrobial action of of raw and Shodhita (Processed) Guñja seeds. METHODS: Guñja seeds after being processed with Godugdha (cow's milk), Nimbu swarasa (Lemon juice), Kañji (Sour gruel) and water, as the media, were evaluated for its antibacterial effect against clinically important bacterial strains using agar well diffusion method. RESULTS: Aqueous extracts of raw seeds of Guñja exert its antibacterial effect on both Gram positive, as well as Gram negative bacteria but none of the Sodhita Guñja seeds showed any bactericidal effect on any bacterial strains. Chloroform extracts of all the Sodhita Guñja seed extracts could inhibit bacterial growth but with variations. CONCLUSION: The study displayed that chloroform extracts of raw and sodhita samples for bacterial study were much sensitive than the aqueous extracts.