Untargeted metabolomics of fermented onion (Allium cepa L) using UHPLC Q-TOF MS/MS reveals anti-obesity metabolites and in vivo efficacy in Caenorhabditis elegans.

The bioconversion of onion extracts with P. acidilactici MNL5 enhances the metabolites and has a synergistic lipid-reduction impact that is beneficial for anti-obesity studies. The 48 h fermented onion extracts (FOE) demonstrated an enhanced inhibitory activity against pancreatic lipase (89.5 ± 1.25 %) as compared to the raw onion extract (ROE) (33.4 ± 0.86 %). The antioxidant properties of FOE significantly increased compared to the ROE inhibitory effect on DPPH (99.5 ± 2.40 mg vitamin C equiv./mg, DW FOE), and ABTS (104.5 ± 2.32 mg vitamin C equiv./mg, DW FOE). Based on FOE's higher antioxidant activity, UHPLC-Q-TOF-MS/MS demonstrated dramatic changes in the untargeted metabolite profile as compared to ROE. Moreover, C. elegans supplemented with FOE and quercetin exhibited an enhanced lifespan activity, lipid reduction, and decreased triglycerides. FOE can lower cholesterol and enhance quercetin to promote pancreatic lipase activity for synergistic anti-obesity effects.

Antibacterial, Antioxidant, Larvicidal and Anticancer Activities of Silver Nanoparticles Synthesized Using Extracts from Fruits of Lagerstroemia speciose and Flowers of Couroupita guianensis.

The present study aimed to analyze the in vitro antibacterial, antioxidant, larvicidal and cytotoxicity properties of green synthesized silver nanoparticles (Ag NPs) using aqueous extracts from fruits of Lagerstroemia speciosa and flowers of Couropita guinensis. Synthesized Ag NPs were characterized using UV-DRS, FTIR, XRD, DLS, and High-Resolution SEM and TEM analyses. Absorption wavelength was observed at 386 nm by UV-DRS analysis and energy band gap was calculated as 3.24 eV. FTIR analysis showed the existence of various functional groups in the aqueous extract and in the NPs. DLS analysis showed the stability and particle size of the synthesized Ag NPs. SEM analysis revealed that Ag NPs are in a face centered cubic symmetry and spherical shape with a size of 23.9 nm. TEM analysis showed particle size as 29.90 nm. Ag NPs showed antibacterial activity against both Gram-positive and Gram-negative bacteria. DPPH scavenging trait of Ag NPs was ranging from 20.0 ± 0.2% to 62.4 ± 0.3% and observed significant larvicidal activity (LC50 at 0.742 ppm and LC90 at 6.061 ppm) against Culex quinquefasciatus. In vitro cytotoxicity activity of Ag NPs was also tested against human breast cancer (MCF-7) and fibroblast cells (L-929) and found that cells viabilities are ranging (500 to 25 µg/mL) from 52.5 ± 0.4 to 94.0 ± 0.7% and 53.6 ± 0.5 to 90.1 ± 0.8%, respectively. The synthesized Ag NPs have the potential to be used in the various biomedical applications.

Anti-Obesity Efficacy of Pediococcus acidilactici MNL5 in Canorhabditis elegans Gut Model.

In the present study, thirty two lactic acid bacteria (LAB) were isolated from fermented Indian herbal medicine. In comparison to other strains, MNL5 had stronger bile salt hydrolase (BSH) and cholesterol-lowering properties. Furthermore, it can withstand the extreme conditions found in the GI tract, due to, e.g., pepsin, bile salts, pancreatin, and acids. Pediococcus acidilactici MNL5 was identified as a probiotic candidate after sequencing the 16S rRNA gene. The antibacterial activity of P. acidilactici MNL5 cell-free supernatants (CFS) against Escherichia coli, Staphylococcus aureus, Helicobacter pylori, Bacillus cereus, and Candida albicans was moderate. A Caenorhabditis elegans experiment was also performed to assess the effectiveness of P. acidilactici MNL5 supplementation to increase life span compared to E. coli supplementation (DAF-2 and LIU1 models) (p < 0.05). An immense reduction of the lipid droplets of C. elegans was identified through a fluorescent microscope. The drastic alteration of the expression of fat genes is related to obesity phenotypes. Hence, several paths are evolutionary for C. elegans; the results of our work highlight the nematode as an important model for obesity.

Polyhydroxybutyrate degradation by biocatalyst of municipal sludge water and degradation efficacy in sequencing batch biofilm reactor.

The main aim of the study was to degrade poly-ß-hydroxybutyrate (P(3HB)) in the sequencing batch biofilm reactor (SBBR) using biocatalyst. Enrichment method was used for the isolation of P(3HB) degrading bacteria. These bacterial strains were isolated from the wastewater sludge sample treated with P(3HB) sheets. A total of 75 bacteria were isolated after 60 days of incubation. The zone of clearance varied between 12 ± 1 mm and 19 ± 2 mm. Two bacterial strains (Nitrobacter vulgaris SW1 and Pseudomonas aeruginosa KS10) showed rapid PHB degradation activity on agar plates. Plate screening experiments confirmed PHB degrading ability of P. aeruginosa KS10 and N. vulgaris SW1. Biodegrading potential improved after 72 h fermentation period. The bacteria produced depolymerase and enzyme activity was maximum after 72 h. The sequencing batch biofilm reactor (SBBR) co-cultured with N. vulgaris SW1 and P. aeruginosa KS10 was operated to remove PHB from the wastewater. Biofilm in the reactor degraded PHB and the production of polyhydroxybutyrate depolymerase influenced on PHB degradation. Polyhydroxybutyrate degradation improved continuously and maximum degradation (95.6%) was achieved after 8 days. The degradation of biopolymers help to reduce environmental pollution associated with the petroleum based polymers.

Carbapenemases producing Klebsiella pneumoniae from the pus of hospitalized patients: In-vitro antibiotic properties of Streptomyces against multidrug resistant infectious bacteria.

BACKGROUND: Klebsiella pneumoniae is predominantly exists in the pus of the human wounds and it creates massive infections in the skin and causes serious health associated infections. Modern antibiotics are highly active in the treatment of wound infections. In this study was aimed to determine resistance of K. pneumoniae screened from wound specimens of patients. Sample was collected from the pus of the patients associated with secondary infection. METHODS: Samples were serially diluted and the isolated bacterial strains were characterized by biochemical tests, colony morphology and Gram's staining methods. Resistance of K. pneumoniae was tested using antibiotics such as, Gentamycin, Ampicillin, Tetracycline, Cefurooxime, Oxacillin, Ofloxacin, Erythromycin, Nalidic acid, Cefepine, Piperacillin, Norfloxacin, Imipenem, Nitrofurantoin, Amikacin, Ciprofloxacin, Vancomycin, Meropeneum and Cefotaxime with Kirby-Bauer disc diffusion method. RESULTS: Among the 73 K. pneumoniae strains, four strains produced AmpC and ESBLs, 42 strains produced ESBLs and 7 bacterial strains synthesized only AmpC enzyme. Four stains produced ESBLs and showed multidrug resistance against various antibiotics. Most of the strains synthesized extracellular polysaccharides and mediated biofilm formation. Among the K. pneumoniae strains, K. pneumoniae PS02 showed multidrug resistant against most of the tested antibiotics. It produced ESBLs and AmpC enzyme. To produce secondary metabolites, actinomycetes were isolated and characterized as Streptomyces sp. AC14. The secondary metabolite was effective against Klebsiella strains. CONCLUSIONS: To conclude, secondary metabolites extracted from Streptomyces sp. AC14 was found to be effective against multidrug resistant bacterium. Further studies are warranted to analyze the drug hydrolyzing pathways of bacteria and to identify the mechanism of action of secondary metabolites from Streptomyces sp. AC14.

Hepatoprotective effect of selected isoandrographolide derivatives on steatotic HepG2 cells and High Fat Diet fed rats.

Non-alcoholic Fatty Liver Disease (NAFLD) is one of the growing epidemics of the globe. This study was aimed to evaluate the anti-NAFLD effect of selected IAN derivatives using in silico, in vitro and in vivo models. In silico tools viz., DataWarrior, SwissADME and Gaussian 09 were used to predict the pharmacokinetic properties and electronic distribution patterns of the derivatives; docking analysis was done with Autodock against PPARα. Toxicities of the derivatives were assessed in HepG2 cells using MTT assay. Anti-NAFLD efficacies of the derivatives were assessed in free fatty acid induced steatotic HepG2 cells. In vivo anti-NAFLD effect of active isoandrographolide (IAN) derivative, 19-propionyl isoandrographolide (IAN-19P) was assessed in High Fat Diet fed rats. In silico and in vitro studies indicated that IAN-19P showed improved drug-likeness and drug score. The toxicity of IAN-19P to HepG2 cells was comparatively less than IAN and other derivatives. In free fatty acid induced steatotic HepG2 cells, treatment with IAN-19P significantly lowered intracellular triglyceride content and leakage of LDH and transaminases. Treating High Fat Diet fed animals with IAN-19P significantly lowered plasma lipids, transaminases, LDH and GGT levels. The treatment with IAN-19P upregulated the expressions of PPARα and CPT-1. IAN-19P did not produce any noticeable adverse effect till 2 g/kg concentration in acute and 250 mg/kg concentration in subacute toxicity studies. This study indicated the beneficial effect of IAN-19P for the treatment of NAFLD; however robust investigations are needed to establish the potential of IAN-19P to treat NAFLD.

Emerging paradigms of viral diseases and paramount role of natural resources as antiviral agents.

In the current scenario, the increasing prevalence of diverse microbial infections as well as emergence and re-emergence of viral epidemics with high morbidity and mortality rates are major public health threat. Despite the persistent production of antiviral drugs and vaccines in the global market, viruses still remain as one of the leading causes of deadly human diseases. Effective control of viral diseases, particularly Zika virus disease, Nipah virus disease, Severe acute respiratory syndrome, Coronavirus disease, Herpes simplex virus infection, Acquired immunodeficiency syndrome, and Ebola virus disease remain promising goal amidst the mutating viral strains. Current trends in the development of antiviral drugs focus solely on testing novel drugs or repurposing drugs against potential targets of the viruses. Compared to synthetic drugs, medicines from natural resources offer less side-effect to humans and are often cost-effective in the productivity approaches. This review intends not only to emphasize on the major viral disease outbreaks in the past few decades and but also explores the potentialities of natural substances as antiviral traits to combat viral pathogens. Here, we spotlighted a comprehensive overview of antiviral components present in varied natural sources, including plants, fungi, and microorganisms in order to identify potent antiviral agents for developing alternative therapy in future.

Molecular diversity and hydrolytic enzymes production abilities of soil bacteria.

Soil is an integral part of ecosystem which is niche for varieties of microflora. The present study was investigated to isolate varied strains of bacteria from soil samples of three different geographical regions of Tamil Nadu (India) and evaluate their hydrolytic enzymes (amylase, cellulase, and inulinase) producing potentialities. Among 72 bacterial cultures isolated from Ambattur Industrial Estate, Neyveli Lignite Corporation, and Arignar Anna Zoological Park regions, 41.66, 38.88, and 36.11% of isolates were observed amylase, cellulase, and inulinase producers, respectively. On the other hand, 20.83% of total bacteria isolated from all three regions exhibited concurrent production of amylase, cellulase, and inulinase. Potent isolates depicting maximum enzyme activities were identified as Bacillus anthracis strain ALA1, Bacillus cereus strain ALA3, Glutamicibacter arilaitensis strain ALA4, and Bacillus thuringiensis strain ALA5 based on molecular characterization tools. Further, the thermodynamics parameters, open reading frames (ORFs) regions, and guanine-cytosine (GC) content were determined by distinct bioinformatics tools using 16S rRNA sequences of strains. Minimum free energy values for strain ALA1, strain ALA3, strain ALA4, and strain ALA5 were calculated as -480.73, -478.76, -496.63, and -479.03 kcal/mol, respectively. Mountain plot and entropy predicted the hierarchical representation of RNA secondary structure. The GC content of sequence for strain ALA1, strain ALA3, strain ALA4, and strain ALA5 was calculated as 53.06, 52.94, 56.78, and 53.06%, respectively. Nine ORFs were obtained for strain ALA1, strain ALA3, and strain ALA5 while 10 ORFs were observed for strain ALA4. Additionally, bootstrap tree demonstrated close resemblance of strains with existing bacteria of similar genus. Findings showed higher variability of bacterial diversity as hydrolytic enzymes producers in the investigated geographical regions.

Purification and characterization of anti-tubercular and anticancer protein from Staphylococcus hominis strain MANF2: In silico structural and functional insight of peptide.

The present context was investigated to purify and characterize anti-tubercular as well as anticancer protein from fermented food associated Staphylococcus hominis strain MANF2. Initially, the anti-tubercular potency of strain MANF2 was assessed against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay which revealed pronounced relative light unit (RLU) reduction of 92.5 ± 1.2%. The anticancer property of strain MANF2 was demonstrated against lung cancer (A549) and colon cancer (HT-29) cell lines using MTT assay which showed reduced viabilities. Anti-tubercular activities of the purified protein were observed to be increased significantly (P < 0.05) ranging from 34.6 ± 0.3 to 71.4 ± 0.4% of RLU reduction. Likewise, the purified protein showed significantly (P < 0.05) reduced viabilities of A549 and HT-29 cancer cells with IC50 values of 46.6 and 48.9 µg/mL, respectively. The nominal mass of the purified protein was found to be 7712.3 Da as obtained from MALDI-TOF MS/MS spectrum. The protein showed the sequence homology with 1-336 amino acids of Glyceraldehyde-3-phosphate dehydrogenase from Staphylococcus sp., thus, categorizing as a new class of Glyceraldehyde-3-phosphate dehydrogenase-like protein. The amino acid sequence of the most abundant peptide (m/z = 1922.12) in the purified protein was obtained as 'KAIGLVIPEIDGKLDGGAQRV' and it was identified as peptide NMANF2. In silico tools predicted significant stereo-chemical, physiochemical, and functional characteristics of peptide NMANF2. In a nutshell, protein purified from strain MANF2 can certainly be used as an ideal therapeutic agent against tuberculosis and cancer (lung and colon).

Green synthesis and characterization of gold nanoparticles using endophytic fungi Fusarium solani and its in-vitro anticancer and biomedical applications.

The present study aimed to explore the anticancer potentials of the gold nanoparticles (NPs) obtained by green synthesis method using an endophytic strain Fusarium solani ATLOY - 8 has been isolated from the plant Chonemorpha fragrans. The formation of the NPs was analyzed by UV, FTIR, SEM and XRD. The synthesized NPs showed pink-ruby red colors and high peak plasmon band was observed between 510 and 560 nm. It is observed that intensity of absorption steadily increases the wavelength and band stabilizes at 551 nm. The XRD pattern revealed the angles at 19, 38.32, 46.16, 57.50, and 76.81° respectively. Interestingly, the FTIR band absorption noted at 1413 cm-1, 1041 cm-1 and 690 cm-1 ascribed the presence of amine II bands of protein, C-N and C-H stretching vibrations of the nanoparticles. SEM analysis indicated that the average diameter of the synthesized nanoparticles was between 40 and 45 nm. These NPs showed cytotoxicity on cervical cancer cells (He La) and against human breast cancer cells (MCF-7) and the NPs exhibited dose dependent cytotoxic effect. IC50 value was 0.8 ± 0.5 µg/mL on MCF-7 cell line and was found to be 1.3 ± 0.5 µg/mL on MCF-7 cell lines. The synthesized NPs induced apoptosis on these cancer cell lines. The accumulation of apoptotic cells decreased in sub G0 and G1 phase of cell cycle in the MCF-7 cancer cells were found to be 55.13%, 52.11% and 51.10% after 12 h exposure to different concentrations. The results altogether provide an apparent and versatile biomedical application for safer chemotherapeutic agent with little systemic toxicity.

Biosynthesis of silver and gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy.

Synthesis of nanoparticles using plant sources as reducing agent has become important, as physical and chemical methods are costlier and affects environment. Hence it is important to develop environment friendly nanoparticle synthesis by avoiding the use of toxic chemicals. The present study aimed to synthesize silver nanoparticles (Ag Nps) and gold nanoparticles (AuNps) using Musa acuminata colla flower and its pharmaceutical activity against extended spectrum beta-lactamase (ESBL) gene producing bacteria and anticancer efficacy. The synthesized Ag and Au NPs were analysed by means of UV-Vis, FTIR, XRD,SEM and EDAX evidenced the bioreduction of Ag+ ions to Ag0 and Au3+ ions to Au0 respectively. Both nanoparticles and flower extracts were studied for antibacterial activity of ESBL gene producing bacteria by disc diffusion and microdilution (Resazurin) method. In vitro anticancer efficacy (MCF-7) and toxicity (VERO) of AgNPs, AuNPs, aqueous extract and ethanol extract of flowers were performed by MTT assay. IC50 value for DPPH analysis was at 390 µg and 460 µg for ethanol and aqueous extract respectively. Total antioxidant content was found be 740 µg/mg and 460 µg/mg for ethanol and aqueous extract. GCMS analysis authenticated the existence of the compounds namely, 9,12-octadecadienoic acid(z,z)- and n-hexadecanoic acid in the crude extract of the samples. Among the samples, AgNPs had best antibacterial activity. AgNPs and AuNPs were confirmed by colour change to reddish brown and ruby red. Further ƛmax were obtained at 474 and 540 nm by UV - visible spectrum. SEM analysis revealed the particle size ranges from 12.6 to 15.7 nm for silver and 10.1 to 15.6 nm for gold nanoparticles. The EDAX spectrum shows a strong signal for elemental Ag and Au at ~ 3 keV and 1.5 keV. The XRD patterns for silver and gold nanoparticles at 36.701, 42.900, 63.281 and 76.398 corresponding to the lattice planes 2.4467, 2.1064, 1.46839, 1.24564 nm and 27.32, 36.7228, 39.56, 42.888, 63.253, 63.253, 65.02 and 76.383 corresponding to the lattice planes 3.262, 2.44530, 2.276, 2.1070, 1.46897, 1.4332 and 1.24585 nm. The IC50 values for MCF-7 and VERO cells were 30.0 µg/ml and 55.0 µg/ml respectively.

Gut Microbiome Modulation Based on Probiotic Application for Anti-Obesity: A Review on Efficacy and Validation.

The growing prevalence of obesity has become an important problem worldwide as obesity has several health risks. Notably, factors such as excessive food consumption, a sedentary way of life, high sugar consumption, a fat-rich diet, and a certain genetic profile may lead to obesity. The present review brings together recent advances regarding the significance of interventions involving intestinal gut bacteria and host metabolic phenotypes. We assess important biological molecular mechanisms underlying the impact of gut microbiota on hosts including bile salt metabolism, short-chain fatty acids, and metabolic endotoxemia. Some previous studies have shown a link between microbiota and obesity, and associated disease reports have been documented. Thus, this review focuses on obesity and gut microbiota interactions and further develops the mechanism of the gut microbiome approach related to human obesity. Specifically, we highlight several alternative diet treatments including dietary changes and supplementation with probiotics. The future direction or comparative significance of fecal transplantation, synbiotics, and metabolomics as an approach to the modulation of intestinal microbes is also discussed.

Isolation and characterization of Aspergillus sp. for the production of extracellular polysaccharides by response surface methodology.

In this study, Aspergillus sp. was isolated for the production of extracellular polysaccharide. The process parameters were initially optimized by traditional methods. The cheap substrate, wheat bran was used for the production of extracellular polysaccharide in solid state fermentation. Supplementation of (1%, w/w) maltose, gelatin enhanced EPS production (5.36 mg/g). The salts such as, Cu2+ (4.9 mg/g), Ca2+ (3.5 mg/g), Zn2+ (2.9 mg/g), Mn2+ (3.4 mg/g) and Mg2+ (1.8 mg/g) stimulated EPS production. In two level full factorial experimental designs, the EPS yield varied from 3.18 to 11.65 mg/g wheat bran substrate with various combinations of the components supplemented with wheat bran substrate. Among these selected factors in central composite design, maltose significantly influenced on extracellular polysaccharide production.

Anti-inflammatory activity of rhein isolated from the flowers of Cassia fistula L. and possible underlying mechanisms.

OBJECTIVE: Anti-inflammatory activity of rhein in animal models with potential mechanism of actions. METHODS: Rhein was isolated from Cassia fistula L. flowers collected in Chennai, Tamil Nadu, India. Its anti-inflammatory activity was then investigated in Wistar rats and mice using carrageenan-induced hind paw oedema, croton oil-induced ear oedema, cotton pellet-induced granuloma and acetic acid-induced vascular permeability models. RESULTS: Administration of rhein (10, 20, 40 mg/kg) significantly (p < 0.05) inhibited carrageenan-induced paw oedema in rats and croton oil-induced ear oedema in mice in dose-dependent manners. Continual administration of rhein to rats using implanted cotton pellets significantly (p < 0.05) reduced granuloma formation (20 mg/kg: 17.24%; 40 mg/kg: 36.12%) compared to control group animals. Administration of rhein increased the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and decreased the levels of nitrite, interleukin-6 (IL-6), interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA) and vascular endothelial growth factor (VEGF) compared to control animals. Western blotting results revealed that rhein diminished carrageenan-induced cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) and increased heme oxygenase (HO)-1, nuclear factor erythroid 2-related factor 2 (Nrf2), peroxisome proliferator-activated receptor gamma (PPAR)-γ and heat shock protein (HSP)-72 expression after 6 h in the paw oedema model. CONCLUSION: The anti-inflammatory mechanisms of rhein might be related to decrease in the levels of MDA, iNOS and COX-2 and the stimulation of HO-1, PPAR-γ and Nrf2 expression via increases in the activities of CAT, SOD and GSH-px through the suppression of nitrite, TNF-α, IL-6 and IL-1ß.

Rapid biosynthesis and characterization of silver nanoparticles from the leaf extract of Tropaeolum majus L. and its enhanced in-vitro antibacterial, antifungal, antioxidant and anticancer properties.

Eco-friendly biosynthesis of nanoparticles from medicinal plants as reducing agent has gained importance due to its potential therapeutic uses. In the present study Silver nanoparticles (AgNPs) were eco-friendly synthesized using the leaf extracts of the medicinal plant Tropaeolum majus. The obtained AgNPs were characterized by UV - visible spectrum, FTIR, SEM and XRD which clearly showed the reduction of Ag+ ions to Ag0. In addition, the aqueous and ethanolic extracts were analyzed for phytochemicals and its antioxidant activities. GC-MS spectrum showed the presence of 25 compounds with benzeneacetic acid as the dominant contents. The synthesized AgNPs revealed maximum absorption spectrum at 463 nm and FTIR vibrational peaks at 3357.46, 21,966.52, 2118.42, 1637.27, 658.571 and 411.728 cm-1 respectively. SEM and XRD studies evidenced the nature of nanocrystalline with face centered cubic (fcc) crystal structure. Both AgNPs and plant extracts showed more inhibition activity against Pseudomonas aeroginosa compared to other bacteria with MIC value of 6.25 µg/ml. Antifungal activities was higher for Penicilium notatum with MIC value 31.2 µg/ml. The IC50 values for MCF7 for aqueous extract were found to be 4.68 µg/ml, ethanol extract 7.5 µg/ml, AgNPs 2.49 µg/ml, and doxorubicin 1.4 µg/ml. The IC50 values for VERO cell line for aqueous extract was 8.1 µg/ml, ethanol extract with 6.8 µg/ml, silver nanoparticles 5.3 µg/ml and doxorubicin 2.6 µg/ml respectively. Conclusively, the antibacterial, antifungal, antioxidant and anticancer properties of the synthesized AgNPs from Tropaeolum majus act as major therapeutic drug for microbial infectious disease and other health associated disorders.

Carboxymethyl cellulase production optimization from Glutamicibacter arilaitensis strain ALA4 and its application in lignocellulosic waste biomass saccharification.

In this context, carboxymethyl cellulase (CMCase) production from Glutamicibacter arilaitensis strain ALA4 was initially optimized by one factor at a time (OFAT) method using goat dung as proficient feedstock. Two-level full factorial design (25 factorial matrix) using first-order polynomial model revealed the significant (p < 0.05) influence of pH, moisture, and peptone on CMCase activity. Central composite design at N = 20 was further taken into account using a second-order polynomial equation, and thereby liberated maximum CMCase activity of 4925.56 ± 31.61 U/g in the goat dung medium of pH 8.0 and 100% moisture containing 1% (w/w) peptone, which was approximately two fold increment with respect to OFAT method. Furthermore, the partially purified CMCase exhibited stability not only at high pH and temperature but also in the presence of varied metal ions, organic solvents, surfactants, and inhibitors with pronounced residual activities. The enzymatic hydrolysis using partially purified CMCase depicted the maximum liberation of fermentable sugars from alkali pretreated lignocellulosic wastes biomass in the order of paddy straw (13.8 ± 0.15 mg/g) > pomegranate peel (9.1 ± 0.18 mg/g) > sweet lime peel (8.37 ± 0.16 mg/g), with saccharification efficiency of 62.1 ± 0.8, 40.95 ± 0.4, and 37.66 ± 0.4%, respectively after 72 hr of treatment.

Chemical composition, antioxidant activity and antibacterial mechanism of action from Marsilea minuta leaf hexane: methanol extract.

BACKGROUND: In the present study, hexane: methanol (50:50) leaf extract of Marisela minuta has been evaluated for its chemical composition, antioxidant effect and the antimicrobial mechanism of action against food borne pathogenic bacteria. RESULTS: The phytochemical evaluation of extract by GC/MS revealed the major abundance of benzoic acid-4-ethoxyethyl ester (43.39%) and farnesol acetate (18.42%). The extract exhibited potential antioxidant and free radical scavenging properties with promising antibacterial activities against the test pathogens with Pseudomonas aeruginosa being the most susceptible with maximum inhibition zone (17 mm) and IC50 value of 125 µg, respectively. The significant (p < 0.05) increase in intracellular super oxide dismutase (SOD), protein leakage, extracellular alkaline phosphatase and lactate dehydrogenase in treated test pathogens suggested an increase in oxidative stress reveling the mechanism of action of phytochemicals. Scanning electron microscopy analysis of treated pathogens also showed swollen and distorted cells. The bioactive molecules in the extract were efficiently docked with virulent enzymes and farnesol acetate showed best energy value of - 5.19 and - 4.27 kcal/mol towards Topoisomerase IV and SHV-2 respectively. Benzoic acid-4-ethoxyethyl ester showed best binding against TEM-72 with low binding energy value of - 4.35 kcal/mol. CONCLUSION: Due to its antioxidant and antibacterial properties, the leaf extract of M. minuta may act as promising natural additives to prevent food spoilage bacteria.

Enhancement of anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array and Box-Behnken design.

The prime focus of the present investigation was to optimize statistically the anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array (OA) and Box-Behnken design (BBD). The anti-tubercular activity of strain MANF2 was determined against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay. Among varied media examined, the isolate exhibited impressive anti-tubercular activity with paramount relative light unit reduction of >90% in de Man Rogose Sharpe (MRS) broth. Primarily, the anti-tubercular activity and biomass of strain MANF2 were estimated in MRS broth by optimizing eight diversified parameters using one factor at a time (OFAT) method after working out a series of experiments. The most significant contributing factors selected through OFAT tool were optimized using Taguchi approach with a standard OA layout of L18 (22 × 36). Results demonstrated the significant (P ≤ 0.05) influence of pH, temperature, yeast extract, magnesium sulphate, and glycerol on response variables. These controlled variables were further optimized using BBD matrix at N = 46 by second-order polynomial equation. The fermentation medium of pH 6.5 constituting yeast extract (0.5% w/v), magnesium sulphate (0.1% w/v), and glycerol (1.5% v/v), being further incubated at 30 °C showed enhanced anti-tubercular activity (98.7%) and approximately 4 fold increment in the bacterial biomass yield (8.3 mg/mL) with respect to traditional OFAT method. Three-dimensional response plots of the quadratic model showed interdependent interaction between the significant variables. In conclusion, the present study revealed the first report on the optimization of anti-tubercular activity and biomass of S. hominis via Taguchi OA as well as BBD design, and thus, paved a path for its proficient applications in pharmaceutical industries as dynamic mycobactericidal agent in future.

In vitro investigation on probiotic, anti-Candida, and antibiofilm properties of Lactobacillus pentosus strain LAP1.

OBJECTIVE: To investigate the probiotic characteristics, anti-Candida activity, and antibiofilm attributes of Hentak derived Lactobacillus pentosus strain LAP1. DESIGN: The probiotic properties of strain LAP1 were depicted by adapting standard protocols. The anti-Candida and antibiofilm properties of isolate were determined using agar well diffusion assay and ELISA reader test, respectively. The time-kill assay was performed using viable colony count assay. Further, the co-aggregation property of strain LAP1 was determined based on standard methodology. RESULTS: Strain LAP1 exhibited not only tolerance to acidic pH but also showed resistivity (P ≤ 0.05) to simulated gastric juice exposure. Similarly, the strain was able to tolerate bile salt, showed hyperproteolytic activity, and also depicted susceptibility to most of the antibiotics tested. Auto-aggregation phenomenon (37.5-60%), hydrophobicity nature (42.85%), and survival potentiality of strain LAP1 under freeze-dried condition (9.0 ±â€¯0.01 log CFU/ml) made the isolate a promising probiotic candidate. Cell-free neutralized supernatant (CFNS) of strain LAP1 exhibited potent antifungal activities against C. albicans, C. tropicalis, and C. krusei with arbitrary unit of 150 ±â€¯4.34, 200 ±â€¯5.21, and 130 ±â€¯5.13 AU/ml, respectively and depicted remarkable reduction in the biofilm formation of respective Candida sp. in a concentration dependent manner. Moreover, time-kill assay data provided the growth inhibition of all Candida sp. in a time dependent manner. Additionally, strain LAP1 revealed significant co-aggregate percentage with C. albicans, C. tropicalis, and C. krusei. CONCLUSIONS: L. pentosus strain LAP1 exhibited a good probiotic characteristics, potent anti-Candida activity, and significant antibiofilm property that could be undoubtedly recommended for its vast applications not only in food industries but also as biotherapeutic agent against Candida infections in pharmaceutical industries.

Anti-tubercular and probiotic properties of coagulase-negative staphylococci isolated from Koozh, a traditional fermented food of South India.

In the last few years, the demand for the tremendous therapeutic applications of indigenous probiotic bacteria from diversified fermented food products has surged. In view of this, the present study was documented to evaluate the anti-tubercular and probiotic properties of coagulase-negative staphylococci (CNS) indigenous to Koozh, a traditional fermented food product of South India. A total of 18 isolates were purified from Koozh, and tested for anti-tubercular activity against Mycobacterium tuberculosis H37Rv using luciferase reporter phage (LRP) assay. Among them, six isolates revealed higher percentage (>90%) of relative light unit (RLU) reduction. These six isolates were further evaluated for their in vitro probiotic attributes using standard protocols. All six staphylococci strains disclosed good probiotic properties. Moreover, Staphylococcus hominis strain MANF2 showed high cell survival percentage (92.2%) at pH 2.0 as well as towards simulated gastric juice (88.51%). Furthermore, strain MANF2 was found to be resistant to bile salt after 24 h of incubation with maximal viability of 5.71 ± 0.02 log cfu/mL, and depicted the deconjugation of bile salt as well. All the isolates exhibited strong auto-aggregation capacity (44.4 ± 1.2-68.1 ± 1.5%), and hydrophobicity against toluene (55.0 ± 1.2-72.0 ± 1.1%). Additionally, strain MANF2 was observed to be highly resistant to phenol (6.27 ± 0.01 log cfu/mL) and lysozyme (81.1 ± 1.6% viability). Most importantly, all six isolates depicted good hypocholesterolemic effect, slight ß-galactosidase activity, and moderate proteolytic property. The strains were sensitive to all the tested conventional antibiotics, except Nalidixic acid. In addition to this, all staphylococci strains demonstrated significant DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging, hydrogen peroxide tolerance, and hydroxyl radical scavenging activity in a dose dependent manner, thereby exhibiting the potent antioxidative properties of isolates. The negative results obtained from haemolytic, DNase, and gelatinase tests revealed the non-pathogenicity and safety aspect of these strains. In a nutshell, the present investigation divulges the persuasive anti-tubercular and probiotic properties of staphylococci, particularly strain MANF2, and recommended the further exploitation of Koozh associated CNS in pharmaceutics.