Investigation of Osteomyelitis Inducing Methicillin-Resistant Staphylococcus aureus Inhibition Effect by Strontium-Substituted Borate Bioactive Glasses.

Borate glass transforms into hydroxycarbonate apatite more rapidly than silicate glass. This research aims to evaluate strontium's structural and biological effects on borate bioactive glass (BBG) and the influence of strontium concentrations (0%, 5%, 10%, and 15% Sr) prepared via the sol-gel method. The study reveals significant findings related to the physicochemical properties of the glass. Immersion of the glass powders in a simulated body fluid (SBF) led to the development of a hydroxyapatite (HAP) layer on the glass surfaces. This transformation was verified through X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared (FTIR) analyses. In particular, 5% strontium exhibited gradual degradation, resulting in particle sizes below 100 nm. The BBG-15%Sr demonstrates heightened pathogenic activity as it shows a significant inhibition zone of 14 mm at 250 µg/mL, surpassing other substituted BBGs. It effectively combats Gram-positive bacteria, completely inhibiting MRSA growth at 50 µg/mL. This underscores its robust biofilm disruption capabilities, eradicating biofilms, even at minimal concentrations after prolonged exposure. C. elegans when subjected to BBG-15%Sr shows less ROS production when compared with the others. Moreover, the results suggest that the modified glass could be a potential material for the treatment of osteomyelitis-affected bone repair.

Fabrication and characterisation of human gut microbiome derived exopolysaccharide mediated silver nanoparticles - An in-vitro and in-vivo approach of Bio-Pm-AgNPs targeting Vibrio cholerae.

Utilising bacteria to produce silver nanoparticles was highly favoured due to its ability to minimise costs and mitigate any potential negative environmental impact. Exopolysaccharides (EPS) extracted from the human gut microbe have demonstrated remarkable efficacy in combating various bacterial infections. Exopolysaccharide (EPS), a naturally occurring biomolecule found in the human gut isolate Proteus mirabilis DMTMMR-11, was characterised using analytical techniques such as Fourier transform infrared spectroscopy (FTIR), 1H-nuclear magnetic resonance, 13C-nuclear magnetic resonance (NMR), and chemical composition analysis, which confirms the presence of carbohydrates (81.03 ± 0.23), proteins (4.22 ± 1.2), uronic acid (12.1 ± 0.12), and nucleic acid content (2.44 ± 0.15) in exopolysaccharide. The one factor at a time (OFAT) and response surface methodology (RSM) - central composite design (CCD) approaches were used to optimise the production of Bio-Pm-AgNPs, leading to an increase in yield of up to 1.86 g/l. The Bio-Pm-AgNPs were then subjected to Fourier transform infrared spectroscopy (FTIR) which determines the functional groups, X-ray diffractometer confers that Bio-Pm-AgNPs are crystalline in nature, field emission-scanning electron microscopy (FE-SEM) reveals the morphology of Bio-Pm-AgNPs, energy dispersive X-ray spectroscopy (EDX) confirms the presence of elements like Ag, C and O, high-resolution transmission electron microscopy (HR-TEM) determines that the Bio-Pm-AgNPs are sphere-shaped at 75 nm. Dynamic light scattering (DLS) and zeta potential analysis were also carried out to reveal the physiological nature of Bio-Pm-AgNPs. Bio-Pm-AgNPs have a promising effect on the inhibitory mechanism of Vibrio cholerae cells at a MIC concentration of 20 µg/ml which significantly affects cellular respiration and energy metabolism through glycolysis and TCA cycles by deteriorating the enzyme responsible for ATP and NADH utilisation. The action of Bio-Pm-AgNPs reduces the purity and concentration of nucleic acids, which leads to higher DNA damage. In-vivo analysis reveals that the treatment of Bio-Pm-AgNPs decreased the colonisation of V. cholerae and improved the survival rates in C. elegans.

Characterization of phenyl propiolic acid from Proteus mirabilis DMTMMR-11 and Evaluation of its mode of action against Yersinia enterocolitica (MTCC-840) an in-Vitro and in-Vivo based approach.

Foodborne illnesses are pervasive in raising public health concerns in both developed and developing nations. Yersinia enterocolitica a zoonotic bacterial species that causes food-transmitted infections, and gastroenteritis, is its most prevalent clinical manifestation. This study aims to investigate the differences, dependencies, and inhibitory mechanisms between the host and the microbiome. Proteus mirabilis DMTMMR-11, the bacterium found in the human gastrointestinal tract was used for the extraction of intracellular metabolite, because of its beneficial effects on the normal flora of the human gut. Phenyl propiolic acid was identified as the dominant compound in the metabolite after characterization using FT-IR, NMR, and LC-MS-MS. To assess its inhibitory mechanism against Yersinia enterocolitica, the pathogen was subjected to biological characterization by MBC and MIC, resulting in the rate of inhibition at 50 µg/ml. Anti-bacterial curve supports the inhibited growth of Y. enterocolitica. Mechanism of inhibition at its cellular level was indicated by the increase in alkaline phosphate content, which drastically reduced the cell membrane and cell wall potential expanding its permeability by intruding the membrane proteins, which was observed in SEM Imaging. Phenyl propiolic acid efficiently disrupts the biofilm formation by reducing the adherence and increasing the eradication property of the pathogen by exhibiting 65% of inhibition at the minimal duration of 12h. In-vivo study was carried out through host-pathogen interaction in C. elegans, an efficient model organism assessed for its life-span, physiological, and behavioral assays.

Liposome encapsulated surfactant abetted copper nanoparticles alleviates biofilm mediated virulence in pathogenic Pseudomonas aeruginosa and MRSA.

In the present study lipopeptide biosurfactant with high emulsification capacity produced by human skin bacterium Paenibacillus thiaminolyticus was purified and subjected to FTIR and NMR spectral analysis which gave evidence of the active characteristics of the surfactant. To augment the antivirulent potential further, the mixer of copper and copper oxide nanoparticles (CuNPs) was synthesized, and characterized by UV-Visible spectroscopy, SEM-EDAX, TEM, and Zeta analysis. Here, we attempted to enhance the antimicrobial and antibiofilm activity with the assistance of encapsulated preparation of lipopeptide and CuNPs in multilamellar liposomes. The proposed mechanism of action of lipopeptide and CuNPs liposomal preparation negatively influences the cell metabolism, secreted virulence such as staphyloxanthin, pyocyanin, and extracellular polysaccharides. The significant decline in the growth of MRSA and P. aeruginosa in both planktonic form and biofilm by lipopeptide and CuNPs treatment were visualized using scanning electron microscopy and High content screening imaging system. In vivo studies revealed that treatment with lipopeptide and CuNPs in multilamellar liposomes extended the lifespan of infected Caenorhabditis elegans by about 75%. Therefore, this study typifies lipopeptide and CuNPs could credibly be a substantial substitute over conventional antibiotics in averting the biofilm associated pathogenesis of MRSA and P. aeruginosa.

Anti-listerial activity of microalgal fatty acid methyl esters and their possible applications as chicken marinade.

Fatty acid methyl esters (FAMEs) from marine microalgae have been reported to possess antimicrobial activities against several Gram positive and Gram negative bacteria, but a majority of them needs to be explored. The objective of this study was to investigate the antibacterial activity, mechanism of FAMEs from selected marine microalgae against Listeria monocytogenes, and to elucidate its efficacy in food model. The minimum inhibitory concentration of FAMEs was calculated to be 155 µg/mL for Chromulina sp. and 162 µg/mL for Nannochloropsis sp. against L. monocytogenes. Time-killing kinetics showed that FAMEs efficiently inhibited the growth of L. monocytogenes in a time and concentration dependent manner. The mechanism of action of FAMEs was studied by analysing its effects at a MIC on the cellular metabolism, membrane permeability, and membrane integrity of L. monocytogenes. Transmission Electron Microscopy (TEM) results showed that cells exposed to FAMEs showed damaged cell membrane structure with leakage of the internal contents in the cells of L. monocytogenes. Fluorescence microscopy images showed that L. monocytogenes cells treated with FAMEs showed high dead cell population corresponding with propidium iodide positive cells. Furthermore, FAMEs significantly down regulated quorum sensing and biofilm related genes (DegU, FlaE, and FlaD). In vivo therapeutic potential of FAMEs revealed improved Caenorhabditis elegans survival and reduced intestinal colonization during L. monocytogenes infection. Growth of listeria was abolished in chicken meat during the cold storage of 9 days when the samples were pre-treated with FAMEs. These results suggest anti-L. monocytogenes activity of FAMEs and elucidated its use in food control of chicken meat at refrigerated conditions.

Protective effects of fucoidan against 4-nitroquinolin-1-oxide provoked genetic damage in mouse bone marrow cells.

Fucoidan is a unique bioactive and dietary polymer enriched mainly in the cell wall matrix of the brown seaweeds. This present study was intended to reveal the antigenotoxicity effect of fucoidan on 4-nitroquinolin-1-oxide (4-NQO) induced genetics damage and apoptosis in mice bone marrow cells. The 4-NQO caused genetic damages in the form of chromosome/chromatic breakage was estimated by micronuclei assay whereas apoptosis by annexin-V FITC kit and DNA damage by comet assay kit. In addition, oxidative damage in terms of plasma lipid peroxidation (LPO) and 8-OHdG was also estimated. In the experimental regime, six groups with each in five either sex of mice were used. Fucoidan constituted (50,100,200 mg/kg bwt) by orally for 5 days consequently and on 6th day, 4-NQO was administered (7.5 mg/kg bwt) by i.p. The results clearly show that negative control (H2O) and fucoidan alone constituted mice were not exhibited significant effect on LPO, genetic damages whereas positive control group (4-NQO 7.5 mg/kg bwt, i.p.) showed significant effect on genetic damage by showing increased level of LPO (6.25 vs 1.3 µM MDA), 8-OHdG (12 vs 4%), micronuclei about six-fold, 5-fold of comet, and 4-fold of apoptosis when compared with negative control, 11.6 ± 2.07, 5.00 ± 1.58, and 4.14 ± 0.65 respectively. Fucoidan pretreatment significantly protected the 4-NQO-induced genetic damage by 77% decreased level of micronuclei and 96% comet at dose of 200 mg/kg bwt over the positive control whereas LPO, 8-OHdG, and apoptosis were restored as equal to negative control. This study found as fucoidan possessing significant antigenotoxicity property by protecting 4-NQO-induced genetic damage in mice bone marrow cells as dose dependent manner suggest as valuable food supplements and medicine for mankind from environmental toxicants.

Hitherto Unknown Terpene Synthase Organization in Taxol-Producing Endophytic Bacteria Isolated from Marine Macroalgae.

Taxol is a successful anti-cancer drug, which extensively studied in Taxus spp. However, microbial endophytes also reported as taxol producers, and especially fungal endophytes extensively studied for the taxol biosynthesis pathway. Although it was well considered, the taxol biosynthesis pathway remains undisclosed since its discovery in bacteria. To decipher this gap, we isolated and identified the endophytic bacteria such as Bacillus flexus strain DMTMMB08, Bacillus licheniformis strain DMTMMB10, and Oceanobacillus picturae strain DMTMMB24, which are unprecedented for taxol production. Subsequently, the genome annotation of these bacteria exhibited the isoprene biosynthesis pathway and terpene synthase profile. Feasibly, this is the very first report on taxol-producing endophytic bacteria from the non-Taxus host and solitary investigation on its genome analysis. The genomic insight into the bacterial system for taxol biosynthesis leads to understanding the terpene synthesis and evolution. This piece of work could expand our perception of the diversity of terpenes and their related natural products.

Development of chitin cross-linked enzyme aggregates of L-methioninase for upgraded activity, permanence and application as efficient therapeutic formulations.

In this study, L-methioninase (METs) was precipitated from Pseudomonas putida MTCC 9782 and was cross-linked with a cross-linking agent glutaraldehyde to obtain a catalytically active insoluble enzyme. Among the various precipitants tested, ammonium sulfate displayed the highest precipitating (80%) efficiency. A double-response statistical concept, software that provides 20 different runs, was employed to assess the role of precipitant, concentration of cross-linking agent, and duration of cross linking. From the different 20 runs performed, the highest enzyme activity was observed in run 6 (88.17 U): the aggregate size was 11.57 µm, the concentration of saturated ammonium sulfate was 80% and glutaraldehyde 2 mM, and the incubation period was 12 h. R2 values of 0.9754 (enzyme activity) and 0.9203 (aggregate size) were obtained, which showed an enhanced association between the experimental and predicted values of enzyme activity. Enzyme molecules covalently cross-linked with chitin beads showed increased activities compared to free enzymes and enzymes cross-linked with glutaraldehyde. FTIR spectra confirmed the secondary structural alterations between CLEA-METs and chitin-cross-linked CLEA-METs. Thermal stability assays showed that chitin cross-linked CLEA-METs and CLEA-METs retained maximum enzyme activities of 95% and 80% at temperatures 55 °C and 60 °C, respectively. Storage stability assays showed that CLEA-METs retained 65% of their initial activity and chitin-immobilized CLEAs retained 88% of their activity. Moreover, scanning electron microscopy, transmission electron microscopy, and high content screening imaging technique revealed that chitin-immobilized CLEA-MET microspheres showed good monodispersity and mesoporous structure with the amorphous clusters of CLEA with few pores. Cytotoxicity analysis demonstrated that chitin-immobilized CLEA-MET significantly inhibited the proliferation of A549 cells up to 96.66% compared to free enzyme (72%) and CLEA-METs (76%).

Effect of biosurfactant derived from Vibrio natriegens MK3 against Vibrio harveyi biofilm and virulence.

Vibrio harveyi is a marine luminous pathogen, which causes biofilm-mediated infections, pressures the search for an innovative alternate approach to strive against vibriosis in aquaculture. This study anticipated to explore the effect of glycolipid biosurfactant as an antipathogenic against V. harveyi to control vibriosis. In this study, 27 bacterial strains were isolated from marine soil sediments. Out of these, 11 strains exhibited surfactant activity and the strain MK3 showed high emulsification index. The potent strain was identified as Vibrio natriegens and named as V. natriegens MK3. The extracted biosurfactant was purified using high-performance liquid chromatography and it was efficient to decrease the surface tension of the growth medium up to 21 mN/m. The functional group and composition of the biosurfactant were determined by Fourier-transform infrared spectroscopy and nuclear magnetic resonance spectroscopy spectral studies and the nature of the biosurfactant was identified as glycolipid. The surfactant was capable of reducing the biofilm formation, bioluminescence, extracellular polysaccharide synthesis, and quorum sensing in marine shrimp pathogen V. harveyi. The antagonistic effect of biosurfactant was evaluated against V. harveyi-infected brine shrimp Artemia salina. This study reveals that biosurfactant can be considered for the management of biofilm-related aquatic infections.

Decrease of growth, biofilm and secreted virulence in opportunistic nosocomial Pseudomonas aeruginosa ATCC 25619 by glycyrrhetinic acid.

The present study elucidates the antibiofilm and antivirulent capability of glycyrrhetinic acid (GRA) against Pseudomonas aeruginosa ATCC 25619. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of GRA against P. aeruginosa were found to be 160 µg/mL and 420 µg/mL respectively. In an acclimatization resistance analysis using P. aeruginosa, no resistance towards GRA was observed during the habituation period. Adequate penetration of GRA over the biofilm matrix was proposed with the membrane penetration model assembly constructed with the preformed biofilm exhibited the prospective penetration of GRA above the mature biofilm. Furthermore, GRA resulted in the attenuation of virulence factors such as motility, biofilm formation, pyocyanin secretion, secreted proteases with its sub MIC concentrations. The antibiofilm property of GRA was assessed with the light microscopy and high content screening fluorescent imaging system, which clearly demonstrates, the thickness of P. aeruginosa biofilm was reduced to 11.33 ±â€¯2.08 µm from 39 ±â€¯2.51 µm. Transmission Electron Microscopy (TEM) images depicted the morphological changes in cells such as disaggregation of colonies, cell disruption with loss of intracellular material, cytolytic damage, the process of morphological transformation, bacteriolysis indicating the potential effect of GRA.