Alginate-Capped Silver Nanoparticles as a Potent Anti-mycobacterial Agent Against Mycobacterium tuberculosis

Tuberculosis (TB) is a leading cause of death from a single infectious agent, Mycobacterium tuberculosis (Mtb). Although progress has been made in TB control, still about 10 million people worldwide develop TB annually and 1.5 million die of the disease. The rapid emergence of aggressive, drug-resistant strains and latent infections have caused TB to remain a global health challenge. TB treatments are lengthy and their side effects lead to poor patient compliance, which in turn has contributed to the drug resistance and exacerbated the TB epidemic. The relatively low output of newly approved antibiotics has spurred research interest toward alternative antibacterial molecules such as silver nanoparticles (AgNPs). In the present study, we use the natural biopolymer alginate to serve as a stabilizer and/or reductant to green synthesize AgNPs, which improves their biocompatibility and avoids the use of toxic chemicals. The average size of the alginate-capped AgNPs (ALG-AgNPs) was characterized as nanoscale, and the particles were round in shape. Drug susceptibility tests showed that these ALG-AgNPs are effective against both drug-resistant Mtb strains and dormant Mtb. A bacterial cell-wall permeability assay showed that the anti-mycobacterial action of ALG-AgNPs is mediated through an increase in cell-wall permeability. Notably, the anti-mycobacterial potential of ALG-AgNPs was effective in both zebrafish and mouse TB animal models in vivo. These results suggest that ALG-AgNPs could provide a new therapeutic option to overcome the difficulties of current TB treatments.

Microorganisms Photocatalytic Inactivation on Ag3PO4 Sub-microcrystals Under WLEDs Light Source

In this article, we report the silver orthophosphate (Ag3PO4) photocatalytic inactivation properties on strains Gram-positive Saprophyte B. subtilis, a diploid fungus Candida albicans and Gram-negative Pseudomonas aeruginosa, using as irradiation source white-light-emitting diodes (WLEDs) with luminous flux (Φ v) of 1.3 x 103 Lumens and relative power density of 15 mW m− 2. Microorganisms death curves and the kinetic constants (Kd) indicated that the inhibitory effect of the Ag3PO4 under WLEDs irradiation is well pronounced to C. albicans (6.6 x 10− 2 min− 1) in relation to P. augenosas (4.6 x 10− 2 min− 1) and B. subitilis (2.5 x 10− 2 min− 1). Decimal reduction time (Dt) were 34.4 min to C. albicans, 50.1 min P. augenosas and 92.1 min to B.subitilis The micrographs obtained by scanning electron microscopy with field emission gun (SEM-FEG) demonstrated that there was cell wall permeability and consequently total rupture in the C. albicans, suggesting the lipid peroxidation phenomenon and protein oxidation promoted by the presence of reactive oxygen species (ROS). Furthermore, it was observed the Ag3PO4 when irradiated by WLEDs demonstrates important sporicidal activity in related to B. subtilis, promoting the endospore wall rupture.

Cell Wall Permeability of Pinto Bean Cotyledon Cells Regulates In Vitro Fecal Fermentation and Gut Microbiota

Processing induced structural changes of whole foods on regulation of colonic fermentation rate and microbiota composition are least understood and often overlooked. In the present study, intact cotyledon cells from...

PERMEABILIZATION, CELL WALL ULTRASTRUCTURE, AND GERMINATION OF BASIDIOSPORES OF THE ECTOMYCORRHIZAL FUNGUS Pisolithus microcarpus TREATED WITH DIFFERENT COMMERCIAL BRANDS OF BLEACH

ABSTRACT Basidiospores of the ectomycorrhizal fungus Pisolithus microcarpus have an impermeable cell wall, a characteristic that is possibly related to the low germination percentages of these propagules, which makes it difficult to obtain monokaryons and use these spores in inoculants. The objective of this study was to evaluate the effect of different concentrations of commercial bleach on the permeabilization of P. microcarpus basidiospores and to analyze the alterations caused in the cell wall ultrastructure and the viability and germination capacity of these propagules. Fungal basidiospores were collected in eucalyptus plantations and permeabilized using different bleach concentrations and exposure times. The basidiospores were then analyzed by scanning and transmission electron microscopy. The percentage of permeabilized basidiospores varied with the commercial brand, bleach concentration, and exposure time. Basidiospores of different basidiocarps differed in susceptibility to permeabilization treatment with bleach. Changes in the ultrastructure of permeabilized basidiospores were observed at bleach concentrations of 15 and 50 % for an exposure time of 40 s, with surface changes and loss of the spicules of the outermost layer of the wall. After permeabilization with 5 % bleach for 40 s, 80 % of the permeabilized spores were viable, resulting in the production of fungal colonies after 15 days of incubation of these propagules in the presence of Corymbia citriodora. However, the germination percentage obtained, 0.001 %, was similar to that of non-permeabilized basidiospores, indicating that other factors, besides cell wall permeability, are determinant for the germination process.

Potensi Ekstrak Biji Pangi (Pangium edule Reinw) sebagai Pengawet Alami Pada Ikan Mujair (Oreochromis mossambicus)

Mujair fish (Oreochromis mossambicus) is a freshwater fish that is commonly consumed. This is due that they are easily found and obtrained and with a price in fresh markets that are relatively cheap. Unfortunately, this fish happens to be a very perishable commodity, due to it’s sensitiveness to spoilage because of its high protein content and moisture content. Pangi seeds (Pangium edule) is known to contain tanines, natural polyphenols that inhibits microorganisms through their cell wall permeability. The purpose of this research was to test the potential of pangi seeds crude extract as natural pangi seed toxiti preservatives on mujair fish, shelf life potential on mujair fish. This was achieved by marinating the fish in crude pangi seed extracts at concentrations of 0% (control), 5% and 10% and then store for 4 days at room temperature. Parameters tested on this research includes cyanide identification BSLT toxicity testing and fish spoilage (degradation) through protein changes and pH. Results of this research indicated that pangi seeds crude extract does not contain any cyanide of whatsoever, pangi seed crude extract toxicity testing indicated thatit was not toxic to a concentration of 128233mg/L, meanwhile protein analysis indicated that the optimum concentration of pangi seed crude extract to inhibit fish spoilage is 10% while in terms of pH degradation, the optimum concentration was 5%.

Mycobacterium Von Willebrand factor protein MSMEG_3641 is involved in biofilm formation and intracellular survival

Aim: Mycobacterium tuberculosis in vitro biofilm is associated with the virulence and persistence capability. Our aim is to delineate factors involved in biofilms development. Materials & methods: We performed transposon mutants screen and found that mutation of MSMEG_3641, a homolog of M. tuberculosis Rv1836c, can change M. smegmatis colony morphology and biofilm. Results: MSMEG_3641 contains a vWA domain that is highly conserved among Mycobacteria. The phenotypes of MSMEG_3641 mutants include disrupted biofilm, weakened migration ability and changed colony morphology. All phenotypes might be contributed to the enhanced cell wall permeability and declined cell aggregation ability. Conclusion: To our knowledge, this is the first report concerning the mycobacteria Von Willebrand factor domain function, especially in colony morphology and biofilm development.

Mycobacterium tuberculosis Rv0341 Promotes Mycobacterium Survival in In Vitro Hostile Environments and within Macrophages and Induces Cytokines Expression

Mycobacterium tuberculosis represents an ancient deadly human pathogen that can survive and multiply within macrophages. The effectors are key players for the successful pathogenesis of this bacterium. M. tuberculosis open reading frame (ORF) Rv0341, a pathogenic mycobacteria-specific gene, was found to be upregulated in macrophages isolated from human tuberculosis granuloma and inside the macrophages during in vitro infection by M. tuberculosis. To understand the exact role of this gene, we expressed the Rv0341 gene in M. smegmatis, which is a non-pathogenic Mycobacterium. We found that Rv0341 expression can alter colony morphology, reduce the sliding capability, and decrease the cell wall permeability of M. smegmatis. Furthermore, Rv0341 remarkably enhanced M. smegmatis survival within macrophages and under multiple in vitro stress conditions when compared with the control strain. Ms_Rv0341 significantly induced expression of TNF-α, IL-1β, and IL-10 compared with M. smegmatis harboring an empty vector. In summary, these data suggest that Rv0341 is one of the M. tuberculosis virulence determinants that can promote bacilli survival in harsh conditions and inside macrophages.

The Sterol Carrier Hydroxypropyl-β-Cyclodextrin Enhances the Metabolism of Phytosterols by Mycobacterium neoaurum

ABSTRACT Androst-4-ene-3,17-dione (AD) and androst-1,4-diene-3,17-dione (ADD) are valuable steroid pharmaceutical intermediates obtained by soybean phytosterol biotransformation by Mycobacterium. Cyclodextrins (CDs) are generally believed to be carriers for phytosterol delivery and can improve the production of AD and ADD due to their effects on steroid solubilization and alteration in cell wall permeability for steroids. To better understand the mechanisms of CD promotion, we performed proteomic quantification of the effects of hydroxypropyl-β-CD (HP-β-CD) on phytosterol metabolism in Mycobacterium neoaurum TCCC 11978 C2. Perturbations are observed in steroid catabolism and glucose metabolism by adding HP-β-CD in a phytosterol bioconversion system. AD and ADD, as metabolic products of phytosterol, are toxic to cells, with inhibited cell growth and biocatalytic activity. Treatment of mycobacteria with HP-β-CD relieves the inhibitory effect of AD(D) on the electron transfer chain and cell growth. These results demonstrate the positive relationship between HP-β-CD and phytosterol metabolism and give insight into the complex functions of CDs as mediators of the regulation of sterol metabolism. IMPORTANCE Phytosterols from soybean are low-cost by-products of soybean oil production and, owing to their good bioavailability in mycobacteria, are preferred as the substrates for steroid drug production via biotransformation by Mycobacterium. However, the low level of production of steroid hormone drugs due to the low aqueous solubility (below 0.1 mmol/liter) of phytosterols limits the commercial use of sterol-transformed strains. To improve the bioconversion of steroids, cyclodextrins (CDs) are generally used as an effective carrier for the delivery of hydrophobic steroids to the bacterium. CDs improve the biotransformation of steroids due to their effects on steroid solubilization and alterations in cell wall permeability for steroids. However, studies have rarely reported the effects of CDs on cell metabolic pathways related to sterols. In this study, the effects of hydroxypropyl-β-CD (HP-β-CD) on the expression of enzymes related to steroid catabolic pathways in Mycobacterium neoaurum were systematically investigated. These findings will improve our understanding of the complex functions of CDs in the regulation of sterol metabolism and guide the application of CDs to sterol production.