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1.
ACS Infect Dis ; 10(2): 746-762, 2024 02 09.
Article in English | MEDLINE | ID: mdl-38232080

ABSTRACT

Pseudomonas aeruginosa, a vivid biofilm-producing bacterium, is considered a dreadful opportunistic pathogen, and thus, management of biofilm-associated infections due to multidrug resistant strains by traditional drugs currently is of great concern. This study was aimed to assess the impact of trigonelline hydrochloride, a pyridine alkaloid, on P. aeruginosa PAO1, in search of an alternative therapeutant. The effect of trigonelline on colony morphology and motility was studied along with its role on biofilm and expression virulence factors. Trigonelline influenced the colony structure, motility, biofilm architecture, and the production of virulence factors in a dose-dependent manner. Alterations in quorum sending (QS)-regulated gene expression after treatment and molecular docking analysis for certain regulator proteins confirmed its effect on the QS-system network by affecting Las, Rhl, and Pqs signaling pathways and as possible molecular targets. Thus, trigonelline might be considered as a potential chemical lead to manage biofilm-associated pathogenesis or to develop other analogues with enhanced pharmacokinetic actions.


Subject(s)
Alkaloids , Anti-Infective Agents , Virulence , Pseudomonas aeruginosa , Molecular Docking Simulation , Quorum Sensing , Biofilms , Alkaloids/pharmacology , Virulence Factors/metabolism , Anti-Infective Agents/pharmacology
2.
Pharm Nanotechnol ; 10(4): 310-326, 2022 11 15.
Article in English | MEDLINE | ID: mdl-36017866

ABSTRACT

BACKGROUND: Copper nanoparticle (CuNP) has well-established antimicrobial activity. Instability in an aqueous medium due to aggregation into larger particles, conversion into metal ions, and oxidation into metal oxides are the major limitations of its practical use against bacterial infections. OBJECTIVE: Development of CuNP Conjugated Chitosan Microparticles as a reservoir that will release CuNP effective against notorious bacteria like Methicillin-resistant Staphylococcus aureus. METHODS: CuNP conjugated chitosan microparticles (CNCCM) were synthesized using a simple twostep process. In the first step, a solution of chitosan in 2% (w/v) ascorbic acid was added dropwise in copper sulphate solution to prepare Cu ion conjugated chitosan beads. In the second step, these beads were soaked in sodium hydroxide solution to get the CNCCM. The dried CNCCM were characterized thoroughly for surface conjugation of CuNP, and the release of CuNP in a suitable medium. The physicochemical properties of release CuNP were further verified with the in silico modelled CuNP. The Antimicrobial and antibiofilm activities of released CuNp were evaluated against methicillin-resistant Staphylococcus aureus (MRSA). RESULTS: 2% (w/v) ascorbic acid solution (pH 3.5) was the optimum medium for the release of ~2 nm CuNP from CNCCM. The CuNP had an optical band gap of ~ 2 eV. It inhibited the cell wall synthesis of MRSA. The minimum inhibitory concentration was 200 nM. At 100 nM dose, the CuNP caused ~73% reduction in biofilm development after 24 h of growth. The cytotoxic effect of CuNP on the human cell line (HEK 293) was significantly less than that on MRSA. The 48 h IC50 value against HEK 293 was 3.45-fold higher than the MIC value against MRSA after 24 h treatment. CONCLUSION: CuNP Conjugated Chitosan Microparticle has been developed. It works as a stable reservoir of ~2 nm CuNP. The CuNP is released in an aqueous medium containing 2% (w/v) ascorbic acid (pH 3.5). The released CuNP has a bacteriostatic effect against MRSA at a concentration safe for human cells.


Subject(s)
Anti-Infective Agents , Chitosan , Methicillin-Resistant Staphylococcus aureus , Nanoparticles , Humans , Chitosan/chemistry , Methicillin/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Copper , HEK293 Cells , Nanoparticles/chemistry , Water/pharmacology
3.
Environ Sci Pollut Res Int ; 28(40): 57440-57448, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34482463

ABSTRACT

Biogeochemical release of soil-bound arsenic (As) governs mobilization of the toxic metalloid into the groundwater. The present study has examined AsV-reduction ability of bacteria from anoxic aquatic sediments that might contribute to arsenic mobilization in the Bengal Delta. Arsenic-reducing bacteria from deep layers of pond sediment were enriched and isolated in anaerobic environments and AsV reduction was assessed in culture medium. The pond sediment enrichments harboured AsV-reducing bacteria belonging to the phyla Firmicutes and Proteobacteria with dominance of Paraclostridium benzoelyticum and P. bifermentans. Among total 17 isolates, the respiratory reductase genes were not detected by the most common primers and only 3 strains had arsenic reductase ArsC gene suggesting involvement of resistance and some unknown mechanisms in AsV reduction. Presence of high levels of organic matter, As, and As-reducing bacteria might make deep aquatic sediments a hot spot of As mobilization and aquifer contamination.


Subject(s)
Arsenic , Bacteria/genetics , Clostridiales , Geologic Sediments
4.
Int J Med Microbiol ; 310(3): 151410, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32057619

ABSTRACT

The study aims to explore the combinatorial effect of naringin with antibiotics, ciprofloxacin and tetracycline on Pseudomonas aeruginosa biofilms. The antibiofilm efficacy of selected treatment regimes against P. aeruginosa biofilm were quantified by crystal violet assay, MTT assay, Congo red binding assay, and were visualized by confocal laser scanning microscopy and scanning electron microscopy. All the assays reflected antibiofilm activities, however, combinatorial performances of naringin with antibiotics were found to be more significant. A significant reduction in swimming and swarming motilities along with pellicle formation and altered colony morphology were observed as a result of combinatorial effect. The cytotoxicity of naringin and its antibiotic combinations was assayed on murine macrophage cell line. The applicability of such combinations was tested for their relative eradication against pre-formed biofilm on urinary catheter surface. This finding indicated that naringin potentiates the efficacy of both ciprofloxacin and tetracycline on P. aeruginosa biofilm in comparison to their solo treatment. The finding would help to open hitherto unexplored possibilities of establishing naringin as a potential antibiofilm agent and suggest on the possibility of its use in drug-herb combinations for managing biofilm-associated bacterial infections.


Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Ciprofloxacin/pharmacology , Flavanones/pharmacology , Pseudomonas aeruginosa/drug effects , Tetracycline/pharmacology , Animals , Cell Line , Macrophages/drug effects , Mice , Microbial Sensitivity Tests , Pseudomonas Infections/microbiology
5.
Microb Pathog ; 138: 103790, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31605761

ABSTRACT

This study investigated the effects of reserpine, the main bioactive compound of Rauwolfia serpentina, on biofilm formation and biofilm-associated virulence factors production in a Gram-positive pathogen, Staphylococcus aureus. Crystal violet assay, MTT assay, Congo red binding, CLSM studies were performed to assess the antibiofilm activity. Molecular docking was performed to explain the possible mode of action, catheter model was used to evaluate its application potential and the combinatorial study was performed in search of an improved therapeutic formulation. Reserpine affected biofilm formation, EPS production, biofilm cell viability and virulence factor production. It could eradicate 72.7% biofilm at ½â€¯× MIC dose and could also stop the metabolic activity of 50.6% bacterial cells in a biofilm. Staphylococcus aureus biofilm- and virulence-regulatory proteins like AgrA, AtlE, Bap, IcaA, SarA and SasG were found to interact with reserpine which might lead to the attenuation of its pathogenicity. Reserpine along with other commercial antibiotics could generate a hightened antibiofilm response, and also eradicated a good percentage of bacterial biofilm from a urinary catheter model. These findings suggested reserpine as a good alternative entity to generate new improved therapeutic formulations.


Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Reserpine/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Virulence/drug effects , Anti-Bacterial Agents/chemistry , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Chemical Phenomena , Dose-Response Relationship, Drug , Drug Synergism , Hemolysis , Microbial Sensitivity Tests , Microbial Viability/drug effects , Models, Molecular , Protein Binding , Reserpine/chemistry , Staphylococcal Infections/drug therapy , Structure-Activity Relationship
6.
Biofouling ; 34(3): 320-334, 2018 03.
Article in English | MEDLINE | ID: mdl-29482361

ABSTRACT

This study aimed to evaluate the effect of reserpine, a plant-derived indole-alkaloid, against Pseudomonas aeruginosa PAO1 biofilms. The anti-biofilm activity of reserpine was evaluated by crystal violet staining, MTT assay, confocal laser scanning microscopy and scanning electron microscopy. Reserpine effects were also assessed by qRT-PCR of quorum sensing (QS)-regulated genes and biochemical quantification of the QS-mediated virulence factors pyocyanin, rhamnolipids, proteases and elastases. Reserpine reduced biofilm formation, cell motility, virulence factor production, and QS-controlled gene expression. Additionally, molecular docking analysis for AHL synthase LasI and QS transcriptional regulators LasR/MvfR revealed a plausible molecular mechanisms of reserpine QS inhibition. These findings provide insights into the underlying mode of action of reserpine, which may be useful in the development of new drugs against biofilm-related infections.


Subject(s)
Bacterial Proteins/drug effects , Pseudomonas aeruginosa/drug effects , Quorum Sensing/drug effects , Reserpine/pharmacology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Glycolipids , Ligases/drug effects , Molecular Docking Simulation , Pseudomonas aeruginosa/metabolism , Pseudomonas aeruginosa/physiology , Virulence Factors
7.
Microb Pathog ; 113: 85-93, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29042302

ABSTRACT

Quorum-sensing (QS) is known to play an essential role in regulation of virulence factors and toxins during Pseudomonas aeruginosa infection which may frequently cause antibiotic resistance and hostile outcomes of inflammatory injury. Therefore, it is an urgent need to search for a novel agent with low risk of resistance development that can target QS and inflammatory damage prevention as well. Andrographis paniculata, a herbaceous plant under the family Acanthaceae, native to Asian countries and also cultivated in Scandinavia and some parts of Europe, has a strong traditional usage with its known antibacterial, anti-inflammatory, antipyretic, antiviral and antioxidant properties. In this study, three different solvent extracts (viz., chloroform, methanol and aqueous) of A. paniculata were examined for their anti-QS and anti-inflammatory activities. Study was carried out to assess the effect on some selected QS-regulatory genes at transcriptional level using Real Time-PCR. In addition, ability to attenuate MAPK pathways upon P. aeruginosa infection was performed to check its potential anti-inflammatory activity. Chloroform and methanol extracts showed significant reduction (p < 0.05) of the QS-controlled extracellular virulence factors in P. aeruginosa including the expression of pyocyanin, elastase, total protease, rhamnolipid and hemolysin without affecting bacterial viability. They also significantly (p < 0.05) reduced swarming motility and biofilm formation of P. aeruginosa. The chloroform extract, which was found to be more effective, decreased expression of lasI, lasR, rhlI and rhlR by 61%, 75%, 41%, and 44%, respectively. Moreover, chloroform extract decreased activation of p-p38 and p-ERK1/2 expression levels in MAPK signal pathways in P. aeruginosa infected macrophage cells. As the present study demonstrates that A. paniculata extracts inhibit QS in P. aeruginosa and exhibit anti-inflammatory activities, therefore it represents itself as a prospective therapeutic agent against P. aeruginosa infection.


Subject(s)
Andrographis/metabolism , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Plant Extracts/pharmacology , Pseudomonas aeruginosa/pathogenicity , Quorum Sensing/drug effects , Virulence Factors/biosynthesis , Animals , Cells, Cultured , Macrophages/immunology , Mice , Microbial Sensitivity Tests , Movement/drug effects , Prospective Studies , Pseudomonas aeruginosa/drug effects
8.
Acta Trop ; 176: 58-67, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28739368

ABSTRACT

African sleeping sickness is a parasitic disease in humans and livestock caused by Trypanosoma brucei throughout sub-Saharan Africa. Absence of appropriate vaccines and prevalence of drug resistance proclaim that a new way of therapeutic interventions is essential against African trypanosomiasis. In the present study, we have looked into the effect of andrographolide (andro), a diterpenoid lactone from Andrographis paiculata on Trypanosoma brucei PRA 380. Although andro has been recognized as a promosing anti-cancer drug, its usefulness against Trypanosoma spp remained unexplored. Andro showed promising anti-trypanosomal activity with an IC50 value of 8.3µM assessed through SYBR Green cell viability assay and also showed no cytotoxicity towards normal murine macrophages. Cell cycle analysis revealed that andro could induce sub-G0/G1 phase arrest. Flow cytometric analysis also revealed that incubation with andro caused exposure of phosphatidyl serine to the outer leaflet of plasma membrane in T. brucei PCF. This event was preceded by andro-induced depolarization of mitochondrial membrane potential (Δym) and elevation of cytosolic calcium. Andro also caused elevation of intracellular reactive oxygen species (ROS) as well as lipid peroxidation level, and depletion in reduced thiol levels. Taken together, these data indicate that andro has promising antitrypanosomal activity mediated by promoting oxidative stress and depolarizing the mitochondrial membrane potential and thereby triggering an apoptosis-like programmed cell death. Therefore, this study merits further investigation to the therapeutic possibility of using andro for the treatment of African trypanosomiasis.


Subject(s)
Antiprotozoal Agents/pharmacology , Diterpenes/pharmacology , Trypanosoma brucei brucei/drug effects , Animals , Cell Death/drug effects , Humans , Lipid Peroxidation/drug effects , Oxidative Stress , Reactive Oxygen Species/metabolism
9.
Folia Microbiol (Praha) ; 62(3): 237-244, 2017 May.
Article in English | MEDLINE | ID: mdl-28097636

ABSTRACT

Increasing bacterial resistance to common drugs is a major public health concern for the treatment of infectious diseases. Certain naturally occurring compounds of plant sources have long been reported to possess potential antimicrobial activity. This study was aimed to investigate the antibacterial activity and possible mechanism of action of andrographolide (Andro), a diterpenoid lactone from a traditional medicinal herb Andrographis paniculata. Extent of antibacterial action was assessed by minimal bactericidal concentration method. Radiolabeled N-acetyl glucosamine, leucine, thymidine, and uridine were used to determine the effect of Andro on the biosyntheses of cell wall, protein, DNA, and RNA, respectively. In addition, anti-biofilm potential of this compound was also tested. Andro showed potential antibacterial activity against most of the tested Gram-positive bacteria. Among those, Staphylococcus aureus was found to be most sensitive with a minimal inhibitory concentration value of 100 µg/mL. It was found to be bacteriostatic. Specific inhibition of intracellular DNA biosynthesis was observed in a dose-dependent manner in S. aureus. Andro mediated inhibition of biofilm formation by S. aureus was also found. Considering its antimicrobial potency, Andro might be accounted as a promising lead for new antibacterial drug development.


Subject(s)
Anti-Bacterial Agents/pharmacology , Diterpenes/pharmacology , Gram-Positive Bacteria/drug effects , Microbial Viability/drug effects , Plant Extracts/pharmacology , Andrographis/chemistry , Biofilms/drug effects , Biosynthetic Pathways/drug effects , DNA/biosynthesis , Diterpenes/isolation & purification , Gram-Positive Bacteria/physiology , Humans , Microbial Sensitivity Tests , Plant Extracts/isolation & purification
10.
Can J Microbiol ; 63(2): 169-178, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28099040

ABSTRACT

The goal of this study was to evaluate the antibiofilm and antimicrobial activities of Bacoside A, a formulation of phytochemicals from Bacopa monnieri, against Staphylococcus aureus and Pseudomonas aeruginosa, which are known to form biofilms as one of their virulence traits. The antimicrobial effects of Bacoside A were tested using the minimum inhibitory concentration and minimum bactericidal concentration assays. A cell membrane disruption assay was performed to find its possible target site. MTT assay, crystal violet assay, and microscopic studies were performed to assess the antibiofilm activity. Bacoside A showed antimicrobial activity against both test organisms in their planktonic and biofilm states. At a subminimum inhibitory concentration of 200 µg·mL-1, Bacoside A significantly removed ∼88%-93% of bacterial biofilm developed on microtiter plates. Biochemical and microscopic studies suggested that the eradication of biofilm might be due to the loss of extracellular polymeric substances and to a change in cell membrane integrity of the selected bacterial strains treated with Bacoside A. These results indicate that Bacoside A might be considered as an antimicrobial having the ability to disrupt biofilms. Thus, either alone or in combination with other therapeutics, Bacoside A could be useful to treat biofilm-related infections caused by opportunistic bacterial pathogens.


Subject(s)
Biofilms/drug effects , Pseudomonas aeruginosa/drug effects , Saponins/pharmacology , Staphylococcus aureus/drug effects , Triterpenes/pharmacology , Anti-Infective Agents/pharmacology , Biofilms/growth & development , Microbial Sensitivity Tests
11.
Pol J Microbiol ; 65(1): 115-8, 2016.
Article in English | MEDLINE | ID: mdl-27282003

ABSTRACT

Nickel resistant bacterial strain Enterobacter asburiae KUNi5 was isolated and showed resistance up to 15 mM and could remove Ni optimally better at 37 degrees C and pH 7. Maximum removal was found at initial concentration of 0.5 to 2 mM, however, growth and Ni removal were affected by other heavy metals. Major amount of the metal was accumulated in the membrane fractions and certain negatively charged groups were found responsible for Ni binding. KUNi5 could also produce 1-aminocyclopropane-1-carboxylate deaminase, indole-acetic acid and siderophore. It seems that KUNi5 could be a possible candidate for Ni detoxification and plant growth promotion in Ni-contaminated field.


Subject(s)
Biodegradation, Environmental , Enterobacter/classification , Enterobacter/metabolism , Environmental Pollutants/metabolism , Nickel/metabolism , Dose-Response Relationship, Drug , Enterobacter/drug effects , Environmental Pollutants/chemistry , Nickel/chemistry
12.
J Biomed Sci ; 23: 40, 2016 Apr 16.
Article in English | MEDLINE | ID: mdl-27084510

ABSTRACT

BACKGROUND: Breast cancer is considered as an increasing major life-threatening concern among the malignancies encountered globally in females. Traditional therapy is far from satisfactory due to drug resistance and various side effects, thus a search for complementary/alternative medicines from natural sources with lesser side effects is being emphasized. Andrographis paniculata, an oriental, traditional medicinal herb commonly available in Asian countries, has a long history of treating a variety of diseases, such as respiratory infection, fever, bacterial dysentery, diarrhea, inflammation etc. Extracts of this plant showed a wide spectrum of therapeutic effects, such as anti-bacterial, anti-malarial, anti-viral and anti-carcinogenic properties. Andrographolide, a diterpenoid lactone, is the major active component of this plant. This study reports on andrographolide induced apoptosis and its possible mechanism in highly proliferative, invasive breast cancer cells, MDA-MB-231 lacking a functional p53 and estrogen receptor (ER). Furthermore, the pharmacokinetic properties of andrographolide have also been studied in mice following intravenous and oral administration. RESULTS: Andrographolide showed a time- and concentration- dependent inhibitory effect on MDA-MB-231 breast cancer cell proliferation, but the treatment did not affect normal breast epithelial cells, MCF-10A (>80 %). The number of cells in S as well as G2/M phase was increased after 36 h of treatment. Elevated reactive oxygen species (ROS) production with concomitant decrease in Mitochondrial Membrane Potential (MMP) and externalization of phosphatidyl serine were observed. Flow cytometry with Annexin V revealed that the population of apoptotic cells increased with prolonged exposure to andrographolide. Activation of caspase-3 and caspase-9 were also noted. Bax and Apaf-1 expression were notably increased with decreased Bcl-2 and Bcl-xL expression in andrographolide-treated cells. Pharmacokinetic study with andrographolide showed the bioavailability of 9.27 ± 1.69 % with a Cmax, of 0.73 ± 0.17 µmol/L and Tmax of 0.42 ± 0.14 h following oral administration. AG showed rapid clearance and moderate terminal half lives (T1/2) of 1.86 ± 0.21 and 3.30 ± 0.35 h following IV and oral administration respectively. CONCLUSION: This investigation indicates that andrographolide might be useful as a possible chemopreventive/chemotherapeutic agent for human breast cancers.


Subject(s)
Apoptosis/drug effects , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Cell Cycle Checkpoints/drug effects , Cytotoxins/pharmacology , Diterpenes/pharmacology , Animals , Apoptosis/genetics , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Cell Cycle Checkpoints/genetics , Female , Humans , MCF-7 Cells , Mice , Mice, Inbred BALB C
13.
Ecotoxicol Environ Saf ; 107: 236-44, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25011120

ABSTRACT

Arsenic (As) contamination of soil and water has been considered as a major global environmental issue during last few decades. Among the various methods so far reported for reclamation of As contaminated rhizosphere soil, bioremediation using bacteria has been found to be most promising. An As resistant bacterial isolate Brevibacillus sp. KUMAs2 was obtained from As contaminated soil of Nadia, West Bengal, India, which could resist As(V) and As(III) a maximum of 265mM and 17mM, respectively. The strain could remove ~40 percent As under aerobic culture conditions. As resistant property in KUMAs2 was found to be plasmid-borne, which carried both As oxidizing and reducing genes. The strain could promote chilli plant growth under As contaminated soil environment by decreasing As accumulation in plant upon successful colonization in the rhizosphere, which suggests the possibility of using this isolate for successful bioremediation of As in the crop field.


Subject(s)
Arsenic/metabolism , Brevibacillus/isolation & purification , Brevibacillus/physiology , Soil Pollutants/metabolism , Arsenic/isolation & purification , Biodegradation, Environmental , Drug Resistance, Bacterial/genetics , Genes, Bacterial , India , Microscopy, Electron, Transmission , Oxidation-Reduction , Plasmids , Rhizosphere , Soil , Soil Microbiology , Soil Pollutants/isolation & purification , Spectrometry, X-Ray Emission
14.
J Hazard Mater ; 260: 1073-82, 2013 Sep 15.
Article in English | MEDLINE | ID: mdl-23892173

ABSTRACT

The present study endeavours to assess the toxic effect of synthesized CdS nanoparticles (NPs) on Escherichia coli and HeLa cells. The CdS NPs were characterized by DLS, XRD, TEM and AFM studies and the average size of NPs was revealed as ∼3 nm. On CdS NPs exposure bacterial cells changed morphological features to filamentous form and damage of the cell surface was found by AFM study. The expression of two conserved cell division components namely ftsZ and ftsQ in E. coli was decreased both at transcriptional and translational levels upon CdS NPs exposure. CdS NPs inhibited proper cell septum formation without affecting the nucleoid segregation. Viability of HeLa cells declined with increasing concentration of CdS NPs and the IC50 value was found to be 4 µg/mL. NPs treated HeLa cells showed changed morphology with condensed and fragmented nuclei. Increased level of reactive oxygen species (ROS) was found both in E. coli and HeLa cells on CdS NPs exposure. The inverse correlation between declined cell viabilities and elevated ROS level suggested that oxidative stress seems to be the key event by which NPs induce toxicity both in E. coli and HeLa cells.


Subject(s)
Cadmium Compounds/toxicity , Escherichia coli/drug effects , Nanoparticles/toxicity , Sulfides/toxicity , Bacterial Proteins/metabolism , Cadmium Compounds/chemistry , Cell Division/drug effects , Cell Line, Tumor , Cell Membrane/drug effects , Cytoskeletal Proteins/metabolism , Dose-Response Relationship, Drug , Escherichia coli Proteins/metabolism , Gene Expression Profiling , Gene Expression Regulation, Bacterial/drug effects , Gene Expression Regulation, Neoplastic/drug effects , HeLa Cells , Humans , Membrane Proteins/metabolism , Microscopy, Atomic Force , Microscopy, Electron, Transmission , Nanoparticles/chemistry , Oxidative Stress , Reactive Oxygen Species , Spectrometry, Fluorescence , Sulfides/chemistry , Time Factors , X-Ray Diffraction
15.
Langmuir ; 28(48): 16614-22, 2012 Dec 04.
Article in English | MEDLINE | ID: mdl-23137198

ABSTRACT

This Article deals with the toxicological study of synthesized CdO nanoparticles (NPs) on Escherichia coli . Characterization of the CdO NPs was done by DLS, XRD, TEM, and AFM studies, and the average size of NPs was revealed as 22 ± 3 nm. The NPs showed bactericidal activity against E. coli. When NPs were added at midlog phase of growth, complete growth inhibitory concentration was found as 40 µg/mL. Bacterial cells changed morphological features to filamentous form with increasing CdO NPs exposure time, and thereafter resulted in filamentation-associated clumping. From AFM study, severe damage of the cell surface was found in CdO NPs-treated cells. CdO NPs were found to interfere with the expression level of two conserved cell division components, ftsZ and ftsQ, in E. coli at both transcriptional and translational levels. Interference of CdO NPs in proper septum formation without affecting the nucleoid segregation was also observed in confocal micrographs. The elevated intracellular oxidative stress due to CdO NPs exposure seems to be one of the reasons for the changes in cell morphology and expression of division proteins in E. coli.


Subject(s)
Cadmium Compounds/chemistry , Cadmium Compounds/toxicity , Cell Division/drug effects , Escherichia coli/cytology , Escherichia coli/growth & development , Nanoparticles/toxicity , Oxides/chemistry , Oxides/toxicity , Toxicity Tests , Cell Survival/drug effects , Escherichia coli/drug effects , Escherichia coli Proteins/metabolism , Intracellular Space/drug effects , Intracellular Space/metabolism , Reactive Oxygen Species/metabolism
16.
Ecotoxicol Environ Saf ; 86: 54-9, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23063080

ABSTRACT

This article deals with toxicological study of cadmium (Cd) as CdCl(2) on the growth and cell morphology of Escherichia coli K-12 MG1655. The minimum inhibitory concentration of Cd was 15µM. When cadmium was added at mid-log phase, growth was completely inhibited at 0.6mM and 50% of the bacterial growth retardation was found at 0.3mM concentration. At sublethal dose of Cd (0.2mM), majority of the cells showed filamentous form, suggested the possible effect of Cd on cell division. AFM study of bacterial cell morphology revealed severe surface damage of the treated cells in comparison to untreated cells. The expression of FtsZ decreased both at transcriptional and translational levels with the time of Cd exposure, thus cell division was affected and as a result cells took filamentous form. Due to Cd exposure, the nucleoid segregation remained unaffected, but improper Z-ring formation was observed. Activities of peroxidase and superoxide dismutase significantly decreased in treated cells with exposure time, which might elevate intracellular ROS level, as a consequence metabolic dysfunction and toxic effect were resulted.


Subject(s)
Cadmium/toxicity , Environmental Pollutants/toxicity , Escherichia coli K12/drug effects , Cell Division/drug effects , Enzyme Activation/drug effects , Microbial Sensitivity Tests , Oxidation-Reduction , Superoxide Dismutase/metabolism
17.
Pol J Microbiol ; 59(3): 185-90, 2010.
Article in English | MEDLINE | ID: mdl-21033582

ABSTRACT

Microbial enzymatic reduction of a toxic form of chromium [Cr(VI)] has been considered as an effective method for bioremediation of this metal. This study reports on the in vitro reduction of Cr(VI) using cell-free extracts from a Cr(VI) reducing Bacillus firmus KUCr1 strain. Chromium reductase was found to be constitutive and its activity was observed both in soluble cell fractions (S12 and S150 and membrane cell fraction (P150). The reductase activity of S12 fraction was found to be optimal at 40 microM Cr(VI) with enzyme concentration equivalent to 0.493 mg protein/ml. Enzyme activity was dependent on NADH or NADPH as electron donor; optimal temperature and pH for better enzyme activity were 70 degrees C and 5.6, respectively. The Km value of the reductase was 58.33 microM chromate having a V(max) of 11.42 microM/min/mg protein. The metabolic inhibitor like sodium azide inhibited reductase activity of membrane fraction of the cell-free extract. Metal ions like Cu2+, Co2+, Ni2+ and As3+ stimulated the enzyme but others, such as Ag+, Hg2+, Zn2+, Mn2+, Cd2+ and Pb2+, inhibited Cr(VI) reductase activity.


Subject(s)
Bacillus/metabolism , Bacterial Proteins/metabolism , Cell Extracts/chemistry , Chromium/metabolism , Oxidoreductases/metabolism , Bacillus/chemistry , Cell Fractionation , Chromium/chemistry , Hydrogen-Ion Concentration , NAD , NADP , Oxidoreductases/chemistry , Temperature
18.
Braz. j. microbiol ; 40(3): 655-662, Sept. 2009. tab
Article in English | LILACS | ID: lil-522496

ABSTRACT

A cadmium (8 mM) resistant Pseudomonas aeruginosa strain KUCd1 exhibiting high Cd accumulation under in vitro aerobic condition has been reported. The isolate showed a significant ability to remove more than 75 percent and 89 percent of the soluble cadmium during the active growth phase from the growth medium and from Cd-amended industrial wastewater under growth supportive condition. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDXS) suggest the presence of Cd in the cells from mid-stationary phase. The cell fractionation study revealed membrane and periplasm to be the major accumulating site in this strain. The chemical nature of the accumulated Cd was studied by X-ray powder diffraction analysis.


Descreve-se a cepa Pseudomonas aeruginosa KUCd1 resistente a cádmio (8mM) que apresenta elevado acumulo de cádmio em condições de aerobiose in vitro. Durante a fase ativa de multiplicação, a cepa apresentou capacidade de remover mais de 75 por cento e 89 por cento de cádmio solúvel do meio de cultura e da água residual industrial contendo Cd. A microscopia eletrônica de transmissão (TEM) e espectroscopia de raio X energia dispersiva indicaram a presença de Cd nas células na fase estacionária intermediária. O fracionamento celular indicou serem a membrana e o periplasma os pontos de maior acúmulo. A natureza química do Cd acumulado foi avaliada através de análise de difração de Raios X.

19.
Braz J Microbiol ; 40(3): 655-62, 2009 Jul.
Article in English | MEDLINE | ID: mdl-24031411

ABSTRACT

A cadmium (8 mM) resistant Pseudomonas aeruginosa strain KUCd1 exhibiting high Cd accumulation under in vitro aerobic condition has been reported. The isolate showed a significant ability to remove more than 75% and 89% of the soluble cadmium during the active growth phase from the growth medium and from Cd-amended industrial wastewater under growth supportive condition. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDXS) suggest the presence of Cd in the cells from mid stationary phase. The cell fractionation study revealed membrane and periplasm to be the major accumulating site in this strain. The chemical nature of the accumulated Cd was studied by X-ray powder diffraction analysis.

20.
Pol J Microbiol ; 57(4): 327-32, 2008.
Article in English | MEDLINE | ID: mdl-19275047

ABSTRACT

A chromium resistant bacterial strain KUCr1 exhibiting potential Cr(VI) reducing ability under in vitro aerobic condition is reported. The bacterial strain showed varied degree of resistance to different heavy metals. The MIC of chromium to this strain was found to be 950 mM under aerobic culture condition in complex medium. The factors affecting Cr(VI) reduction by this strain under culture condition were evaluated. Maximal Cr(VI) reduction was observed at the pH 8 to 10 and at a temperature of 35 degrees C. Higher concentration of Cr(VI) slowed down the reduction, eventually all the metal could be reduced with longer incubation time. Different toxic metals showed differential effect on reduction. Cadmium and zinc were found to inhibit reduction. Cr(VI) reduction and bioremediation were found to be related to the growth supportive condition in terms of carbon, phosphorous and nitrogen supply in wastewater fed with tannery effluent indicating cell mass dependency of Cr(VI) reduction. Through biochemical characterization and 16S rDNA sequence analysis, the strain KUCr1, as the name given to it, was identified as a strain of Bacillus firmus.


Subject(s)
Bacillus/growth & development , Bacillus/isolation & purification , Bacillus/metabolism , Chromium/metabolism , DNA, Bacterial , RNA, Ribosomal, 16S/genetics , Arsenic/metabolism , Biodegradation, Environmental , Cadmium/metabolism , Cobalt/metabolism , Culture Media , Hydrogen-Ion Concentration , Industrial Waste/analysis , Nickel/metabolism , Oxidation-Reduction , Sequence Analysis, DNA , Soil Microbiology , Temperature , Zinc/metabolism
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