scholarly journals Chitosan Stabilized Silver Nanoparticles for the Electrochemical Detection of Lipopolysaccharide: A Facile Biosensing Approach for Gram-Negative Bacteria

Micromachines ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 413 ◽  
Author(s):  
Muhammad Imran ◽  
Christopher J. Ehrhardt ◽  
Massimo F. Bertino ◽  
Muhammad R. Shah ◽  
Vamsi K. Yadavalli

Negatively charged lipopolysaccharide (LPS), a major endotoxin and component of the outer membrane of several Gram-negative bacteria, provides a useful biomarker for the indirect detection of these pathogens. For instance, Escherichia coli (E. coli) is a pathogenic bacterium that causes infections in almost all age groups, and has been implicated in food and water contamination. Current diagnostic and detection methods tend to be labor-intensive or expensive, necessitating the need for an easy, sensitive, rapid, and low-cost method. We report on the synthesis and use of positively charged chitosan stabilized silver nanoparticles (Chi-AgNPs) as a sensitive electrochemical nanobiosensor for the detection of LPS. Chi-AgNPs were synthesized through a facile, single step protocol, and characterized for size, charge, and morphology. Glassy carbon electrodes modified with Chi-AgNPs resulted in an enhancement of signal in the presence of both LPS and E. coli. Detection was accomplished over a large concentration range (several orders of magnitude) of 0.001–100 ng/mL and 10–107 CFU/mL. The biosensors can reliably detect LPS and E. coli at very low concentrations. Chi-AgNPs have potential as low cost, sensitive nanobiosensors for Gram-negative bacteria due to strong electrostatic interaction with LPS present in their outer membranes.

2018 ◽  
Vol 10 (5) ◽  
pp. 209
Author(s):  
Shyla M. Haqq ◽  
Himanshu Pandey ◽  
Manju Gerard ◽  
Amit Chattree

Objective: In the present research work silver nanoparticles were synthesized using the flower extract of Chrysanthemum coronarium and their in-vitro antibacterial activity was evaluated against both the gram-positive S. aureus and gram-negative bacteria E. coli. The flower extract acted both as a reducing as well as a capping agent.Methods: Silver nanoparticles were verified using various spectroanalytical techniques such as visible ultraviolet spectroscopy, zeta potential, fourier transform infrared spectroscopy and particle size analyser. The antibacterial activity was evaluated against both the gram-positive bacteria S. aureus and gram-negative bacteria E. coli using the agar well diffusion method.Results: The silver nanoparticles synthesized were confirmed by the visual colour change. The ultraviolet, visible spectroscopy showed a surface plasmon resonance at 430 nm. Zetapotential was found to be around-15.6mV where the negative value indicated that the synthesized silver nanoparticles are stable. Fourier transform infrared spectroscopy showed the functional groups responsible for the stabilization of the nanoparticles. Particle size analyser showed that the size of the nanoparticles ranged from 5-50 nm. The antibacterial activity of the silver nanoparticles which was performed against S. aureus and E. coli showed good inhibition against both the bacteria. Better antibacterial activity was found for E. coli in comparison to S. aureus as the zone of inhibition for E. coli was found to be at 12 mm at 50ug/ml whereas the zone of inhibition against S. aureus was found to be at 10 mm.Conclusion: The silver nanoparticles were successfully synthesized using a green approach and can be used as a potential resource for therapeutic purpose.


2021 ◽  
pp. 1330-1341
Author(s):  
Mbarga M. J. Arsène ◽  
I. V. Podoprigora ◽  
Anyutoulou K. L. Davares ◽  
Marouf Razan ◽  
M. S. Das ◽  
...  

Background and Aim: The gradual loss of efficacy of conventional antibiotics is a global issue. Plant material extracts and green-synthesized nanoparticles are among the most promising options to address this problem. Therefore, the aim of this study was to assess the antibacterial properties of aqueous and hydroalcoholic extracts of grapefruit peels as well as their inclusion in green-synthesized silver nanoparticles (AgNPs). Materials and Methods: Aqueous and hydroalcoholic extracts (80% v/v) were prepared, and the volume and mass yields were determined. The synthesis of AgNPs was done in an eco-friendly manner using AgNO3 as a precursor. The nanoparticles were characterized by ultraviolet–vis spectrometry and photon cross-correlation spectroscopy. The antibacterial activity of the extracts was tested on three Gram-positive bacteria (Staphylococcus aureus ATCC 6538, clinical Enterococcus faecalis, and S. aureus) and two Gram-negative bacteria (two clinical Escherichia coli) using various concentrations of extracts (100, 50, 25, 12, and 5 mg/mL and 5% dimethyl sulfoxide as negative control). Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the microdilution method. Modulation of cefazoline and ampicillin on resistant E. coli and S. aureus strains was added to the mixture design response surface methodology with extreme vertices design, with the diameters of inhibition and the fractional inhibitory concentration index as responses and factors, respectively. The antibiotic, the ethanolic extract, and water varied from 0.1 MIC to 0.9 MIC for the first two and from 0 to 0.8 in proportion for the third. Validating the models was done by calculating the absolute average deviation, bias factor, and accuracy factor. Results: The volume yield of the EE and aqueous extract (AE) was 96.2% and 93.8% (v/v), respectively, whereas their mass yields were 7.84% and 9.41% (m/m), respectively. The synthesized AgNPs were very uniform and homogeneous, and their size was dependent on the concentration of AgNO3. The antibacterial activity of the two extracts was dose-dependent, and the largest inhibition diameter was observed for the Gram-positive bacteria (S. aureus ATCC 6538; AE, 12; EE, 16), whereas AgNPs had a greater effect on Gram-negative bacteria. The MICs (mg/mL) of the AEs varied from 3.125 (S. aureus ATCC 6538) to 12.5 (E. coli 1 and E. coli 2), whereas the MICs of the EEs varied from 1.5625 (S. aureus 1, S. aureus ATCC 6538, and E. faecalis) to 6.25 (E. coli 1). There was a significant difference between the MICs of AEs and EEs (p=0.014). The MBCs (mg/mL) of the AEs varied from 12.5 (S. aureus ATCC 6538) to 50 (S. aureus 1), whereas those of the EEs varied from 6.25 (S. aureus 1) to 25 (E. coli 1 and E. faecalis). Ethanolic grapefruit extracts demonstrated an ability to modulate cefazolin on E. coli and S. aureus but were completely indifferent to ampicillin on E. coli. Conclusion: Grapefruit peel extracts and their AgNPs exhibit antibacterial properties that can be exploited for the synthesis of new antimicrobials and their EEs may be efficiently used synergistically with other antibiotics against bacteria with intermediate susceptibility.


2019 ◽  
Vol 10 ◽  
pp. 1864-1872
Author(s):  
Prof. Teodora P. Popova

The effect of ionized aqueous solutions (anolytes and catholyte) in the processing of fruits (cherries, morellos, and strawberries) for decontamination has been tested. Freshly prepared analytes and catholyte without the addition of salts were used, as well as stored for 7 months anolytes, prepared with 0.5% NaCl and a combination of 0.5% NaCl and 0.5% Na2CO3. The anolyte prepared with a combination of 0.5% NaCl and 0.5% Na2CO3, as well as the anolyte obtained with 0.5% NaCl, exhibit high antimicrobial activity against the surface microflora of strawberries, cherries, and sour cherries. They inactivate E. coli for 15 minutes. The other species of the fam. Enterobacteriaceae were also affected to the maximum extent, as is the total number of microorganisms, especially in cherries and sour cherries. Even stored for 7 months, they largely retain their antimicrobial properties. Anolyte and catholyte, obtained without the addition of salts, showed a lower effect on the total number of microorganisms, but had a significant effect on Gram-negative bacteria, and especially with regard to the sanitary indicative E. coli.


2020 ◽  
Vol 16 (4) ◽  
pp. 481-488
Author(s):  
Heli Sanghvi ◽  
Satyendra Mishra

Background: Curcumin, one of the most important pharmacologically significant natural products, has gained significant consideration among scientists for decades since its multipharmacological activities. 1, 3-Dicarbonyl moiety of curcumin was found to be accountable for the rapid degradation of curcumin molecule. The aim of present work is to replace 1, 3-dicarbonyl moiety of curcumin by pyrazole and phenylpyrazole derivatives with a view to improving its stability and to investigate the role of substitution in N-phenylpyrazole curcumin on its antibacterial activity against both Gram-positive as well as Gram-negative bacteria. Methods: Pyrazole derivatives of curcumin were prepared by heating curcumin with phenyhydrazine/ substituted phenyhydrazine derivatives in AcOH. The residue was purified by silica gel column chromatography. Structures of purified compounds were confirmed by 1H NMR and Mass spectroscopy. The synthesized compounds were evaluated for their antibacterial activity by the microdilution broth susceptibility test method against gram positive (S. aureus) and gram negative (E. coli). Results: Effects of substitution in N-phenylpyrazole curcumin derivatives against S. aureus and E. coli were studied. The most active N-(3-Nitrophenylpyrazole) curcumin (12) exhibits twenty-fold more potency against S. aureus (MIC: 10μg/mL)) and N-(2-Fluoroophenylpyrazole) curcumin (5) fivefold more potency against E. coli (MIC; 50 μg/mL) than N-phenylpyrazole curcumin (4). Whereas, a remarkable decline in anti-bacterial activity against S. aureus and E. coli was observed when electron donating groups were incorporated in N-phenylpyrazole curcumin (4). Comparative studies of synthesized compounds suggest the effects of electron withdrawing and electron donating groups on unsubstituted phenylpyrazole curcumin (4). Conclusion: The structure-activity relationship (SAR) results indicated that the electron withdrawing and electron donating at N-phenylpyrazole curcumin played key roles for their bacterial inhibitory effects. The results of the antibacterial evaluation showed that the synthesized pyrazole derivatives of curcumin displayed moderate to very high activity in S. aureus. In conclusion, the series of novel curcumin derivatives were designed, synthesized and tested for their antibacterial activities against S. aureus and E. coli. Among them, N-(3-Nitrophenylpyrazole curcumin; 12) was most active against S. aureus (Gram-positive) and N-(2-Fluoroophenylpyrazole) curcumin (5) against E. coli (Gram-negative) bacteria.


2017 ◽  
Vol 6 (04) ◽  
pp. 5347 ◽  
Author(s):  
Omar B. Ahmed* ◽  
Anas S. Dablool

Several methods of Deoxyribonucleic acid (DNA) extraction have been applied to extract bacterial DNA. The amount and the quality of the DNA obtained for each one of those methods are variable. The study aimed to evaluate bacterial DNA extraction using conventional boiling method followed by alcohol precipitation. DNA extraction from Gram negative bacilli was extracted and precipitated using boiling method with further precipitation by ethanol. The extraction procedure performed using the boiling method resulted in high DNA yields for both E. coli and K. pneumoniae bacteria in (199.7 and 285.7μg/ml, respectively) which was close to control method (229.3 and 440.3μg/ml). It was concluded that after alcohol precipitation boiling procedure was easy, cost-effective, and applicable for high-yield quality of DNA in Gram-negative bacteria.


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 339
Author(s):  
Denise Dekker ◽  
Frederik Pankok ◽  
Thorsten Thye ◽  
Stefan Taudien ◽  
Kwabena Oppong ◽  
...  

Wound infections are common medical problems in sub-Saharan Africa but data on the molecular epidemiology are rare. Within this study we assessed the clonal lineages, resistance genes and virulence factors of Gram-negative bacteria isolated from Ghanaian patients with chronic wounds. From a previous study, 49 Pseudomonas aeruginosa, 21 Klebsiellapneumoniae complex members and 12 Escherichia coli were subjected to whole genome sequencing. Sequence analysis indicated high clonal diversity with only nine P. aeruginosa clusters comprising two strains each and one E. coli cluster comprising three strains with high phylogenetic relationship suggesting nosocomial transmission. Acquired beta-lactamase genes were observed in some isolates next to a broad spectrum of additional genetic resistance determinants. Phenotypical expression of extended-spectrum beta-lactamase activity in the Enterobacterales was associated with blaCTX-M-15 genes, which are frequent in Ghana. Frequently recorded virulence genes comprised genes related to invasion and iron-uptake in E. coli, genes related to adherence, iron-uptake, secretion systems and antiphagocytosis in P. aeruginosa and genes related to adherence, biofilm formation, immune evasion, iron-uptake and secretion systems in K. pneumonia complex. In summary, the study provides a piece in the puzzle of the molecular epidemiology of Gram-negative bacteria in chronic wounds in rural Ghana.


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 257
Author(s):  
Florian Turbant ◽  
David Partouche ◽  
Omar El Hamoui ◽  
Sylvain Trépout ◽  
Théa Legoubey ◽  
...  

Hfq is a bacterial regulator with key roles in gene expression. The protein notably regulates translation efficiency and RNA decay in Gram-negative bacteria, thanks to its binding to small regulatory noncoding RNAs. This property is of primary importance for bacterial adaptation and survival in hosts. Small RNAs and Hfq are, for instance, involved in the response to antibiotics. Previous work has shown that the E. coli Hfq C-terminal region (Hfq-CTR) self-assembles into an amyloid structure. It was also demonstrated that the green tea compound EpiGallo Catechin Gallate (EGCG) binds to Hfq-CTR amyloid fibrils and remodels them into nonamyloid structures. Thus, compounds that target the amyloid region of Hfq may be used as antibacterial agents. Here, we show that another compound that inhibits amyloid formation, apomorphine, may also serve as a new antibacterial. Our results provide an alternative in order to repurpose apomorphine, commonly used in the treatment of Parkinson’s disease, as an antibiotic to block bacterial adaptation to treat infections.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tessa B. Moyer ◽  
Ashleigh L. Purvis ◽  
Andrew J. Wommack ◽  
Leslie M. Hicks

Abstract Background Plant defensins are a broadly distributed family of antimicrobial peptides which have been primarily studied for agriculturally relevant antifungal activity. Recent studies have probed defensins against Gram-negative bacteria revealing evidence for multiple mechanisms of action including membrane lysis and ribosomal inhibition. Herein, a truncated synthetic analog containing the γ-core motif of Amaranthus tricolor DEF2 (Atr-DEF2) reveals Gram-negative antibacterial activity and its mechanism of action is probed via proteomics, outer membrane permeability studies, and iron reduction/chelation assays. Results Atr-DEF2(G39-C54) demonstrated activity against two Gram-negative human bacterial pathogens, Escherichia coli and Klebsiella pneumoniae. Quantitative proteomics revealed changes in the E. coli proteome in response to treatment of sub-lethal concentrations of the truncated defensin, including bacterial outer membrane (OM) and iron acquisition/processing related proteins. Modification of OM charge is a common response of Gram-negative bacteria to membrane lytic antimicrobial peptides (AMPs) to reduce electrostatic interactions, and this mechanism of action was confirmed for Atr-DEF2(G39-C54) via an N-phenylnaphthalen-1-amine uptake assay. Additionally, in vitro assays confirmed the capacity of Atr-DEF2(G39-C54) to reduce Fe3+ and chelate Fe2+ at cell culture relevant concentrations, thus limiting the availability of essential enzymatic cofactors. Conclusions This study highlights the utility of plant defensin γ-core motif synthetic analogs for characterization of novel defensin activity. Proteomic changes in E. coli after treatment with Atr-DEF2(G39-C54) supported the hypothesis that membrane lysis is an important component of γ-core motif mediated antibacterial activity but also emphasized that other properties, such as metal sequestration, may contribute to a multifaceted mechanism of action.


Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 875
Author(s):  
Tomasz Bogiel ◽  
Mateusz Rzepka ◽  
Eugenia Gospodarek-Komkowska

Non-fermenting Gram-negative rods are one of the most commonly isolated bacteria from human infections. These microorganisms are typically opportunistic pathogens that pose a serious threat to public health due to possibility of transmission in the human population. Resistance to beta-lactams, due to carbapenemases synthesis, is one of the most important antimicrobial resistance mechanisms amongst them. The aim of this study was to evaluate the usefulness of the Carbapenem Inactivation Method (CIM), and its modifications, for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. This research involved 81 strains of Gram-negative rods. Of the tested strains, 55 (67.9%) synthesized carbapenemases. For non-fermenting rods, 100% sensitivity and specificity was obtained in the version of the CIM test using imipenem discs and E. coli ATCC 25922 strain. The CIM test allows for differentiation of carbapenems resistance mechanisms resulting from carbapenemase synthesis from other resistance types. It is a reliable diagnostic method for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. Application of imipenem discs and P. aeruginosa ATCC 27853 reference strain increases CIM results sensitivity, while imipenem discs and E. coli ATCC 25922 strain use maintains full precision of the test for non-fermenting rods.


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