Gram-positive Research Articles

In vitro antibacterial acitivity of Naproxen and its combination with ciprofloxacin

Naproxen, a nonsteroidal anti-inflammatory drug (NSAID), is commonly used to reduce fever, and to treat pain and inflammation caused by several conditions. Previously, naproxen was evaluated for its antimicrobial potency in various studies. In our study, we aimed to demonstrate the antibacterial and synergistic activities of naproxen and ciprofloxacin against various Gram-positive and Gram-negative bacteria including, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 700603. The results showed promising antibacterial activity against tested Gram-positive bacteria. However, there was no effect on Gram-negative bacteria. Additionally, checkerboard assays did not reveal any additive or synergistic activity when combined with Ciprofloxacin. Collectively, our study's data show naproxen's selectivity against Gram-positive bacteria. This result suggests that naproxen can be further used as a potential source of antibiotics against Gram-positive bacteria.

In vitro antibacterial acitivity of Naproxen and its combination with ciprofloxacin

Naproxen, a nonsteroidal anti-inflammatory drug (NSAID), is commonly used to reduce fever, and to treat pain and inflammation caused by several conditions. Previously, naproxen was evaluated for its antimicrobial potency in various studies. In our study, we aimed to demonstrate the antibacterial and synergistic activities of naproxen and ciprofloxacin against various Gram-positive and Gram-negative bacteria including, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 700603. The results showed promising antibacterial activity against tested Gram-positive bacteria. However, there was no effect on Gram-negative bacteria. Additionally, checkerboard assays did not reveal any additive or synergistic activity when combined with Ciprofloxacin. Collectively, our study's data show naproxen's selectivity against Gram-positive bacteria. This result suggests that naproxen can be further used as a potential source of antibiotics against Gram-positive bacteria.

Antibacterial potency of Ibuprofen and its interaction with ciprofloxacin against Gram positive and Gram negative bacteria

Ibuprofen, a nonsteroidal anti-inflammatory drug (NSAID), works by reducing hormones for the treatment of fever, inflammation, and pain. Previously, it was only shown that the Ibuprofen inhibits the effect of various bacteria that are stimulated by bacterial infections and not directly on bacterial cells. In this study, we aimed to investigate antibacterial and synergistic activities of Ibuprofen against Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 700603. The results revealed promising antibacterial activity against tested Gram-positive bacteria, but there was no effect on Gram-negative bacteria. Furthermore, checkerboard assays did not reveal any additive or synergistic activity when combined with ciprofloxacin against tested Gram-positive bacteria. Collectively, our data reveal the selective antibacterial activity of ibuprofen against Gram-positive bacteria which suggests that ibuprofen can be further investigated as a potential source for new therapeutic options.

Simultaneous removal of diclofenac sodium, cadmium and bacterial inactivation from aqueous solutions by activated MgO nanoparticles

This study investigated whether MgO nanoparticles (NPs) produced via the sol–gel process can simultaneously remove the pharmaceutical diclofenac sodium (DCF) and cadmium ions (Cd2+) from water. The UV spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX) were used to confirm the composition, morphology and particle size of the MgO NPs. The adsorption performance of MgO NPs was compared to that of bentonite-nano zero valent iron (B-nZVI) and activated carbon (AC). The impact of contact time, adsorbate concentration, pH, and temperature on the removal efficacy of DCF and Cd2+ via MgO NPs in aqueous solutions were investigated, and the removal data was best described by the kinetic model of pseudo-second-order and the Langmuir isotherm model (R 2 > 0.996). The maximum removal capacity of MgO for DCF and Cd2+ at pH 7 was 66.2 mg.g−1 (%R = 90) and 50.8 mg.g−1 (%R = 80), respectively. The average adsorption enthalpy for DCF and Cd2+ (ΔH° 50 kJ. mol−1) demonstrates the occurrence of physical adsorption process, while the negative value of ΔG° reveals the spontaneous nature of DCF and Cd2+ adsorption on the surface of MgO NPs. The DCF and Cd2+ adsorbed MgO surfaces were reused against various gram-positive bacteria. Finally, it was shown that MgO and sand column were more effective for removal of persistent substances and heavy metal residues than a column packed by a mixture of B-nZVI and sand.

Poly Lactic-co-Glycolic Acid (PLGA) Loaded with a Squaraine Dye as Photosensitizer for Antimicrobial Photodynamic Therapy

Antimicrobial Photodynamic Therapy (aPDT) is an innovative and promising method for combating infections, reducing the risk of antimicrobial resistance compared to traditional antibiotics. Squaraine (SQ) dyes can be considered promising photosensitizers (PSs) but are generally hydrophobic molecules that can self-aggregate under physiological conditions. To overcome these drawbacks, a possible solution is to incorporate SQs inside nanoparticles (NPs). The present work deals with the design and development of innovative nanophotosensitizers based on poly lactic-co-glycolic acid (PLGA) NPs incorporating a brominated squaraine (BrSQ) with potential application in aPDT. Two designs of experiments (DoEs) based on the single emulsion and nanoprecipitation methods were set up to investigate how different variables (type of solvent, solvent ratio, concentration of PLGA, stabilizer and dye, sonication power and time) can affect the size, zeta (ζ)-potential, yield, entrapment efficiency, and drug loading capacity of the SQ-PLGA NPs. SQ-PLGA NPs were characterized by NTA, FE-SEM, and UV-Vis spectroscopy and the ability to produce reactive oxygen species (ROS) was evaluated, proving that ROS generation ability is preserved in SQ-PLGA. In vitro antimicrobial activity against Gram-positive bacteria in planktonic state using Staphylococcus aureus was conducted in different conditions and pH to evaluate the potential of these nanophotosensitizers for aPDT in the local treatment of infections.

Cast Extruded Films Based on Polyhydroxyalkanoate/Poly(lactic acid) Blend with Herbal Extracts Hybridized with Zinc Oxide

The aim of the presented work was to functionalize a blend based on polyhydroxyalkanoate (PHA): poly(hydroxybutyrate (PHB) with poly(lactic acid) (PLA) and a mixture of three selected herb extracts, namely, Hypericum L., Urtica L. and Chelidonium L., (E), zinc oxide (ZnO) and a combined system (EZnO), produced via extrusion. Before processing with bioresin, the natural modifiers were characterized using thermal analysis, FTIR and antimicrobial tests. The results revealed interactions between the extracts and the filler, leading to higher thermal stability in EZnO than when using E alone. Moreover, the mixture of extracts exhibited antimicrobial properties toward both Gram-negative (S. aureus) as well as Gram-positive bacteria (E. coli). Modified regranulates were transformed into films by cast extrusion. The influence of the additives on thermal (DSC, TGA and OIT), mechanical, barrier (WVTR and OTR), morphological (FTIR) and optical properties was investigated. The EZnO additive had the highest impact on the mechanical, barrier (OTR and WVTR) and optical properties of the bioresin. The microbial test results revealed that PHA-EZnO exhibited higher activity than PHA-ZnO and PHA-E and also reduced the number of S. aureus, E. coli and C. albicans cells. The findings confirmed the synergistic effect between the additive components. Modified polyester films did not eliminate the phi6 bacteriophage particles completely, but they did decrease their number, confirming moderate antiviral effectiveness.

A General Strategy for Enhanced Photodynamic Antimicrobial Therapy with Perylenequinonoid Photosensitizers Using a Macrocyclic Supramolecular Carrier.

Perylenequinonoid natural products are a class of photosensitizers (PSs) that exhibit high reactive oxygen species (ROS) generation and excellent activity for Type I/Type II dual photodynamic therapy. However, their limited activity against gram-negative bacteria and poor water solubility significantly restrict their potential in broad-spectrum photodynamic antimicrobial therapy (PDAT). Herein, a general approach to overcome the limitations of perylenequinonoid photosensitizers (PQPSs) in PDAT by utilizing a macrocyclic supramolecular carrier is presented. Specifically, AnBox·4Cl, a water-soluble cationic cyclophane, is identified as a universal macrocyclic host for PQPSs such as elsinochrome C, hypocrellin A, hypocrellin B, and hypericin, forming 1:1 host-guest complexes with high binding constants (≈107 m -1) in aqueous solutions. Each AnBox·4Cl molecule carries four positive charges that promote strong binding with the membrane of gram-negative bacteria. As a result, the AnBox·4Cl-PQPS complexes can effectively anchor on the surfaces of gram-negative bacteria, while the PQPSs alone cannot. In vitro and in vivo experiments demonstrate that these supramolecular PSs have excellent water solubility and high ROS generation, with broad-spectrum PDAT effect against both gram-negative and gram-positive bacteria. This work paves a new path to enhance PDAT by showcasing an efficient approach to improve PQPSs' water solubility and killing efficacy for gram-negative bacteria.

Aloe vera-assisted biogenic synthesis of tunable magnesium oxide nanoparticles for enhanced photocatalytic and antibacterial applications

In this study, magnesium oxide (MgO) nanoparticles (NPs) tuned via doping with transition metal ions, Co2+/Cu2+/Zn2+ have been synthesized biologically using Aloe vera leaf extract as an encapsulating agent. These pure and doped MgO NPs have been characterized by using various physico-chemical characterization techniques. X-ray diffraction (XRD) analysis confirms the formation of face-centered cubic structures of pure and doped MgO NPs with their average crystallite sizes ranging from 8.11-17.83 nm. X-ray photoelectron spectroscopic (XPS) analysis confirmed the presence of doped element in their +2 oxidation state. UV-Visible spectral analysis revealed a significant reduction in band gap energies in nano-dimensions which further gets modified upon doping (1.76 − 4.9 eV). Our findings also demonstrated the engagement of phytochemicals as capping agents in Fourier transform infrared (FTIR) examination. Scanning electron microscopic (SEM) and Transmission electron microscopic (TEM) analysis provided insights into the morphologies and particle sizes. Elemental analysis by Energy dispersive X-ray (EDX) confirms the presence of Co, Cu, and Zn elements in doped MgO NPs. The photocatalytic degradation of dyes has confirmed that Co and Cu doping substantially enhance the photocatalytic activity compared to both undoped and Zn-doped MgO NPs. Furthermore, these nanoparticles demonstrate superior efficiency in degrading the cationic dye Malachite green (MG) compared to the anionic dye Methyl orange (MO). Regarding their antibacterial properties, only Cu doping induces a notable antibacterial effect in MgO NPs, particularly against gram-positive bacteria. Additionally, these nanoparticles exhibit substantial antioxidant activities. These tailored MgO nanoparticles exhibit potential for applications in wastewater treatment.

Green synthesis of zinc oxide nanoparticles (ZnO-NPs) by Streptomyces baarnensis and its active metabolite (Ka): a promising combination against multidrug-resistant ESKAPE pathogens and cytotoxicity

Various eco-friendly techniques are being researched for synthesizing ZnO-NPs, known for their bioactivity. This study aimed at biosynthesizing ZnO-NPs using Streptomyces baarnensis MH-133, characterizing their physicochemical properties, investigating antibacterial activity, and enhancement of their efficacy by combining them with a water-insoluble active compound (Ka) in a nanoemulsion form. Ka is a pure compound of 9-Ethyl-1,4,6,9,10-pentahydroxy-7,8,9,10-tetrahydrotetracene-5,12-dione obtained previously from our strain of Streptomyces baarnensis MH-133. Biosynthesized ZnO-NPs employing Streptomyces baarnensis MH-133 filtrate and zinc sulfate (ZnSO4.7H2O) as a precursor were purified and characterized by physicochemical investigation. High-resolution-transmission electron microscopy (HR-TEM) verified the effective biosynthesis of ZnO-NPs (size < 12 nm), whereas dynamic light scattering (DLS) analysis showed an average size of 17.5 nm. X-ray diffraction (XRD) exhibited characteristic diffraction patterns that confirmed crystalline structure. ZnO-NPs efficiently inhibited both Gram-positive and Gram-negative bacteria (MICs: 31.25–125 µg/ml). The pure compound (Ka) was combined with ZnO-NPs to improve effectiveness and reduce dose using checkerboard microdilution. Niteen treatments of Ka and ZnO-NPs combinations obtained by checkerboard matrix inhibited Klebsiella pneumonia. Eleven combinations had fractional inhibitory concentration index (FICi) between 1.03 and 2, meaning indifferent, another five combinations resulted from additive FICi (0.625–1) and only one combination with FICi of 0.5, indicating synergy. In the case of methicillin-resistant S. aureus (MRSA), Ka-ZnO-NPs combinations yielded 23 treatments with varying degrees of interaction. The results showed eleven treatments with indifferent interaction, eight additive interactions, and two synergies with FICi of 0.5 and 0.375. The combinations that exhibited synergy action were transformed into a nanoemulsion form to improve their solubility and bioavailability. The HR-TEM analysis of the nanoemulsion revealed spherical oil particles with a granulated core smaller than 200 nm and no signs of aggregation. Effective dispersion was confirmed by DLS analysis which indicated that Ka-ZnO-NPs nanoemulsion droplets have an average size of 53.1 nm and a polydispersity index (PI) of 0.523. The killing kinetic assay assessed the viability of methicillin-resistant Staphylococcus aureus (MRSA) and K. pneumonia post-treatment with Ka-ZnO-NPs combinations either in non-formulated or nanoemulsion form. Results showed Ka-ZnO-NPs combinations show concentration and time-dependent manner, with higher efficacy in nanoemulsion form. The findings indicated that Ka-ZnO-NPs without formulation at MIC values killed K. pneumonia after 24 h but not MRSA. Our nanoemulsion loaded with the previously mentioned combinations at MIC value showed bactericidal effect at MIC concentration of Ka-ZnO-NPs combination after 12 and 18 h of incubation against MRSA and K. pneumonia, respectively, compared to free combinations. At half MIC value, nanoemulsion increased the activity of the combinations to cause a bacteriostatic effect on MRSA and K. pneumonia after 24 h of incubation. The free combination showed a bacteriostatic impact for 6 h before the bacteria regrew to increase log10 colony forming unit (CFU)/ml over the initial level. Similarly, the cytotoxicity study revealed that the combination in nanoemulsion form decreased the cytotoxicity against kidney epithelial cells of the African green monkey (VERO) cell line. The IC50 for Ka-ZnO-NPs non-formulated treatment was 8.17/1.69 (µg/µg)/ml, but in nano-emulsion, it was 22.94 + 4.77 (µg/µg)/mL. In conclusion, efficient Ka-ZnO-NPs nanoemulsion may be a promising solution for the fighting of ESKAPE pathogenic bacteria according to antibacterial activity and low toxicity.

Screening of Secondary Metabolites, Antibacterial and Antioxidative Capacity of Paederia Foetida L. Leaf Extracts

Plant resources have been utilized since ancient times for the healing of various illnesses of humankind. This study investigated the secondary metabolites, antibacterial, and antioxidative capability of Paederia foetida L. (P. foetida) leaf extract. Qualitative tests were done to determine the presence of secondary metabolites. The disc diffusion method was used for the determination of the antibacterial activity of the extract. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging assay, reducing power assay, total phenolic, and total flavonoid content methods were followed for the evaluation of antioxidative potentials of the leaf extract. The leaf extract was found to show significant (p &lt; 0.05) antibacterial activity against Gram-positive bacteria compared to Gram-negative ones. In DPPH free radical scavenging assay, the inhibitory concentration (IC50) value for methanol crude extract was found to be fairly significant (162.929 μg/mL) in comparison with that (1.57 µg/mL) of the reference standard ascorbic acid. The maximum absorbance for reducing power assay was 0.823 compared to 1.891 for standard ascorbic acid. The extract was found to have a total phenolic content of 8.03 ±1. 05 mg/g gallic acid equivalent and flavonoid content 134.74 mg/g quercetin equivalent. The results demonstrated a moderate antibacterial and promising antioxidative activity of P. foetida leaf extract, but it is necessary to further analyze prospective pharmaceutical preparation. The Chittagong Univ. J. B. Sci.,Vol. 10(1 &amp;2):121-137, 2020

Metabolic Profile of the Genome-Reduced Bacillus subtilis Strain IIG-Bs-27-39: An Attractive Chassis for Recombinant Protein Production.

The Gram-positive bacterium Bacillus subtilis is extensively used in the industry for the secretory production of proteins with commercial value. To further improve its performance, this microbe has been the subject of extensive genome engineering efforts, especially the removal of large genomic regions that are dispensable or even counterproductive. Here, we present the genome-reduced B. subtilis strain IIG-Bs-27-39, which was obtained through systematic deletion of mobile genetic elements, as well as genes for extracellular proteases, sporulation, flagella formation, and antibiotic production. Different from previously characterized genome-reduced B. subtilis strains, the IIG-Bs-27-39 strain was still able to grow on minimal media. We used this feature to benchmark strain IIG-Bs-27-39 against its parental strain 168 with respect to heterologous protein production and metabolic parameters during bioreactor cultivation. The IIG-Bs-27-39 strain presented superior secretion of difficult-to-produce staphylococcal antigens, as well as higher specific growth rates and biomass yields. At the metabolic level, changes in byproduct formation and internal amino acid pools were observed, whereas energetic parameters such as the ATP yield, ATP/ADP levels, and adenylate energy charge were comparable between the two strains. Intriguingly, we observed a significant increase in the total cellular NADPH level during all tested conditions and increases in the NAD+ and NADP(H) pools during protein production. This indicates that the IIG-Bs-27-39 strain has more energy available for anabolic processes and protein production, thereby providing a link between strain physiology and production performance. On this basis, we conclude that the genome-reduced strain IIG-Bs-27-39 represents an attractive chassis for future biotechnological applications.

Metabolic Profile of the Genome-Reduced Bacillus subtilis Strain IIG-Bs-27-39: An Attractive Chassis for Recombinant Protein Production.

The Gram-positive bacterium Bacillus subtilis is extensively used in the industry for the secretory production of proteins with commercial value. To further improve its performance, this microbe has been the subject of extensive genome engineering efforts, especially the removal of large genomic regions that are dispensable or even counterproductive. Here, we present the genome-reduced B. subtilis strain IIG-Bs-27-39, which was obtained through systematic deletion of mobile genetic elements, as well as genes for extracellular proteases, sporulation, flagella formation, and antibiotic production. Different from previously characterized genome-reduced B. subtilis strains, the IIG-Bs-27-39 strain was still able to grow on minimal media. We used this feature to benchmark strain IIG-Bs-27-39 against its parental strain 168 with respect to heterologous protein production and metabolic parameters during bioreactor cultivation. The IIG-Bs-27-39 strain presented superior secretion of difficult-to-produce staphylococcal antigens, as well as higher specific growth rates and biomass yields. At the metabolic level, changes in byproduct formation and internal amino acid pools were observed, whereas energetic parameters such as the ATP yield, ATP/ADP levels, and adenylate energy charge were comparable between the two strains. Intriguingly, we observed a significant increase in the total cellular NADPH level during all tested conditions and increases in the NAD+ and NADP(H) pools during protein production. This indicates that the IIG-Bs-27-39 strain has more energy available for anabolic processes and protein production, thereby providing a link between strain physiology and production performance. On this basis, we conclude that the genome-reduced strain IIG-Bs-27-39 represents an attractive chassis for future biotechnological applications.

AutoBioTech─A Versatile Biofoundry for Automated Strain Engineering.

The inevitable transition from petrochemical production processes to renewable alternatives has sparked the emergence of biofoundries in recent years. Manual engineering of microbes will not be sufficient to meet the ever-increasing demand for novel producer strains. Here we describe the AutoBioTech platform, a fully automated laboratory system with 14 devices to perform operations for strain construction without human interaction. Using modular workflows, this platform enables automated transformations of Escherichia coli with plasmids assembled via modular cloning. A CRISPR/Cas9 toolbox compatible with existing modular cloning frameworks allows automated and flexible genome editing of E.coli. In addition, novel workflows have been established for the fully automated transformation of the Gram-positive model organism Corynebacterium glutamicum by conjugation and electroporation, with the latter proving to be the more robust technique. Overall, the AutoBioTech platform excels at versatility due to the modularity of workflows and seamless transitions between modules. This will accelerate strain engineering of Gram-negative and Gram-positive bacteria.

Pheromone cCF10 inhibits the antibiotic persistence of Enterococcus faecalis by modulating energy metabolism

IntroductionBacterial resistance presents a major challenge to both the ecological environment and human well-being, with persistence playing a key role. Multiple studies were recently undertaken to examine the factors influencing the formation of persisters and the underlying process, with a primary focus on Gram-negative bacteria and Staphylococcus aureus (Gram-positive bacteria). Enterococcus faecalis (E. faecalis) is capable of causing a variety of infectious diseases, but there have been few studies of E. faecalis persisters. Previous studies have shown that the sex pheromone cCF10 secreted by E. faecalis induces conjugative plasmid transfer. However, whether the pheromone cCF10 regulates the persistence of E. faecalis has not been investigated.MethodsAs a result, we investigated the effect and potential molecular mechanism of pheromone cCF10 in regulating the formation of persisters in E. faecalis OG1RF using a persistent bacteria model.Results and discussionThe metabolically active E. faecalis OG1RF reached a persistence state and temporarily tolerated lethal antibiotic concentrations after 8 h of levofloxacin hydrochloride (20 mg/mL) exposure, exhibiting a persistence rate of 0.109 %. During the growth of E. faecalis OG1RF, biofilm formation was a critical factor contributing to antibiotic persistence, whereas 10 ng/mL cCF10 blocked persister cell formation. Notably, cCF10 mediated the antibiotic persistence of E. faecalis OG1RF via regulating metabolic activity rather than suppressing biofilm formation. The addition of cCF10 stimulated the Opp system and entered bacterial cells, inhibiting (p)ppGpp accumulation, thus maintaining the metabolically active state of bacteria and reducing persister cell generation. These findings offer valuable insights into the formation, as well as the control mechanism of E. faecalis persisters.

Pheromone cCF10 inhibits the antibiotic persistence of Enterococcus faecalis by modulating energy metabolism

IntroductionBacterial resistance presents a major challenge to both the ecological environment and human well-being, with persistence playing a key role. Multiple studies were recently undertaken to examine the factors influencing the formation of persisters and the underlying process, with a primary focus on Gram-negative bacteria and Staphylococcus aureus (Gram-positive bacteria). Enterococcus faecalis (E. faecalis) is capable of causing a variety of infectious diseases, but there have been few studies of E. faecalis persisters. Previous studies have shown that the sex pheromone cCF10 secreted by E. faecalis induces conjugative plasmid transfer. However, whether the pheromone cCF10 regulates the persistence of E. faecalis has not been investigated.MethodsAs a result, we investigated the effect and potential molecular mechanism of pheromone cCF10 in regulating the formation of persisters in E. faecalis OG1RF using a persistent bacteria model.Results and discussionThe metabolically active E. faecalis OG1RF reached a persistence state and temporarily tolerated lethal antibiotic concentrations after 8 h of levofloxacin hydrochloride (20 mg/mL) exposure, exhibiting a persistence rate of 0.109 %. During the growth of E. faecalis OG1RF, biofilm formation was a critical factor contributing to antibiotic persistence, whereas 10 ng/mL cCF10 blocked persister cell formation. Notably, cCF10 mediated the antibiotic persistence of E. faecalis OG1RF via regulating metabolic activity rather than suppressing biofilm formation. The addition of cCF10 stimulated the Opp system and entered bacterial cells, inhibiting (p)ppGpp accumulation, thus maintaining the metabolically active state of bacteria and reducing persister cell generation. These findings offer valuable insights into the formation, as well as the control mechanism of E. faecalis persisters.

A novel antibiotic: the antimicrobial effects of CFBSA and its application on electronspun wound dressing

N-chloro-N-fluorobenzenesulfonylamide (CFBSA), was a novel chlorinating reagent, which exhibits potential antibacterial activities. In this study, CFBSA was confirmed as a wide-broad antimicrobial and bactericidal drug against different gram-negative bacteria, gram-positive bacteria and fungi, while it was found to have low cytotoxicity for eukaryotic cells. In addition, microorganism morphology assay and oxidative stress test was used to determine the antimicrobial mechanisms of CFBSA. According to the results, CFBSA probably had a target on cell membrane and killed microorganism by disrupting its cell membrane. Then, CFBSA was first combined with poly(L-lactide-co-caprolactone) (PLCL)/SF via electrospinning and applied in wound dressings. The characterization of different PLCL/SF of CFBSA-loaded nanofibrous mats was investigated by SEM, water contact angle, Fourier transform infrared spectroscopy, cell compatibility and antimicrobial test. CFBSA-loaded PLCL/SF nanofibrous mats showed excellent antimicrobial activities. In order to balance of the biocompatibility and antibacterial efficiency, SP-2.5 was selected as the ideal loading concentration for further application of CFBSA-loaded PLCL/SF. In conclusion, the electrospun CFBSA-loaded PLCL/SF nanofibrous mat with its broad-spectrum antimicrobial and bactericidal activity and good biocompatibility showed enormous potential for wound dressing.

AutoBioTech─A Versatile Biofoundry for Automated Strain Engineering.

The inevitable transition from petrochemical production processes to renewable alternatives has sparked the emergence of biofoundries in recent years. Manual engineering of microbes will not be sufficient to meet the ever-increasing demand for novel producer strains. Here we describe the AutoBioTech platform, a fully automated laboratory system with 14 devices to perform operations for strain construction without human interaction. Using modular workflows, this platform enables automated transformations of Escherichia coli with plasmids assembled via modular cloning. A CRISPR/Cas9 toolbox compatible with existing modular cloning frameworks allows automated and flexible genome editing of E.coli. In addition, novel workflows have been established for the fully automated transformation of the Gram-positive model organism Corynebacterium glutamicum by conjugation and electroporation, with the latter proving to be the more robust technique. Overall, the AutoBioTech platform excels at versatility due to the modularity of workflows and seamless transitions between modules. This will accelerate strain engineering of Gram-negative and Gram-positive bacteria.

Complete genome sequence of Enterococcus faecalis B05.

Enterococcus faecalis, a gram-positive bacterium, is the main intestinal bacteria in humans and animals. We report here the complete genome sequence of E. faecalis B05, whose genome length is 2,745,741 bp, containing 2,503 protein-coding genes, with a GC content of 37.64%.

Comparable Antibacterial Effects of Silver and Iron Oxide Nanoparticles on Urinary Calculi Obtained After Percutaneous Nephrolithotomy

Background: In recent years, science and industry have focused on preparing nanoparticles (NPs). Due to the resistance of bacteria to antibiotics, there is an overwhelming need to find effective antimicrobials with fewer side effects. Objectives: The aim of this study was to synthesize silver and iron oxide NPs and investigate their antimicrobial effects on bacteria isolated from urinary stones. Methods: Cases of urolithiasis from 45 patients with staghorn stones, extracted through percutaneous nephrolithotomy (PCNL), were included in the study. Urinary stone cultures were performed, and the isolation and identification of bacteria were done using standard microbiological techniques. Silver and iron oxide NPs were synthesized, and the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) were evaluated for the isolates. Results: Seventeen (60.7%) out of 28 positive cultured cases of the isolated bacteria were gram-negative, and 11 (39.3%) were gram-positive. The most abundant isolated bacterium was Escherichia coli, with 13 cases (46.4%). The antibacterial effects of silver and iron oxide NPs revealed that the maximum inhibitory zone for Ag NPs at 1000 ppm was 23 mm, and for Fe3O4 NPs, it was 18.5 mm. The MIC was 180 µg/mL for Ag and 250 µg/mL for Fe3O4. The MBC was 228 µg/mL for Ag and 300 µg/mL for Fe3O4. Conclusions: Nanoparticles exhibited antibacterial effects on the bacteria studied in a concentration-dependent manner. Ag NPs showed a more pronounced bactericidal effect than Fe3O4 NPs. As a result, these two NPs demonstrated effective antibacterial activity against both gram-positive and gram-negative bacteria.

Polydopamine modified colloidal gold nanotag-based lateral flow immunoassay platform for highly sensitive detection of pathogenic bacteria and fast evaluation of antibacterial agents

Bacterial infection is a great threat to human health. Lateral flow immunoassays (LFIAs) with the merits of low cost, quick screening, and on-site detection are competitive technologies for bacteria detection, but their detection limits depend on the optical performance of the adopted nanotags. Herein, we presented a LFIA platform for bacteria detection using polydopamine (PDA) functionalized Au nanoparticles (denoted as Au@PDA) as the nanotag. The introduction of PDA could provide enhanced light absorption of Au, as well as numerous functional groups for conjugation. Small recognition molecules i.e. vancomycin (Van) and p-mercaptophenylboronic acid (PMBA) were covalently anchored to Au@PDA, and selected as the specific probes towards Gram-positive (G+) and Gram-negative (G—) bacteria, respectively. Taken Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) as the representative targets of G+ and G— bacteria, two LFA strips were successfully constructed based on the immuno-sandwich principle. They could quantitatively detect S. aureus and E. coli both down to 102 cfu/mL, a very competitive detection limit in comparison with other colorimetric or luminescent probes-based LFIAs. Furthermore, the proposed two strips were applied for the quantitative, accurate, and rapid detection of S. aureus and E. coli in food and human urine samples with good analytical results obtained. In addition, they were integrated as a screening platform for quick evaluation of diverse antibacterial agents within 3 h, which is remarkably shortened compared with that of the two traditional methods i.e. bacterial culture and plate-counting.