Bacterial Viability Assay Research Articles

Antimicrobial Potential of Strontium-Functionalized Titanium Against Bacteria Associated With Peri-Implantitis.

To explore the antimicrobial potential of strontium (Sr)-functionalized wafers against multiple bacteria associated with per-implant infections, in both mono- and multispecies biofilms. The bactericidal and bacteriostatic effect of silicon wafers functionalized with a strontium titanium oxygen coating (Sr-Ti-O) or covered only with Ti (controls) against several bacteria, either grown as a mono-species or multispecies biofilms, was assessed using a bacterial viability assay and a plate counting method. Mono-species biofilms were assessed after 2 and 24 h, while the antimicrobial effect on multispecies biofilms was assessed at Days 1, 3, and 6. The impact of Sr functionalization on the total percentage ofPorphyromonas gingivalis in the multispecies biofilm, using qPCR, and gingipain activity was also assessed. Sr-functionalized wafers, compared to controls, were associated with statistically significant less viable cells in both mono- and multispecies tests. The number of colony forming units (CFUs) within the biofilm was significantly less in Sr-functionalized wafers, compared to control wafers, for Staphylococcus aureus at all time points of evaluation and forEscherichia coli at Day 1. Gingipain activity was less in Sr-functionalized wafers, compared to control wafers, and the qPCR showed that P. gingivalis remained below detection levels at Sr-functionalized wafers, while it consisted of 15% of the total biofilm on control wafers at Day 6. Sr functionalization displayed promising antimicrobial potential, possessing bactericidal and bacteriostatic ability against bacteria associated with peri-implantitis grown either as mono-species or mixed in a multispecies consortium with several common oral microorganisms.

Heparin mimicking sulfonated polyester synergistically promotes bacterial Infected-Wound healing with ROS responsive cinnamaldehyde based prodrugs

Excessive antibiotic usage has resulted in antibiotic resistance. This work introduces a novel ROS-responsive, cinnamaldehyde (CA)-based sulfonated all-polyester prodrug (RCSAP) to combat antibiotic resistance. RCSAP is a unique amphiphilic pseudo triblock copolymer comprising of two hydrophilic, heparin-mimicking sulfonated polyester (HSP) blocks and a hydrophobic polymeric block containing ROS-responsive CA thioacetal. RCSAP nanoparticles (NPs) not only achieve self-amplifying degradation and release, but orchestrate a “dual wielding” synergistic antibacterial mechanism. HSP shell competes with bacteria for host cell surface binding, hindering its internalization and infection. Simultaneously, ROS-responsive core liberates CA, boosting bacterial membrane permeability. The minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined. RCSAP NPs display exceptional antibacterial efficacy against both E. coli and S. aureus, as confirmed by survival analysis, colony-forming unit (CFU) quantification, microbial growth curve analysis, and live/dead bacterial viability assays. Importantly, RCSAP NPs effectively eradicated methicillin-resistant Staphylococcus aureus (MRSA). In addition, RCSAP NPs possess the unique ability to bind to S. aureus, aggregate and precipitate it. They competitively hinder S. aureus infection in both ARPE-19 and 293 T cells. Molecular simulation further verified the electrostatic interactions between HSP and the S. aureus SpA protein, which may mediate the binding of RCSAP NPs to S. aureus. In vivo assessments demonstrated that RCSAP NPs significantly accelerates wound healing in S. aureus-infected models, with excellent biocompatibility and minimal systemic toxicity. This work paves the way for all-polyester-based nanomedicines harnessing the power of natural antibacterial and heparin-mimicking agents synergistically for enhanced disinfection and wound healing, curbing drug resistance.

Synergism with Shikimic Acid Restores β-Lactam Antibiotic Activity against Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) has evolved into a dangerous pathogen resistant to beta-lactam antibiotics (BLAs) and has become a worrisome superbug. In this study, a strategy in which shikimic acid (SA), which has anti-inflammatory and antibacterial activity, is combined with BLAs to restart BLA activity was proposed for MRSA treatment. The synergistic effects of oxacillin combined with SA against oxacillin resistance in vitro and in vivo were investigated. The excellent synergistic effect of the oxacillin and SA combination was confirmed by performing the checkerboard assay, time-killing assay, live/dead bacterial cell viability assay, and assessing protein leakage. SEM showed that the cells in the control group had a regular, smooth, and intact surface. In contrast, oxacillin and SA or the combination treatment group exhibited different degrees of surface collapse. q-PCR indicated that the combination treatment group significantly inhibited the expression of the mecA gene. In vivo, we showed that the combination treatment increased the survival rate and decreased the bacterial load in mice. These results suggest that the combination of oxacillin with SA is considered an effective treatment option for MRSA, and the combination of SA with oxacillin in the treatment of MRSA is a novel strategy.

An innovative tunable bimodal porous PCL/gelatin dressing fabricated by electrospinning and 3D printing for efficient wound healing and scalable production

This study presents the development of tunable scaffolds with bimodal porosity comprising poly(ε-caprolactone) (PCL) micro-meshes and PCL/gelatin/ε-polylysine (ε-PL) fibrous layers. Pure PCL scaffolds were prepared using the fused deposition modeling technique featuring grid geometry and interconnected micro-pores, followed by electrospinning to produce PCL/gelatin/ε-PL nanofibrous layers. Field emission scanning electron microscopy was employed to investigate the morphological features of the scaffolds, while the physicomechanical properties were studied using tensile and contact angle tests. Antibacterial performance and skin cell toxicity of the scaffolds were determined by bacterial disc diffusion and cell viability assays, respectively. Morphological analysis showed the presence of micro-to nano-sized pores in the developed scaffolds. The mechanical test results revealed that the prepared scaffolds exhibited Young's modulus values similar to the human skin with higher strain. The nanocomposite scaffolds were cytocompatible and effectively eradicated common bacteria associated with cutaneous wounds. In light of the aforementioned results along with facile fabrication, the tunable PCL/gelatin/ε-PL porous scaffolds hold great promise for applications in skin wound repair.

Automated and miniaturized screening of antibiotic combinations via robotic-printed combinatorial droplet platform

Antimicrobial resistance (AMR) has become a global health crisis in need of novel solutions. To this end, antibiotic combination therapies, which combine multiple antibiotics for treatment, have attracted significant attention as a potential approach for combating AMR. To facilitate advances in antibiotic combination therapies, most notably in investigating antibiotic interactions and identifying synergistic antibiotic combinations however, there remains a need for automated high-throughput platforms that can create and examine antibiotic combinations on-demand, at scale, and with minimal reagent consumption. To address these challenges, we have developed a Robotic-Printed Combinatorial Droplet (RoboDrop) platform by integrating a programmable droplet microfluidic device that generates antibiotic combinations in nanoliter droplets in automation, a robotic arm that arranges the droplets in an array, and a camera that images the array of thousands of droplets in parallel. We further implement a resazurin-based bacterial viability assay to accelerate our antibiotic combination testing. As a demonstration, we use RoboDrop to corroborate two pairs of antibiotics with known interactions and subsequently identify a new synergistic combination of cefsulodin, penicillin, and oxacillin against a model E. coli strain. We therefore envision RoboDrop becoming a useful tool to efficiently identify new synergistic antibiotic combinations toward combating AMR.

Effect of β-cyclodextrin on the CO release kinetics and antimicrobial activity of [NEt4][Mo(CO)5Br

In the present work the possibility of improving the solubility, bioavailability and bactericidal activity of the carbon monoxide releasing molecule (CORM) [Mo(CO)5Br]− (as its tetraethylammonium salt) (1) by solvent-free co-grinding with β-cyclodextrin (βCD) in a planetary ball mill was investigated. Data obtained by FT-IR spectroscopy, Raman spectroscopy, powder X-ray diffraction and thermogravimetric analysis showed that, other than a small decrease in the crystallinity of 1, the co-grinding process produced a finely dispersed physical mixture of crystalline CORM and crystalline excipient. The aqueous solubility of 1 in the 1-βCD product was enhanced with respect to sparingly soluble pure 1, which might be ascribed to increased wettability and a CD-CORM interaction in solution. Investigation of the CO release kinetics by the standard myoglobin assay showed that the half-life of CO release increased from ca. 6 min for 1 to ca. 19 min for 1-βCD, while the number of equivalents released decreased from 3.2 to 1.8. The antibacterial properties of 1 and 1-βCD were evaluated using the broth microdilution method to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against the model Gram-negative bacterium, Escherichia coli. The compounds showed similar growth inhibitory (MIC values of 200 μM) and bactericidal (MBC/MIC ≈ 2) effects. Bacterial viability assays corroborated the MBC/MIC studies, showing 3 logs (99.9% of relative light units - RLU) reduction in viable cell count after 15 min exposure to 2 × MIC. Although the CORM-CD system displays a lengthening of the half-life of CO release and a decrease in the CO release efficiency relative to 1, the co-grinding with βCD does not affect the bactericidal activity of the CORM. Overall, the βCD could be a suitable excipient for the development of immediate-release formulations of CORMs like the pentacarbonyl complex 1.

Synergy with farnesol rejuvenates colistin activity against Colistin-resistant Gram-negative bacteria in vitro and in vivo

Colistin (COL) is considered the last line of treatment against infections due to multidrug-resistant (MDR) Gram-negative bacteria (GNB). However, the increasing number of colistin-resistant (COL-R) bacteria is a great threat to public health. In this study, a strategy of combining farnesol (FAR), which has anti-inflammatory and antitumor properties, with COL to restart COL activity was proposed. The synergistic effect of FAR combined with COL against COL-R GNB in vivo and in vitro were investigated. The excellent synergistic antibacterial activity of the COL–FAR combination was confirmed by performing the checkerboard assay, time-killing assay, and LIVE/DEAD bacterial cell viability assay. Crystal violet staining and scanning electron microscopy results showed that COL–FAR prevented biofilm formation and eradicated pre-existing mature biofilm. Cytotoxicity assay showed that FAR at 64 µg/mL was not cytotoxic to RAW264.7 cells. In vivo infection experiments showed that COL–FAR increased the survival rate of infected Galleria mellonella and decreased the bacterial load in a mouse thigh infection model. These results indicate that COL–FAR is a potentially effective therapeutic option for combating COL-R GNB infections.

Chitosan can improve antimicrobial treatment independently of bacterial lifestyle, biofilm biomass intensity and antibiotic resistance pattern in non-aureus staphylococci (NAS) isolated from bovine clinical mastitis.

Bovine mastitis is the most frequent and costly disease that affects dairy cattle. Non-aureus staphylococci (NAS) are currently one of the main pathogens associated with difficult-to-treat intramammary infections. Biofilm is an important virulence factor that can protect bacteria against antimicrobial treatment and prevent their recognition by the host's immune system. Previously, we found that chronic mastitis isolates which were refractory to antibiotic therapy developed strong biofilm biomass. Now, we evaluated the influence of biofilm biomass intensity on the antibiotic resistance pattern in strong and weak biofilm-forming NAS isolates from clinical mastitis. We also assessed the effect of cloxacillin (Clx) and chitosan (Ch), either alone or in combination, on NAS isolates with different lifestyles and abilities to form biofilm. The antibiotic resistance pattern was not the same in strong and weak biofilm producers, and there was a significant association (p ≤ 0.01) between biofilm biomass intensity and antibiotic resistance. Bacterial viability assays showed that a similar antibiotic concentration was effective at killing both groups when they grew planktonically. In contrast, within biofilm the concentrations needed to eliminate strong producers were 16 to 128 times those needed for weak producers, and more than 1,000 times those required for planktonic cultures. Moreover, Ch alone or combined with Clx had significant antimicrobial activity, and represented an improvement over the activity of the antibiotic on its own, independently of the bacterial lifestyle, the biofilm biomass intensity or the antibiotic resistance pattern. In conclusion, the degree of protection conferred by biofilm against antibiotics appears to be associated with the intensity of its biomass, but treatment with Ch might be able to help counteract it. These findings suggest that bacterial biomass should be considered when designing new antimicrobial therapies aimed at reducing antibiotic concentrations while improving cure rates.

Effects of Structural Variations on Antibacterial Properties for Conjugated Diynes Generated through Glaser Hay Couplings.

Antibiotic resistance is a growing problem facing global societies today. Many new antibiotics are derivatized versions of already existing antibiotics, which allows for antibiotic resistance to arise. To combat this issue, new antibiotics with different core structures need to be elucidated. Asymmetrical polyacetylenes have been isolated from natural products and they have previously been demonstrated to exhibit antimicrobial and antibacterial activity; however, their synthetic preparation has not made them easily amenable to rapid derivatization for SAR studies. Using a combination of solution and solid-supported chemistries, an array of diynes inspired by a known natural product were prepared and assessed for antibacterial activity. Ultimately, several compounds were identified with improved activity in bacterial viability assays. Moreover, some compounds were discovered that displayed a degree of specificity for E. coli over P. fluorescens and vice versa. These new compounds show promise, and further investigation is needed to pinpoint the specific structural components that elicit biological activity.

Measuring Single Bacterial Viability in Optical Traps with a Power Sweeping Technique.

Assessing bacterial viability is crucial in public health, food safety, environmental microbiology, and other relevant fields. The classical agar plate counting method and the popular dye-based assays have shown their strengths, but they also have limitations including high time consumption, relatively complex sample preparations, and cytotoxicity. In this work, we present a new bacterial viability assay based on optical tweezers utilizing a power sweeping strategy. By monitoring and analyzing bacterial nanomotion in optical traps under different trapping laser powers, the slope of the proportionality between the quantified extent of motion and the trapping laser power was defined as the mobility restriction coefficient (MRC) to quantify bacterial viability. We first established a firm correlation between the viability and MRC by measuring alive and dead Escherichia coli and Photobacterium phosphoreum. Then the capability of real-time long-term characterization of the assay was validated by measuring the viability of individual P. phosphoreum while regulating the viability with an inactivation light. Notably, a 'spinning-induced stabilization' mechanism was proposed to explain the surprising increase of apparent bacterial mobility after inactivation. Overall, the assay was proved to be a reliable label-free bacterial viability assay at a single-cell level, which holds potential in antibiotic susceptibility testing, drug screening, and rapid diagnostics.

Novel Antimicrobial Peptide "Octoprohibitin" against Multidrug Resistant Acinetobacter baumannii.

Octoprohibitin is a synthetic antimicrobial peptide (AMP), derived from the prohibitin-2 gene of Octopus minor. It showed substantial activity against multidrug resistant (MDR) Acinetobacter baumannii with a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 200 and 400 µg/mL, respectively. Time-kill kinetics and bacterial viability assays confirmed the concentration-dependent antibacterial activity of octoprohibitin against A. baumannii. The morphology and ultrastructure of A. baumannii were altered by treatment with octoprohibitin at the MIC and MBC levels. Furthermore, propidium iodide-fluorescein diacetate (PI-FDA) staining and 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) staining of octoprohibitin-treated A. baumannii revealed membrane permeability alterations and reactive oxygen species (ROS) generation, respectively. Agarose gel retardation results confirmed the DNA-binding ability of octoprohibitin to the genomic DNA of A. baumannii. Furthermore, octoprohibitin showed concentration-dependent inhibition of biofilm formation and eradication. The minimum biofilm inhibition concentration (MBIC) and minimum biofilm eradication concentration (MBEC) of octoprohibitin were 1000 and 1460 µg/mL, respectively. Octoprohibitin produced no significant cytotoxicity up to 800 µg/mL, and no hemolysis was observed up to 400 µg/mL. Furthermore, in vivo analysis in an A. baumannii-infected zebrafish model confirmed the effective bactericidal activity of octoprohibitin with higher cumulative survival percent (46.6%) and fewer pathological signs. Histological analysis showed reduced alterations in the gut, kidney, and gill tissues in the octoprohibitin-treated group compared with those in the phosphate-buffered saline (PBS)-treated group. In conclusion, our results suggest that octoprohibitin is a potential antibacterial and antibiofilm agent against MDR A. baumannii.

Quantification and Degradation of 2,2-Dibromo-3-Nitrilopropionamide (DBNPA) in Bioethanol Fermentation Coproducts

2,2-Dibromo-3-nitrilopropionamide (DBNPA) has been used as a biocide in industrial water applications due to its instantaneous antimicrobial activity and rapid chemical breakdown. In this study, DBNPA is considered a potential alternative for antibiotics used for bacterial control during corn-to-ethanol fermentation. A method using LC/MS/MS was developed to accurately quantify DBNPA in water. When this method was applied to quantify DBNPA concentration in a fermentation matrix, DBNPA was found to be unstable and to decay rapidly, preventing validation of the method or quantitation. This method was then used to evaluate the degradation rate of DBNPA in whole stillage, which is the nonvolatile residue produced by removal of ethanol from corn-based fermentation beer by distillation through the relative decrease in measured signal. In addition, a method was developed and validated to quantify bromide, one of the degradation products of DBNPA, in whole stillage using LC/MS/MS. The degradation rate of DBNPA in whole stillage was found to display first-order kinetics with a calculated half-life of 85 min. Laboratory analytical chemistry results on DBNPA degradation were confirmed based on a bacterial viability assay in field trials.

Disulfiram Enhances the Activity of Polymyxin B Against Klebsiella pneumoniae by Inhibiting Lipid A Modification

BackgroundThe use of antibiotic adjuvants is a complementary strategy to the development of new antibiotics. The essential role of the ArnA dehydrogenase domain (ArnA_DH) in the addition of 4-amino-L-arabinose (L-Ara4N) to lipid A makes it a potential target in polymyxin adjuvant design.PurposeThis study aimed to identify a dehydrogenase inhibitor that enhances the antibacterial effect of polymyxin B (PB) and to further understand the mechanism of this drug combination.MethodsA susceptible K. pneumoniae strain, ATCC13883, was used to screen a dehydrogenase inhibitor library based on 3-(4,5)-dimethylthiazol(-z-y1)-2,5-diphenyltetrazolium bromide (MTT) and chequerboard assays. The protein- and cell-based effects of disulfiram (DSF) on ArnA activity were assessed, and the transcription levels of genes in the arn operon were evaluated by quantitative real-time polymerase chain reaction (qRT–PCR). Lipid A was isolated, and a structural analysis was performed. The cell wall function was evaluated through membrane integrity and bacterial viability assays. The in vivo antibacterial activity was evaluated using a mouse pulmonary infection model.ResultsWe screened a dehydrogenase inhibitor library and found that the anti-alcoholism drug DSF significantly enhanced the antibacterial activity of PB in vitro and in vivo. The protein-based enzyme activity assay showed that DSF exerted no direct effect on the dehydrogenase activity of ArnA. Treatment with the combination of DSF and PB but not with PB alone decreased both the transcription level of genes in the arn operon and the modification level of lipid A. DSF also strengthened the disruption of the cell membrane integrity of PB. Moreover, the enhanced PB antibacterial activity was effective against clinical PB-resistant strains.ConclusionWe identified a new drug combination that can be used to reduce the necessary dosage of PB and overcome PB resistance, and this drug combination has good prospects for clinical application.

Characterization of the selective binding of modified chitosan nanoparticles to Gram-negative bacteria strains

Chitosan is a nature-sourced polysaccharide widely used in numerous applications. The antibacterial potential of chitosan has attracted researchers to further develop and utilize this polymer for the formation of biocompatible antibacterial agents for both the food and healthcare industries. The tested hypothesis in this study is that modified N-alkylaminated chitosan nanoparticles (CNPs) have selective binding properties to Gram-negative bacteria strains that result in bacterial aggregation. Various bacterial strains were tested of five Gram-negative bacteria including Erwinia carotovora, Escherichia coli, Pseudomonas aeruginosa, Salmonella, and Serratia marcescens, as well as three Gram-positive bacteria strains including Bacillus licheniformis, Bacillus megaterium, and Bacillus subtilis. The fluorescence microscopy characterization showed that the presence of CNPs caused the aggregation of Escherichia coli bacteria cells, where modified CNPs with a shorter chain length of the substituent caused a higher aggregation effect. Moreover, it was found that the CNPs exhibited a selective binding behavior to Gram-negative as compared to Gram-positive bacteria strains, mainly to Escherichia coli and Salmonella. Also, the scanning electron microscopy characterization showed that CNPs exhibited selective binding to Gram-negative bacteria, which was especially understood when both Gram-negative and Gram-positive bacteria strains were within the same sample. In addition, the bacterial viability assay suggests that CNPs with a lower degree of substitution have a higher inhibitory effect on bacterial growth. CNPs with longer side chains had a less inhibitory effect on the bacterial growth of Gram-negative strains, where a concentration-dependent response pattern was only seen for the cases of Gram-negative strains, and not for the case of Gram-positive strain. To conclude, the further understanding of the selective binding of CNPs to Gram-negative bacteria strains may produce new opportunities for the discovery and characterization of effective antibacterial agents.

Select Phytochemicals Reduce Campylobacter jejuni in Postharvest Poultry and Modulate the Virulence Attributes of C. jejuni.

Consumption or handling of poultry and poultry products contaminated with Campylobacter species are a leading cause of foodborne illness in humans. Current strategies employed to reduce Campylobacter in live chickens provide inconsistent results indicating the need for an alternative approach. This study investigated the efficacy of phytochemicals, namely, turmeric, curcumin, allyl sulfide, garlic oil, and ginger oil, to reduce Campylobacter jejuni in postharvest poultry and sought to delineate the underlying mechanisms of action. Two experiments were conducted on the thigh skin of the chicken, and each experiment was repeated twice. Samples were inoculated with 50 μl (∼107 CFU/sample) of C. jejuni strain S-8 and allowed to adhere for 30 min. Skin samples were dipped into their respective prechilled treatment solutions (0.25 and 0.5% in experiments 1 and 2, respectively) at 4°C for an hour to simulate chilling tank treatment, followed by plating to enumerate C. jejuni (n = 3 samples/treatment/trial). The mechanisms of action(s) were investigated using subinhibitory concentration (SIC) in adhesion, quorum sensing, and gene expression analyses. Adhesion assay was conducted on the monolayers of ATCC CRL-1590 chicken embryo cells challenged with C. jejuni and incubated in the presence or absence of phytochemicals for 1.5 h, followed by plating to enumerate adhered C. jejuni. The effects of phytochemicals on quorum sensing and cell viability were investigated using Vibrio harveyi bioluminescence and LIVE/Dead BacLightTM bacterial viability assays, respectively. In addition, droplet digital PCR determined the gene expression analyses of C. jejuni exposed to phytochemicals. Data were analyzed by GraphPad Prism version 9. C. jejuni counts were reduced by 1.0–1.5 Log CFU/sample with garlic oil or ginger oil at 0.25 and 0.5% (p < 0.05). The selected phytochemicals (except curcumin) reduced the adhesion of C. jejuni to chicken embryo cells (p < 0.05). In addition, all the phytochemicals at SIC reduced quorum sensing of C. jejuni (p < 0.05). The cell viability test revealed that cells treated with 0.25% of phytochemicals had compromised cell membranes indicating this as a mechanism that phytochemicals use to damage/kill C. jejuni. This study supports that the application of phytochemicals in postharvest poultry would significantly reduce C. jejuni in poultry meat.

Octopromycin: Antibacterial and antibiofilm functions of a novel peptide derived from Octopus minor against multidrug-resistant Acinetobacter baumannii

The emergence of carbapenem-resistant Acinetobacter baumannii has increased the risk of nosocomial infections, which pose a huge health threat. There is an urgent need to develop alternative therapies, including broad-spectrum antimicrobial peptides. In this study, we designed, characterized, and studied the antibacterial, antibiofilm effects and possible mode of actions of a novel synthetic peptide Octopromycin, derived from the proline-rich protein 5 of Octopus minor. Octopromycin consists of 38 amino acids, (+5) net positive charge, high hydrophobic residue ratio (36%), and two α-helix secondary structures. The minimum inhibitory concentration and minimum bactericidal concentration against A. baumannii were 50 and 200 μg/mL, respectively. Time-kill kinetics and bacterial viability assays confirmed the concentration-dependent antibacterial activity of Octopromycin. Field emission scanning electron microscopy images clearly showed ultrastructural alterations in Octopromycin-treated A. baumannii cells. Propidium iodide penetrated into Octopromycin-treated A. baumannii cells, demonstrating the loss of cell membrane integrity. Octopromycin treatment increased the production of reactive oxygen species in a concentration-dependent manner, and it inhibited the biofilm formation and showed biofilm eradication activity against A. baumannii. In vitro and in vivo safety evaluation revealed that Octopromycin was nontoxic to HEK293T and Raw 264.7 cells (<400 μg/mL), as well as mice red blood cells (<300 μg/mL), and zebrafish embryos (<4 μg/mL). An in vivo study results revealed that the A. baumannii-infected fish treated with Octopromycin exhibited a significantly higher relative percent survival (37.5%) than the infected mock-treated fish with PBS (16.6%). Furthermore, a decreased bacterial load and fewer alterations in histological analysis confirmed the successful control of A. baumannii by Octopromycin in vivo. Collectively, the results indicate that the antibacterial peptide Octopromycin may achieve rapid control of A. baumannii through multi-target interactions; it presents a desirable therapeutic option for the prevention and control of the infections.

Physicochemical and antibacterial characterization of Aspergillus sp. filtrate-reduced graphene oxide

Graphene-based nanomaterials are rapidly emerging as antimicrobial agents against a range of pathogenic microorganisms. In the present study, we have demonstrated the synthesis of reduced graphene oxide (RGO) using Aspergillus sp. filtrate as a reducing agent. The obtained Aspergillus sp. filtrate-RGO (AF-RGO) was characterized by ultraviolet–visible absorption spectroscopy, dynamic light scattering, X-ray diffraction, Fourier-transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, and atomic force spectroscopy to obtain details about the size, composition, structure, and morphology. The characterization confirmed the reduction of graphene oxide, the restoration of sp2 conjugation, and the formation of few-layered RGO sheets. The surface charge and stability of the AF-RGO in aqueous media were examined by zeta potential measurements. The thermal stability of AF-RGO was increased significantly due to the restoration of the graphitic sp2 network. The antibacterial activity of AF-RGO was evaluated using the agar well diffusion method, broth microdilution method, and Live/Dead bacterial viability assay. AF-RGO exhibited good antibacterial activities against pathogenic Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 15 and 30 mg/ml, respectively. Therefore, fungal filtrate-mediated synthesis of RGO is a promising antibacterial carbon nanomaterial for a range of biomedical and nano(bio)technological applications.

Synthesis, characterization and application of Co/Co3O4 nanocomposites as an effective photocatalyst for discoloration of organic dye contaminants in wastewater and antibacterial properties

Contamination of surface water with dye chemical compounds and/or biological substances, even in small amounts, can affect the health of humans and other organisms. The photocatalytic oxidation process has been considered as a commercial technique to remove environmental pollutants. In the current study, we reported the synthesis of Co/Co3O4 nanocomposites investigated for their photocatalytic and antimicrobial activities. The affecting parameters (various surfactants and calcination) on the synthesis process were investigated. The synthesis of Co/Co3O4 nanocomposites was confirmed via methodical characterization such as SEM, FT-IR, XRD, VSM, EDX, CV and DRS investigations. Well diffusion assay and bacterial cell viability assay were executed against clinical pathogens to prepare the antibacterial activity of synthesized Co/Co3O4 nanocomposites. Also, the photocatalytic activity of nano-catalysts was concluded against the organic colors (acid blue 92 and acid red 151). Cobalt oxide nanoparticles (NPs) synthesized in the presence of SDBS as an anionic template showed the highest decolorization of 93% over acid red 151 after 120 min of illumination. The results showed a minimum bacterial inhibitory concentration for bacteria P. aeruginosa, and B. subtilis is about 31.25 µg/mL and 125 µg/mL, respectively. The Co/Co3O4 nanocomposites exhibited vigorous antibacterial activity against gram-negative microorganisms mentioned like Pseudomonas aeruginosa.

Sustained Antibacterial Effect and Wear Behavior of Quaternary Ammonium Contact-Killing Dental Polymers after One-Year of Hydrolytic Degradation

This study intended to investigate the long-term antibacterial effect, mechanical performance, and surface topography of new anticaries dental composites. While most artificial aging studies of dental resins lasted for 30–90 days, this study prolonged the water-aging to one year to be more clinically relevant. The base resin was loaded with dimethylaminohexadecyl methacrylate (DMAHDM) at 3 or 5 wt.% and nano-sized amorphous calcium phosphate (NACP) at 20 wt.%. Composites were subjected to one-year water storage and wear. Following water aging, samples were evaluated for flexural strength, elastic modulus, and microbiological assays. Biofilm plate counting method, metabolic assay, colorimetric quantification of lactic acid, and Baclight bacterial viability assay were measured after one year. Topography changes (ΔRa, ΔRq, ΔRv, ΔRt) were examined after wear and observed by scanning electron microscopy. Biofilm assays and topography changes data were analyzed via one-way ANOVA and Tukey’s tests. Mechanical properties and normalized data were verified using a t-test. The flexural strength values for the formulations that contained 5% DMAHDM-20% NACP, 3% DMAHDM, and 5% DMAHDM were reduced significantly (p &lt; 0.05) in relation to the baseline but the values were still above the ISO standards. No significant differences were observed between the groups concerning the topography changes, except for the ΔRt, where there was a significant increase in the 5% DMAHDM-20% NACP group. All the groups demonstrated robust biofilm-inhibition, with slightly reduced antibacterial properties following water aging. The aged samples reduced the total microorganisms, total streptococci, and mutans streptococci by 1.5 to 3-log, compared to the experimental control. The new formulations containing DMAHDM and NACP were able to sustain the antibacterial performance after one-year of aging. Mechanical properties and surface topography were slightly affected over time.

Paracetamol modulates biofilm formation in Staphylococcus aureus clonal complex 8 strains

Staphylococcus aureus biofilms are a major problem in modern healthcare due to their resistance to immune system defenses and antibiotic treatments. Certain analgesic agents are able to modulate S. aureus biofilm formation, but currently no evidence exists if paracetamol, often combined with antibiotic treatment, also has this effect. Therefore, we aimed to investigate if paracetamol can modulate S. aureus biofilm formation. Considering that certain regulatory pathways for biofilm formation and virulence factor production by S. aureus are linked, we further investigated the effect of paracetamol on immune modulator production. The in vitro biofilm mass of 21 S. aureus strains from 9 genetic backgrounds was measured in the presence of paracetamol. Based on biofilm mass quantity, we further investigated paracetamol-induced biofilm alterations using a bacterial viability assay combined with N-Acetylglucosamine staining. Isothermal microcalorimetry was used to monitor the effect of paracetamol on bacterial metabolism within biofilms and green fluorescent protein (GFP) promoter fusion technology for transcription of staphylococcal complement inhibitor (SCIN). Clinically relevant concentrations of paracetamol enhanced biofilm formation particularly among strains belonging to clonal complex 8 (CC8), but had minimal effect on S. aureus planktonic growth. The increase of biofilm mass can be attributed to the marked increase of N-Acetylglucosamine containing components of the extracellular matrix, presumably polysaccharide intercellular adhesion. Biofilms of RN6390A (CC8) showed a significant increase in the immune modulator SCIN transcription during co-incubation with low concentrations of paracetamol. Our data indicate that paracetamol can enhance biofilm formation. The clinical relevance needs to be further investigated.