Minimum Bactericidal Concentration Research Articles

Antibacterial and Antibiofilm Potential of Bacterial Cellulose Hydrogel Containing Vancomycin against Multidrug-Resistant Staphylococcus aureus and Staphylococcus epidermidis.

The present study aimed to evaluate the in vitro antibacterial and antibiofilm activity of bacterial cellulose hydrogel produced by Zoogloea sp. (HYDROGEL) containing vancomycin (VAN) against bacterial strains that cause wound infections, such as multidrug-resistant (MDR) Staphylococcus aureus and Staphylococcus epidermidis. Initially, HYDROGEL was obtained from sugar cane molasses, and scanning electron microscopy (SEM) was performed to determine morphological characteristics. Then, VAN was incorporated into HYDROGEL (VAN-HYDROGEL). The antibacterial activity of VAN, HYDROGEL, and VAN-HYDROGEL was assessed using the broth microdilution method to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) against methicillin-sensitive S. aureus (MSSA) ATCC 25923, methicillin-resistant S. aureus (MRSA) ATCC 33591, S. epidermidis INCQS 00016 (ATCC 12228), five clinical isolates of MRSA, and nine clinical isolates of methicillin-resistant S. epidermidis, following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Additionally, the antibacterial activity of VAN, HYDROGEL, and VAN-HYDROGEL was studied using the time-kill assay. Subsequently, the antibiofilm activity of VAN, HYDROGEL, and VAN-HYDROGEL was evaluated using crystal violet and Congo red methods, as well as SEM analysis. VAN and VAN-HYDROGEL showed bacteriostatic and bactericidal activity against MRSA and methicillin-resistant S. epidermidis strains. HYDROGEL did not show any antibacterial activity. Analysis of the time-kill assay indicated that HYDROGEL maintained the antibacterial efficacy of VAN, highlighting its efficiency as a promising carrier. Regarding antibiofilm activity, VAN and HYDROGEL inhibited biofilm formation but did not demonstrate biofilm eradication activity against methicillin-resistant S. aureus and S. epidermidis strains. However, it was observed that the biofilm eradication potential of VAN was enhanced after incorporation into HYDROGEL, a result also proven through images obtained by SEM. From the methods carried out in this study, it was possible to observe that HYDROGEL preserved the antibacterial activity of vancomycin, aside from exhibiting antibiofilm activity and enhancing the antibiofilm effect of VAN. In conclusion, this study demonstrated the potential of HYDROGEL as a candidate and/or vehicle for antibiotics against MDR bacteria that cause wound infections.

Biosynthesis, characterization and antimicrobial activities of zinc oxide nanoparticles from leaf and seed extracts of Malva neglecta Wallr

Recent years have witnessed a growing embrace of green chemistry for nanoparticle manufacturing due to its time and energy efficiency compared to conventional methods. This research aimed to biosynthesize zinc oxide nanoparticles (ZnO NPs) using environmentally friendly aqueous extracts from Malva neglecta leaves and seeds as reducing and stabilizing agents. Various analytical methods, including UV–Vis spectrophotometry, FTIR, XRD, TGA, EDX, and TEM, confirmed the synthesis and characterization of ZnO NPs. Their antioxidant activity was assessed using the DPPH assay, while antibacterial properties were examined against S. aureus and E. coli, along with determining minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. FTIR spectra indicated ZnO bonding and surfactant molecule adsorption on ZnO NPs’ surface. TEM revealed spherical and hexagonal ZnO NPs with an average size of 40–50 nm. This green synthesis method with Malva neglecta extracts is eco-friendly, cost-effective, and safe, producing antibacterial nanoparticles without adverse effects.

Evaluation of the mechanistic basis for the antibacterial activity of ursolic acid against Staphylococcus aureus.

The antibiotics are generally regarded as the first choice approach to treat dairy mastitis, targeting the public health problems associated with the food safety and the emergence of antibioticresistant bacteria. The objective of the study was to evaluate the antibacterial efficacy of ursolic acid (UA) when used to treat Staphylococcus aureus and other isolates associated with bovine mastitis and to clarify the mechanistic basis for these effects. The bacteriostatic properties of UA extracted from Rosmarinus officinalis L. at four different purity levels were assessed by calculating minimum inhibitory concentration (MIC) values, while the synergistic effects of combining 98% UA with antibiotics were evaluated by measuring the fractional inhibitory concentration index (FICI). Changes in biofilm formation and the growth curves of the clinical isolates were assessed to clarify the bacteriostatic effect of UA. Furthermore, the cell wall integrity, protein synthesis, and reactive oxygen species (ROS) production were assessed to determine the antibacterial mechanism of UA treatment. Ultimately, UA was revealed to exhibit robust activity against Gram-positive bacteria including S. aureus (ATCC 25923), Streptococcus dysgalactiae (ATCC27957), Streptococcus agalactiae (ATCC13813), Enterococcus faecalis (ATCC29212), and Streptococcus mutans (ATCC25175). However, it did not affect Escherichia coli (ATCC 25922). The MIC values of UA preparations that were 98, 50, 30, and 10% pure against S. aureus were 39, 312, 625, and 625 μg/mL, respectively, whereas the corresponding MIC for E. coli was >5,000 μg/mL. The minimum bactericidal concentrations of 98% UA when used to treat three clinical S. aureus isolates (S4, S5, and S6) were 78, 78, and 156 μg/mL, respectively. Levels of biofilm formation for clinical S. aureus isolates decreased with increasing 98% UA concentrations. Above the MIC dose, UA treatment resulted in the dissolution of bacterial cell walls and membranes, with cells becoming irregularly shaped and exhibiting markedly impaired intracellular protein synthesis. S. aureus treated with 98% UA was able to rapidly promote intracellular ROS biogenesis. Together, these data highlight the promising utility of UA as a compound that can be used together with other antibiotics for the treatment of infections caused by S. aureus.

Treatment of subclinical mastitis in dairy goats experimentally infected with Staphylococcus aureus using natural therapeutic protocol.

In herds of dairy goats, mastitis represents a major health and economic problem due to the multiresistance of some microorganisms. In this context, the study aimed to determine the potential of antimicrobial action and antibiofilm of the crude ethanolic extract (CEE) of Hymenaea martiana (jatobá) leaves, as well its fractions, on Staphylococcus sp isolated from bacterial cultures of goat milk. In vitro assays were performed to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC), as well as tests of the effect of CEE on biofilm formation and quantification and the consolidated biofilm. The experimental infection was performed in two groups, each consisting of five goat. Experimental Group 1 (G1) consisted of five females treated with an intramammary ointment based on the CEE, at a concentration of 5%. Experimental Group 2 (G2) consisted of five females treated with a commercial intramammary ointment based on gentamicin, once a day, for six consecutive days. The diagnosis of mastitis was performed using a bacterial culture. The dichloromethane fraction of CEE was the one with the lowest concentrations of MBC, ranging from 195.3 to 781µg / ml. Concerning to the biofilm, interference of the tested extract was observed for two isolates. In the present study, the ointment prepared from H. martiana extract (jatobá) was able to reduce bacterial infection in mammary glands experimentally infected with S. aureus. Antibacterial activity may be related to the classes of secondary metabolites found.

Silibinin-loaded chitosan-capped silver nanoparticles exhibit potent antimicrobial, antibiofilm, and anti-inflammatory activity against drug-resistant nosocomial pathogens

Nanoparticles capped with natural products can be a cost-effective alternative to treat drug-resistant nosocomial infections. Therefore, silibinin-loaded chitosan-capped silver nanoparticles (S-C@AgNPs) were synthesized to evaluate their antimicrobial and anti-inflammatory potential. The S-C@AgNPs plasmon peak was found at 430 nm and had a particle size distribution of about 130 nm with an average hydrodynamic diameter of 101.37 nm. The Scanning Electron Microscopy images showed the presence of sphere-shaped homogeneous nanoparticles. The Fourier Transform Infrared Spectroscopy analysis confirmed the loading of silibinin and chitosan on the AgNPs surface. The minimum inhibitory concentration of the S-C@AgNPs was reported between 3.12 μg/ml to 12.5 μg/ml and a minimum bactericidal concentration between 6.25 μg/ml to 25 μg/ml against drug-resistant nosocomial pathogens. Moreover, concentration-dependent significant inhibition of the biofilm formation was reported against P. aeruginosa (70.21%) and K. pneumoniae (71.02%) at 30 μg/ml, and the highest destruction of preformed biofilm was observed at 100 μg/ml against P. aeruginosa (89.74%) and K. pneumoniae (77.65%) as compared to individual bacterial control. Additionally, the fluorescence live/dead assay for bacterial biofilm confirmed that 100 µg/ml effectively inhibits the biofilm formed by these pathogens. S-C@AgNPs also showed anti-inflammatory activity, which is evident by the significant decrease in the proinflammatory cytokines and chemokines level in THP1 cells treated with LPS. This study concluded that S-C@AgNPs have potent antimicrobial, antibiofilm, and anti-inflammatory properties and could be a potential option for treating drug resistant nosocomial infections.

Biomimetic protein structural transitions regulate activation and inhibition of the broad-spectrum bactericidal activity of cationic nanoparticles

The development of cationic polymers as alternative materials to antibiotics necessitates addressing the challenge of balancing their antimicrobial activity and toxicity. Here we propose a precise switching strategy inspired by biomimetic voltage-gated ion channels, enabling controlled activation and inhibition of cationic antimicrobial functions through protein conformational transitions in diverse physiological environments. Following thermodynamic studies on the specific recognition between mannose end groups on polycations and concanavalin A (ConA), we synthesized a type of ConA-polycation nanoparticle. The nanoparticle was inhibited under neutral conditions, with cationic moieties shielded by ConA's β-sheet. This shielding suppresses their antimicrobial activity, thereby ensuring satisfactory biocompatibility. In mildly acidic environments, however, the transition of a portion of ConA to an α-helix conformation exposed cations at the particle periphery, activating antibacterial functionality. Compared to inhibited nanoparticles, those in the activated state exhibited a 32–256 times reduction in the minimum bactericidal concentration against bacteria and fungi (2–16 µg/mL). In a murine acute pulmonary infection model, intravenous administration of inhibited nanoparticles effectively reduced bacterial counts by 4-log within 12 h. The biomimetic design, regulating cationic antimicrobial functionality through the alteration in protein secondary structure, significantly retards bacterial resistance development, holding great promise for intelligent antimicrobial materials. Statement of significanceCationic antimicrobial polymers exhibit advantages distinct from antibiotics due to their lower propensity for resistance development. However, the presence of cationic moieties also poses a threat to healthy cells and tissues, significantly constraining their potential for clinical applications. To address this challenge, we propose a biomimetic strategy that mimics voltage-gated ion channels to activate the antimicrobial functionality of cations selectively in bacterial environments through the conformational transitions of proteins between β–sheets and α–helices. In healthy tissues, the antimicrobial functionality is inhibited, ensuring satisfactory biocompatibility. Antimicrobial cationic materials capable of intelligent switching between an activated state and an inhibited state in response to environmental changes offer an effective strategy to prevent the development of resistance and mitigate potential side effects.

Diacetylcurcumin: a novel strategy against Enterococcus faecalis biofilm in root canal disinfection.

Aim: We evaluated Diacetylcurcumin (DAC), a derivative of curcumin, for its antibacterial and antibiofilm properties against Enterococcus faecalis. Methods: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration were determined, along with antibiofilm potential and toxicity in Galleria mellonella. Additionally, in silico computational analysis was performed to understand its mechanisms of action. Results & conclusion: DAC demonstrated significant antibacterial effects, with MIC and MBC values of 15.6 and 31.25μg/ml, respectively, and reduced biofilm formation. A synergistic effect, reducing biofilm by 77%, was observed when combined with calcium hydroxide. G. mellonella toxicity tests confirmed DAC's safety at tested concentrations, suggesting its potential for use in root canal disinfection products.

Antibacterial activity of aqueous and methanol extracts of <i>Jatropha tanjorensis</i> and <i>Adansonia digitata</i> against selected clinical bacteria pathogens

The powdered plant parts (bark, stem, and leaf) were extracted with methanol, and cold and hot water using the maceration method. A modified agar well diffusion technique was used for the assessment of the antibacterial activities of the aqueous and methanol extracts of J. tanjorensis and A. digitata. The methanol extract of A. digitata bark gave the highest yield of 75 % while the lowest yield was observed with the cold water extract of A. digitata bark of 33 %. The phytochemical screening showed an abundance of alkaloids, terpernoids, saponins, flavonoids, tannin, glycoside, and phenol. The bark, leaf, and stem of J. tanjorensis and A. digitata showed varying degrees of antibacterial activities. against the Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Proteus spp, and Klebsiella spp) and Gram-positive bacteria (Staphylococcus aureus, Streptococcus pyogenes and Streptococcus pneumonia). The bark, stem, and leaf extract of J. tanjorenis significantly (p < 0.05) inhibited all the pathogens except P. aeruginosa and S. aureus.The result of the minimum bactericidal concentration MBC for the combined extracts of A. digitata and J. tanjorenis subfractions showed that the extracts have greater antibacterial activities at concentrations not lesser than 100 mg/ml. Thus, J. tanjorensis and A. digitata could be used as potent herbal remedies to mitigate the adverse effects of Gram-negative and positive clinical pathogens. Thus, the aim of this study was to evaluate the antibacterial activity of aqueous and methanol extracts of Jatropha tanjorensis and Adansonia digitata against selected Clinical bacteria pathogens.

Magnetic separation, sunlight-driven photocatalytic activity, and antibacterial studies of Sm-doped Co0.33Mg0.33Ni0.33Fe2O4 nanoparticles.

Magnetic nanoparticles have emerged as a promising tool for wastewater treatment due to their unique properties. In this regard, Co0.33Mg0.33Ni0.33SmxFe2-xO4 (0.00 x 0.08) nanoparticles were prepared to examine their magnetic separation efficiency (MSE), photocatalytic, antibacterial, and antibiofilm performances. Pure nanoparticles, having the highest saturation magnetization (Ms = 31.87emu/g), exhibit the highest MSE, where 95.6% of nanoparticles were separated after 20min of applying a magnetic field of 150 mT. The catalytic performance of the prepared samples is examined by the photodegradation of rhodamine B (RhB) dye exposed to direct sunlight radiation. Improved photocatalytic activity is exhibited by Co0.33Mg0.33Ni0.33Sm0.04Fe1.96O4 nanoparticles, labeled as Sm0.04, where the rate of the degradation reaction is enhanced by 4.1 times compared to pure nanoparticles. Rising the pH and reaction temperature improves the rate of the photodegradation reaction of RhB. The incorporation of 15 wt% reduced graphene oxide (rGO) with Sm0.04 enhanced the rate of the reaction by 1.7 and 2.4 times compared with pure Sm0.04 sample and rGO, respectively. The antibacterial and antibiofilm activities against Escherichia coli, Leclercia adecarboxylata, Staphylococcus aureus, and Enterococcus faecium are assessed by the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) broth microdilution, the agar well diffusion, the time-kill assays, the biofilm formation, and destruction assays. The bacteria used in these assessments are isolated from wastewater. The nanoparticles exhibit a bacteriostatic activity, with a better effect against the Gram-positive isolates. Co0.33Mg0.33Ni0.33SmxFe2O4 (x = 0.00) nanoparticles have the best effect. The effect is exerted after 2-3h of incubation. Gram-positive biofilms are more sensitive to nanoparticles.

Identification of Secondary Metabolites from Actinomycetes Isolated from the Hilly Region of Nepal

Actinomycetes are a unique class of Gram-positive bacteria with a fungus-like filamentous mycelium. They are often called ‘antibiotics factories’ because they can produce many potent secondary metabolites. At present, multidrug-resistant (MDR) pathogens are the main problem for humans and animals and to overcome this problem, more potent antibiotics are urgently required. Fortunately, Nepal has great geographical and climatic diversity serving as a home for various actinomycetes. This study identified several actinomycetes using morphological and biochemical assays from seven soil samples collected from different elevations in Nepal. The primary objective of this study was to screen the potent metabolites-producing actinomycetes. For this purpose, antimicrobial assays of seven isolates were carried out against Escherichia coli ATCC 25922, Salmonella typhi ATCC 14028, Shigella sonnei ATCC 25931, and Staphylococcus aureus ATCC 43300. The sample BT36 demonstrated the highest zone of inhibition against the tested microorganisms among others. Similarly, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of all isolates were carried out. Among them, the MIC of samples BT36, PT7, and AB1 were found to be 1.952 mg/mL, 0.012 mg/mL, and 0.029 mg/mL, respectively. Ethyl acetate extracts of resulting potent isolates PT7 and AB1 were subjected to mass spectrometric analysis. A total of 22 bioactive compounds were annotated from liquid chromatography-high resolution mass spectroscopy (LC-HRMS), including epopromycin A, myxopyronin B, gilvocarcin HE, and okilactomycin A. It is concluded that this study will be useful in setting up a strategy to isolate useful secondary metabolites from soil microbes from the hilly region of Nepal.

Antimicrobial and antioxidant activities of lignin by-product from sugarcane leaf conversion to levulinic acid and hydrochar

To enhance the sustainability of lignocellulosic biomass utilization process, a significant attention is given to a high-value lignin byproduct of this process. Herein, antimicrobial, antioxidant, and physicochemical properties of lignin in black liquor extracted from sugarcane leaf (SCL) for levulinic acid (LA) and hydrochar production were investigated. The lignin was extracted from SCL through an alkaline pretreatment process using 5–25 wt% of either NaOH or KOH at 120–160 °C. Disk diffusion susceptibility tests, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) revealed that the SCL lignin is effective against Gram-positive bacteria (S. aureus) rather than Gram-negative bacteria (E. coli and S. typhimurium). Different alkaline reagents marginally affected the antimicrobial and antioxidant activities of the lignin. Py-GC/MS analysis results determined that the SCL lignin contained mainly p-hydroxyphenyl (H) and guaiacyl (G) with a small amount of syringyl (S) lignin. Furthermore, the structural alterations and linkages of SCL lignin during alkaline pretreatment were examined using 2D-HSQC NMR, providing valuable insights into the transformation processes. The conversion of delignified SCL to LA through a catalytic hydrothermal process using various acid catalysts indicated that the alkaline pretreatment could enhance LA yield with a maximum yield of 33.4 wt%. Additionally, fuel property characterization of the hydrochar co-product from LA production determined that the hydrochar can be used as a coal substitute.

Unraveling the antimicrobial activity of nutmeg and turmeric essential oils against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella spp.

The consumer demand for a reduction in the use of synthetic additives in food has been providing a greater search and incentive for the food industries to use new alternatives for food preservation. Among them, there is the initiative to use essential oils (EOs) due to their antimicrobial properties, coming from specific compounds in their compositions. However, in view of limitations related to the use of EOs, as well as their susceptibility to oxidation and degradation, the possibility arises of employing protection methods such as microencapsulation to minimize the impairment of the benefits associated with the application of EOs. This study aimed to examine the antimicrobial effect of turmeric and nutmeg EOs against microbial strains of Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella spp., as well as the microparticles of the EOs involved. Analyses of the minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC), and the developed microparticles, as well as the verification of the synergistic inhibitory action between these oils, were carried out. For free oils, antimicrobial action was evidenced against the vast majority of microorganisms tested, with free nutmeg EO having a better antimicrobial effect than free turmeric oil. In contrast, for encapsulated oils, only antimicrobial action was noted against strains of Listeria. Furthermore, the synergism of free oils did not potentiate the antimicrobial action. Regarding the alternative of microencapsulation of EOs, it was obtained that the results in which chitosan was used as wall material were more promising than when gelatin was used as wall material.

Evaluation of Mediterranean Tree Leaves as Valuable Biomass of Digestive Enzymes and Bacterial Inhibitors in the Concept of Circular Bioeconomy

This study aspires to evaluate the antibacterial and inhibitory effects of carbohydrate digestive enzymes in tree leaves that are widely distributed in the Mediterranean region. Leaves were sequentially extracted with solvents of increasing polarity. The results demonstrated a wide range of phenolic (3.5–770.7 mg gallic acid equivalent g−1) and flavonoid (0.2–321.3 mg catechin equivalent g−1) contents in leaf extracts. The minimum inhibitory and bactericidal concentration of leaf extracts was determined for six bacteria using the broth microdilution method. The polar extracts of carob, lentisk, and white mulberry leaves exerted strong antibacterial potency against Gram-positive bacteria, while the susceptibility of Escherichia coli on relative apolar extracts of carob, fig, and olive leaves was also observed. In parallel, the inhibitory effects of leaf extracts on carbohydrate digestive enzymes were evaluated. A robust inhibition of α-glucosidase was found for carob and lentisk leaf extracts, followed by extracts produced by white mulberry and olive leaves. Carob and lentisk leaves also act as a-amylase inhibitors at high concentrations. Overall, this study provides valuable data for the nutraceutical value of the “forgotten” treasure of Mediterranean tree leaves and assesses these plants as potential sources of antibacterial and carbohydrate digestive enzyme inhibitory agents for drug discovery.

In vitro assessment of the effect of magnetic fields on efficacy of biosynthesized selenium nanoparticles by Alborzia kermanshahica

Cyanobacteria represent a rich resource of a wide array of unique bioactive compounds that are proving to be potent sources of anticancer drugs. Selenium nanoparticles (SeNPs) have shown an increasing potential as major therapeutic platforms and led to the production of higher levels of ROS that can present desirable anticancer properties. Chitosan–SeNPs have also presented antitumor properties against hepatic cancer cell lines, especially the Cht-NP (Chitosan–NPs), promoting ROS generation and mitochondria dysfunction. It is proposed that magnetic fields can add new dimensions to nanoparticle applications. Hence, in this study, the biosynthesis of SeNPs using Alborzia kermanshahica and chitosan (CS) as stabilizers has been developed. The SeNPs synthesis was performed at different cyanobacterial cultivation conditions, including control (without magnetic field) and magnetic fields of 30 mT and 60 mT. The SeNPs were characterized by uv-visible spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), Dynamic light scattering (DLS), zeta potential, and TEM. In addition, the antibacterial activity, inhibition of bacterial growth, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC), as well as the antifungal activity and cytotoxicity of SeNPs, were performed. The results of uv-visible spectrometry, DLS, and zeta potential showed that 60 mT had the highest value regarding the adsorption, size, and stabilization in compared to the control. FTIR spectroscopy results showed consistent spectra, but the increased intensity of peaks indicates an increase in bond number after exposure to 30 mT and 60 mT. The results of the antibacterial activity and the inhibition zone diameter of synthesized nanoparticles showed that Staphylococcus aureus was more sensitive to nanoparticles produced under 60 mT. Se-NPs produced by Alborzia kermanshahica cultured under a 60 mT magnetic field exhibit potent antimicrobial and anticancer properties, making them a promising natural agent for use in the pharmaceutical and biomedical industries.

Inhibition and Mechanism of Citral on Bacillus cereus Vegetative Cells, Spores, and Biofilms.

Bacillus cereus causes food poisoning by producing toxins that cause diarrhea and vomiting and, in severe cases, endocarditis, meningitis, and other diseases. It also tends to form biofilms and spores that lead to contamination of the food production environment. Citral is a potent natural antibacterial agent, but its antibacterial activity against B. cereus has not been extensively studied. In this study, we first determined the minimum inhibitory concentrations and minimum bactericidal concentrations, growth curves, killing effect in different media, membrane potential, intracellular adenosine triphosphate (ATP), reactive oxygen species levels, and morphology of vegetative cells, followed by germination rate, morphology, germination state of spores, and finally biofilm clearance effect. The results showed that the minimum inhibitory concentrations and minimum bactericidal concentrations of citral against bacteria ranged from 100 to 800 μg/mL. The lag phase of bacteria was effectively prolonged by citral, and the growth rate of bacteria was slowed down. Bacteria in Luria-Bertani broth were reduced to below the detection limit by citral at 800 μg/mL within 0.5 h. Bacteria in rice were reduced to 3 log CFU/g by citral at 4000 μg/mL within 0.5 h. After treatment with citral, intracellular ATP concentration was reduced, membrane potential was altered, intracellular reactive oxygen species concentration was increased, and normal cell morphology was altered. After treatment with citral at 400 μg/mL, spore germination rate was reduced to 16.71%, spore morphology was affected, and spore germination state was altered. It also had a good effect on biofilm removal. The present study showed that citral had good bacteriostatic activity against B. cereus vegetative cells and its spores and also had a good clearance effect on its biofilm. Citral has the potential to be used as a bacteriostatic substance for the control of B. cereus in food industry production.

Antibacterial activity of angelica essential oil, its mechanism against Pseudomonas fluorescens, and its application in the preservation of chilled fresh beef

The mechanism by which Angelica essential oil (AEO) can act as an antimicrobial agent at cellular and metabolic levels to inhibit Pseudomonas fluorescens (P. fluorescens), and its effectiveness in beef preservation, were investigated and evaluated in this study. Our results showed that the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AEO to P. fluorescens were 12.5 μL/mL and 25 μL/mL, respectively, at which AEO could effectively inhibit P. fluorescens reproduction and retard its growth (p < 0.05). Severe morphological damage to the AEO-treated cells of the microorganism, including cell walls and membranes, were observed via scanning electron microscopy (SEM). AEO treatment was also seen to lead to an increase in alkaline phosphatase (AKPase) activity and conductivity, and a significant leakage of intracellular macromolecular components (such as nucleic acids and proteins), providing further evidence of damage to the integrity of cell wall and cell membrane. Metabolomics analysis revealed that the differential metabolites in the AEO hindered nucleic acid synthesis and expression in the P. fluorescens, leading to tricarboxylic acid (TCA) cycle blockage and amino acid restriction, energy metabolism disorders and membrane damage, mainly through nucleotide metabolism, amino acid metabolism, carbohydrate metabolism and membrane transport. Moreover, application testing demonstrated the efficacy of AEO in slowing the increase of beef pH and total volatile basic nitrogen (TVB-N), extending the shelf life of chilled beef by 5.39 d, and effectively maintaining the organoleptic quality of beef. These results lay a foundation for the development of AEO as a natural bacteriostatic agent.

Antibacterial and antibiofilm activities of selenium nanoparticles-antibiotic conjugates against anti-multidrug-resistant bacteria

The crucial demand to overcome the issue of multidrug resistance is required to refine the performance of antibiotics. Such a process can be achieved by fastening them to compatible nanoparticles to obtain effective pharmaceuticals at a low concentration. Thus, selenium nanoparticles (Se NPs) are considered biocompatible agents that are applied to prevent infections resulting from bacterial resistance to multi-antibiotics. The current evaluated the effectiveness of Se NPs and their conjugates with antibiotics such as amikacin (AK), levofloxacin (LEV), and piperacillin (PIP) against Pseudomonas aeruginosa (P. aeruginosa). In addition, the study determined the antibacterial and antibiofilm properties of Se NPs and their conjugates with LEV against urinary tract pathogens such as Staphylococcus aureus (S. aureus), Enterococcus faecalis (E. faecalis), P. aeruginosa, and Escherichia coli (E. coli). The result of minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for eight isolates of P. aeruginosa revealed that the conjugation of Se NPs with AK, LEV, and PIP resulted in a reduction in the concentration of antibiotic-conjugated Se NPs. The concentration was found to be about 10–20 times lower than that of bare antibiotics. The MIC of the Se NPs with LEV (i.e., Se NPs:LEV) for S. aureus, E. faecalis, P. aeruginosa, and E. coli was found to be 1.4:0.5, 0.7:0.25, 22:8, and 11:4 µg/mL, respectively. The results of the half-maximal inhibitory concentration (IC50) demonstrated that Se NPs:LEV conjugate have inhibited 50 % of the mature biofilms of S. aureus, E. faecalis, P. aeruginosa, and E. coli at a concentration of 27.5 ± 10.5, 18.8 ± 3.1, 40.6 ± 10.7, and 21.6 ± 3.3 µg/mL, respectively compared to the control. It has been suggested that the antibiotic-conjugated Se NPs have great potential for biomedical applications. The conjugation of Se NPs with AK, LEV, and PIP increases the antibacterial potency against resistant pathogens at a low concentration.

Innovative optimization for maximum magnetic nanoparticles production by Trichoderma asperellum with evaluation of their antibacterial activity, and application in sustainable dye decolorization

A pioneer microbial-based synthesis of various nanoparticles is gaining more attention due to their potent applications in various industries. The magnetic nanoparticles (M-nps) were fabricated for the first-time using the cell free extract of Trichoderma asperellum. The M-nps bioprocess optimization was performed using Response Surface Methodology and Artificial Neural Network.The bioprocess was significantly (P < 0.05) affected by pH, stirring speed, and incubation period. The M-nps displayed a strong antibacterial efficiency against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus in a dose-dependent manner. The minimum inhibitory concentration (MIC) for E. coli, P. aeruginosa, and S. aureus were respectively found to be 25.0, 6.25, and 3.125 µg/ml. While the minimum bactericidal concentration (MBC) was 25.0, 12.5, and 6.25 µg/ml. The highest inhibition zone of 22 mm was recorded for B. subtilis at 80 µg/ml with MIC and MBC of 3.125 µg/ml. The sodium alginate/M-nps microbeads was successfully prepared without any crosslinkers. The microbeads exhibited the highest adsorptive efficiency (84.32%) for malachite green at pH 8.0, dose 0.1 g, and time 60 min. The experimental data were superlative fitted to Langmuir’s isotherm, hinting monolayer adsorption of MG molecules with 117.65 mg g‐−1 adsorptive capacity. The regenerated beads continued up-to six consecutive cycles and maintained more than 90%, indicating the multiple time use of the prepared material. Hence, the green magnetic nanoparticles display promising application in combating bacterial pathogens. The magnetic microbeads can be employed as an alternative eco-friendly, highly efficient, sustainable, and low-cost biosorbent in removing contaminants from wastewater.

Antibacterial activity of silver and gold nanoparticles that have been synthesized by curcumin

The green synthesis of metal nanoparticles with curcumin which is a compound with antibiotic effects, was considered for the synthesis of metal nanoparticles. Gold and silver nanoparticles were synthesized with an average diameter of 19.8 ± 6.2 nm and 29.8 ± 6.4 nm respectively. Both particles represent stability in the physiological environment and after a series of washing procedures to remove unreacted precursors. The antimicrobial efficacy of the synthesized nanoparticles was investigated in different techniques such as disk diffusion, agar well diffusion, minimum inhibitory concentration, and minimum bactericidal concentration tests. The results of both curcumin-coated metal nanoparticles in the disk diffusion test were negative. However, in the agar dilution and MIC tests of antibacterial effect of both nanoparticles on the tested bacteria was observed. The silver nanoparticles had an inhibitory effect on all selected bacteria in the agar dilution (8-256 µg/ml). However, gold nanoparticles had a lower inhibitory effect on the growth of some of the selected bacteria (64-512 µg/ml). In conclusion, silver nanoparticles were more effective than gold nanoparticles in most of the investigated methods.

Enhancing the antioxidant and anti-inflammatory potentials of mulberry-derived postbiotics through submerged fermentation with B. subtilis H4 and B. amyloliquefaciens LFB112

Mulberry-derived postbiotics (MDP) have been scientifically proven to possess beneficial properties for human health, making them potential nutraceuticals. This study aims to enhance the antioxidant and anti-inflammatory effect of MDP via submerged fermentation with Bacillus subtilis H4 (H4) and Bacillus amyloliquefaciens LFB112 (LFB112) bacterial strains. Our results demonstrated compelling evidence that the combination of two strains with the optimized time (24 h), inoculum size (8%), and substrate size (10%) significantly increased the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities. When compared to the non-fermented mulberry (NFM), the MDP with the above-optimized conditions showed substantially higher antioxidant activities such as DPPH, 2,2′-Azino-bis (3-ethylebenzothiazoline-6-sulfonic acid) (ABTS), hydroxyl scavenging activities, and reducing power capabilities. Moreover, the MDP showed a significant decrease in the lipopolysaccharides (LPS)-induced reactive oxygen species (ROS) generated in RAW264.7 cells. The antibacterial assays such as minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) showed that MDP possesses antibacterial activity against various pathogenic bacterial strains with the lowest MIC recorded as 32.5 mg/ml against S. pullorum CVCC519, while the highest MIC observed was 40 mg/ml against P. aeruginosa. The lowest and highest MBCs observed were 40 and 55 mg/ml against S. aureus ATCC43300 and S. pullorum CVCC519 respectively. Furthermore, the MDP showed good ant-inflammatory activities by significantly reducing the nitric oxide (NO) concentration (p<0.0001) and mRNA expression of the proinflammatory cytokines in response to the LPS-induced inflammation in the RAW264.7 macrophage cell line. To sum up, our findings indicate that MDP as a novel postbiotics can be utilized both as a functional food and a safe feed additive, possibly benefiting gut and metabolic health.