Minimum Inhibitory Concentration Range Research Articles

Detection of Macrolide-Resistant Streptococcus pneumoniae Genes and Its Clinical Outcomes in a Tertiary Teaching Hospital in Malaysia.

Streptococcus pneumoniae is one of the leading causes of mortality and morbidity worldwide. The dramatic increase in in-vitro resistance of antimicrobial agents, particularly beta-lactams and macrolides, makes pneumococcal infections difficult to treat. The aim of this study was to describe the drug resistance rate, assess the prevalence of macrolide-resistant genes and review the clinical complications of pneumococcal infections among patients presented to Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia. This is a descriptive cross-sectional study. All S. pneumoniae isolates collected from clinical specimens within a 1-year period were subjected to selected antimicrobial susceptibility testing using E-test strips. Polymerase chain reaction (PCR) analysis was conducted to detect macrolide-resistant determinants. The patient's clinical data were obtained from clinical notes. A total of 113 patients with a positive growth of S. pneumoniae were included in the study. The most common predisposing factors among them were bronchopulmonary diseases (15.9%). The penicillin-resistant rate was 7.1%, with minimal inhibitory concentration (MIC) ranging between 0.012 μg/mL and >32 μg/mL, and the erythromycin-resistant rate was 26.5%, with a MIC range of 0.03 μg/mL-> 256 μg/mL. Most of the erythromycin-resistant isolates were found to have the mef(A) gene (50.4%) and the erm(B) gene (20%); 16.7% had a combination of genes mef(A) and erm(B), and 13.3% had none of the two genes. Community-acquired pneumonia is the predominant type of pneumococcal infection. There was no significant association between the presence of macrolide resistance determinants and mortality (P = 0.837) or complications (P > 0.999 for empyema and cardiac complication; P = 0.135 for subdural abscess). The majority of erythromycin-resistant isolates were found to have the mef(A) gene, followed by the erm(B) gene and a combination of genes mef(A) and erm(B).

The characteristics of fungal responses to uranium mining activities and analysis of their tolerance to uranium

The influence of uranium (U) mining on the fungal diversity (FD) and communities (FC) structure was investigated in this work. Our results revealed that soil FC richness and FD indicators obviously decreased due to U, such as Chao1, observed OTUs and Shannon index (P<0.05). Moreover, the abundances of Mortierella, Gibberella, and Tetracladium were notably reduced in soil samples owing to U mining activities (P<0.05). In contrast, the abundances of Cadophora, Pseudogymnoascus, Mucor, and Sporormiella increased in all soil samples after U mining (P<0.05). Furthermore, U mining not only dramatically influenced the Plant_Pathogen guild and Saprotroph and Pathotroph modes (P<0.05), but also induced the differentiation of soil FC and the enrichment of the Animal_Pathogen-Soil_Saprotroph and Endophyte guilds and Symbiotroph and Pathotroph Saprotroph trophic modes. In addition, various fungal populations and guilds were enriched to deal with the external stresses caused by U mining in different U mining areas and soil depths (P<0.05). Finally, nine U-tolerant fungi were isolated and identified with a minimum inhibitory concentration range of 400–600 mg/L, and their adsorption efficiency for U ranged from 11.6% to 37.9%. This study provides insights into the impact of U mining on soil fungal stability and the response of fungi to U mining activities, as well as aids in the screening of fungal strains that can be used to promote remediation of U mining sites on plateaus.

Design, Synthesis and In Silico Molecular Docking Evaluation of New Series of 1,2,3‐Triazole Derivatives as Potent Antimicrobial Agents

AbstractChalcone and triazole scaffolds have demonstrated a crucial role in the advancement of science and technology. Due to their significance, research has proceeded on the design and development of novel benzooxepine connected to 1,2,3‐triazolyl chalcone structures. The new chalcone derivatives produced by benzooxepine triazole methyl ketone and different aromatic carbonyl compounds. Product structures were established by various spectral techniques such as IR, 1H NMR, 13C NMR, and mass specrometric analysis. Among the tested compounds, two compounds exhibited exceptional antibacterial susceptibilities with MIC (Minimum inhibitory concentration) range of 3.59–10.30 μM against the tested S. aureus strain. Few of the compounds displayed superior antifungal activity against F. oxysporum with MIC value range from 3.25‐4.89 μM, when compared to fluconazole (MIC=3.83 μM). In addition, few derivatives demonstrated equivalent antitubercular action against H37Rv strain with MIC range of 2.16–4.90 μM. The capacity of one of the ligand to form a stable compound on the active site of CYP51 from M. tuberculosis (1EA1) was confirmed by docking studies. Additionally, the chalcone‐1,2,3‐triazole hybrids ADME (absorption, distribution, metabolism, and excretion), molecular characteristics, estimation of toxicity, and bioactivity scores were assessed.

Durlobactam, a Diazabicyclooctane β-Lactamase Inhibitor, Inhibits BlaC and Peptidoglycan Transpeptidases of Mycobacterium tuberculosis.

Peptidoglycan synthesis is an underutilized drug target in Mycobacterium tuberculosis (Mtb). Diazabicyclooctanes (DBOs) are a class of broad-spectrum β-lactamase inhibitors that also inhibit certain peptidoglycan transpeptidases that are important in mycobacterial cell wall synthesis. We evaluated the DBO durlobactam as an inhibitor of BlaC, the Mtb β-lactamase, and multiple Mtb peptidoglycan transpeptidases (PonA1, LdtMt1, LdtMt2, LdtMt3, and LdtMt5). Timed electrospray ionization mass spectrometry (ESI-MS) captured acyl-enzyme complexes with BlaC and all transpeptidases except LdtMt5. Inhibition kinetics demonstrated durlobactam was a potent and efficient DBO inhibitor of BlaC (KIapp 9.2 ± 0.9 μM, k2/K 5600 ± 560 M-1 s-1) and similar to clavulanate (KIapp 3.3 ± 0.6 μM, k2/K 8400 ± 840 M-1 s-1); however, durlobactam had a lower turnover number (tn = kcat/kinact) than clavulanate (1 and 8, respectively). KIapp values with durlobactam and clavulanate were similar for peptidoglycan transpeptidases, but ESI-MS captured durlobactam complexes at more time points. Molecular docking and simulation demonstrated several productive interactions of durlobactam in the active sites of BlaC, PonA1, and LdtMt2. Antibiotic susceptibility testing was conducted on 11 Mtb isolates with amoxicillin, ceftriaxone, meropenem, imipenem, clavulanate, and durlobactam. Durlobactam had a minimum inhibitory concentration (MIC) range of 0.5-16 μg/mL, similar to the ranges for meropenem (1-32 μg/mL) and imipenem (0.5-64 μg/mL). In β-lactam + durlobactam combinations (1:1 mass/volume), MICs were lowered 4- to 64-fold for all isolates except one with meropenem-durlobactam. This work supports further exploration of novel β-lactamase inhibitors that target BlaC and Mtb peptidoglycan transpeptidases.

New strain Brevibacillus laterosporus TSA31-5 produces both brevicidine and brevibacillin, exhibiting distinct antibacterial modes of action against Gram-negative and Gram-positive bacteria.

The growing prevalence of antibiotic resistance has made it imperative to search for new antimicrobial compounds derived from natural products. In the present study, Brevibacillus laterosporus TSA31-5, isolated from red clay soil, was chosen as the subject for conducting additional antibacterial investigations. The fractions exhibiting the highest antibacterial activity (30% acetonitrile eluent from solid phase extraction) were purified through RP-HPLC. Notably, two compounds (A and B) displayed the most potent antibacterial activity against both Escherichia coli and Staphylococcus aureus. ESI-MS/MS spectroscopy and NMR analysis confirmed that compound A corresponds to brevicidine and compound B to brevibacillin. Particularly, brevicidine displayed notable antibacterial activity against Gram-negative bacteria, with a minimum inhibitory concentration (MIC) range of 1-8 μg/mL. On the other hand, brevibacillin exhibited robust antimicrobial effectiveness against both Gram-positive bacterial strains (MIC range of 2-4 μg/mL) and Gram-negative bacteria (MIC range of 4-64 μg/mL). Scanning electron microscopy analysis and fluorescence assays uncovered distinctive morphological alterations in bacterial cell membranes induced by brevicidine and brevibacillin. These observations imply distinct mechanisms of antibacterial activity exhibited by the peptides. Brevicidine exhibited no hemolysis or cytotoxicity up to 512 μg/mL, comparable to the negative control. This suggests its promising therapeutic potential in treating infectious diseases. Conversely, brevibacillin demonstrated elevated cytotoxicity in in vitro assays. Nonetheless, owing to its noteworthy antimicrobial activity against pathogenic bacteria, brevibacillin could still be explored as a promising antimicrobial agent.

In vitro antimicrobial and antibiofilm screening of eighteen Iranian medicinal plants

BackgroundNatural products are one of the best candidates for controlling drug-resistant pathogens, the advantages of which include low production costs and low side effects. In this study, as potential antimicrobials, the anti-bacterial and antibiofilm activities of several Iranian native medicinal plants were screened.MethodsThe antibacterial/antifungal and anti-biofilm activities of 18 medicinal plants including Reseda lutea L., Nepeta sintenisii Bunge., Stachys turcomanica Trautv., Stachys lavandulifolia Vahl, Diarthron antoninae (Pobed.) Kit Tan., Ziziphora clinopodioides Lam., Euphorbia kopetdaghi Prokh, Euphorbia serpens Kunth., Hymenocrater calycinus Benth., Scutellaria pinnatifida A.Ham., Viola tricolor L., Hypericum helianthemoides (Spach) Boiss., Hypericum scabrum L., Convolvulus lineatus L., Scabiosa rotata M.Bieb Greuter & Burdet, Delphinium semibarbatum Bien. Ex Boiss., Glycyrrhiza triphylla Fisch. & C.A.Mey., and Ziziphus jujuba Mill., against two Gram-positive bacteria, Staphylococcus aureus, Bacillus cereus, as well as two Gram-negative bacteria, Pseudomonas aeruginosa, Escherichia coli; and Candida albicans as a fungal strain, were evaluated. The minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) values of the extracts against tested microorganisms were reported and we investigated their effect on the biofilm inhibition of Pseudomonas aeruginosa PAO1, Staphylococcus epidermis, Staphylococcus aureus and Streptococcus mutans. In addition, the effect of the extracts on the eradication of the biofilms of these bacteria was evaluated.ResultsIn this study, H. scabrum was found to exhibit potentially significant activity against Gram-positive bacteria with the MIC range of 6.25–25 µg/mL. This extract also showed a significant effect on inhibiting the biofilm of S. aureus, S. mutans, and S. epidermidis and eradicating the biofilm of S. epidermidis DSMZ 3270. In addition, Hymenocrater calycinus root extract had moderate antibacterial activity against B. cereus with the MIC and MBC 62.5 µg/mL, respectively.ConclusionsThe results of this study showed that the root extracts of two plants, Hypericum scabrum and Hymenocrater calycinus, had antimicrobial and anti-biofilm effects. Based on the observed anti-biofilm effects, these two plants may be considered in future studies to find responsible antimicrobial compounds.

Fungus-mediated synthesis of Se-BiO-CuO multimetallic nanoparticles as a potential alternative antimicrobial against ESBL-producing Escherichia coli of veterinary origin.

Bacterial infections emerge as a significant contributor to mortality and morbidity worldwide. Emerging extended-spectrum β-lactamase (ESBL) Escherichia coli strains provide a greater risk of bacteremia and mortality, are increasingly resistant to antibiotics, and are a major producer of ESBLs. E. coli bacteremia-linked mastitis is one of the most common bacterial diseases in animals, which can affect the quality of the milk and damage organ functions. There is an elevated menace of treatment failure and recurrence of E. coli bacteremia necessitating the adoption of rigorous alternative treatment approaches. In this study, Se-Boil-CuO multimetallic nanoparticles (MMNPs) were synthesized as an alternate treatment from Talaromyces haitouensis extract, and their efficiency in treating ESBL E. coli was confirmed using standard antimicrobial assays. Scanning electron microscopy, UV-visible spectroscopy, and dynamic light scattering were used to validate and characterize the mycosynthesized Se-BiO-CuO MMNPs. UV-visible spectra of Se-BiO-CuO MMNPs showed absorption peak bands at 570, 376, and 290 nm, respectively. The average diameters of the amorphous-shaped Se-BiO-CuO MMNPs synthesized by T. haitouensis extract were approximately 66-80 nm, respectively. Se-BiO-CuO MMNPs (100 μg/mL) showed a maximal inhibition zone of 18.33 ± 0.57 mm against E. coli. Se-BiO-CuO MMNPs also exhibited a deleterious impact on E. coli killing kinetics, biofilm formation, swimming motility, efflux of cellular components, and membrane integrity. The hemolysis assay also confirms the biocompatibility of Se-BiO-CuO MMNPs at the minimum inhibitory concentration (MIC) range. Our findings suggest that Se-BiO-CuO MMNPs may serve as a potential substitute for ESBL E. coli bacteremia.

Comparative study of volatile oil constituents, anti-microbial properties, and antibiofilm activities in Eucalyptus camaldulensis and Eucalyptus globulus: insights from central Saudi Arabia

Eucalyptus is a fragrant plant with a highly volatile oil content grown worldwide. This study investigated the phytochemical and biological features of two Eucalyptus species, E. camaldulensis and E. globulus, cultivated in Central Saudi Arabia. Phytochemical analysis revealed 17 compounds in E. camaldulensis and 13 in E. globulus, with α-pinene and 1,8-cineole being both plants’ most abundant volatile components. The antimicrobial activity revealed that Salmonella typhimurium ATCC 13311 and Candida albicans ATCC 10231 are the most susceptible to E. camaldulensis, with the zone of inhibition diameters (ZIDs) of 17.0 ± 0.2 mm and 22.0 ± 0.3 mm, respectively. In contrast, for E. globulus, methicillin-resistant Staphylococcus aureus and Candida albicans ATCC 10231 were the most susceptible, with ZIDs of 13.0 ± 0.2 mm and 20.0 ± 0.2 mm, respectively. E. camaldulensis had minimum inhibitory concentration (MIC) range from 1.56 to 12.5 μL/mL, minimum biocidal concentration (MBC) range from 3.125 to 25.0 μL/mL, minimum biofilm inhibitory concentration (MBIC) range from 3.125 to 25.0 μL/mL, and minimum biofilm eradication concentration (MBEC) from 6.25 to 50.0 μL/mL, while E. globulus had MIC range from 3.125 to 12.5 μL/mL, MBC range from 6.25 to 25.0 μL/mL, MBIC range from 6.25 to 25.0 μL/mL, and MBEC range from 12.5 to 50.0 μL/mL, for the tested organisms. E. camaldulensis demonstrated better antimicrobial activity than E. globulus against the tested organisms, making it a promising antimicrobial drug candidate. Further research is required to determine its effectiveness in treating different human infections.

In vitro effects of different ozone preparations on microorganisms responsible for endometritis in the mare

Infectious endometritis is considered one of the major causes of infertility and it can affect up to 60% of barren mares. It is characterized by the presence of one or more microorganisms in the reproductive tract and it is treated with the administration of antibiotics, ecbolic agents and uterine lavages. Ozone, thanks to its antimicrobial properties that are based on its high oxidative potential, could represent an effective alternative treatment for endometritis. The aim of this study was to test in vitro the bactericidal and fungicidal properties of different ozone formulations, either as gas (experiment 1) or dissolved in two liquid matrices (experiment 2), specifically distilled water or oil (Neozone 4000, Cosmoproject, Parma, Italy), onto 6 different species of microorganisms isolated from mares with clinical endometritis, namely Escherichia coli, Staphylococcus aureus, Streptococcus equi subsp. Zooepidemicus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans. In the first experiment, 3 clinical antibiotic-resistant strains per each species were exposed to different conditions: to O2O3 gas mixtures (15 and 40 μg/ml for 1, 3 and 5 min), to 100 % O2 or left untreated. The results showed a reduction of the microbial count of over 99,9% for every pathogen, time and concentration of O2O3 gas mixtures tested. Furthermore, gaseous ozone showed both a time-dependant effect (5 vs 3 vs 1 min of exposure) and a concentration-dependant effect (40 vs 15 μg/ml) at 1 and 3 min, while after 5 min no differences were observed. In the second experiment, minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC, MFC) of ozonated distilled water and ozonated oil were evaluated. Ozonated oil showed a bactericidal/fungicidal activity against all the strains tested (MIC range 12.5–25 % v/v, MBC/MFC range 12.5–50 % v/v) while ozonated distilled water didn't show an observable antimicrobial effect, discouraging its use as an antimicrobial agent for the treatment of endometritis.The results of this in vitro study indicate that both gaseous ozone and ozonated oil exerted remarkable antimicrobial activities and are promising alternative treatments for infectious endometritis, even when caused by antibiotic-resistant bacteria, and encourage further experiments in an effort to scale down or even prevent the use of antibiotics in equine reproduction.

Staurosporine-producing Streptomyces sp. strain 11 × 1 cell-free culture filtrates control diseases caused by the oomycete plant pathogens Pythium myriotylum and Phytophthora sojae

ABSTRACT Oomycetes are devastating soil-borne plant pathogens, and cause numerous losses on diverse crops worldwide. In this study, our aim was to identify actinomycetes with antimicrobial activity towards oomycetes from the rhizosphere of soybean. Strong growth inhibition of P. myriotylum was found in dual culture with a strain called 11 × 1, and also from using the 11 × 1 cell-free culture filtrate and EtOAc extract. The P. myriotylum growth-inhibiting activity of the 11 × 1 culture filtrates was stable at high temperatures, and acidic and alkaline conditions. The 11 × 1 strain was identified as a Streptomyces strain with the highest 16S rDNA sequence identity to S. scabiei, and phylogenomic analysis also showed clustering with S. scabiei strains. Applying the 11 × 1 culture filtrates to ginger and soybean leaves inhibited the necrosis caused by P. myriotylum and P. sojae, respectively. Pot assay experiments showed a strong control effect of the 11 × 1 culture filtrate against P. myriotylum-caused soft-rot disease of ginger, and against P. sojae caused root-rot of soybean. Analysis of the biosynthetic gene clusters from the genome of 11 × 1 predicted the production of staurosporine, and staurosporine was subsequently identified from LC-MS analysis of the 11 × 1 extract. The concentrations of staurosporine quantified in the 11 × 1 extracts were in the range of the staurosporine minimum inhibitory concentrations for P. myriotylum, supporting the contribution of staurosporine to the biocontrol effect of 11 × 1. The use of 11 × 1 culture filtrate to control both ginger and soybean diseases could be beneficial in an intercropping system of ginger and the leguminous nitrogen-fixing soybean.

Uncovering the antimicrobial activity of G-type lysozyme 2 derived from Cyprinus carpio mucus against bacterial and fungal pathogens

This study aimed to explore the antimicrobic potential of mucus samples collected from Cyprinus carpio and identify the specific antimicrobial peptides responsible for its activity. The crude extract was tested against various bacterial and fungal pathogens, and its protein content and profile were analyzed. Purification steps, including gel filtration chromatography, were employed to isolate the most active fraction (peak IV), which was further identified via liquid chromatography and mass spectroscopy. The results revealed varying degrees of antimicrobial activity of the crude extract against different bacterial and fungal strains, with Leclercia adecarboxylata, Candida glabrata, and Candida parapsilosis showing the highest susceptibility. SDS-PAGE analysis demonstrated the existence of multiple low molecular weight protein bands in the crude extract, while fraction IV obtained from gel filtration chromatography exhibited the strongest antimicrobial activity. Peak IV displayed a range of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) values against the tested pathogens, spanning from 0.038 to 4.960 mg/mL. Further investigation identified the purified peptide derived from peak IV as G-type lysozyme 2, characterized by a molecular weight of 21 kDa. These findings shed light on the existence of a highly effective antimicrobial peptide, G-type lysozyme 2, within the mucus of Cyprinus carpio. This peptide demonstrates notable activity against diverse bacterial and fungal pathogens. The insights from this study enhance our understanding of the fish's antimicrobial defense mechanisms and hold promise for developing novel antimicrobial agents.

Determining Susceptibility and Potential Mediators of Resistance for the Novel Polymyxin Derivative, SPR206, in Acinetobacter baumannii.

With the increase in carbapenem-resistant A. baumannii (CRAB) infections, there has been a resurgence in the use of polymyxins, specifically colistin (COL). Since the reintroduction of COL-based regimens in treating CRAB infections, several COL-resistant A. baumannii isolates have been identified, with the mechanism of resistance heavily linked with the loss of the lipopolysaccharide (LPS) layer of the bacterial outer membrane through mutations in lpxACD genes or the pmrCAB operon. SPR206, a novel polymyxin derivative, has exhibited robust activity against multidrug-resistant (MDR) A. baumannii. However, there is a dearth of knowledge regarding its efficacy in comparison with other A. baumannii-active therapeutics and whether traditional polymyxin (COL) mediators of A. baumannii resistance also translate to reduced SPR206 activity. Here, we conducted susceptibility testing using broth microdilution on 30 A. baumannii isolates (17 COL-resistant and 27 CRAB), selected 14 COL-resistant isolates for genomic sequencing analysis, and performed time-kill analyses on four COL-resistant isolates. In susceptibility testing, SPR206 demonstrated a lower range of minimum inhibitory concentrations (MICs) compared with COL, with a four-fold difference observed in MIC50 values. Mutations in lpxACD and/or pmrA and pmrB genes were detected in each of the 14 COL-resistant isolates; however, SPR206 maintained MICs ≤ 2 mg/L for 9/14 (64%) of the isolates. Finally, SPR206-based combination regimens exhibited increased synergistic and bactericidal activity compared with COL-based combination regimens irrespective of the multiple resistance genes detected. The results of this study highlight the potential utility of SPR206 in the treatment of COL-resistant A. baumannii infections.

In vitro activity of ceftazidime/avibactam, cefiderocol, meropenem/vaborbactam and imipenem/relebactam against clinical strains of the Stenotrophomonas maltophilia complex.

Infections caused by Stenotrophomonas maltophilia and related species are increasing worldwide. Unfortunately, treatment options are limited, whereas the antimicrobial resistance is increasing. We included clinical isolates identified as S. maltophilia by VITEK 2 Compact. Ceftazidime/avibactam, meropenem/vaborbactam, imipenem/relebactam, cefiderocol, quinolones, and tetracycline family members were evaluated by broth microdilution method and compared with first-line treatment drugs. Minimum inhibitory concentrations (MICs) were reported for all antibiotics. We sequenced the Whole Genome of cefiderocol resistant strains (CRSs) and annotated their genes associated with cefiderocol resistance (GACR). Presumptive phylogenetic identification employing the 16S marker was performed. One hundred and one clinical strains were evaluated, sulfamethoxazole and trimethoprim, levofloxacin and minocycline showed susceptibilities of 99.01%, 95.04% and 100% respectively. Ceftazidime was the antibiotic with the highest percentage of resistance in all samples (77.22%). Five strains were resistant to cefiderocol exhibiting MIC values ≥ 2 μg/mL (4.95%). The β-lactamase inhibitors meropenem/vaborbactam and imipenem/relebactam, failed to inhibit S. maltophilia, preserving both MIC50 and MIC90 ≥64 μg/mL. Ceftazidime/avibactam restored the activity of ceftazidime decreasing the MIC range. Tigecycline had the lowest MIC range, MIC50 and MIC90. Phylogeny based on 16S rRNA allowed to identify to cefiderocol resistant strains as putative species clustered into Stenotrophomonas maltophilia complex (Smc). In these strains, we detected GARCs such as Mutiple Drug Resistance (MDR) efflux pumps, L1-type β-lactamases, iron transporters and type-1 fimbriae. Antimicrobial resistance to first-line treatment is low. The in vitro activity of new β-lactamase inhibitors against S. maltophilia is poor, but avibactam may be a potential option. Cefiderocol could be considered as a potential new option for multidrug resistant infections. Tetracyclines had the best in vitro activity of all antibiotics evaluated.

Daptomycin MICs by Broth Microdilution Method Against Staphylococcus aureus in Clinical Specimens: A Cross-Sectional Study from Pakistan.

To explore the distribution of daptomycin (DAP) minimum inhibitory concentrations (MICs) in Staphylococcus aureus isolated from complicated skin, soft tissue, and bloodstream infections collected from the Pakistani population using broth microdilution (BMD). Descriptive, cross-sectional study. Place and Duration of the Study: Department of Pathology and Laboratory Medicine, The Aga Khan University Hospital, from May to October 2021. Through consecutive sampling techniques, 169 Staphylococcus aureus (S. aureus) isolated from clinical specimens including pus, tissue, and blood were collected. Patients' data including age, gender, geographical location, specimen site, and methicillin susceptibility were collected from the laboratory data. BMD was used to determine MICs of clinical isolates and S. aureus ATCC 29213. DAP MIC ≤1.0 µg/ml was considered susceptible according to the Clinical and Laboratory Standards Institute M100. Among all the clinical isolates, 144 (85%) and 25 (15%) were from skin and soft tissue and blood, respectively. All isolates were susceptible to DAP with MIC50, MIC90, and MIC range of 0.25 µg/ml, 0.5 µg/ml, and 0.06 - 0.5 µg/ml, respectively. These study findings demonstrated low in-vitro MICs for DAP against S. aureus in tested isolates from a diverse variety of patient specimens from across Pakistan. Daptomycin, Staphylococcus aureus, Broth microdilution, Minimum inhibitory concentrations.

Antimicrobial Activity of African Walnut (&lt;em&gt;Tetracarpidium conophorum&lt;/em&gt;) Oil against Bacterial and Fungal Species Causing Food Spoilage and Food Poisoning Diseases

This study evaluated the antibacterial and antifungal activity of African walnut (Tetracarpidium conophorum) oil against species causing food poisoning and food spoilage. African walnut oil was extracted using the solvent extraction method. The fatty acid profile of the oil was determined using gas chromatography mass spectrometry (GC-MS). The agar disc diffusion method was used to determine the antibacterial activity of the oil before using it for further tests. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were also determined using the disc diffusion assay. The result indicates that palmitic (1.53%), linoleic (13.5%), linolenic 80.59%, stearic (4.41%), and eicosenoic acid (0.42%) were present in the oil with higher polyunsaturated fatty acid values. African walnut oil showed remarkable activity against all the Gram positive organisms (Bacillus cereus ATCC 33018, Staphylococcus aureus ATCC 25923, Clostridium perfringes NCTC 8799, and Listeria monocytogenes ATCC 13932) with the inhibition zones ranging from 21.6 ± 0.27 mm to 26.3 ± 0.23 while the Gram negative organisms (Pseudomonas fluorescens ATCC 13883, Pseudomonas fragi ATCC 4973, Escherichia coli ATCC 12806, Salmonella enterica subsp. enterica serovar Typhimurium strain ATCC 14028) were less sensitive and resistant to the oil chromatography-mass with inhibition zones ranging from 6.3 ± 0.31 to 9.9 ± 0.21 mm. Out of the tested oil concentrations from 100 – 0.39 mg/mL, MIC obtained for the Gram positive organisms was 1.56 mg/mL while it was 3.13 mg/mL for Gram negative bacteria (except L. monocytogenes). The antifungal test reveals the MIC range of 100 - 0.78 mg/mL in all the fungal isolates tested. African walnut oil has proved to be potentially active against the Gram positive bacterial and fungal species tested indicating that it can be used as a natural alternative to control food spoilage and food poisoning diseases.

Synthesis and characterization of Fe3O4@SiO2@PDA@Ag core–shell nanoparticles and biological application on human lung cancer cell line and antibacterial strains

Novel magnetic and metallic nanoparticles garner much attention of researchers due to their biological, chemical and catalytic properties in many chemical reactions. In this study, we have successfully prepared a core–shell Fe3O4@SiO2@PDA nanocomposite wrapped with Ag using a simple synthesis method, characterised and tested on small cell lung cancer and antibacterial strains. Incorporating Ag in Fe3O4@SiO2@PDA provides promising advantages in biomedical applications. The magnetic Fe3O4 nanoparticles were coated with SiO2 to obtain negatively charged surface which is then coated with polydopamine (PDA). Then silver nanoparticles were assembled on Fe3O4@SiO2@PDA surface, which results in the formation core–shell nanocomposite. The synthesised nanocomposite were characterized using SEM-EDAX, dynamic light scattering, XRD, FT-IR and TEM. In this work, we report the anticancer activity of silver nanoparticles against H1299 lung cancer cell line using MTT assay. The cytotoxicity data revealed that the IC50 of Fe3O4@SiO2@PDA@Ag against H1299 lung cancer nanocomposites cells was 21.52 µg/mL. Furthermore, the biological data of nanocomposites against Gram-negative ‘Pseudomonas aeruginosa’ and Gram-positive ‘Staphylococcus aureus’ were carried out. The range of minimum inhibitory concentration was found to be 115 µg/mL where gentamicin was used as a standard drug. The synthesized AgNPs proves its supremacy as an efficient biomedical agent and AgNPs may act as potential beneficial molecule in lung cancer chemoprevention and antibacterial strains.

Designing and Synthesis of an Anti-Candida parapsilosis Thymoquinone Lipid Formulation

Background: Candida parapsilosis complex, as an opportunistic pathogen, can cause various infections, especially in immunocompromised patients. Objectives: This study investigated a novel thymoquinone-liposomal nanoparticle to increase the stability of thymoquinone with antifungal effects against C. parapsilosis isolates. Methods: The thymoquinone was encapsulated in liposomal nanoparticles using a thin-film hydration technique and then analyzed by transmission electron microscopy (TEM), particle size, zeta potential, and UV-visible spectrophotometer. Also, the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was used on peripheral blood mononuclear cells (PBMCs) for cell metabolic activity. The antifungal activity of thymoquinone-liposomal nanoparticle against 15 clinical isolates of C. parapsilosis and the reference strain was examined based on the M27-A3 guideline. Results: The synthesized thymoquinone-liposomal nanoparticle was approved by the TEM, particle size, zeta potential, and UV-Vis. The thymoquinone-liposomal nanoparticle showed no toxic effect on PBMCs following the MTT assay. The minimum inhibitory concentration (MIC) range of free thymoquinone and liposomal formulations with inhibitory effects on Candida isolates was 50 to 6.25 and 150 to 18.75 µg/mL, respectively. The MIC50 and MIC90 values of free thymoquinone and thymoquinone-liposomal nanoparticle were 25 and 50, and 75 and 150 µg/mL. Conclusions: The synthesized thymoquinone-liposomal nanoparticle shows significant antifungal activity against C. parapsilosis isolates compared to free thymoquinone. However, because of its hydrophilicity and hydrophobicity, biocompatibility, particle size, non-toxic effect, and higher cell viability, the thymoquinone-liposomal nanoparticle is a more effective approach for treating fungal infections.

Biological Synthesis and Characterization of Silver-Doped Nanocomposites: Antibacterial and Mechanistic Studies

The development of antimicrobial resistance has increased the prevalence of infectious diseases, causing a global health problem that accounts for over 4.95 million deaths worldwide annually. The side effects associated with current antibiotics prompt a crucial need to search for effective and safe antimicrobial agents. In this study, silver nanoparticles (AgNPs) were prepared by chemical reduction method using silver nitrates as a metallic precursor and Croton macrostachyus bark aqueous extract as a reducing and capping agent. The nanoparticles were further functionalized using C. macrostachyus-based activated carbon (CAC) to generate nanocomposites (CAC-AgNPs). The nanomaterials were characterized by ultraviolet-visible (UV–vis) absorption spectra and Fourier transform infrared (FTIR) spectra. The antibacterial activity of the as-prepared nanomaterials was evaluated against an array of bacterial strains by microdilution method, whereas their cytotoxicity profile was evaluated using Vero cells (human mammalian cells). Antibacterial mechanistic studies of active nanomaterials were carried out through bacterial growth kinetics, nucleic acid leakage tests, and catalase inhibition assays. As a result, the as-prepared nanomaterials exhibited antibacterial activity against an array of bacterial strains (minimum inhibitory concentration (MIC) range: 62.5 to 500 µg/mL), the most susceptible being Escherichia coli and Staphylococcus aureus. Cytotoxicity studies of the nanomaterials on Vero cells revealed that the nanocomposite (median cytotoxic concentration (CC50): 213.6 µg/mL) was less toxic than its nanoparticle (CC50 value: 164.75 µg/mL) counterpart. Antibacterial mechanistic studies revealed that the nanomaterials induced (i) bacteriostatic activity vis à vis E. coli and S. aureus and (ii) inhibition of catalase in these bacteria. This novel contribution regarding the antibacterial mechanisms of action of silver nanocomposites from C. macrostachyus-based activated carbon may contribute to our understanding of the antibacterial action of these biomaterials. Nevertheless, more chemistry and in vivo experiments as well as in depth antibacterial mechanistic studies are warranted for the successful utilization of these antibacterial biomaterials.

Synergistic Antifungal Bioassay of Neem Leaves Extract with Essential Oils against Dermatophytes

Introduction: The neem plant is also known as a panacea in dentistry due to its antimicrobial properties and plays vital role in the cosmetic industry as it is extensively used in cosmetic preparations for hair and skin care. Objective: The present study was carried out to evaluate the synergistic activity of essential oils in combination with neem leaves ethanolic extracts. Methods: The antifungal activity of neem leaves (Azadirachta indica) ethanolic extracts in combination with A. indica, Eucalyptus citriodora and Cymbopogon martini essential oils was conducted by agar well diffusion method followed by M38-A2 broth microdilution method on pathogenic dermatophytes strains; Microsporum gypseum, Trichophyton mentagrophytes and T. rubrum. Neem leaves extract was obtained by treating the matured green leaves with ethanol. Moreover, HPLC analysis was performed to narrate the terpenoid content having extensively noted antifungal spectrum. The potent antimycotic fluconazole taken as a positive control. Result: The result exhibited that there was overall growth retardation of the dermatophytes at minimum inhibitory concentration between 125 to 250 μg/mL for leaves extract and between 0.078 to 5.0 μL for selected essential oils. The range of minimum inhibitory concentration (MIC) of fluconazole was found between 0.25 and 0.50 μg/mL. Conclusion: Neem leaves ethanolic extracts analysis showed the possible existence of terpenoids in extracts that are recognized having extensive biological action. The outcomes of this study showed a novel report on the therapeutic potential of neem leaves extract with A. indica, E. citriodora and C. martini to control dermatophytosis. The neem leaf extract and antifungal herbal oil in combination have been proven to improve and broad-spectrum antifungal activity and they can be formulated in suitable formulations for the effective treatment of mycosis.

Antifungal susceptibility testing following the CLSI M27 document, along with the measurement of MFC/MIC ratio, could be the optimal approach to detect amphotericin B resistance in Clavispora (Candida) lusitaniae. Susceptibility patterns of contemporary isolates of this species.

Antifungal susceptibility testing (AST) is crucial in clinical settings to guide appropriate therapy. Nevertheless, discrepancies between treatment response and some results still persist, particularly in detecting resistance to amphotericin B (AMB) in Clavispora (Candida) lusitaniae. This study aimed to assess the susceptibility patterns of 48 recent isolates of C. lusitaniae to 9 antifungal agents and explore the feasibility of using a CLSI reference-based method to identify AMB resistance. Microdilution techniques revealed a wide range of minimal inhibitory concentration (MIC) values for azole antifungals, while echinocandins and AMB exhibited a narrow range of MIC values, with all strains considered wild-type for the tested polyene and echinocandins. However, when agar diffusion (ellipsometry) was employed for AST, certain strains displayed colonies within the inhibition ellipse, indicating potential resistance. Interestingly, these strains did not respond to AMB treatment and were isolated during AMB treatment (breakthrough). Moreover, the evaluation of AMB minimum fungicidal concentrations (MFCs) indicated that only the strains with colonies inside the ellipse had MFC/MIC ratios ≥ 4, suggesting reduced fungicidal activity. In conclusion, this study confirms the effectiveness of ellipsometry with RPMI-1640 2% glucose agar for detecting AMB resistance in C. lusitaniae. Additionally, the proposed approach of culturing "clear" wells in the microdilution method can aid in uncovering resistant strains. The findings highlight the importance of appropriate AST methods to guide effective treatment strategies for deep-seated candidiasis caused by C. lusitaniae. Further collaborative studies are warranted to validate these findings and improve the detection of AMB clinical resistance.