Mono-/Bis-Alkenoic Acid Derivatives From an Endophytic Fungus Scopulariopsis candelabrum and Their Antifungal Activity

During a screening for antifungal secondary metabolites, six new mono-/bis-alkenoic acid derivatives (2–7) and one known alkenoic acid derivative (1) were isolated from an endophytic fungi Scopulariopsis candelabrum. Their chemical structures were identified by 1H-NMR, 13C-NMR, 2D NMR, and high-resolution mass spectrometry, as well as comparisons with previously reported literatures. Among them, fusariumesters C‒F (2–5) are bis-alkenoic acid derivatives dimerized by an ester bond, while acetylfusaridioic acid A (6) and fusaridioic acid D (7) are alkenoic acid monomers. All the isolates were submitted to an antifungal assay against Candida albicans and the corn pathogen Exserohilum turcicum using the filter paper agar diffusion method. As a result, only compound 1 decorating with β-lactone ring turned out to be active against these two tested fungi. The broth microdilution assay against Candida albicans showed the minimum inhibitory concentration (MIC) value of 1 to be 20 μg/ml, while the minimum inhibitory concentration value of the positive control (naystatin) was 10 μg/ml. And the half maximal inhibitory concentration (IC50) value (21.23 μg/ml) of 1 against Exserohilum turcicum was determined by analyzing its inhibition effect on the mycelial growth, using cycloheximide (IC50 = 46.70 μg/ml) as the positive control.

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria

High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-1) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.

Simulation of inference test performance for minimum inhibitory concentration censored data

The estimation of the minimum inhibitory concentration is usually performed by a method of serial dilutions by a factor of 2, introducing the overestimation of antimicrobial efficacy, quantified by a simulation model that shows that the variability of the bias is higher for the standard deviation, being dependent on the metric distance to the values of the concentrations used. We use a methodological approach through modeling and simulation for the measurement error of physical variables with censored information, proposing a new inference method based on the calculation of the exact probability for the set of possible samples from nmeasurements that allows quantifying the p-value in one or two independent sample tests for the comparison of censored data means. Tests based on exact probability methods offer a reasonable solution for small sample sizes, with statistical power varying according to the hypothesis evaluated, providing insight into the limitations of censored data analysis and providing a tool for decision making in the diagnosis of antimicrobial efficacy.

Phytochemical screening and antibacterial activity of Vernonia amygdalina (bitter leaf) on some selected bacterial isolates

Plants with medicinal value produce certain chemical elements known as phytochemicals that have antibacterial activity. The study was aimed at determining the antibacterial activity of Vernonia amygdalina against bacterial isolates using agar well diffusion method. In addition, the phytochemicals analysis of the extracts was also determined. The phytochemical analysis showed the presence of saponins, steroids, terpenoids, tannins, alkaloids, and flavonoids. The result of Vernonia amygdalina showed that the average zones of inhibitions observed against these bacterial ranges from 6-22mm. The highest zone is also exhibited against E. coli with average diameter of zone of inhibition of 22mm. At 100mg/ml concentration for Samonella, the zone of inhibition was recorded to be 21mm while at 12.5mg/ml there was no inhibition. At 25mg/ml and 12.5mg/ml, against Pseudomonas there was no inhibition. In other to further confirm the activity of these plant extracts, the minimum inhibitory concentration and minimum bactericidal concentration was determined and the result showed that the extract exerted good antibacterial activity on all the test organisms at different concentration. The result of minimum inhibitory concentration ranges from 10 to 12.5mg/ml and that of MBC ranges from 5 to 20mg/ml. It is worthy to note that MBC values is greater than that of minimum inhibitory concentration. The study provides insight into the antibacterial activities of the plant extracts and its use in the treatment of bacterial infections.

SYNTHESIS AND ANTIMICROBIAL EVALUATION OF SOME NEW OXAZINE DERIVATIVES DERIVED FROM COUMARINYL CHALCONES

A series of novel substituted 3-(2-amino-6-phenyl-6H-1,3-oxazin-4-yl)-2H-chromen-2-ones(AO1-AO10) was synthesized upon refluxing ethanolic solution of substituted 3-cinnamoyl-2H chromen-2-one with urea in presence of 20 % NaOH. The intermediate chalcones, substituted 3-cinnamoyl-2H-chromen2-ones, were synthesized by condensing 3-acetyl coumarin with various substituted benzaldehydes in presence of 20 % NaOH. The structures of the final synthesized compounds were characterized by UV, IR, 1 H NMR and mass spectra. The antibacterial activity of the synthesized compounds was estimated by tube dilution method. The turbidity was observed for the synthesized compounds. Few compounds showed promising minimum inhibitory concentration by tube dilution method.

Klebsiella pneumoniae and Colistin Susceptibility Testing: Performance Evaluation for Broth Microdilution, Agar Dilution and Minimum Inhibitory Concentration Test Strips and Impact of the “Skipped Well” Phenomenon

The emergence of multidrug resistant Gram-negative pathogens, particularly carbapenemase producers, has forced clinicians to use last line antibiotics, such as colistin. Since colistin susceptibility testing presents several challenges, this study aimed at evaluating the performance of two alternative susceptibility methods for Klebsiella pneumoniae, namely, agar dilution (AD) and MIC test strips (MTS). These approaches were compared with the reference method, broth microdilution (BMD), and provide a quantitative description for the “skipped well” (SW) phenomenon. Colistin susceptibility was evaluated by BMD and AD in parallel and triplicate, using 141 K. pneumoniae clinical isolates while MTS performance was evaluated only for a subset (n = 121). Minimum inhibitory concentration analysis revealed that a substantial part (n = 26/141; 18.4%) of the initial isolates was deemed undetermined by BMD due to the following: discordance between replicates (1.4%); presence of multiple SWs (7.8%); and the combination of both events (9.2%). Both AD and MTS revealed a high number of false-susceptible strains (“very major errors”), 37.5% and 68.8%, respectively. However, AD agreement indices were reasonably high (EA = 71.3% and CA = 94.8%). For MTS these indices were lower, in particular EA (EA = 41.7% and CA = 89.6), but the approach enabled the detection of distinct sub-populations for four isolates. In conclusion, this study provides the most comprehensive study on the performance of AD and MTS for colistin susceptibility testing in K. pneumoniae, highlighting its limitations, and stressing the importance of sample size and composition. Further, this study highlights the impact of the SW phenomenon associated with the BMD method for K. pneumoniae.

Phenotypical and Genotypical Comparison of Clostridium difficile Isolated from Clinical Samples: Homebrew DNA Fingerprinting versus Antibiotic Susceptibility Testing (AST) and Clostridial Toxin Genes

Background. Clostridium (Clostridioides) difficile is recognized as the major cause of healthcare antibiotic-associated diarrhea. We surveyed a molecular epidemiological correlation between the clinical isolates from two general hospitals in Iran through clustering toxigenic types and antibiotic susceptibility testing (AST) accuracy. Methods. Study population included 460 diarrhoeic specimens from inpatients with a history of antibiotic therapy. All samples underwent enriched anaerobic culture, confirmed by detection of gluD gene with PCR. Toxin status and AST were assessed by the disk diffusion method (DDM) and minimal inhibitory concentrations (MICs) of metronidazole, vancomycin, and rifampin. C. difficile outbreak was analyzed through conventional PCR by tracing toxin genes and Homebrew pulsed-field gel electrophoresis (PFGE) for characterizing isolates within our healthcare systems. Results. A total of 29 C. difficile strains were isolated by enriched anaerobic culture from the clinical samples. Among them, 22 (4.8%) toxigenic profiles yielded toxins A and B (tcdA, tcdB) and binary toxins (cdtA, cdtB). The minimum inhibitory concentration (MIC) was 18.1% and 9% for vancomycin and metronidazole, and all isolates were susceptible to rifampicin and its minimum inhibitory concentration was at <0.003 μg/mL. The most dominant toxigenic and antibiotic-resistant “pulsotype F” was detected through PFGE combined with multiple Clostridial toxigenic pattern and AST. Conclusions. DNA fingerprinting studies represent a powerful tool in surveying hypervirulent C. difficile strains in clinical settings. Resistance to vancomycin and metronidazole, as first-line antibiotics, necessitate accomplishment of proper control strategies and also prescription of tigecycline as a more appropriate option.

Studies on meropenem and cefixime metal ion complexes for antibacterial activity

Background The metal ion complexes of meropenem and cefixime with cadmium, silver, palladium, zinc, nickel, cobalt and copper were synthesized and characterized by UV, FTIR and H1-NMR spectrophotometry. The antibacterial effects of the complexes were studied using cup and plate method against S. aureus, B. subtilis, E. coli, P. aeruginosa and K. pneumoniae for normal and resistant strains of bacteria. The minimum inhibitory concentration of the metal ion complexes was determined by broth dilution method. Results UV spectroscopic studies suggested that meropenem ligand form complex with different metal ions and FTIR spectrum confirmed the proposed structure. Similarly, UV spectrum of cefixime metal ion complexes at λmax 202–295 nm and meropenem metal ion complexes at λmax 249–304 nm was observed in all the complexes. FTIR peaks for a proposed structure were observed in all the meropenem and cefixime metal ion, indicating the formation of complexes, and retained the functional groups of drugs. Meropenem as well as cefixime metal ion complexes exhibited more antibacterial activity against all the selected bacterial strains. Specifically, the lowest minimum inhibitory concentration against P. aeruginosa and K. pneumoniae was observed to be 100 and 150 μg/ml, respectively. Conclusion The present study concluded that the meropenem and cefixime metal complexes can exhibit the better treatment than individual drug on normal as well as resistant bacteria.

An integrated approach to evaluate different tetracycline derivatives for formulary decisions

Disclaimer In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. Purpose Stenotrophomonas maltophilia has emerged as a critical opportunistic pathogen associated with significant morbidity and mortality. Tetracycline derivatives have been recognized as alternative treatment options, but they have varied pharmacokinetic properties. An integrated approach to different tetracycline derivatives for formulary decisions is reported. Methods The minimum inhibitory concentration (MIC) data from clonally diverse bloodstream S. maltophilia isolates were examined, along with the pharmacokinetic profiles of 4 tetracycline derivatives, to predict achievable pharmacodynamic exposures with standard intravenous dosing regimens. Antimicrobial therapy was assessed using the ratio of daily drug acquisition cost relative to the ratio of the free-drug area under the time-concentration curve (fAUC) to the 90th percentile for minimum inhibitory concentration (MIC) values for isolates (fAUC/MIC90). Results In our analysis, minocycline had the greatest fAUC/MIC90. Doxycycline was the most financially preferred agent, as calculated using 2020 average wholesale price for base-case estimates of drug acquisition cost. Conclusion An integrated evaluation for antimicrobial formulary decision-making addressed local susceptibility data, pharmacokinetics, pharmacodynamics, dosing regimens, and drug acquisition costs. This comprehensive method is more objective than the conventional approach and warrants validation.