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Objective: The objective of this work is to develop a stability-indicating HPLC method for the quantification of posaconazole (PCZ) in tablet formulation using an Analytical Quality by Design (AQbD) approach., Materials and Methods: The development process involved the Design of Experiments (DOE) utilizing distinctive constraints mixture design for mobile phase ratio optimization and a 2-level factorial design for selection of extraction diluent compositions. Key responses measured included % assay and system suitability parameters. Method operable design regions (MODR) were determined, and final optimum conditions were selected. Forced degradation studies were conducted to assess method stability., Results: The optimized HPLC method employed a Zorbax C18 column with a mobile phase consisting of pH 3.5 10mM phosphate buffer, acetonitrile, and methanol in a ratio of 30:53:17% v/v/v. The method demonstrated stability-indicating capabilities, with PCZ degradation observed in acidic and oxidative environments, while remaining stable in alkali. Peak purity analysis from Empower software confirmed the absence of interaction with degradants. Validation according to ICH Q2 (R2) guidelines showed precision, linearity over the range of 0.25 to 376μg/mL, and accuracy demonstrated through recovery studies from 50 to 150%., Conclusion: The developed HPLC method utilizing AQbD approach is specific, robust, precise, and accurate for the quantification of PCZ in tablet formulations, thus suitable for routine analysis., (Copyright © 2024. Published by Elsevier Masson SAS.)

Significant pharmacokinetic variation occurs in critically ill patients, leading to underexposure to antibiotics and poor prognosis. In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LCMS/MS) platform for the simultaneous quantification of 8 antibacterial and 2 antifungal drugs, which is optimally suited for clinically efficient, real-time therapeutic drug monitoring (TDM). Multiple reaction monitoring (MRM) mass spectrometry was used in this method, and samples were prepared via protein precipitation with methanol. Chromatographic separation was accomplished on a BGIU Column-U02 (2.1ｘ50 mm, 3 µm), with six stable isotopes and one analog as an internal standard. The overall turnaround time of the assay was 5 minutes. All the drugs tested (piperacillin, cefoperazone, meropenem, levofloxacin, moxifloxacin, daptomycin, linezolid, vancomycin, fluconazole and voriconazole) were linear in the test concentration range (r ≥ 0.9900), the accuracy was 95 %-111 %, the precision variation coefficient was greater than or equal to 10 %, the lower limit of quantitation was 0.31-7.51 mg/L, and the coefficient of variation of the matrix factor was less than 10 %. The recovery rates ranged from 85 % to 115 %, and the antibiotics were stable at 4°C and -20°C for 6 days, with an offset of greater than or equal to 15 %. This method was successfully applied to routine TDM in 252 elderly critically ill patients., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Forest species are characterized by their wealth of essential oils (EOs), which play key requested for alternative control methods against weeds, fungi and pests. This study reports the chemical composition and highlight the antioxidant, antifungal and phytotoxic properties of the EOs obtained from Eucalyptus populifolia Desf, Eucalyptus woollsiana and Eucalyptus exserta . The EOs were analyzed by gas chromatography coupled to mass spectrometry (GC/MS). Their antioxidant, antifungal and phytotoxic properties were investigated. All EOs displayed a specific richness in eucalyptol (75.94-7.01%), camphene (6.97-0.4%) globulol (5.20-5.83%) and α-pinene (0.71-12.82 %). EOs isolated from E. populifolia exhibited significant antioxidant activity. Furthermore, all investigated EOs displayed significant antifungal properties against four phytopathogenic fungi belonging to Fusarium genus. The phytotoxic activity against five weeds showed varying effectiveness on seed germination and seedling growth, which exhibited greater effectiveness compared to glyphosate. Eucalyptus EOs have the potential to be used in the formulation of biopesticides.

The effect of internal and external magnetic fields on the separation of antifungal drugs by centrifugal acceleration thin-layer chromatography was reported for the first time. External and internal magnetic fields were applied using neodymium magnets and CoFe 2 O 4 @SiO 2 ferromagnetic nanoparticles. Separation of ketoconazole and clotrimazole was performed using a mobile phase consisting of n-hexane, ethyl acetate, ethanol, and ammonia (2.0:2.0:0.5:0.2, v/v). The influence of the magnetic field on the entire chromatographic system led to changes in the properties of the stationary and mobile phases and the analytes affecting the retention factor, shape, and width of the separated rings. The extent of this impact depended on the structure of the analyte and the type and intensity of the magnetic field. In the presence of the external magnetic field, there were more significant changes in the chromatographic parameters of the drugs, especially the width of the separated rings, and ketoconazole was more affected than clotrimazole. The changes are conceivably due to the effect of the magnetic field on the analyte distribution between the stationary and mobile phases, which is also caused by the possibility of the magnetic field affecting the viscosity, surface tension, and surface free energy between the stationary and mobile phases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Voriconazole is a broad-spectrum triazole antifungal agent recommended for invasive fungal diseases, including invasive aspergillosis. Therapeutic drug monitoring via voriconazole target trough concentration is important to ensure efficacy while preventing toxicity. Our aim was to determine the stability of voriconazole as adapted and measured by an immunoassay., Methods: Plasma from patient samples (n = 45) evaluated by a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was compared against an ARK immunoassay method, adapted and optimized on the Abbott Alinity c analyzer. Stability of voriconazole and analytical performance of ARK immunoassay was assessed, including functional sensitivity, limit of blank (LoB), limit of detection (LoD), and limit of quantification (LoQ), linearity, and precision., Results: ARK voriconazole immunoassay was highly correlated (Pearson R = 0.988) to the LC-MS/MS method, with an average bias of 0.09 mg/L (2%). CV at LoQ of 0.5 mg/L was 3.7% while the functional sensitivity was established at 0.05 mg/L. Overall imprecision with liquid quality control material obtained from ARK was 5.0%, 6.3%, and 5.9% at 1 mg/L, 5 mg/L, and 10 mg/L, respectively. Limit of blank and LoD were 0.02 mg/L and 0.05 mg/L, respectively. Voriconazole in lithium heparin plasma separator tube declines over time, with a decrease that is more evident near or above toxic concentrations., Conclusion: Voriconazole collected in gel separation tubes declines over time, possibly due to absorptive properties. Voriconazole measurements by immunoassay and LC-MS/MS demonstrated acceptable comparability with sufficient level of sensitivity and precision., (© Association for Diagnostics & Laboratory Medicine 2024. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Azole compounds are utilized to combat fungal infections in plants to protect them and also used for treating mycosis in humans. The LC-MS/MS method is a technique that combines liquid chromatography with tandem mass spectrometry for analysis of twelve azole compounds from wastewater (influent, effluent) and sewage sludge. The compounds were isolated from waste water using automatic extraction in the solid phase. Sludge samples were dried by lyophilization, after which they were subjected to ultrasound extraction with methanol. The quantification limits ranged from 0.3 ng/L (clotrimazole-CLO and prochloraz-PRO) to 1.5 ng/L (tetraconazole-TEB and penconazole-PEN), for wastewater samples and for sewage sludge, the LOQs ranged from 0.1 ng/g to 0.6 ng/g. High concentrations of climbazole-CLI (207-391 ng/L), tebuconazole (92-424 ng/L), and clotrimazole (6.9-93-ng/L) were observed in influent samples of the 8 urban wastewater treatment plants, followed by fluconazole (49.3-76.8 ng/L), and prochloraz (7.3-72 ng/L). The ∑Azoles had a maximum of 676 ng/L in the Galati effluent, followed by the Bucharest station 357 ng/L, and 345 ng/L in the Braila effluent. The highest value of the daily mass loading (input) level was observed for climbazole, 265 mg/day/1000 in Iasi station, followed by tebuconazole, 238 mg/day/1000 people in the Bucharest station, and 203 mg/day/1000 people for climbazole in the Targoviste station. The daily mass emission presented values between 0.7 and 247 mg/day/1000 people. The highest emissions were observed for climbazole, 247 mg/day/1000 people in Braila station; 174 mg/day/1000 people in the Iasi station and 129 mg/day/1000 people in the Bucharest station. The concentrations of climbazole detected in the effluent can present a high risk for the plants Lemna minor and Navicula pelliculosa. Clotrimazole may present a high risk to the plant Desmodesmus subspicatus and to the invertebrate Daphnia magna. PRO may present high risk to the invertebrate Mysidopsis Bahia., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Vasile IANCU reports financial support was provided by Romanian Ministry of Research Innovation and Digitalization. Vasile IANCU reports a relationship with Romanian Ministry of Research Innovation and Digitalization, that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This research was performed to evaluate the antimicrobial activity of methanol extract of Lannea coromandelica bark against fruit damage causing microbes such as fungi: Alternaria sp., Aspergillus sp., Botrytis sp., Cladosporium sp., Fusarium sp., Penicillium sp., Phytophthora sp., and Trichoderma sp. The bacteria: such as Chromobacter sp., Enterobacter sp., Erwinia sp., Flavobacterium sp., Lactobacillus sp., Pseudomonas sp., and Xanthomonas sp. was investigated. Furthermore, their biocompatibility nature was determined through animal (rat) model study and their fruit preserving potential was determined by edible coating preparation with chitosan and other substances. Interestingly, the extract showed dose dependent (1000 μg mL -1 ) activity against these microbes in the following order: Enterobacter sp. (26.4 ± 1.5) > Chromobacter sp. (25.4 ± 1.6) > Pseudomonas sp. (24.5 ± 1.3) > Flavobacterium sp. (24.3 ± 1.4) > Xanthomonas sp. (23.6 ± 1.6) > Erwinia sp. (23.6 ± 1.6) > Lactobacillus sp. (19.6 ± 1.3). Similarly, the antifungal activity was found as Penicillium sp. (32.6 ± 1.3) > Cladosporium sp. (32.6 ± 1.5) > Alternaria sp. (30.3 ± 1.2) > Aspergillus sp. (29.9 ± 1.8) > Botrytis sp. (29.8 ± 1.2) > Fusarium sp. (28.6 ± 1.5) > Trichoderma sp. (19.8 ± 1.4) > Phytophthora sp. (16.2 ± 1.1). The acute toxicity and histopathological study results revealed that the extract possesses biocompatible in nature. The illumination transmittance and active functional groups involved in interaction among test methanol extract and chitosan investigated by UV-vis and Fourier-transform infrared spectroscopy (FTIR) analyses and found average light transmittance and few vital functional groups accountable for optimistic interaction to creak edible coating. Approximately four (set I-IV) treatment sets were prepared, and it was discovered that all of the coated Citrus maxima fruit quality characteristics including total soluble solids (TSS), weight loss (%), pH of fruit pulp juice, and decay percentage were significantly (p>0.05) better than uncoated fruit., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Anidulafungin has poor oral bioavailability, with hardly any available information on how it affects breast milk, oral absorption, or gastrointestinal side effects in the infant. Case Presentation: A 40-year-old woman who recently gave birth to a healthy infant was treated for a period of 14 days for a C andida glabrata with 100 mg anidulafungin once a day. The department of clinical pharmacy was consulted to provide advice on how long the patient had to wait after ceasing anidulafungin before it was safe to start breastfeeding, with regard to preventing possible side effects of the drug to the infant, such as diarrhea or cholestasis and increase in liver enzyme values. The advice of the hospital pharmacist was pragmatic: to start breastfeeding within 2 days after the medication was discontinued based on half-time. Results: Owing to this lack of information, we measured anidulafungin concentrations in breast milk and found low levels. Conclusion: We concluded that anidulafungin is detectable in breast milk until 32 hours after anidulafungin treatment was stopped, and that no side effects were observed by the infant.

Herein we describe the in situ inhibitory activity of three hybrid films (FL1, FL2, and FL3) against two wild strains of Colletotrichum gloeosporioides and Penicillium digitatum as causal agents of rot in Persian limes. The films FL2 and FL3 contained 1.0 and 1.3 % weight/volume Litsea glaucescens essential oil (LgEO) and significantly (p<0.05) delayed rot emergence in Persian limes caused by both pathogens up to 10 days. The physicochemical properties of LgEO and hybrid films were obtained, whereas detailed HPLC profiling revealed that fruit covered with these films significantly (p<0.01) preserved reducing sugars (sucrose, fructose, and glucose), organic acids (citric acid, ascorbic acid, malic acid, and oxalic acid), and flavonoids with nutraceutical activity (hesperidin, eriocitrin, naruritin, neohesperidin, diosmin, vitexin, rutin, and quercetin). This evidence sustains that the composites generated in this investigation improve the shelf life of Persian limes and conserve their nutraceutical content., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

(1) Background: Previous studies reported the promising inhibitory effect of cold atmospheric plasma (CAP) on Candida albicans . However, the exact mechanisms of CAP's action on the fungal cell are still poorly understood. This study aims to elucidate the CAP effect on C. albicans cell wall, by evaluating the alterations on its structure and biochemical composition; (2) Methods: C. albicans cells treated with Helium-CAP were analyzed by atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) in order to detect morphological, topographic and biochemical changes in the fungal cell wall. Cells treated with caspofungin were also analyzed for comparative purposes; (3) Results: Expressive morphological and topographic changes, such as increased roughness and shape modification, were observed in the cells after CAP exposure. The alterations detected were similar to those observed after the treatment with caspofungin. The main biochemical changes occurred in polysaccharides content, and an overall decrease in glucans and an increase in chitin synthesis were detected; (4) Conclusions: Helium-CAP caused morphological and topographic alterations in C. albicans cells and affected the cell wall polysaccharide content.

The present study offers detailed insights into the antifungal and anti-mycotoxigenic potential of a biofilm forming lactic acid bacterium (Pediococcus pentosaceus) against one atoxigenic (Aspergillus flavus) and two toxigenic (Aspergillus nomius and Fusarium verticillioides) fungal strains. The antifungal effect of P. pentosaceus LBM18 strain was initially investigated through comparative analysis of fungi physiology by macroscopic visual evaluations and scanning electron microscopy examinations. The effects over fungal growth rate and asexual sporulation were additionally accessed. Furthermore, analytical evaluations of mycotoxin production were carried out by HPLC-MS/MS to provide insights on the bacterial anti-mycotoxigenic activity over fungal production of the aflatoxins B1, B2, G1 and G2 as well as fumonisins B1 and B2. Finally, reverse transcription quantitative real-time PCR (RT-qPCR) analysis was employed at the most effective bacterial inoculant concentration to evaluate, at the molecular level, the down-regulation of genes aflR, aflQ and aflD, related to the biosynthesis of aflatoxins by the strain of Aspergillus nomius. The effects over mycotoxin contamination were thought to be result of a combination of several biotic and abiotic factors, such as interaction between living beings and physical-chemical aspects of the environment, respectively. Several possible mechanisms of action were addressed along with potentially deleterious effects ascribing from P. pentosaceus misuse as biopesticide, emphasizing the importance of evaluating lactic acid bacteria safety in new applications, concentrations, and exposure scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cymbopogon flexuosus is a highly valued botanical species with significant applications in the food and food supplement industries, medicine, and cosmetics. The effects of four extraction techniques, supercritical CO 2 , microwave-ultrasonic, steam distillation, and hydrodistillation techniques, on the yield, phytochemical constituents, and antifungal activity against nine fungal species of Cymbopogon flexuosus aromatic oil (AO) were explored in this investigation. Gas chromatography connected with a mass spectrometry apparatus was employed for the qualitative and quantitative analyses of the investigated plant AOs. In addition, using the broth microdilution method, minimum inhibitory concentrations (MICs) were calculated for several fungi species. The supercritical CO 2 method gave the highest yield of AO (11.62 ± 0.03 ( w / w )) followed by the microwave-ultrasonic method (1.55 ± 0.05% ( w / w )) and the steam distillation method (1.24 ± 0.04% ( w / w )), while the hydrodistillation methods gave the lowest yield (1.17 ± 0.01 ( w / w )). In addition, eighteen molecules were specified in the AOs obtained with the supercritical CO 2 , microwave-ultrasonic, steam distillation, and hydrodistillation techniques, which constituted 99.36, 98.6, 98.21, and 98.31% ( v / v ) of the total oils, respectively. Additionally, linalyl acetate was the trending molecule in the microwave-ultrasonic and steam distillation methods, representing 24.61 and 24.34% ( v / v ), respectively, while geranial was the dominant molecule in the AOs extracted with the hydrodistillation and supercritical CO 2 extraction techniques (27.01 and 25.6% ( v / v ), respectively). The antifungal screening results revealed that the tested C. flexuosus AOs have potential antifungal effects against all the screened fungi species. The antifungal effect of the AOs extracted with the steam distillation and microwave-ultrasonic methods was remarkable compared with that of the commercial antifungal drug Fluconazole. However, the AOs extracted with these two methods have a more potent antifungal effect against Candida parapsilosis than that of Fluconazole with MICs of 3.13 ± 0.01, 3.13 ± 0.01, and 6.25 ± 0.91 µg/mL, respectively. The same effects were also observed against Trichophyton rubrum with MICs of 6.25 ± 0.91 µg/mL, respectively. The results of this investigation demonstrated that the steam distillation and microwave-ultrasonic methods are promising processes for the extraction of C. flexuosus AO with a potent antifungal effect. This may be an advantage for the utilization of C. flexuosus AO over some antifungal synthetic agents commonly utilized as medicines, preservatives, food additives, cosmetics, and nutrient supplements.

The antifungal and antiaflatoxigenic effects of four distinct plant species against Aspergillus flavus and Aspergillus parasiticus were investigated. Essential oils and methanolic extracts were prepared from aerial parts of Lippia javanica, Ocimum gratissimum, Satureja punctata, and stem barks of Toddalia asiatica by hydro-distillation and maceration, respectively. The poisoned food method was used to confirm the antifungal activity of essential oils and methanolic extracts from four different plant species against Aspergillus flavus and Aspergillus parasiticus, and high-performance liquid chromatography was used to quantify the antiaflatoxigenic activity. The essential oils of Satureja punctata and Lippia javanica showed the highest antiaflatoxigenic activity against the fungi strains tested at concentrations of 1.25, 2.5, and 5 µL/mL, followed by Ocimum gratissimum essential oil while Toddalia asiatica essential oil exerted moderate antiaflatoxigenic activity. Meanwhile, the methanolic extracts showed a wide spectrum of low to high antifungal and antiaflatoxigenic activities at concentrations of 125, 250, and 500 µg/mL against A. flavus and A. parasiticus. This study has indicated that the essential oils of Satureja punctate, Lippia javanica, and Ocimum gratissimum had substantial antifungal and antiaflatoxigenic activities compared to their methanolic extracts, while Toddalia asiatica methanolic extract had a moderate antifungal activity compared to its essential oil., (© 2023. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: Lantana trifolia L. (Verbenaceae) is a shrubby plant. In folk medicine, its leaves are used in the form of infusions and syrups to treat angina, coughs, and colds; they are also applied as tranquilizer. Previous studies have reported the antimicrobial potential of the compounds present in L. trifolia leaves., Objectives: To report the anti-Candida activities of the fractions obtained from the fruits and leaves of two L. trifolia specimens., Methods: The L. trifolia fractions were submitted to UFLC-DAD-(+)-ESI-MS/MS, and the data were analyzed by using multivariate statistical tools (PCA, PLS-DA) and spectral similarity analyses based on molecular networking, which aided dereplication of the bioactive compounds. Additionally, NMR analyses were performed to confirm the chemical structure of some of the major compounds in the fractions., Results: The ethyl acetate fractions presented MIC values lower than 100 µg mL -1 against the three Candida strains evaluated herein (C. albicans, C. tropicalis, and C. glabrata). Fractions FrPo AcOEt, FrPe AcOEt, and FrPe nBut had MIC values of 1.46, 2.93, and 2.93 µg mL -1 against C. glabrata, respectively. These values resembled the MIC value of amphotericin B, the positive control (0.5-1.0 µg mL -1 ), against this same strain. Cytotoxicity was measured and used to calculate the selectivity index., Conclusion: On the basis of our data, the most active fractions in the antifungal assay were more selective against C. glabrata than against non-infected cells. The analytical approach adopted here allowed us to annotate 29 compounds, nine of which were bioactive (PLS-DA results) and belong to the class of phenolic compounds., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Aptameric receptors are important biosensor components, yet our ability to identify them depends on the target structures. We analyzed the contributions of individual functional groups on small molecules to binding within 27 target-aptamer pairs, identifying potential hindrances to receptor isolation-for example, negative cooperativity between sterically hindered functional groups. To increase the probability of aptamer isolation for important targets, such as leucine and voriconazole, for which multiple previous selection attempts failed, we designed tailored strategies focused on overcoming individual structural barriers to successful selections. This approach enables us to move beyond standardized protocols into functional group-guided searches, relying on sequences common to receptors for targets and their analogs to serve as anchors in regions of vast oligonucleotide spaces wherein useful reagents are likely to be found.

Fungal infections represent a relevant issue in agri-food and biomedical fields because they could compromise quality of food and humans' health. Natural extracts represent a safe alternative to synthetic fungicides and in the green chemistry and circular economy scenario, agro-industrial wastes and by-products offer an eco-friendly source of bioactive natural compounds. In this paper, phenolic-rich extracts from Olea europaea L. de-oiled pomace, Castanea sativa Mill. wood, Punica granatum L. peel, and Vitis vinifera L. pomace and seeds were characterized by HPLC-MS-DAD analysis. Finally, these extracts were tested as antimicrobial agents against pathogenic filamentous fungi and dermatophytes such as Aspergillus brasiliensis , Alternaria sp., Rhizopus stolonifer , and Trichophyton interdigitale . The experimental results evidenced that all extracts exhibited a significant growth inhibition for Trichophyton interdigitale . Punica granatum L., Castanea sativa Mill., and Vitis vinifera L. extracts showed a high activity against Alternaria sp. and Rhizopus stolonifer . These data are promising for the potential applications of some of these extracts as antifungal agents in the food and biomedical fields.

Grindelia squarrosa is an arid lands herb that has been used in Native American traditional medicine, is a potential source of pharmacologically active compounds, and has been explored as a source of biofuel. The purpose of this work was to examine the essential oil composition of G. squarrosa from southern Idaho. Gas chromatographic methods revealed the essential oil of G. squarrosa var. serrulata to be rich in monoterpenoids, α-pinene (21.9%), limonene (17.1%), terpinolene (10.6%), and borneol (6.5%). The essential oil composition of G. squarrosa from Idaho is similar to that previously reported from specimens collected from Montana and confirms the volatile phytochemistry of plants growing in North America. The major essential oil components were screened for antimicrobial activity against respiratory and dermal pathogens. (-)-β-Pinene showed strong antibacterial activity against Streptococcus pneumoniae (MIC 39.1 μg/mL) and (-)-borneol showed strong activity against Staphylococcus aureus (MIC 78.1 μg/mL).

Antifungals are used widely in clinical and agricultural practice to control fungal growth, either treating or preventing infection. There are reports of increasing prevalence of resistance to antifungals in human pathogens and concern that their use in agriculture is driving clinical resistance in patients. While crop protection products are the most obvious source in agriculture, a further source may be biosolids from wastewater treatment. In the UK, these are applied to land to provide nutrients and improve soil structure for crops. In this study, biosolids from ten sites in England and one in Wales were analysed for clinical antifungals. Ketoconazole and miconazole were detected in all samples with a median concentration of 0.87 and 0.54 mg kg -1 dry weight (DW), respectively. Clotrimazole was detected at seven of eleven sites at a median level of 1.32 mg kg -1 DW and its absence at four others was considered treatment related. Two prescription-only and systemic medications, itraconazole and posaconazole, were frequently detected with median concentrations of 0.14 mg kg -1 DW and 0.09 mg kg -1 DW, respectively. The biosolid levels of itraconazole found in this study were two orders of magnitude higher than an indicative Predicted No Effect Concentration for resistance selection (PNEC-R) in soil. Neither fluconazole, griseofulvin, and voriconazole nor flucytosine and nystatin were found above the limit of detection of 0.01 or 0.1 mg kg -1 as received, respectively. The findings show that biosolids represent a viable pathway for antifungal agents to reach soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2023. Published by Elsevier B.V. All rights reserved.)

The purpose of this study was to evaluate the phytochemical profiles of the seeds, sprouts, leaves, flowers, roots and herb of Salvia hispanica and to demonstrate their significant contribution to antioxidant and antimicrobial activities. Applied methods were: HPLC-DAD coupled with post-column derivatization with ABTS reagent, untargeted metabolomics performed by LC-Q-Orbitrap HRMS, and two-fold micro-dilution broth method, which involved suspending a solution of tested compounds dissolved in DMSO in Mueller-Hinton broth for bacteria or Mueller-Hinton broth with 2% glucose for fungi. Metabolomic profiling using LC-Q-Orbitrap HRMS used in this study yielded the identification and preliminary characterization of one hundred fifteen compounds. The dominant class of compounds was terpenoids (31 compounds), followed by flavonoids (21 compounds), phenolic acids and derivatives (19 compounds), organic acids (16 compounds) and others (fatty acids, sugars and unidentified compounds). The organic and phenolic acids were the most abundant classes in terms of total peak area, with distribution depending on the plant raw materials obtained from S. hispanica. The main compound among this class for all types of extracts was rosmarinic acid which was proven to be the most abundant for antioxidant potential. All tested extracts exhibited considerable antibacterial and antifungal activity. The strongest bioactivity was found in leaf extracts, which presented bactericidal activity against Gram-positive bacteria ( S. aureus , S. epidermidis , M. luteus and E. faecalis ). The work represents the first compendium of knowledge comparing different S. hispanica plant raw materials in terms of the profile of biologically active metabolites and their contribution to antioxidant, antimicrobial and antifungal activity.

The present study involves investigation of Azadiracta Indica flowers with respect to its pharmacognostic properties, phytochemical screening, and its application as anti-oxidant, anti-biofilm, and anti-microbial agent. The Pharmacognostic characteristics were evaluated with respect to moisture content, total ash content, acid, and water-soluble ash content, swelling index, foaming index, and metal content. The macro and micronutrient content of the crude drug was estimated by AAS and Flame photometric methods and it gives the quantitative estimation of minerals, where calcium is present in abundance (88.64 mg/L). Soxhlet extraction was carried out in the increasing order of polarity of the solvent viz Petroleum Ether (PE), Acetone (AC), and Hydroalcohol (20 %) (HA) to extract the bioactive compounds. The characterization of the bioactive compounds of all the three extract have been carried out using gcms and lcms. The presence of 13 major compounds have been identified in PE extract and 8 compounds in AC extract using gcms studies. The HA extract is found to contain polyphenols, flavanoids, and glycosides. The antioxidant activity of the extracts was evaluated by DPPH, FRAP, and Phosphomolybdenum assay. This reveals that HA extract shows good scavenging activity than PE and AC extracts which is well correlated with the bioactive compounds, especially phenols which are present as a major component in the extract. The anti-microbial activity was investigated via Agar well diffusion method for all the extracts. Among all the extracts HA extract shows good antibacterial activity with MIC of 25 μg/mL and AC extract shows good anti-fungal activity with MIC of 25 μg/mL. The antibiofilm assay confirms that the HA extract shows good biofilm inhibition about 94 % among other extracts when tested on human pathogens. The results confirm that the HA extract of A. Indica flowers will be an excellent source of natural anti-oxidant and also antimicrobial agents. This paves the way for its potential uses in herbal product formulation., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

The main objective of our study was to investigate the possible differences in the chemical composition of extractives from the bark of silver fir ( Abies alba ) with respect to the location of the bark sample on the tree, viz. differences in extract composition between stem bark and branch bark samples. Extractives in the bark samples from branches, depending on the distance of the sample from the trunk, were also analysed, and the stem bark samples were analysed with respect to their inner and outer parts. The results of the chemical analysis of extractives were supported by information about their antifungal and antioxidant effects. After felling and sampling silver fir trees, the collected bark samples were ground and freeze-dried. Extraction of bark samples was followed by a system of accelerated extraction using only water as a solvent. The extracts were analysed chemically using gravimetry, spectrophotometry and chromatography. Free-radical-scavenging activity was measured using the DPPH method, and the antifungal effect towards three moulds and three wood-decaying fungi was investigated with antifungal assay using the agar well diffusion method. It was found that the moisture content in bark samples decreased intensively just after the bark samples were peeled off the stem. Detailed chromatographic analysis showed that the bark extracts contained 14 compounds, among which phenolic acids, flavonoids and lignans were found to be the characteristic ones. The content of hydrophilic extractives in the branch bark samples decreased with increasing distance of the sample location from the tree stem. The largest amounts of phenolic extractives were measured in stem bark, followed by branch bark sampled at the point at which the branch entered the tree. Analysis of the separated parts of the bark showed that the outer layers of stem bark contained larger amounts of phenolic extractives, as well catechin and epicatechin, compared to the inner layers. Concentrated extracts of branch bark showed the largest free-radical-scavenging activity among the investigated samples, while strong antifungal effects of the bark extract were not found.

Fungal infestation of cereals seriously affects human health and the economy. Protecting cereals from fungal infestation is meaningful. Therefore, safe and economical antifungal agents must be explored. Oregano essential oil (OEO) has broad-spectrum antifungal activity but is unstable and poorly water-dispersible. CoAl-LDH ultrathin nanosheets (CoAl-LDH UNs) are a biosafe carrier with peroxidase-like activity and good water dispersibility. A new nanocomposite was constructed by integrating OEO and CoAl-LDH UNs named OEO/CoAl-LDH UNs. The antifungal activity of OEO was improved for the synergetic effect of OEO and CoAl-LDH UNs. OEO/CoAl-LDH UNs showed excellent antifungal activity against Fusarium moniliforme, Fusarium graminearum and Aspergillus flavus. A polyvinyl alcohol film containing OEO/CoAl-LDH UNs was prepared as a food package and effectively inhibited fungal infestation on cereals. This work can provide some insights for the exploration of other new antifungal materials. The good biosafety of this antifungal material can be effectively applied to food packaging., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In the present study, six commercial pesticide formulations with antifungal activity were characterized. Thus, two complementary injection methods based on gas chromatography were employed: direct injection (DI) and headspace (HS), both coupled to high resolution mass spectrometry (GC-Q-Orbitrap-MS). The combination of both injection modes allowed the tentatively identification of potential co-formulants. Available analytical standards were acquired for their confirmation, and 21 compounds were successfully confirmed. Finally, the concentration of these co-formulants was calculated, finding the highest value in one of the pesticide formulation, at 218.22 g L -1 for cyclohexanone. Results clearly show that this methodology is suitable for the reliable identification of co-formulants in pesticide formulations, offering high sensitivity, and highlighting that five co-formulants were detected by both injection techniques (DI and HS). Moreover, one of the main advantages of the proposed methods was the great capacity for the elucidation of compounds with similar molecular formula, bearing in mind that up to 8 co-formulants with the same molecular formula C 10 H 14 , were well differentiated by retention times between 8.46 (1-methyl-3-propylbenzene) and 10.98 min (1,2,3,4- tetramethylbenzene) in one of the pesticide formulation. Toxicity to human health and the environment has been evidenced for the co-formulants detected, finding compounds with relatively high toxicity, as naphthalene and cyclohexanone., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

During the postharvest period, citrus fruits are exposed to Penicillium italicum , Penicillium digitatum, and Geotrichum candidum . Pesticides such as imazalil (IMZ), thiabendazole (TBZ), orthophenylphenol (OPP), and guazatine (GUA) are commonly used as antifungals. Glyphosate (GP) is also used in citrus fields to eliminate weed growth. The sensitivity of fungal pathogens of citrus fruit to these pesticides and 1,8-cineole was evaluated, and the effect of GP on the development of cross-resistance to other chemicals was monitored over a period of 3 weeks. IMZ most effectively inhibited the mycelial growth and spore germination of P. digitatum and P. italicum , with minimum inhibitory concentrations (MICs) of 0.01 and 0.05 mg/mL, respectively, followed by 1,8-cineole, GP, and TBZ. 1,8-Cineole and GP more effectively inhibited the mycelial growth and spore germination of G. candidum , with minimum inhibitory concentrations (MICs) of 0.2 and 1.0 mg/mL, respectively, than OPP or GUA. For the spore germination assay, all substances tested showed a total inhibitory effect. Subculturing the fungal strains in culture media containing increasing concentrations of GP induced fungal tolerance to GP as well as to the fungicides. In soil, experiments confirmed that GP induced the tolerance of P. digitatum to TBZ and GP and the tolerance of P. italicum to IMZ, TBZ, and GP. However, no tolerance was recorded against 1,8-cineole. In conclusion, it can be said that 1,8-cineole may be recommended as an alternative to conventional fungicides. In addition, these results indicate that caution should be taken when using GP in citrus fields.

Aspergillus westerdijkiae may produce large amounts of ochratoxin A (OTA) in dry-cured meat products. Natural strategies to control ochratoxigenic moulds using biocontrol agents (BCAs) are currently in the spotlight. The aim of this study was to test the effects of Debaryomyces hansenii and its combination with rosemary derivatives and with a commercial antifungal preparation composed by natamycin and potassium sorbate (AP) against A. westerdijkiae in a dry-cured fermented sausage based-medium. The yeast and rosemary leaves were added to the medium, and rosemary essential oil and AP were added on the casings put on the medium surface to simulate the real product. The growth rate, OTA production and comparative proteomics were analysed. The mould growth in the presence of each treatment was not indicative of their efficiency on OTA repression. The treatment with AP did not affect to the OTA concentration, maybe as consequence of the stressful stimulation of the subinhibitory doses used. D. hansenii added alone or with rosemary showed the best results, decreasing the OTA production >80 %, suggesting that it can be useful as preservative agent during industrial processing. Attending to the proteomic results, its antifungal activity seems to be based on the reduction in abundance of proteins involved in OTA biosynthesis and in the cell wall integrity pathway., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Plants produce a variety of high-value chemicals (e.g., secondary metabolites) which have a plethora of biological activities, which may be utilised in many facets of industry (e.g., agrisciences, cosmetics, drugs, neutraceuticals, household products, etc.). Exposure to various different environments, as well as their treatment (e.g., exposure to chemicals), can influence the chemical makeup of these plants and, in turn, which chemicals will be prevalent within them. Essential oils (EOs) usually have complex compositions (>300 organic compounds, e.g., alkaloids, flavonoids, phenolic acids, saponins and terpenes) and are obtained from botanically defined plant raw materials by dry/steam distillation or a suitable mechanical process (without heating). In certain cases, an antioxidant may be added to the EO (EOs are produced by more than 17,500 species of plants, but only ca. 250 EOs are commercially available). The interesting bioactivity of the chemicals produced by plants renders them high in value, motivating investment in their production, extraction and analysis. Traditional methods for effectively extracting plant-derived biomolecules include cold pressing and hydro/steam distillation; newer methods include solvent/Soxhlet extractions and sustainable processes that reduce waste, decrease processing times and deliver competitive yields, examples of which include microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), subcritical water extraction (SWE) and supercritical CO2 extraction (scCO2). Once extracted, analytical techniques such as chromatography and mass spectrometry may be used to analyse the contents of the high-value extracts within a given feedstock. The bioactive components, which can be used in a variety of formulations and products (e.g., displaying anti-aging, antibacterial, anticancer, anti-depressive, antifungal, anti-inflammatory, antioxidant, antiparasitic, antiviral and anti-stress properties), are biorenewable high-value chemicals.

Aim: To investigate hand-dug well water used for drinking and domestic purposes in a rural community in Southwest Nigeria for water safety and fungal presence as well as to determine the antifungal resistance and aflatoxigenic potentials of isolated fungi., Methods and Results: Water samples were analysed for risk of contamination, bacteriological and mycological parameters using a standard sanitary survey checklist and microbiological culturing. Isolates were identified and subjected to antifungal resistance profiling using the diffusion method for susceptibility testing of filamentous fungi. Multidrug-resistant strains were confirmed with DNA barcoding identification. Fungal isolates were screened for aflatoxigenic potentials by culture methods and confirmed by densitometric analysis. From the 23 hand-dug wells assessed, 56.52% had a high risk of contamination (ROC) score, nitrate >50 mg/L (73.9%), and the presence of total coliforms (100%), Escherichia coli (43.48%) and fungi (91.3%). Spearman rank correlation coefficient gave a positive and strong correlation between Total Fungi and Faecal Coliform (r = 0.701; p = 0.016; n = 23) at 0.05 significance level (2-tailed). Aspergillus sp. (34%), Penicillium sp. (18%) and Rhizopus sp. (17%) were the most dominant fungal genera. Isolates were resistant to fluconazole (76.19%), ketoconazole (73.80%), clotrimazole (92.86%), griseofulvin (88.09%) and nystatin (100%). Penicillium and Aspergillus (50%) were positive for cultural mycotoxin screening. A strain of antifungal-resistant A. flavus produced aflatoxin B1 (752 ppb) and B2 (15 ppb)., Significance of the Study: The existence of antifungal-resistant and aflatoxigenic fungi in water used for drinking and domestic purposes shows that filamentous fungi constitute greater threats than previously recognized and this call for a paradigm shift from the perceived safety of untreated hand-dug well-water., (© 2022 Society for Applied Microbiology.)

Tetradenia riparia is known for its richness in essential oil which has been widely investigated due to its biological activities such as antimicrobial, insecticidal, trypanocidal, antimalarial and antioxidant. The objective of this work was to chemically analyze and evaluate the antifungal and antimycotoxigenic activity of the essential oil and the crude extract of leaves, flower buds and stems of T. riparia from the northwest region of the state of Paraná. The essential oil was obtained by hydrodistillation using a Clevenger-type apparatus. To obtain the crude extract, the leaves, flower buds and stems were pulverized and subjected to a dynamic maceration process using 70% v v -1 ethyl alcohol. Chemical analysis of the essential oil was performed by GC/MS, and chemical identification of the crude extract by UHPLC-ESI/qTOF. Antifungal activity ( Rhizopus oryzae , Aspergillus flavus , Aspergillus ochraceus , Penicillium verrucosum and Fusarium graminearum ) was performed by broth microdilution and the antimycotoxigenic assay was performed with A. ochraceus and P. verrucosum . Ochratoxin A was extracted by partition with chloroform and quantified by HPLC-FL. The oil yield was 0.29% for leaves, 0.34% for stems and 0.38% for flower buds, and the major compounds were fenchone, β-caryophyllene, α-cadinol, 14-hydroxy-9- epi-caryophyllene, 9β,13β-epoxy-7-abietene, α-cadinol and 6-7-dehydroroyleanone. The main chemical compounds identified in the crude extract were terpenes, anthocyanins, flavonoids, tannins and phenolic acids. The minimum inhibitory concentration (MIC) of oils from leaves, flower buds and stems for the strains tested ranged from 0.87   mg mL -1 to 33.3 mg mL -1 , while the minimum fungicidal concentration (MFC) ranged from 6.94 mg mL -1 and 33.3 mg mL -1 . The MIC and MFC for ketoconazole, tebuconazole, sorbate and nitrite ranged from 0.05 to 33.3 mg mL -1 . The oil and crude extract of leaves, stems and flower buds showed an inhibition of ochratoxin A production for P. verrucosum of approximately 100%.

The study evaluated the antifungal activities of the 70% ethanol extracts of Sclerocarya birrea leaves (SBL) and stem bark (SBB) against C. albicans strains and fluconazole-resistant isolates, their antifungal effects in combination with conventional antifungals as well as their effects on the biofilms of the C. albicans strains and isolates. UPLC-QTOF-MS/MS analysis was then carried out to investigate the metabolite profile of the extracts and UPLC fingerprints developed for their routine identification as part of quality control measures. The extracts exhibited considerable antifungal activity with MIC ranging from 12.21 to 97.66  μ g/mL and MFC from 12.21 to 390.63  μ g/mL against the C. albicans strains and isolates. The antifungal activity of the stem bark extract was higher than the leaf extract. SBL and SBB also significantly inhibited biofilm formation (IC 50 = 12.49 to 164.42  μ g/mL) and the mature biofilms (IC 50 = 91.50 to 685.20  μ g/mL) of the strains and isolates of the C. albicans and demonstrated potential for their use in combination therapies with currently used antifungals especially the stem bark extract with nystatin. Metabolite profiling identified the presence of polyphenolic compounds in both leaves and stem bark mostly flavonoids, their derivatives, and proanthocyanidins, which contribute in part to the bioactivity of the plant. Whereas flavonoids like quercetin, myricetin, and their derivatives were abundant in the leaves, epicatechin monomers with their condensed tannins, including procyanidin B2 and procyanidin C, were abundant in the stem bark. Fingerprints of SBL and SBB were developed and validated and could be used as qualitative tools to authenticate the plant. The outcomes of the study show the promise of the leaf and stem bark extracts of S. birrea to be studied further and developed as antifungal agents., Competing Interests: The authors declare no conflict of interest., (Copyright © 2022 Benjamin Kingsley Harley et al.)

Lupinus plants are well-recognized due to their significant alkaloid content, which has made them the subject of several studies. However, the lack of chemical and biological information on the Colombian Lupinus species remains a fact. Therefore, the alkaloidal fractions from the leaves of L. mirabilis obtained by conventional solvent and ultrasound-assisted extraction (CSE and UAE, respectively) at different time frames were analyzed. Sparteine ( 2 ) was the main component in all cases; however, its relative abundance showed large variability, ranging from 64.7% to 80.6%. Minor constituents were also affected by the extraction conditions. In general, prolonged times gave a higher proportion of alkaloids under CSE, while only a slight decrease was observed under UAE. Both the method and extraction time appeared to equally affect the ratios of particular alkaloids, leading to variations in their effect on the mycelial growth of Fusarium oxysporum . Holistic analysis through multiple-covariate statistical methods as an approach to integrating chemical and bioactivity datasets allowed inferring the compounds most likely responsible for the changes in mycelial growth inhibition. 13α-Hydroxylupanine ( 12 ) might represent a promising compound to be included in further studies against this phytopathogen.

Luliconazole (LCZ) is a novel antifungal imidazole with broad-spectrum and high susceptibility of Aspergillus and Fusarium are the dominant species of fungal keratitis, may potentially be a new medical treatment option for ocular fungal infection. To evaluate LCZ distribution in ocular tissues after topical application for the development of ophthalmic delivery system, it is important to have a bioanalytical method for measuring the drug concentrations in different ocular tissues and aqueous humor (AH). A selective and sensitive ultrahigh performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was developed for the quantification of LCZ in rabbit ocular tissues, including conjunctiva, cornea, AH, iris, lens, vitreous humor (VH), retinal choroid and sclera, using lanoconazole as internal standard (IS). Chromatographic separation was achieved on a Xterra MS, C18 column (2.1 × 50 mm, 3.5 μm) using mobile phase with formic acid solution (0.2%, v/v): acetonitrile (50:50, v/v) at a flow rate of 0.2 ml/min, and the run time was 2.5 min. Detection was performed using the transitions 354.1 → 150.3 m/z for LCZ and 320.1 → 150.3 m/z for IS by positive ion electrospray ionization in multiple reaction monitoring (MRM) mode. Method validation was conducted in accordance with U.S. Food and Drug Administration's regulatory guidelines for bioanalytical method validation. The calibration curves were linear over the concentration range from 2.80 ng/ml to 2038 ng/ml for conjunctiva, cornea and sclera, 2.09 ng/ml to 1019 ng/ml for AH, 2.09 ng/ml to 509.5 ng/ml for iris, 2.09 ng/ml to 203.8 ng/ml for retinal choroid and VH, 2.04 ng/ml to 101.9 ng/ml for lens, with all the squared correlation coefficients (r 2 ) more than 0.99. The accuracy of the method was within the acceptable limit of 89.34%∼112.78% at the lower limit of quantification and other concentrations, Inter-day and intra-day precision values, expressed in terms of RSD (%), in all tissues were within 15% at all concentrations. The mean recoveries of LCZ in rabbit ocular tissues was 84.85%∼100.52%. No interference was found due to matrix components. Luliconazole was stable during the stability studies, including autosampler stability, benchtop stability, freeze/thaw stability and long-term stability. The method was successfully applied to the ocular pharmacokinetic and tissues distribution studies of LCZ in rabbit after topical administration of LCZ ophthalmic drug delivery system., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The utilization of fermented foods with health-promoting properties is becoming more popular around the world. Consequently, kefir, a fermented milk beverage made from kefir grains, was shown in numerous studies to be a probiotic product providing significant health benefits. Herein, we assessed the antibacterial and antifungal potential of kefir against a variety of pathogenic bacteria and fungi. This study also showed the effectiveness of kefir in healing wounds in human gastric epithelial cells (GES-1) by (80.78%) compared with control (55.75%) within 48 h. The quantitative polymerase chain reaction (qPCR) results of kefir-treated HCV- or HBV- infected cells found that 200 µg/mL of kefir can eliminate 92.36% of HCV and 75.71% of HBV relative to the untreated infected cells, whereas 800 µg/mL (the highest concentration) completely eradicated HCV and HBV. Moreover, the estimated IC 50 values of kefir, at which HCV and HBV were eradicated by 50%, were 63.84 ± 5.81 µg/mL and 224.02 ± 14.36 µg/mL, correspondingly. Kefir can significantly suppress the elevation of TNF-α and upregulate IL-10 and INF-γ in both treated HCV- and HBV-infected cells. High-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analysis of kefir revealed the presence of numerous active metabolites which mainly contribute to the antimicrobial, antiviral, and immunomodulatory activities. This study demonstrated, for the first time, the anti-HBV efficacy of kefir while also illustrating the immunomodulatory impact in the treated HBV-infected cells. Accordingly, kefir represents a potent antiviral agent against both viral hepatitis C and B, as well as having antimicrobial and wound healing potential.

Background: Peanut stem rot is a serious plant disease that causes great economic losses. At present, there are no effective measures to prevent or control the occurrence of this plant disease. Biological control is one of the most promising plant disease control measures. In this study, Pseudomonas chlororaphis subsp. aurantiaca strain zm-1, a bacterial strain with potential biocontrol properties isolated by our team from the rhizosphere soil of Anemarrhena asphodeloides, was studied to control this plant disease., Methods: We prepared extracts of Pseudomonas chloroaphis zm-1 extracellular antibacterial compounds (PECEs), determined their antifungal activities by confrontation assay, and identified their components by UPLC-MS/MS. The gene knockout strains were constructed by homologous recombination, and the biocontrol efficacy of P. chlororaphis zm-1 and its mutant strains were evaluated by pot experiments under greenhouse conditions and plot experiments, respectively., Results: P. chlororaphis zm-1 could produce extracellular antifungal substances and inhibit the growth of Sclerotium rolfsii, the main pathogenic fungus causing peanut stem rot. The components of PECEs identified by UPLC-MS/MS showed that three kinds of phenazine compounds, i.e., 1-hydroxyphenazine, phenazine-1-carboxylic acid (PCA), and the core phenazine, were the principal components. In particular, 1-hydroxyphenazine produced by P. chlororaphis zm-1 showed antifungal activities against S. rolfsii, but 2-hydroxyphenazine did not. This is quite different with the previously reported. The extracellular compounds of two mutant strains, ΔphzH and ΔphzE, was analysed and showed that ΔphzE did not produce any phenazine compounds, and ΔphzH no longer produced 1-hydroxyphenazine but could still produce PCA and phenazine. Furthermore, the antagonistic ability of ΔphzH declined, and that of ΔphzE was almost completely abolished. According to the results of pot experiments under greenhouse conditions, the biocontrol efficacy of ΔphzH dramatically declined to 47.21% compared with that of wild-type P. chlororaphis zm-1 (75.63%). Moreover, ΔphzE almost completely lost its ability to inhibit S. rolfsii (its biocontrol efficacy was reduced to 6.19%). The results of the larger plot experiments were also consistent with these results., Conclusions: P. chlororaphis zm-1 has the potential to prevent and control peanut stem rot disease. Phenazines produced and secreted by P. chlororaphis zm-1 play a key role in the control of peanut stem rot caused by S. rolfsii. These findings provide a new idea for the effective prevention and treatment of peanut stem rot., (© 2021. The Author(s).)

Treatment of invasive fungal infections with Caspofungin is used as the first-line antifungal agents. The minimum inhibitory concentration value is a test which indicates the degree of sensitivity of a strain regarding a drug. However, no value of minimum inhibitory concentration for caspofungin is available because very variable value is obtained. In this work, we study the link with the adsorption phenomenon of CSF previously described in literature and the lack of minimum inhibitory concentration value. A systematic study of the impact of different parameters on CSF adsorption is reported. The effect of the nature of container material, the aqueous solution pH and the organic solvent proportion was studied. In addition, the possibility of using a coating agent to minimize the adsorption was assayed and evaluated. Results obtained showed the importance of the material used during the manipulation of CSF. The use of acidic pH aqueous solution or the addition of acetonitrile or methanol proportions (50 % and 70 %, respectively) were found efficient to avoid adsorption of CSF on glassware material, which is the relevant strategy for analytical samples of caspofungin. The treatment of HPLC glass vials and 96-well plates with N-(2-aminoethyl)-3-aminopropyltrimethoxysilane reduced the adsorption. The significant adsorption observed in this work especially with plastic materials, questions the results obtained before in different assays and explained the absence of MIC value., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

The current study aimed to explore the crude oils obtained from the n-hexane fraction of Scutellaria edelbergii and further analyzed, for the first time, for their chemical composition, in vitro antibacterial, antifungal, antioxidant, antidiabetic, and in vivo anti-inflammatory, and analgesic activities. For the phytochemical composition, the oils proceeded to gas chromatography-mass spectrometry (GC-MS) analysis and from the resultant chromatogram, 42 bioactive constituents were identified. Among them, the major components were linoleic acid ethyl ester (19.67%) followed by ethyl oleate (18.45%), linolenic acid methyl ester (11.67%), and palmitic acid ethyl ester (11.01%). Tetrazolium 96-well plate MTT assay and agar-well diffusion methods were used to evaluate the isolated oil for its minimum inhibitory concentrations (MIC), minimum bactericidal concentration (MBC), half-maximal inhibitory concentrations (IC 50 ), and zone of inhibitions that could determine the potential antimicrobial efficacy's. Substantial antibacterial activities were observed against the clinical isolates comprising of three Gram-negative bacteria, viz., Escherichia coli , Klebsiella pneumoniae , and Pseudomonas aeruginosa , and one Gram-positive bacterial strain, Enterococcus faecalis . The oils were also effective against Candida albicans and Fusarium oxysporum when evaluated for their antifungal potential. Moreover, significant antioxidant potential with IC 50 values of 136.4 and 161.5 µg/mL for extracted oil was evaluated through DPPH (1,1-Diphenyl-2-picryl-hydrazyl) and ABTS assays compared with standard ascorbic acid where the IC50 values were 44.49 and 67.78 µg/mL, respectively, against the tested free radicals. The oils was also potent, inhibiting the α-glucosidase (IC50 5.45 ± 0.42 µg/mL) enzyme compared to the standard. Anti-glucosidase potential was visualized through molecular docking simulations where ten compounds of the oil were found to be the leading inhibitors of the selected enzyme based on interactions, binding energy, and binding affinity. The oil was found to be an effective anti-inflammatory (61%) agent compared with diclofenac sodium (70.92%) via the carrageenan-induced assay. An appreciable (48.28%) analgesic activity in correlation with the standard aspirin was observed through the acetic acid-induced writhing bioassay. The oil from the n-hexane fraction of S. edelbergii contained valuable bioactive constituents that can act as in vitro biological and in vivo pharmacological agents. However, further studies are needed to uncover individual responsible compounds of the observed biological potentials which would be helpful in devising novel drugs.

Pharmaceuticals and heavy metals are contaminants present in groundwaters, which are the main source of drinking water in most parts of the world. In the northwest region of Spain, Galicia, groundwater harvesting is a common practice for drinking water supply, crop irrigation, cattle watering, as well as recreational use such as filling pools. In order to assess the quality of Galician groundwaters, the presence of 21 pharmaceuticals and 10 heavy metals was analysed by UPLC-MS/MS and ICP/MS methods, respectively, in a total of 118 groundwater samples from private wells. Seventeen of the 21 compounds studied were detected in 28% of the samples, with the highest presence of pharmaceuticals belonging to the antimicrobial group (52%), specifically the sulphonamides group in a range of concentration between 21 and 14.9 ng/L. In addition, 30% of the samples contained at least one heavy metal (Mn, As and Fe) above the legally permitted levels. Evaluation of the risk associated with the consumption of the analysed groundwater indicated no human risk for any of the detected pharmaceuticals but high cancer risk for children due to Cd, Cr and As concentrations was observe.

A fully automated sequential injection system was tested in terms of its application in liberation testing, and capabilities and limitations were discussed for clotrimazole liberation from three semisolid formulations. An evaluation based on kinetic profiles obtained in short and longer sampling intervals and steady-state flux values were applied as traditional methods. The obtained clotrimazole liberation profile was faster in the case of Delcore and slower for Clotrimazol AL and Canesten cream commercial formulations. The steady-state flux values for the tested formulations were 52 µg cm -2 h -1 for Canesten, 35 µg cm -2 h -1 for Clotrimazol AL, and 7.2 µg cm -2 h -1 for Delcore measured in 4 min sampling intervals. A simplified approach for the evaluation of the initial rate based on the gradient between the second and third sampling points was used for the first time and was found to correspond well with the results of the conventional methods. A comparison based on the ratio of the steady-state flux and the initial rate values for Canesten and Clotrimazol AL proved the similarity of the obtained results. The proposed alternative was successfully implemented for the comparison of short-term kinetic profiles. Consequently, a faster and simpler approach for dissolution/liberation testing can be used.

Aims: Compare and characterize Chaetomium strains with special regard to their potentialities as biocontrol agents., Methods and Results: Twelve strains of the fungal genus Chaetomium from diverse ecological niches were identified as belonging to six different species. Large differences were observed between the strains with regard to temperature requirements for mycelial growth and pigmentation of culture filtrates. Culture filtrates and ethyl acetate extracts were assayed for fungicidal effects against important phytopathogens both on agar media and in multiwell plates. The samples from Chaetomium globosum were particularly active against Botrytis cinerea, Pyrenophora graminea and Bipolaris sorokiniana, while those from C. cochliodes and C. aureum were inhibitory towards Phytophthora infestans, and P. infestans and Fusarium culmorum respectively. To narrow down the active principle, the most promising extracts were separated by preparative HPLC and the resulting fractions tested in bioassays. Chaetoglobosins were identified as active compounds produced by C. globosum., Conclusions: The bioassays revealed C. aureum and C. cochliodes as promising candidates for use in biocontrol. Both showed remarkably good activity against the prominent plant pathogen P. infestans., Significance and Impact of the Study: We provide the first systematic study comparing six different Chaetomium species with regard to their use as biocontrol agents., (© 2020 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

The Cistaceae family is well represented in Sardinia, and the Cistus genus is widely used in traditional medicine. Nowadays only few studies have been performed on this genus vegetating in Sardinia in spite of its ethnobotanical importance. Moreover, in the past there have been conflicting opinions among botanists for the exact assignment of the species growing in Sardinia. We started several years ago to carried out studies on this genus and in the present study was to evaluate the in vitro activity of several samples of Cistus salvifolius L., Cistus monspeliensis L., and Cistus albidus L. collected in Sardinia as antimicrobial agents against Escherichia coli, Staphylococcus aureus, and four Candida species and their antioxidant activity using DPPH, ABTS, and FRAP assays. Furthermore, the phenolic content and composition of the extracts were first evaluated. Using statistical multivariate analysis on the complete metabolomics profile of all Cistus species growing wild in Sardinia, we confirmed the botanical classification, and we observed an interesting correlation between metabolomics profile and antioxidant activity., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

Eugenia copacabanensis and Myrciaria tenella are present in restingas of the Atlantic Forest, but little information is available about their chemical and biological potential. In this context, the hexane, dichloromethane, ethyl acetate and butanol fractions from the leaves of methanolic extract were analyzed by GC/MS and HPLC-DAD and the antioxidant potential was determined by DPPH and ABTS assays and using a Saccharomyces cerevisiae model. Dereplication allowed the identification of 68 compounds, 42 and 41 of which, respectively, are first reported here for E. copacabanensis and M. tenella. In vivo results revealed that the ethyl acetate and butanol fractions showed expressive antioxidant protection in the BY4741 and Δgsh1 strains, with greater impact on glutathione-deficient cells. With a high diversity of phenolic compounds, these polar fractions of E. copacabanensis and M. tenella leaves are potential protectors against intracellular oxidative stress., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

The objectives of current investigation are (1) to find out wavelength of maximum absorbance (λ max ) for combined cyclosporin A and etodolac solution followed by selection of mobile phase suitable for the RP-HPLC method, (2) to define analytical target profile and critical analytical attributes (CAAs) for the analytical quality by design, (3) to screen critical method parameters with the help of full factorial design followed by optimization with face-centered central composite design (CCD) approach-driven artificial neural network (ANN)-linked with the Levenberg-Marquardt (LM) algorithm for finding the RP-HPLC conditions, (4) to perform validation of analytical procedures (trueness, linearity, precision, robustness, specificity and sensitivity) using combined drug solution, and (5) to determine drug entrapment efficiency value in dual drug-loaded nanocapsules/emulsions, percentage recovery value in human plasma spiked with two drugs and solution state stability analysis at different stress conditions for substantiating the double-stage systematically optimized RP-HPLC method conditions. Through isobestic point and scouting step, 205 nm and ACN:H 2 O mixture (74:26) were selected respectively as the λ max and mobile phase. The ANN topology (3:10:4) indicating the input, hidden and output layers were generated by taking the 20 trials produced from the face-centered CCD model. The ANN-linked LM model produced minimal differences between predicted and observed values of output parameters (or CAAs), low mean squared error and higher correlation coefficient values in comparison to the respective values produced by face-centered CCD model. The optimized RP-HPLC method could be applied to analyze two drugs concurrently in different formulations, human plasma and solution state stability checking.

Introduction: The absence of a standardized classification scheme for the antifungal potency of compounds screened against Candida species may hinder the study of new drugs. This systematic review proposes a scheme of interpretative breakpoints for the minimum inhibitory concentration (MIC) of bioactive compounds against Candida species in in vitro tests., Materials and Methods: A literature search was conducted in the PubMed, Scopus, Web of Science, Lilacs, and SciFinder databases for the period from January 2015 to April 2020. The following inclusion criterion was used: organic compounds tested by the microdilution technique according to the Clinical and Laboratory Standards Institute protocol against reference strains of the genus Candida . A total of 545 articles were retrieved after removing duplicates. Of these, 106 articles were selected after applying the exclusion criteria and were evaluated according to the number of synthesized molecules and their chemical classes, the type of strain (reference or clinical) used in the antifungal test, the Candida species, and the MIC (in μ g/mL) used., Results: The analysis was performed based on the median, quartiles (25% and 75%), maximum, and minimum values of four groups: all strains, ATCC strains, C. albicans strains, and C. albicans ATCC strains. The following breakpoints were proposed to define the categories: MIC < 3.515  μ g/mL (very strong bioactivity); 3.516-25  μ g/mL (strong bioactivity); 26-100  μ g/mL (moderate bioactivity); 101-500  μ g/mL (weak bioactivity); 500-2000  μ g/mL (very weak bioactivity); and >2000  μ g/mL (no bioactivity)., Conclusions: A classification scheme of the antifungal potency of compounds against Candida species is proposed that can be used to identify the antifungal potential of new drug candidates., Competing Interests: The authors declare no conflict of interest related to publication of this paper., (Copyright © 2021 Danielle da Nóbrega Alves et al.)

Crude extracts and fractions of Cordia sinensis leaves were subjected to gas chromatography-flame ionisation detection (GC-FID), gas chromatography-mass spectrometry (GC-MS) analyses and preliminary screening for biological potentials using antibacterial, antifungal, phytotoxic, cytotoxic, insecticidal, antileishmanicidal and antiglycation bioassays. Overall thirty-one phytochemicals including three hydrocarbons, seven fatty acids, fifteen fatty acid esters, three terpenes, one each of phytosterol, terpenoid, and polyunsaturated aldehyde were identified. n-Hexadecanoic acid (13.2%), methyl hexadecanoate (9.0%), octadec-9 Z -enoic acid (8.3%) and methyl octadec-9 Z ,12 Z ,15 Z -trienoate (7.8%) were the main components. Presumably, this is a first report of twenty, thirteen and twenty phytochemicals from C. sinensis , Cordia and Boraginaceae, respectively. Moreover, ethyl acetate fraction exhibited significant insecticidal and antifungal activity against Sitophilus oryzae and Microsporum canis , respectively. Similarly, n-hexane fraction significantly inhibited (77.4%) advanced glycation end products in antiglycation assay. Conclusively, C. sinensis leaves with bioactive metabolites are a potential source for the development of insecticides, fungicides and pharmaceutically active antidiabetic drugs.

Background: The polyene macrocyclic compound amphotericin B (AmB) is an important antifungal antibiotic for the clinical treatment of invasive fungal infections. To rationally guide the improvement of AmB production in the main producing strain Streptomyces nodosus, comparative metabolomics analysis was performed to investigate the intracellular metabolic changes in wild-type S. nodosus ZJB20140315 with low-yield AmB production and mutant S. nodosus ZJB2016050 with high-yield AmB production, the latter of which reached industrial criteria on a pilot scale., Results: To investigate the relationship of intracellular metabolites, 7758 metabolites were identified in mutant S. nodosus and wildtype S. nodosus via LC-MS. Through analysis of metabolism, the level of 26 key metabolites that involved in carbon metabolism, fatty acids metabolism, amino acids metabolism, purine metabolism, folate biosynthesis and one carbon pool by folate were much higher in mutant S. nodosus. The enrichment of relevant metabolic pathways by gene overexpression strategy confirmed that one carbon pool by folate was the key metabolic pathway. Meanwhile, a recombinant strain with gene metH (methionine synthase) overexpressed showed 5.03 g/L AmB production within 120 h fermentation, which is 26.4% higher than that of the mutant strain., Conclusions: These results demonstrated that comparative metabolomics analysis was an effective approach for the improvement of AmB production and could be applied for other industrially or clinically important compounds as well.

RI76 is a novel 2-thiazolylhydrazone compound with reported antifungal activity. In preclinical drug development, it is fundamental to know the impurity profile and to understand degradation mechanisms of the molecule. In our study, RI76 was subjected to forced degradation conditions, and a stability-indicating HPLC-DAD method was developed and validated. Separation was carried out on a C18 column (150 × 4.6 mm i.d., 5 μm) maintained at 40°C using a 1 mL/min flow rate of 2 mM ammonium acetate with 0.1% formic acid (pH 3.0) and acetonitrile in gradient mode. The method was linear in the range of 0.7-91 μg/mL for RI76 and 0.7-25 μg/mL for its degradation product PD76. The formation of a major degradation product was quickly observed when RI76 was in aqueous solution. The chemical structure of this product, named PD76, was proposed based on LC-UV-MS experiments, synthesized in-house, and confirmed by NMR spectroscopy and chromatographic analysis. In vitro antifungal activity assays demonstrated that this resultant product shows a promising activity against clinically important Candida and Cryptococcus strains, matching or surpassing the activity of its precursor and of well-established antifungal drugs., (© 2020 John Wiley & Sons, Ltd.)

Metastable and rarely reported GO warped tetragonal phase t-lanthanum vanadate nanocomposites (GO@LaVO 4 -NCs) are reported for the sensitive electrochemical determination of antifungal drug Clioquinol (CQ). The hydrothermal method was adopted for synthesis of GO@LaVO 4 -NCs. The electrochemical performance of CQ was examined using cyclic voltammetry (CV) and differential plus voltammetry (DPV) at GO@LaVO 4 -NCs modified glassy carbon electrode (GCE). The electrocatalytic oxidation of CQ at the GO@LaVO 4 -NCs/GCE shows the highest anodic peak current at a potential of +0.51 V vs. Ag/AgCl. The proposed sensor provides excellent sensitivity of 4.1894 μA μM -1  cm -2 , a very low detection limit (LOD) of 2.44 nM, and a wide range of 25 nM to 438.52 μM towards CQ detection. Finally, the detection of CQ in biological media was successfully done using the GO@LaVO 4 -NCs/GCE and possesses recoveries of 94.67-98.0%.

Ching-Hsuan Lin works in the field of Candida biology. In this mSphere of Influence article, he reflects on how the papers "Use of ichip for high-throughput in situ cultivation of uncultivable microbial species" by D. Nichols, N. Cahoon, E. M. Trakhtenberg, L. Pham, et al. (Appl Environ Microbiol 76:2445-2450, 2010, https://doi.org/10.1128/AEM.01754-09) and "A new antibiotic kills pathogens without detectable resistance" by L. L. Ling, T. Schneider, A. J. Peoples, A. L. Spoering, et al. (Nature 517:455-459, 2015, https://doi.org/10.1038/nature14098) made an impact on him by inspiring him to explore new bioactive antimicrobial compounds with his collaborators., (Copyright © 2021 Lin.)

Micafungin is characterized as one of the most active available drugs for candidemia treatment; however, their use is also associated in prophylaxis protocols in cases of invasive fungal infections. The use of this drug is widely appreciated in the medical field due to be the most active echinocandin available for invasive fungal infections. In order to provide important parameters related to the chemical, physical, biological and therapeutic characteristics, this review article gathers important research results that demonstrate the biological potential of this drug, as well as to present analytical methods that can be used to determine the antifungal potential and a monitoring of administered dosages. Important studies about the methods most commonly used in biological activity evaluation and determination/quantification by analytical methods are provided in this review article. With the data provided, the scientific community will have the possibility to choose the analytical methods and biological that can be employed in clinical and scientific research to provide greater safety and reliability of the results to be found.

Candida auris is an emerging fungal pathogen that exhibits resistance to multiple drugs, including the most commonly prescribed antifungal, fluconazole. Here, we use a combinatorial screening approach to identify a bis-benzodioxolylindolinone (azoffluxin) that synergizes with fluconazole against C. auris. Azoffluxin enhances fluconazole activity through the inhibition of efflux pump Cdr1, thus increasing intracellular fluconazole levels. This activity is conserved across most C. auris clades, with the exception of clade III. Azoffluxin also inhibits efflux in highly azole-resistant strains of Candida albicans, another human fungal pathogen, increasing their susceptibility to fluconazole. Furthermore, azoffluxin enhances fluconazole activity in mice infected with C. auris, reducing fungal burden. Our findings suggest that pharmacologically targeting Cdr1 in combination with azoles may be an effective strategy to control infection caused by azole-resistant isolates of C. auris.

In solving the issue of basal stem rot diseases caused by Ganoderma, an investigation of Scytalidium parasiticum as a biological control agent that suppresses Ganoderma infection has gained our interest, as it is more environmentally friendly. Recently, the fungal co-cultivation has emerged as a promising method to discover novel antimicrobial metabolites. In this study, an established technique of co-culturing Scytalidium parasiticum and Ganoderma boninense was applied to produce and induce metabolites that have antifungal activity against G. boninense . The crude extract from the co-culture media was applied to a High Performance Liquid Chromatography (HPLC) preparative column to isolate the bioactive compounds, which were tested against G. boninense . The fractions that showed inhibition against G. boninense were sent for a Liquid Chromatography-Time of Flight-Mass Spectrometry (LC-TOF-MS) analysis to further identify the compounds that were responsible for the microbicidal activity. Interestingly, we found that eudistomin I, naringenin 7-O-beta-D-glucoside and penipanoid A, which were present in different abundances in all the active fractions, except in the control, could be the antimicrobial metabolites. In addition, the abundance of fatty acids, such as oleic acid and stearamide in the active fraction, also enhanced the antimicrobial activity. This comprehensive metabolomics study could be used as the basis for isolating biocontrol compounds to be applied in oil palm fields to combat a Ganoderma infection.