Antifungal Activity Research Articles

A Non-Prescription Nitric Oxide Releasing Gel Demonstrates In Vitro Anti-fungal Activity and Supports Nail Clearance in Patients with Onychomycosis

Background: Onychomycosis is a common fungal nail infection. Nitric oxide has broad-spectrum antifungal activity; NO releasing formulations are in development to support nail wellness. Objective: to examine antifungal properties of NO releasing formulations liquid (NORS) and gel (NORS-gel) to support nail clearance in onychomycosis. Methods: NORS In Vitro antifungal testing: Minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) were used to compare NORS with efinaconazole. Trichophyton tonsurans, Trichophyton mentagrophytes, and Epidermophyton floccosum were selected and MFC was performed. Cidal activity: ≥ 99.9% reduction in CFUs/mL from baseline; Static activity: <99.9% reduction from baseline CFU/mL. (CFU: colony forming units). NORS keratin binding: Six tubes were prepared with 1 mL + 107 conidia/mL of T. mentagrophytes strain (37035) + sterilized cadaver nail; six treatment tubes included: NORS, NORS+Keratin (50mg/mL), Terbinafine (0.002 µg/mL)+keratin (50 mg/mL), Terbinafine (0.002 µg/mL), Growth control (T. mentagrophytes+saline). After 15-min, tubes were centrifuged, supernatant decanted, and pellets resuspended in sterile saline; 10-fold serial dilutions were plated (triplicate). Ex-Vivo in Onychomycosis: Toenail samples from patients with suspected onychomycosis were shredded and either placed onto agar plates incubated at 30°C for 1-month, or, treated with 1.5mL NORS-gel to 50mL tube, placing the clippings onto the gel and then overlaying another 1.5mL NORS-gel, samples were stored in the dark at room temperature for 8-hours, washed, dried, plated and incubated at 25°Cx7days Nail penetration: Underside of cadaver toenails were inoculated with T. mentagrophytes 107 conidia/mL and placed on inert gel agar. A bead of NORS-gel (approx. 0.125mL), control gel and Terbinafine 0.002 µg/mL were spread to cover the topside of the nail. Covered petri dishes were placed for 15-minutes and 8-hours; the nail was removed, washed 3x in sterile saline, and pulverized; serial dilutions with plated samples incubated at 25°Cx7days. Statistics: Log CFU ± standard deviation, analyzed by t-test, p ≤ 0.05. Human Volunteers: Three individuals with confirmed onychomycosis applied daily NORS-gel to infected nails (covered overnight), for one month. Photographs were taken at baseline, day 1 after application, and 4-5 months post-application. Volunteers self-reported nail appearance. Results: NORS In Vitro antifungal testing: NORS showed potent antifungal activity against dermatophytes, including those less susceptible to efinaconazole. NORS keratin binding: NORS with and without keratin demonstrated the lowest fungal burdens with average log CFUs/mL ± SD of 0 ± 0, and 2.45 ± 0.5, respectively, demonstrating significant activity vs control, and terbinafine (P-values < 0.0001). Ex Vivo: NORS-gel demonstrated significant fungicidal activity as indicated by the absence of fungal growth on petri dishes. Nail penetration: NORS-gel treated nail showed no fungal growth (average log CFUs/g ± SD of 0 ± 0) vs. terbinafine–burden of 3.58 ± 0.2 average log CFU/g, at 8 hrs. NORS-gel significantly reduced fungal burden vs. terbinafine and control (P-value < 0.05). (preliminary results; methodology undergoing refinement.) Human Volunteers: Daily application of NORS-gel for one-month improved nail clarity in volunteers with confirmed onychomycosis. Temporary discoloration was observed during treatment. Conclusions: NORS and NORS-gel demonstrated antifungal activity; NORS-gel supported nail clearance and was well tolerated in individuals with onychomycosis.

Growth dynamics, antagonistic activity and acid production of Lactiplantibacillus (=Lactobacillus) plantarum KT-L18/1 and Bacillus subtilis BPT-B1 inoculants in alfalfa silage

Aim. To study growth dynamics, antagonistic activity and acid formation of two inoculant strains, Lactiplantibacillus (=Lactobacillus) plantarum КТ-L18/1 and Ваcillus subtilis ВРТ-В1, that were introduced into alfalfa silage, up to 30 days of fermentation. Methods. A streptomycin-resistant strain of both above-mentioned organisms, obtained in earlier research, was used to achieve the research aims. Alfalfa was artificially wilted and dried to a dry matter content of 39–40 %. Fermentation was conducted under anaerobic conditions in polyethylene bags. Inoculant was added at a concentration of 108 CFU/kg alfalfa. The resulting 12 × 3 = 36 bags were kept at room temperature in a dark place for 30 days. At each examination date one sample per treatment was used and examined after cultivation of the two inoculants on elective De Man-Rogosa-Sharpe (MRS) agar and meat infusion agar (MIA), containing streptomycin. The antibacterial (ABA) and antifungal activities (AFA) were studied by the agar diffusion method. Results. After 30 days of ensiling the alfalfa samples, L. plantarum КТ-L18/1str and В. subtilis ВРТ-В1str were still present at levels of 9.2 lg CFU/g and 6.9 lg CFU/g respectively. In the natural population of LAB, L. plantarum КТ-L18/1str had a dominant position. After 30 days of the alfalfa fermentation L. plantarum КТ-L18/1str and В. subtilis ВРТ-В1str maintained ABA and AFA to the PPB strains (S. aureus, P. aeruginosa, S. typhimurium) and fungal strains tested. However, the antagonistic activities of the two inoculant strains decreased during the fermentation, for L. plantarum КТ-L18/1str from 7–21 %, and for В. subtilis ВРТ-В1str from 16–21 %. Compared to the uninoculated control treatment, the inoculated silage treatments had a lower pH level (5.2–5.3), an increased number of lactic acid bacteria (LAB) (by 32 %), and clostridia were no longer detected. The inoculation impacted fungi down to a level of 102–103 CFU/g that can ensure the aerobic stability of the feed. Conclusions. The inoculant strains and L. plantarum КТ-L18/1str and Ваcillus subtilis ВРТ-В1str showed a high competitiveness with background microbiota in an ensiling experiment with alfalfa. The antagonistic activity of the two strains to PPB after fermentation was preserved at a high level, albeit with a decrease of 7–21 %. Ваcillus subtilis ВРТ-В1str kept its AFA against all three fungal strains tested during the 30 days of fermentation. For L. plantarum КТ-L18/1str this was true for a strain of the Penicillium chrysogenum only. The inoculation of and L. plantarum КТ-L18/1str and Ваcillus subtilis ВРТ-В1str caused a decrease in pH of 8–9 % at the end of the fermentation period, an accumulation of LAB with 32 %, and the inhibition of the growth of clostridia up to undetectability in the fermented alfalfa. For both inoculants, compared to uninoculated control variant, fungi decreased down to 102–103 CFU/g of the feed, which can ensure the aerobic stability of the feed.

Precision management of Fusarium fujikuroi in rice through seed coating with an enhanced nanopesticide using a tannic acid-ZnIIformulation.

Seed coating with fungicides is a common practice in controlling seed-borne diseases, but conventional methods often result in high toxicity to plants and soil. In this study, a nanoparticle formulation was successfully developed using the metal-organic framework UiO-66 as a carrier of the fungicide ipconazole (IPC), with a tannic acid (TA)-ZnII coating serving as a protective layer. The IPC@UiO-66-TA-ZnII nanoparticles provided a controlled release, triggered and regulated by environmental factors such as pH and temperature. This formulation efficiently controlled the proliferation of Fusarium fujikuroi spores, with high penetration into both rice roots and fungal mycelia. The product exhibited high antifungal activity, achieving control efficacy rates of 84.09% to 93.10%, low biotoxicity, and promoted rice growth. Compared to the IPC flowable suspension formula, IPC@UiO-66-TA-ZnII improved the physicochemical properties and enzymatic activities in soil. Importantly, it showed potential for mitigating damage to beneficial soil bacteria. This study provides a promising approach for managing plant diseases using nanoscale fungicides in seed treatment.

Synthesis and Antimicrobial Activity of 3-Alkylidene-2-Indolone Derivatives

The escalating threat of antibiotic-resistant bacteria and fungi underscores an urgent need for new antimicrobial agents. This study aimed to synthesize and evaluate the antimicrobial activities of two series of 3-alkylidene-2-indolone derivatives. We synthesized 32 target compounds, among which 25 exhibited moderate to high antibacterial or antifungal activities. Notably, compounds 10f, 10g, and 10h demonstrated the highest antibacterial activity with a minimum inhibitory concentration (MIC) of 0.5 μg/mL, matching the activity of the positive control gatifloxacin against three Gram-positive bacterial strains: Staphylococcus aureus ATCC 6538, 4220, and Methicillin-resistant Staphylococcus aureus ATCC 43300. Moreover, the three most active compounds 10f, 10g, and 10h were evaluated for their in vitro cytotoxicity in the HepG2 cancer cell line and L-02; only compound 10h was found to exert some level of cytotoxicity. These findings suggest that the synthesized 3-alkylidene-2-indolone derivatives hold potential for further development as antibacterial agents.

Formulation and Evaluation of Polyherbal Soap

The aim of the study is to formulate and evaluate Polyherbal soap using coconut oil, beeswax, NaOH(lye), Glycerine, Neem, aloe vera, Turmeric, Tulsi, Sandalwood powder, orange peel powder, Multani mitti, Reetha, Honey and Vitamin E capsule. The herbal formulation was prepared then evaluation of the analysis of Physical parameter, pH, Foam height, Foam retention time, Moiture content, Alcohol insoluble matter etc. Various study found that, these herbal plant extract proven to anti-bacterial activity, anti-inflammatory activity, anti-fungal activity etc. The finding for the herbal soap reveals that these soaps are cost effective, convenient, natural foam generate on application, not exhibit any skin irritation and less or no side effect. Keywords: Polyherbal soap, Neem, Aloevera, Turmeric, Moisture content.

Targeting Candida albicans O-acetyl-L-homoserine sulfhydrylase (Met15p) in antifungal treatment.

Fungal infections are a serious threat to public health as they are becoming increasingly frequent. A major problem stems also from a rising fungal resistance to currently available antifungal therapies, therefore novel molecular targets are highly desirable. Exploration of enzymes participating in the biosynthesis pathways of essential amino acids such as L-methionine (L-Met) may provide new insights into pharmaceutical development. The MET15 gene from Candida albicans, encoding O-acetyl-L-homoserine sulfhydrylase (Met15p), an enzyme catalyzing the second step in that pathway, was cloned and expressed in two versions: as N and C-terminal oligo-His-tagged fusion proteins. The recombinant enzymes revealed appropriate activity, and catalyzed conversion of O-acetyl-L-homoserine and a sulfide ion to produce L-homocysteine. A new RP-HPLC-DAD method, using the enzymatic reaction product pre-column derivatization with 5,5'-dithio-bis-(2-nitrobenzoic acid) was developed and used by us to determine Met15p activity. Newly synthesized compounds as well as two commercially available exhibited a Met15p inhibitory effect which was related to antifungal activity. Fungal cells' sensitivity to inhibitors depending on the presence or absence of L-Met in the medium clearly indicated Met15p targeting. Moreover, the synergistic effect of the first methionine biosynthetic enzyme affecting inhibitor and Met15p inhibitors indicate that methionine biosynthesis pathway enzymes are promising molecular targets.

Formation and Preparation of Herbal Hair Tonic

Today, people are keenly interested in hair preparations and conditioner materials that includes shampoos, hair tonic, and conditioner formulation with herbal extracts. Hair tonic is the product which is used in styling hair. The objective of the present study involved the preparation of herbal hair tonic through the use of jojoba, tulsi, and coconut oil and its evaluation for antifungal and antibacterial activity. The final preparation of these ingredients is formulated in batches with change in concentration. Each formulation is tested for antibacterial and antifungal activity. The formulation of different concentrations was characterized for proximate analysis including moisture content, total ash, acid insoluble ash, water soluble ash, water insoluble ash, sulphated ash. It means for antifungal and antibacterial activity the formulation gives good results, the formulation having good consistency, good spread ability, homogeneity, appearance, and pH.

The preliminary study of antifungal and antibacterial properties of wine by-products

Background: Nowadays, the search for novel alternative antimicrobials has gained significant popularity. This surge is driven by the vast array of adverse effects of synthetic antimicrobials on human health and the environment. Consequently, the study of new natural sources of comparably ecologically safe and simultaneously effective antimicrobials is extremely important. In this context, the interest in wine by-products is growing due to their diverse beneficial properties. Traditionally considered industrial waste, wine by-products now offer promising opportunities. Notably, the unique climatic and geographical features of Armenian black grape varieties, such as Areni sev, make their by-products an attractive subject for study, with potential applications in agriculture. Objective: The study aims to investigate the antifungal and antibacterial properties of wine by-products obtained from the Areni sev grape variety. Methods: For the extraction of bioactive compounds, frozen and dry samples were homogenized. The maceration was performed in 70% hydroethanolic solution after incubation on a magnetic stirrer at room temperature. The obtained extracts were filtered through a 0.45 µm filter and evaluated for antifungal and antibacterial activity by disco-diffusion method. Results: Hydroethanolic extracts of wine by-products have exhibited antifungal and antibacterial activities. Specifically, the frozen samples demonstrated low antifungal activity against the Penicillium canescens with a 7.5 mm growth inhibition zone. In contrast, dried samples showed no antifungal activity. The highest antifungal activity was observed against Aspergillus niger with growth inhibition zones of 18 mm (frozen samples) and 15 mm (dried samples), respectively. Regarding antibacterial activity, wine by-product hydroethanolic extracts showed extremely low efficacy. Notably, frozen samples exhibited an 8 mm growth inhibition zone against Curtobacterium flaccumfaciens and 6.35 mm against Xanthomonas vesicatoria. Extracts from dried samples demonstrated a 6.35 mm inhibition zone against Curtobacterium flaccumfaciens and a 7 mm growth inhibition zone against Xanthomonas vesicatoria. Conclusion: Summarizing the results of the conducted experiments, it can be concluded that the used wine by-product derived from Areni sev grape has demonstrated notable antimicrobial activities, with a stronger emphasis on antifungal properties. The extracts obtained from frozen samples exhibited greater efficacy than those derived from dried samples. Future research plans include a more detailed investigation of Areni sev grape wine by-products. Keywords: wine by-product, grape pomace, Areni sev, bioactive compounds, antimicrobial properties

Combination of atmospheric and room temperature plasma and ribosome engineering techniques to enhance the antifungal activity of Bacillus megaterium L2 against Sclerotium rolfsii.

Sclerotium rolfsii is an extremely destructive phytopathogenic fungus that causes significant economic losses. Biocontrol strategies utilizing antagonistic microorganisms present a promising alternative for controlling plant pathogens. Bacillus megaterium L2 has been identified as a potential microbial biocontrol agent in our previous study; however, its efficacy in controlling pathogens has yet to meet current demands. This study aims to enhance the antifungal activity of strain L2 against S. rolfsii R-67 through a two-round mutagenesis strategy and to preliminarily investigate the mutagenesis mechanism of the high antifungal activity mutant. We obtained mutant Dr-77 with the strongest antifungal activity against R-67, and its cell-free supernatant significantly reduced the infection potential of R-67 to Amorphophallus konjac corms, which may be attributed to the antimicrobial compound phenylacetic acid (PAA), and PAA content in Dr-77 (5.78 mg/mL) was 28.90 times higher than original strain L2. This compound exhibited strong antifungal ability against R-67, with a half maximal effective concentration (EC50) value of 0.475 mg/mL, significantly inhibiting mycelial growth and destroying the ultrastructure of R-67 at EC50 value. Notably, PAA also exhibited broad-spectrum antifungal activity against six phytopathogens at EC50 value. Moreover, genome analysis revealed nine different gene mutations, including those involved in PAA biosynthesis, and the activities of prephenate dehydratase (PheA) and phenylacetaldehyde dehydrogenase (ALDH) in PAA biosynthesis pathway were significantly increased. These results suggest that the elevated PAA content is a primary factor contributing to the enhanced antifungal activity of Dr-77, and that this mutagenesis strategy offers valuable guidance for the breeding of functional microbial resources. © 2024 Society of Chemical Industry.

Synthesis and Antifungal Activity Evaluation of Novel L-Carvone-Based 1,3,4-Thiadiazole-amide Derivatives as a Potential Succinate Dehydrogenase Inhibitor.

In an effort to explore new-type high-efficiency antifungal agents, 25 novel L-carvone-based 1,3,4-thiadiazole-amide derivatives were designed, synthesized, and structurally characterized by IR, 1H NMR, 13C NMR, and high-resolution mass spectrometry (HRMS) analyses. The antifungal activity of the target compounds was preliminarily assayed at a concentration of 50 μg/mL, and boscalid, a commercialized fungicide identified as a succinate dehydrogenase inhibitor (SDHI), was employed as the positive control. It was found that all of the target compounds showed moderate to potent antifungal activity against the tested fungi compared to boscalid. Surprisingly, compound 4b exhibited broad-spectrum and significant inhibition activity against the growth of eight phytopathogenic strains with inhibitory rates of 67-89%. Further, the results of the EC50 value test suggested that the EC50 values of compound 4b against Physalospora piricola and Colletotrichum orbiculare were 16.33 and 18.06 μg/mL, respectively, and both of them were better than those of boscalid (16.64 and >50). Therefore, compound 4b deserves further study as a lead compound for novel fungicides. In addition, the inhibitory activity of compound 4b against succinate dehydrogenase (SDH) was evaluated as well to prove that compound 4b (IC50 = 3.38 μM) displayed higher SDH-inhibition activity than boscalid (IC50 = 7.02 μM). The binding mode of compound 4b and SDH was simulated by molecular docking and found to be similar to that of boscalid. The structure-activity relationships (SARs) of the target compounds were analyzed by establishing a 3D-QSAR model. Besides, a 4b-loaded complex 4b/CSTA on a reported L-carvone-based nanochitosan carrier CSTA containing the 1,3,4-thiadiazole-amide group was constructed, and its sustained release performance was investigated in the EtOH-H2O system (1:9, v/v). The complex 4b/CSTA exhibited preferred sustained release performance, indicating its potential for developing environmentally friendly nanofungicides.

Development and Biological Evaluation of New Diphenyl Ether Formylhydrazide Compounds as Potent Inhibitors of Succinate Dehydrogenase.

Succinate dehydrogenase (SDH), also recognized as succinate ubiquinone oxidoreductase (SQR), is considered one of the most promising targets for fungicide development, garnering significant international interest. We have focused on the development of highly effective, broad-spectrum-targeted SDH inhibitors. Using an active scaffold combining strategy, we designed and synthesized a series of novel diphenyl ether formylhydrazine derivatives, and most compounds have demonstrated broad-spectrum antifungal activity. Notably, compound M8 exhibited antifungal activity of more than 93% against four tested pathogen types at a concentration of 10 μg/mL, with an EC50 value below 0.3 μg/mL for each pathogen, outperforming boscalid. Additionally, compound M8 exhibited a control efficacy of 83% against Sclerotinia sclerotiorum on rapeseed leaves at a concentration of 200 μg/mL and demonstrated an 87% efficacy in controlling Fusarium graminearum on wheat ears when applied at 400 μg/mL. Structure-activity relationship research suggested that para-substituted benzene rings are more effective, offering stronger and more extensive antifungal potency. Further investigation, including enzyme inhibition assays, mycelial morphology observations, and molecular docking studies, suggests that the antifungal potency of M8 is due to the inhibition of its SDH activity. Therefore, our research positions compound M8 as a highly promising lead compound with broad-spectrum antifungal properties, potentially introducing a new class of fungicide.

The Underrepresented Quinolinone Alkaloids in Genera Penicillium and Aspergillus: Structure, Biology, and Biosynthetic Machinery.

Quinolone alkaloids are N-heterocycles with extensive structural diversity, mainly derived from in fungi from anthranilic acid and amino acids as precursors with a wide range of biological activities as antifungal, antimicrobial, anti-inflammatory, and insecticidal activities. The quinolone basic skeleton comprised of either 2-quinolones or 4-quinolones generated more than one hundred compounds. Several reviews discussed quinolones; particularly, the fluoroquinolones, yet few studies tackled natural quinolones. Many of these quinolones were not assayed for their antimicrobial potential despite their unique stereospecificity, which can supersede synthetic quinolones if their discovery is coupled with OMICS techniques, biochemical and molecular strategies as heterologous expression to maximize their yield. Herein, we conducted a comprehensive review of the quinolone's family in Aspergillus and Penicillium species, the exclusive producers of quinolones whether they are soil, endophytic or marine derived highlighting their isolation, chemical structures, pharmacological effects, structure activity relationships if any, and biosynthetic machinery. We believe that our initiative will pave the way for further development of natural quinolones as future antimicrobial agents.

The distinctive pharmacokinetic profile of rezafungin, a long-acting echinocandin developed in the era of modern pharmacometrics.

Echinocandin drugs are the current first-line therapy for fungal infections caused by Candida spp. Most patients require once-daily intravenous (IV) administration in a hospital or outpatient setting for treatment, which may negatively impact their quality of life and stress healthcare resources. Similar to other echinocandins, the novel FDA-, EMA-, and Medical and Healthcare Products Regulatory Agency-approved echinocandin, rezafungin (CD101), exhibited strong antifungal activity against several fungal pathogens and a low drug-drug interaction liability, which are important for medically complex patients. A pharmacometric-based approach has been adopted throughout the development of rezafungin, which contrasts with older echinocandins where dosing regimens were largely derived empirically, and only recently based on pharmacometric guidance. This state-of-the-art approach used model-based simulations incorporating pre-clinical and clinical data as it became available to optimize the dosing regimen for rezafungin. The enhanced stability of the molecular structure and the safety profile of rezafungin allow for the administration of once-weekly IV doses, compared to the daily dosing requirement for other echinocandin drugs, with this distinctive pharmacokinetic profile of rezafungin resulting in a front-loaded dosing regimen with high exposures early in therapy for enhanced fungal killing. The long shelf-life of rezafungin makes this echinocandin more flexible in terms of storage and manufacturing. Demonstrated across clinical development, rezafungin may provide patients with next-generation first-line antifungal treatment for the treatment of candidaemia and invasive candidiasis.

Biofabrication and Characterisation of Ni‐Doped SnO2 Nanoparticles With Enhanced Cytotoxicity, Antioxidant, Antimicrobial and Photocatalytic Activities

ABSTRACTThe present work focuses on Annona muricata leaf extract–mediated green synthesis of pure and nickel‐doped tin oxide nanoparticles for 3%, 5% and 7% concentrations. The properties of the obtained nanoparticles were investigated through XRD, FTIR, XPS, HRTEM–SAED, SEM–EDX and UV–visible spectroscopy techniques. The XRD pattern reveals all samples have tetragonal structures with high crystallinity. The Sn–O stretching vibration has been confirmed through FTIR spectra. The XPS results demonstrate that Sn0.93Ni0.07O2 nanoparticles have Sn3d, Ni2p and O1s oxidation states. EDX spectra contain Ni, Sn and O elements, which indicate the purity of the samples. UV–visible absorption spectrum shows decreases in optical energy bandgap with increases in nickel dopant concentration. The samples exhibit notable antibacterial activity against P. aeruginosa and S. aureus. Also, Sn0.93Ni0.07O2 nanoparticle has higher antifungal activity against A. niger than against C. albicans. Moreover, SnO2 nanoparticles have considerable antioxidant activity in DPPH scavenging compared to Sn0.93Ni0.07O2 nanoparticles. Furthermore, SnO2 nanoparticles have a better cytotoxic effect for L929 normal fibroblast cell lines with an LD50 value of 146.42 μg/mL. Additionally, the photocatalytic activity of SnO2 and Sn0.93Ni0.07O2 nanoparticles was done against methylene blue dye under direct sunlight irradiation. Overall, environmentally friendly synthetic pure and Ni‐doped SnO2 nanoparticles can be used in wastewater filter technologies as well as in medical aids due to their better cytotoxic, antioxidant and antimicrobial effects.

Antifungal Activity of Novel Isoindoline-2-Yl Putrescines as Potential Autophagy-Activated Fungicide.

The exacerbation of plant fungal diseases necessitates the development of new fungicides to prevent outbreaks. In this study, five novel isoindoline-2-yl putrescines (ISPs) were synthesized, and their synthetic procedures and gram-scale preparation were explored. When tested at 50 μg mL-1, ISPs did not significantly inhibit mycelial growth on agar plates. However, at 100 μg mL-1, they demonstrated remarkable in vivo efficacy in mitigating Botrytis cinerea infection, especially ISP3 showed curative and protective activities of 91.9 % and 92.6 %, respectively. Moreover, ISP3 also effectively halted lesion expansion of gray mold, Sclerotic rot, and Fusarium scabs, while inducing excessive malformed top mycelial branches of B. cinerea and Sclerotinia sclerotiorum, suppressing sclerotia formation in S. sclerotiorum, and triggering autophagic vacuolization with numerous autophagosomes in the mycelia of these fungi. Molecular docking revealed that ISP3 effectively bound to the active site of BcAtg3, forming hydrogen bonds with Ser279, Gly343, Asp370, and Asp13, along with establishing a stable salt bridge with Asp13. Furthermore, ISP3 possessed favorable ADMET properties. These findings highlight ISP3 as a promising antifungal candidate through autophagy activation.

In vitro antifungal activities of medicinal plants used for treatment of candidiasis in Pader district, Northern Uganda.

The emergence of multidrug resistant Candida species to available drugs has led to renewed interest in the use of herbal medicines globally. This study scientifically verified antifungal effectiveness of five commonly used plant species in Pader district, against selected pathogenic candida strains. Powdered roots of Momordica foetida, Sansevieria dawei and Distimake dissectus; and stem barks of Khaya anthotheca and Mitragyna rubrostipulatawere extracted sequentially using petroleum ether and methanol, respectively; and total water extraction at 24.4°C (maceration), 60°C (decoction) and boiling water at 87°C (hot water infusion). Extracts and their combinations, positive controls (amphotericin B, and fluconazole) and negative control (80% dimethyl sulfoxide, verified to be tolerable concentration to the tested Candida species) were screened and verified for their antifungal activity against Candida albicans (ATCC: American Type Culture Collection reference strain 10231, ATCC 90028, 0770a and 0796), C. glabrata (VVc 004, ATCC 2950) and C. tropicalis (ATCC 750 and 0210) using agar well diffusion and broth micro-dilution, respectively. Aqueous extract (24.4°C) of M. rubrostipulata (ZOI: 18.00 ± 1.00 to 38.33 ± 0.17; MIC: 3.13 ± 0.00 to 20.83 ± 4.17; MFC: 12.50 ± 0.00 to 200.00 ± 0.00), methanol extract of K. anthotheca (10.11 ± 0.31 to 15.11 ± 0.65; 1.04 ± 0.26 to 12.50 ± 0.00; 12.50 ± 0.00 to 100.00 ± 0.00), and combination of aqueous extract (60°C) of D. dissectus + methanol extract of K. anthotheca (7.89 ± 0.26 to 19.67 ± 0.37; 0.78 ± 0.00 to 50.00 ± 0.00; 12.50 ± 0.00 to 200.00 ± 0.00) exhibited broad spectrum antifungal activities and were fungistatic against all tested Candida species, which comprised 8 clinical/control and susceptible/resistant strains. None of the conventional drugs used demonstrated broad spectrum antifungal activity across all tested Candida species/strains. Methanol extract of K. anthotheca, aqueous extract (24.4°C) of M. rubrostipulata, and combination of aqueous extract (60°C) of D. dissectus + methanol extract of K. anthotheca could be effective in the treatment of candidiasis. They demonstrated potential broad spectrum antifungal activity against different species and strains of tested Candida than the fluconazole and amphotericin B drugs. Their fungistatic nature showed their ability to inhibit fungal growth. Hence, these extracts/extract combination can offer better treatment option for candidiasis if they are standardized and also their active curative compounds isolated and made into antifungal drugs.

Optimization of Ultrasonication Probe-Assisted Extraction Parameters for Bioactive Compounds from Opuntia macrorhiza Using Taguchi Design and Assessment of Antioxidant Properties

Opuntia macrorhiza, commonly referred to as red prickly pear, is a type of cactus fruit. The Opuntia macrorhiza (OM) fruit is rich in polyphenols and contains a high amount of ascorbic acid and betalains. The fruit peels have demonstrated many biological abilities, including antioxidant, antifungal, and antibacterial activities. Ultrasound probe-assisted extraction (UPAE) is a highly promising method for efficiently extracting valuable molecules from natural sources. The objective of this study is to optimize the parameters of UPAE, including the appropriate solvent, liquid-to-solid ratio, extraction duration, and pulsation level. The aim is to maximize the yield of bioactive compounds (polyphenols, betalains, and ascorbic acid) from OM fruits (pulps and peels) and assess their antioxidant activities using Taguchi design. The optimal extraction conditions through the partial least squares method for OM pulp were determined to be aqueous extraction for 12 min with a liquid-to-solid ratio of 60 mL/g and 48 pulses/min, while for OM peels they were determined to be aqueous extraction for 20 min with a liquid-to-solid ratio of 60 mL/g and a pulsation of 48 pulses/min. The optimum UPAE conditions were compared with the values obtained from the optimum extraction under stirring extraction (STE). Overall, UPAE exhibited higher yields than STE. The obtained total polyphenol content ranged from 10.27 to 13.07 mg gallic acid equivalents/g dry weight, while the betalain content ranged from 974 to 1099 μg/g dry weight. Overall, these fruits demonstrated potential as new components for food and medicinal uses due to their good health effects and lack of toxicity.

Recent Discoveries of Antifungal Activity in Plant Antimicrobial Peptides

Recent Discoveries of Antifungal Activity in Plant Antimicrobial Peptides

Fusarium spp. in Metalworking Fluid Systems: Companions Forever

Water-miscible metalworking fluids (MWFs) are utilized in a variety of metal removal and forming operations. For end-use, formulation concentrates are diluted in water, creating conditions conducive to microbial growth and metabolism, possibly compromising the fluid’s integrity and mechanically obstructing filters or piping systems. Metalworking machines offer additional habitats on surfaces that are in permanent or temporary contact with MWFs. For that reason, biocides have been incorporated into concentrates for years, but legal constraints will restrain their use in the future. While bacterial contamination of MWFs is well documented, fungal contamination is often overseen and infrequently reported in the literature. In this study, we report fungal prevalence in in-use MWFs sampled worldwide over 10 years, and we are convinced that the presence of fungi is the norm rather than the exception. In addition, we evaluated the inhibitory effect of fungicides on fungal growth, sporulation and spore viability using traditional culture-dependent methods and flow cytometry. In essence, we show that the effectiveness of these fungicides is limited and dependent on the chemical construction of the fluid. We think that the ecology created by water-diluted MWFs is of higher importance than the anti-fungal activity of single components.

Comparative Study of Essential Oils Extracted From Foeniculum vulgare Miller Seeds Using Hydrodistillation, Steam Distillation, and Superheated Steam Distillation

ABSTRACTFoeniculum vulgare Miller is a highly valued aromatic and nutritious plant. The unique compositions of its essential oil make it more valuable in the flavor, fragrance, and medicinal industries. However, the potential of superheated steam distillation for obtaining essential oils from its seeds has not been explored in detail. This study assessed the composition, yield, antimicrobial, and antioxidant activities of essential oils distilled from F. vulgare seeds using traditional hydrodistillation, steam distillation, and superheated steam distillation. Superheated steam distillation resulted in the maximum quantity of essential oil (5.24%) compared to steam (3.47%) and hydrodistillation (2.47%). Trans‐anethole, fenchone, estragole, and limonene were the main identified by GC–MS analysis in the essential oils, and these compounds were abundant in the essential oil produced by superheated steam distillation. Essential oil distilled by superheated steam distillation presented the highest antibacterial activity against Staphylococcus aureus, Pastrulla multocida, Bacillus subtilis, and Escherichia coli. The highest antifungal activity against Aspergillus niger, Fusarium solani, Aspergillus flavus, and Alternaria alternate was also demonstrated by the same essential oil. These findings demonstrated the potential of superheated steam distillation as a highly effective and efficient technique for distilling high‐quality essential oils from Foeniculum vulgare Miller seeds. It is suitable for various applications in the food, cosmetic, and pharmaceutical industries.