Natural Antifungal Agent Research Articles

Chemical Profiling and Antifungal Activity of Ziziphora clinopodioides Leaf Essential Oil against the Pathogen Verticillium dahliae.

Essential oils (EOs) are often used as natural antifungal agents to control the growth of phytopathogenic fungi. The aim of this study was to determine the effect of Ziziphora clinopodioides leaf EO against Verticillium dahliae, a pathogenic fungus of cotton. Gas chromatography-mass spectrometry (GC/MS) analysis revealed the presence of 15 compounds of the total of extracted oil, which was consisted of 98.79 % monoterpenes and 0.61 % sesquiterpenes. The major constituents were pulegone (62.17 %), isomenthone (18.42 %), l-menthone (5.55 %) and piperitenone (3.99 %). The mycelial growth of Verticillium dahliae was completely inhibited at 0.24 μL/mL air under vapor phase condition. Considerable morphological variations were also observed in the fungal sclerotia at the contact phase at 3 μL/mL. This study demonstrated for the first time that Z. clinopodioides EO can effectively inhibit the growth of V. dahliae, implying that it has the potential to be explored as an antifungal agent against Verticillium Wilt of cotton.

Antifungal activity of wheat-flour sourdough (Type II) from two different Lactobacillus in vitro and bread

This study aims to evaluate the antifungal capacity and chemical properties of the aqueous extracts (AE) obtained from the fermentation with Lactobacillus acidophilus NRRL B-4495 or L. casei 21/1 of wheat flour (poolish-type sourdough). The antifungal activity of poolish-type sourdoughs was tested against bread spoilage molds (Penicillium chrysogenum, and P. corylophilum) in vitro and bread. In addition, the physicochemical and sensory properties of bread formulated with the poolish-type sourdoughs were assessed. The AEs were obtained by separating the poolish supernatant at the end of fermentation (24, 48, or 72 h). Adding L. acidophilus or L. casei affects the sourdoughs’ pH and titratable acidity (TA), wherein the poolish added with L. casei presented a lower pH and higher acidity. HPLC identified mainly lactic acid in a concentration between 1 and 2% for the sourdoughs fermented with L. acidophilus, while for the poolish fermented by L. casei, lactic acid (1–2%) and acetic acid were identified (0.1–0.2%). Concentrations between 25 and 50% of AEs decreased the radial growth rate of P. crysogenum and P. corylophilum; in particular, the AEs obtained with L. acidophilus showed an effect on the growth of both molds. Bread added with sourdoughs fermented with Lactobacillus had an increase in shelf life compared to bread without them. There was an effect of the addition of poolish sourdoughs on the bread's physicochemical, quality, and sensory properties. But, incorporating L. acidophilus NRRL B-4495 or L. casei 21/1 into poolish-type sourdoughs can be an alternative as natural antifungal agents, useful for the bakery industry.

Discovery of a novel antifungal agent: All-hydrocarbon stapling modification of peptide Aurein1.2.

Aurein1.2 is secreted by the Australian tree frog Litoria aurea and is active against a broad range of infectious microbes including bacteria, fungi, and viruses. Its antifungal potency has garnered considerable interest in developing novel classes of natural antifungal agents to fight pathogenic infection by fungi. However, serious pharmacological hurdles remain, hindering its clinical translation. To alleviate its susceptibility to proteolytic degradation and improve its antifungal activity, six conformationally locked peptides were synthesized through hydrocarbon stapling modification and evaluated for their physicochemical and antifungal parameters. Among them, SAU2-4 exhibited significant improvement in helicity levels, protease resistance, and antifungal activity compared to the template linear peptide Aurein1.2. These results confirmed the prominent role of hydrocarbon stapling modification in the manipulation of peptide pharmacological properties and enhanced the application potential of Aurein1.2 in the field of antifungal agent development.

Synthesis, Antifungal Activity, and Molecular Simulation Study of L-Carvone-Derived 1,3,4-Oxadiazole-Thioether Compounds.

To discover potent antifungal molecules with new and distinctive structures, 20 novel L-carvone-derived 1,3,4-oxadiazole-thioether compounds 5 a-5 t were synthesized through multi-step reaction of L-carvone, and their structures were confirmed by FT-IR, 1 H-NMR, 13 C-NMR, and HR-MS. The antifungal activities of compounds 5 a-5 t were preliminarily tested by in vitro method, and the results indicated that all of the title compounds displayed certain antifungal activities against the eight tested plant fungi, especially for P. piricola. Among them, compound 5 i (R=p-F) with the most significant antifungal activity deserved further study for discovering and developing novel natural product-based antifungal agents. Moreover, two molecular simulation technologies were employed for the investigation of their structure-activity relationships (SARs). Firstly, a reasonable and effective 3D-QSAR model was established by the comparative molecular field (CoMFA) method, and the relationship of the substituents linked with the benzene rings and the inhibitory activities of the title compounds against P. piricola was elucidated. Then, the binding mode of compound 5 i (R=p-F) and its potential biological target (CYP51) was simulated by molecular docking, and it was found that compound 5 i could readily bind with CYP51 in the active site, and the ligand-receptor interactions involved three hydrogen bonds and several hydrophobic effects.

Synthesis, anti-oomycete and anti-fungal activities of novel cinchona alkaloid derivatives bearing carbamate moiety

In order to explore novel natural product-based anti-oomycete and anti-fungal agents, twenty cinchona alkaloid carbamate derivatives (1a-d, 2a-d, 3a-d, 4a-d, and 5a-d) were designed and prepared, and structurally confirmed by 1H NMR, 13C NMR, HRMS, and melting point. The stereochemical configuration of compound 1a was unambiguously confirmed by single-crystal X-ray diffraction. Furthermore, we evaluated the anti-oomycete and anti-fungal activities of these target compounds against Phytophthora capsici and Fusarium graminearum in vitro. The results showed that five compounds 1d, 2d, 3d, 4d, and 5d exhibited prominent anti-oomycete and anti-fungal activities, and the median effective concentration (EC50) values of 1d, 2d, 3d, 4d, and 5d against P. capsici and F. graminearum were 21.1, 16.6, 22.4, 14.4, 13.9 mg/L and 38.2, 19.8, 36.6, 30.3, 27.7 mg/L, respectively. This study suggested that when a specific substituent is introduced into the cinchona alkaloid skeleton, i.e. R1/R2 = Ph/Ph, the corresponding derivatives exhibit significant anti-oomycete and anti-fungal activities. And the configuration of the C8/9 position of the target compound is crucial for its anti-oomycete and anti-fungal activities, and 9S-configuration is optimal.

Anthocyanins as Natural Plant-derived Antifungal Agents

Anthocyanins are polyphenolic compounds found as natural pigments in various parts of plants that share the same diphenyl propane skeleton (C6C3C6) and are collectively known as flavonoids. They are water-soluble pigments responsible for the blue, purple, and red color of various plant parts and are particularly abundant in fruits, flowers, and vegetables. These pigments are natural colorants that are low to non-toxic. They also show antifungal, antibacterial, antioxidant, and food preservative properties. This study aims to perform in vitro analysis of the antifungal activity of anthocyanins extracted from different plant materials. Anthocyanins were extracted in acetone and acidified methanol. For qualitative determination of anthocyanins, thin layer chromatography was used whereas for quantitative determination pH differential method was used. The antifungal potential of anthocyanins was determined using the MIC and agar well diffusion methods against Penicillium sp. and Aspergillus sp. Acetone extract exhibited higher antifungal potential compared to acidified methanol extract against both fungal strains. It is suggested that anthocyanins, being strong antifungal agents, can be used to prevent fungal spoilage and deterioration of processed foods.

Synergistic effects of combined cinnamaldehyde and nonanal vapors against Aspergillus flavus

This study evaluated the synergistic antifungal effects of vapor-phase natural agents against Aspergillus flavus with an aim to prevent fungal contamination in agricultural products. Screening different combinations of natural antifungal vapor agents using the checkerboard assay revealed that the cinnamaldehyde and nonanal (SCAN) blend could exert the strongest synergistic antifungal activities against A. flavus, with a minimum inhibitory concentration (MIC) of 0.03 μL/mL, which caused a 76 % decrease in fungal population compared to when each agent was used separately. Subsequent gas chromatography–mass spectrometry (GC/MS) analysis demonstrated that the cinnamaldehyde/nonanal combination was stable and no effects on their individual molecular structures. SCAN at 2 × MIC completely inhibited the fungal conidia production and mycelial growth. The calcofluor white (CFW) and dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining assays showed that SCAN treatment could accelerate the destruction of cell wall integrity and accumulation of reactive oxygen species (ROS) in A. flavus. Moreover, pathogenicity assay indicated that in contrast to separate treatment with cinnamaldehyde or nonanal, SCAN could cause a decrease in the production of A. flavus asexual spores and AFB1 on peanuts, which verified its potential synergistic activity against fungal propagation. In addition, SCAN effectively preserves the organoleptic and nutritional properties of stored peanuts. Overall, our findings strongly indicated that the cinnamaldehyde/nonanal combination is a potentially significant antifungal agent against A. flavus contamination during the postharvest storage of peanuts.

Uji aktivitas antimikroba ekstrak etanol buah renggak (Amomum dealbatum Roxb.) terhadap Staphylococcus aureus dan Candida albicans

Staphylococcus aureus and Candida albicans are microorganisms that cause infection in humans and have developed resistance to several classes of antibiotics and antifungals. Plants in the genus Amomum have been used in traditional medicine and have scientific potential as alternatives to natural antibacterial and antifungal agents. Renggak (Amomum dealbatum Roxb.) is a species of the genus Amomum that is commonly found in Lombok. Scientific research related to the antibacterial and antifungal activity of the fruit is still limited. The aim of this study was to determine the antibacterial and antifungal activity of ethanol extract of renggak fruit against Staphylococcus aureus and Candida albicans. The extraction method used sonication with 96% ethanol. Testing of antibacterial and antifungal activity by disc diffusion method. The positive control used chloramphenicol for antibacterial testing and ketoconazole for antifungal testing, while the negative control used 10% DMSO solution. The antibacterial and antifungal activity of the extract was measured based on the diameter of the inhibition zone formed around the disc. Data analysis used the Kruskall-Wallis statistical test with a 95% confidence level. The results showed that the ethanol extract had moderate activity against S. aureus at concentrations of 20% w/v & 30%w/v with inhibitory diameters ranging from 5-7 mm, while at concentrations of 40% w/v & 50% w/v. has strong activity with inhibitory diameters ranging from 10-15 mm. The ethanol extract showed no activity at concentrations of 50%, 75%, and 100% in inhibiting the growth of the fungus C. albicans. Statistical test of each extract concentration had a significant effect on the inhibition of growth of S. aureus compared to the negative control. Based on the description above, it can be concluded that the extract of renggak fruit has the potential to inhibit the growth of S. aureus.

A magnetic nanocomposite combined with cinnamic acid for capture–inhibition–separation of Aspergillus fumigatus

AbstractTo prevent the generation of drug‐resistant fungi from long‐term exposure to microorganisms, cinnamic acid (CA), a natural effective antifungal agent, was successfully coupled with poly‐dopamine magnetic nanoparticles (CA–DMPs). Due to the low solubility of CA, the saturated solution of CA (1.61 mg/mL, 45°C) had no antifungal effect. Contrarily, CA–DMPs showed a good antifungal effect. The resulting heat‐stable and reusable antifungal CA–DMP composite particles were superparamagnetic (49.79 emu g−1) and had an average diameter of 25.01 ± 1.36 nm. The novel composites showed good antifungal activity and excellent recycling performance, the sterilization rate of CA–DMPs remained above 96% after seven consecutive running cycles. CA–DMP composites could damage the fungal cell wall and membrane system, leading to the leakage of cell inclusions. Furthermore, transcriptome analysis of Aspergillus fumigatus treated with composites showed that 466 differentially expressed genes were primarily associated with cell wall membrane, membrane transporters, energy metabolism, genetic expression, and oxidation–reduction. The effect of CA–DMPs in inducing mitochondrial membrane dysfunction might result in the disruption of energy metabolism and REDOX homeostasis. Overall, the results reported herein provide new insight into the potential antifungal nanomaterials. In vitro antifungal experiments performed on cherry tomatoes confirm the application potential of the synthesized material in the field of fruit and vegetable preservation.

Antifungal Activities of Essential Oil Obtained from Mentha Spicata var. Crispa against Selected Penicillium Species

Attention of the scientific community has still focused on application of essential oils (EOs) as natural antifungal agents in the food industry to prolong the shelf-life of food products. In this regard, the current study was designed to evaluate chemical composition, antioxidant capacity, and antifungal (in vitro, in situ) activities of spearmint (Mentha spicata var. crispa) essential oil (SEO) commercially obtained from Slovak company against selected Penicillium species. The EO was used in four concentrations (62.5, 125, 250, and 500 µL/L) chosen, and gas chromatography–mass spectrometry, DPPH, agar disc diffusion and vapor phase methods were employed for such analyses. Our results revealed carvone (57.5%) and α-limonene (17.6%) to be the principal constituents in the EO chemical composition. Although only a weak antioxidant capacity (20.40 ± 0.80% free radical-scavenging inhibition) was displayed by the SEO, the highest EO concentration (500 µL/L) was shown to be a moderate growth inhibitor of P. expansum (inhibition zone of 11.46 ± 0.63 mm) and P. crustosum (inhibition zones of 12.93 ± 0.46 mm). The growth of P. citrinum was only weakly inhibited by the SEO (≥ 250 µL/L). Most importantly, the ability of the SEO to inhibit the mycelial growth of three Penicillium spp. tested was pronounced (P < 0.05) for all applied concentrations. Accordingly, the results from the current study complement our previous ones dealing with the possibility of utilizing diverse EOs commercially achieved from the same company in the food sector.

Antifungal Efficacy of Mint Essential Oil Against Penicillium spp. Inoculated on Carrots

Current research aims to investigate the chemical composition, antioxidant, and in vitro and in situ antifungal activities of commercially available Mentha arvensis essential oil (mint, MEO). The identification of the volatile substances was done using Gas chromatography–Mass spectrometry (GC-MS) analysis. A total of 42 components representing 99.5% of the total oil were identified. The main compounds in the oil were menthol (37.3%), menthone (17.4%), neo-menthol (14.1%), and 1,8-cineole (4.9%). Antioxidant assays (1,1-diphenyl-2-picrylhydrazyl radical) demonstrate only weak activity for the MEO in values 195.00 ± 5.30 μg TEAC.mL-1, with 22.8 ± 1.2% free radical-scavenging inhibition. Evaluation of in vitro and in situ antifungal activities of MEO (in four concentrations: 62.5 µL/L, 125 µL/L, 250 µL/L, and 250 µL/L) against three strains of Penicillium (P.) spp. fungi strains (P. expansum, P. citrinum, P. crustosum) were assessed by disc diffusion method and vapor contact method on the carrot as model food, respectively. The suitability of carrots as a substrate for analyzes was verified by determining moisture content (MC) and water activity (aw), which showed values of 82.80 ± 2.33% and 0.959 ± 0.001, respectively. MEO exhibited promising antifungal activity against analyzed strains of test fungi as a diameter of zones of inhibition (from 2.88 ± 0.55 to 12.33 ± 1.14 mm), as well as the effectiveness of this oil was detected on the carrot model (from -5.41 ± 7.35 to 100.00 ± 0.00%). Moreover, it can be concluded that the growth inhibition of fungi strains significantly depends (P < 0.05) on the concentration of the MEO used in both procedures. Our results suggest that MEO, as a promising natural antifungal agent, can be applied in the innovative packaging of food products including carrots.

Revalorization by lactic acid bacterial fermentation of goat whey from cheese industry as a potential antifungal agent

The industry is continuously searching for new methods to reduce the use of chemical preservatives while still produce safer foods for the consumer. Goat whey is a waste from the cheese industry which is primarily treated as a residue. It is rich in different nutrients and has the potential to be used as a matrix for lactic acid bacteria fermentation. This work aims to find a revalorization of whey for the production of a natural origin antifungal agent by a fermentation that can be used as food ingredient. First different whey culture mediums were developed and fermented. Quantification of different antifungal molecules in whey culture mediums in comparation to a fermentation of De Man, Rogosa and Sharpe (MRS) culture medium evidenced that the production of this antifungal metabolites was similar. The in vitro qualitative and quantitative assays evidenced that the whey-based mediums increased the antifungal activity of the CFS in comparation to the MRS. The fermented whey produced bigger inhibition halos (above 1 cm) in the agar diffusion test in comparation to the fermented MRS (bellow 0.6 cm). Alike, in the minimal fungicidal concentration test the lowest antifungal concentrations were detected on the fermented whey mediums, averaging antifungal concentrations lower than 130 g/L. Finally, a multivariable data analysis determined that more antifungal metabolites yet to be detected were affecting in the antifungal activity from the whey samples. The fermented whey has the potential to be a tool in reducing the use of chemical preservatives in foods.

Design, synthesis and antifungal evaluation of novel nopol derivatives as potent laccase inhibitors.

To explore further potential natural product-based antifungal agents, a series of novel nopol-based carboxamide and hydrazide derivatives containing a natural pinene structure were designed, synthesized, and evaluated for their inhibitory activities against seven phytopathogenic fungi and oomycetes. The bioassay results indicated that some compounds exhibited good inhibitory activities against Gibberella zeae, Sclerotinia sclerotiorum, and Phytophthora capsici. Among them, compound 3h displayed excellent in vitro activities against G. zeae, with EC50 values of 1.09 mg L-1 , which was comparable with the commercial fungicides bixafen and carbendazim (median effective concentration [EC50 ]=1.21 and 0.89 mg L-1 , respectively). Notably, in vivo bioassay results suggested that compound 3h also showed prominent protective and curative effects (95.6% and 94.2%) at 200 mg L-1 against G. zeae. The scanning electron microscopy study indicated that compound 3h could destroy the morphological integrity of G. zeae hyphae. The in vitro enzyme inhibitory bioassay revealed that compound 3h exhibited potent inhibitory activity against laccase with median inhibitory concentration (IC50 ) values of 4.93 μm, superior to positive control cysteine (IC50 =35.50 μm), and its binding modes with laccase were elucidated by molecular docking study. In addition, the fluorescent imaging of the dansylamide-labeled derivatives 8 on wheat leaf epidermal cells and the hyphae of G. zeae revealed that this class of hydrazide derivatives could readily permeate into wheat leaves and reached the laccase target in fungal cells. Some nopol-based hydrazide derivatives exhibited excellent anti-G. zeae activity and laccase inhibitory activity, which merits further development as a new fungicide candidate for controlling Fusarium head blight. © 2023 Society of Chemical Industry.

Antifungal molecular details of MNQ-derived novel carbon dots against Penicillium digitatum

It is urgent to develop high-efficiency and low-toxicity natural antifungal agents on green mold caused by Penicillium digitatum. The effect of 2-methoxy-1, 4-naphthoquinone (MNQ) inhibition of P. digitatum was not very satisfactory. MNQ-derived carbon dots (MNQ-CDs) synthesized through a solvothermal route were used as antifungal agents against P. digitatum. The antifungal activity of prepared MNQ-CDswas enhanced compared to MNQ, and the minimum inhibitory concentration was 2.8 μg/mL. A total of 441 genes and 122 metabolites have undergone significant changes. The omics data revealed that MNQ-CDs primarily modified the metabolism of aromatic amino acids and synthesis of the cell membrane in P. digitatum, thereby inhibiting its propagation. Furthermore, compared with MNQ, MNQ-CDs had a better control effect on the green mold of citrus fruits, and could more significantly inhibit the propagation of P. digitatum. This study provides a new idea for the design of new and efficient antifungal materials.

Moroccan Endemic Artemisia herba-alba Essential Oil: GC-MS Analysis and Antibacterial and Antifungal Investigation

In Morocco, the endemic Artemisia herba-alba is well known by its traditional uses and health benefits. The search for natural, safe, and effective antibacterial and antifungal agents from plants is in high demand due to microbial and fungal resistance to conventional synthetic antibiotics and antifungal drugs. In this study, the A. herba-alba was collected from the region of Fez-Boulemane during the periods of March, June, and September. Essential oils (EOs) were extracted from the aerial part of the plant by the hydrodistillation method. The chemical constituents were determined using GC-MS as analytical tools. The antimicrobial activities of different oils were tested using the macrodilution method. The results showed the difference in the yields between the three EOs (0.49, 1.74, 1.30% (mL/100 g)), respectively, as well as in their corresponding chemical compositions. The main constituents revealed by GC-MS are higher contents of oxygenated monoterpenes (84.7, 84.4, 81%), such as cis chrysanthenyl acetate (30, 26.7, 27.6%), β-thujone (23.2, 12.9, 15.4%), camphor (9.76, 14.3, 15.8%), chrysanthenone (2.4, 1, 14%), 1,8-cineole (1.5, 11.7, 11.8%), trans β-dihydroterpineol (7.8, 7.2, 6.9%), α-thujone (4.8, 3, 5.4%), and sesquiterpenic davanone (3.9, 1.5, 1.4%), respectively. The three EOs biological activities’ results showed significant antimicrobial effects against four bacteria tested (E. coli, B. subtilis, S. aureus, M. luteus), with the MIC values ranging from 0.1 to 0.03% (v/v), as well as interesting antifungal effects on both wood rot fungi against four fungi examined (G. trabeum, P. placenta, C. puteana, C. versicolor) and molds against three microorganisms tested (A. niger, P. digitatum, P. expansum), with MIC values ranging from 0.2 to 0.03% (v/v) and 0.4 to 0.03% (v/v), respectively. The June and September EO samples showed more potent activities than those collected during March. Our research findings showed quantitative variability in both EO contents and chemical compositions, which could be due to the phenological stages, climatic conditions of growth, and harvesting periods. The potent results of the antimicrobial/antifungal activities were provided by the EOs of June and September and might be correlated to the contribution and synergism effect of all oxygenated monoterpenes. These results support the possible application of A. herba-alba EOs as natural and safe antibacterial agents, and an effective alternative to synthetic drugs, enabling the prevention and treatment of certain pathogenic infections in food and health, and the preservation of wood alteration against fungi.

Influences of flavonoids from Sedum aizoon L. on the cell membrane of Botrytis cinerea

Botrytis cinerea is a plant pathogen, which affects the quality of fruits. However, chemical antifungal agents are extremely harmful to the environment, hence it's critical to create a natural plant-derived antifungal agent to replace chemical antifungal agents. The study explored the antifungal mechanisms of flavonoids from Sedum aizoon L. (FSAL) against B. cinerea. In the study, cell membrane served as an important breakthrough point. The minimum inhibitory concentration (MIC) of FSAL against B. cinerea was investigated. The propidium iodide (PI) staining observation of B. cinerea was explored. The activities of crucial enzymes involved in the metabolism were studied as well as the phospholipid and fatty acid changes. The expression of genes related to the membrane lipid metabolism was also measured. The results showed that FSAL had a certain inhibitory effect on B. cinerea and the MIC was 1.500 mg/mL. FSAL disrupted the cell membrane of B. cinerea, which decreased the contents of glutathione and proline at a low level as well as the membrane phospholipids. Compared with the control group, the activities of crucial enzymes were decreased, and the expression of key genes was reduced. It indicated that FSAL inhibited B. cinerea by disrupting its cell membrane, offering the possibility of a natural plant-derived antifungal agent for the preservation of fruits.

Towards potential antifungal agents: synthesis, supramolecular self-assembly and in vitro activity of azole mono-, sesqui- and diterpenoids.

Terpenes and their derivatives are natural antifungal and antimicrobial agents. In this paper, potential antifungal agents were developed on the basis of farnesol, geraniol, myrtenol, perillyl alcohol, cedrol and phytol. The synthesized compounds exist in aqueous solutions as stable associates (D = 142-216 nm, PDI 0.04-0.16, Z = +0.9-+46 mV). Formation of stable associates of the compounds in solution promotes compaction and dosed release of the drug. The membranotropic activity of the compounds was also investigated to open up their possible application in the treatment of skin diseases. The relationship between membranotropism and lipophilicity coefficient (log P) has been established. The antifungal and antimicrobial activities of the obtained compounds were studied in vitro on the clinical isolate of Candida sp., Candida albicans, yeast Saccharomyces cerevisiae, Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus, Bifidobacterium bifidum) bacteria. Candida sp. (0.0781 mg mL-1) and Saccharomyces cerevisiae (0.0049 mg mL-1) showed the highest sensitivity to the agents.

In vitro and in vivo antifungal activity of extracts from Jamaica cherry Muntingia calabura l. leaves

Previous studies on M. calabura (Jamaica cherry) leaves reported the presence of polyphenols, flavonoids, glycosides, and tannins. However, the deep research focus on the antifungal activity of Jamaica cherry leaf extract (JCLE) is still limited. Hence, the current study was conducted on JCLE samples by using various solvents, i.e., 70% ethanol, 70% methanol, and 70% acetone, and then evaluating their antifungal activity. Analytical results showed that JCLE could be used as a natural antifungal potential agent against mold-causing spoilage in fruits. Specifically, the ethanolic extract had a strong inhibition of Colletotrichum gloeosporioides and Lasiodiplodia theobromae. While the methanolic and acetonic extracts prevented sharply the growth of Alternaria alternata and Fusarium equiseti, respectively. Generally, the ethanolic extract was able to inhibit the various mold strains better than the other solvents. Furthermore, the results showed that the percentage inhibition of radial growth (PIRG) of JCLE increased gradually as the concentration increased from 4000 to 40000 mg GAE/kg. The highest data obtained for C. gloeosporioides, A. alternata, F. equiseti, and L. theobromae were 53.29%, 57.03%, 54.81%, and 75.03%, respectively. In vivo experiment, the samples were treated with and without JCLE (the control sample) occuring the black spots caused by C. gloeosporioides on mango’s surface after 21 days and 30 days of storage at 12 ± 2 °C, respectively. Whereas the TSS and TA values of both samples were similar. While the visual appearance of the control was more yellow compared to the samples treated with JCLE. In conclusion, JCLE was able to significantly delay the development of pathogenic mold, though it quite inhibited the maturation of mango fruit. Further studies may be needed to evaluate the phtychemical profile of M. calabura extract to gain a deeper understanding of its antifugal activity.

Antifungal Efficacy of Gallic Acid Extracted From Pomegranate Peel Against Trichophyton rubrum: In Vitro Case Study

Objectives: Trichophyton rubrum is one of the main pathogens causing superficial dermatophytosis, producing symptoms such as skin itching and pain, which seriously affects the quality of life of patients. Pomegranate peel extract is rich in gallic acid (GA), which has been reported to have biological effects including antifungal activity. However, the morphological and molecular mechanisms underlying the effects of GA on T rubrum are not well understood. The objectives of this study were to determine the antifungal efficacy of GA extracted from pomegranate peel against T rubrum in vitro, and to explore the underlying molecular mechanisms. Methods: The effects of 0-, 0.5-, and 1 mg/mL GA in pomegranate peel extract on T rubrum was investigated by detecting cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Transmission electron microscopy (TEM) was used to analyze the ultrastructure of T. rubrum, and transcriptome sequencing was used to analyze the enrichment pathway of differentially expressed genes. The identification of biosynthesis-related and key genes in the pathways involved using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) technology. Terbinafine hydrochloride (TERB) as a positive control group. Results: Pomegranate peel extract has a GA content of 1.0 mg/mL. Compared with untreated group, following treatment with 1.0 mg/mL GA content is rich in pomegranate peel extract, and the microstructure of T rubrum is destroyed. TEM results show that the number of lipid droplets in T rubrum was significantly increased, mitochondrial vacuoles degenerated, the serosa were damaged, and the boundary of thallus was unclear. In addition, 1 mg/mL GA can significantly inhibit T rubrum proliferation, and its inhibition ability is better than TERB. Transcriptomics results show that GA can change the gene expression profile of T rubrum, specifically: The biosynthesis was blocked, drug resistance was weakened, the transport of ATP-binding cassette (ABC) drugs transporter was increased, and the mitogen-activated protein kinase (MAPK) pathway was significantly inhibited. Conclusions: Pomegranate peel extract is rich in GA, which strongly inhibited the growth of T rubrum and reduced its drug resistance. This extract is a promising natural antifungal agent for clinical use.

Antifungal activity of selected sea cucumber species from Tukuran, Zamboanga del Sur, Mindanao, Philippines using modified SPOTi assay

Abstract. Ereguero MG, Cordero MA, Dalayap R, Tabugo SR. 2022. Antifungal activity of selected sea cucumber species from Tukuran, Zamboanga del Sur, Mindanao, Philippines using modified SPOTi assay. Biodiversitas 23: 6049-6055. Marine organisms have become essential candidates for discovering novel compounds that can be medically and commercially important. They are believed to yield novel compounds with unique chemical structures and significant biological activities since they managed to survive a milieu of pathogenic microorganisms and essential predators. The quest against fungal infections has been challenging due to acquired resistance to commercially available drugs hence, novel sources to treat infectious diseases caused by fungi have been investigated. Sea cucumbers have been recognized in Asian countries as a rich food source. Apart from that, they have also been studied for their medicinal properties. This study aimed to evaluate the antifungal activity of the selected sea cucumber species found in Brgy. Sugod, Tukuran, Zamboanga del Sur, Mindanao, Philippines using the modified SPOTi assay. Methanol: ethyl acetate crude extracts of Holothuria scabra, Stichopus sp., and Holothuria atra were tested for their antifungal activity against Aspergillus niger and Candida tropicalis. A 96-well plate assay was used, and the growth of fungi was assessed visually for up to 72hrs. The concentration resulting in ‘no fungal growth’ was observed and taken as the Minimum Inhibitory Concentration (MIC) value. The average MIC was calculated. Results showed that sea cucumber crude extracts have MIC values of 0.0333-0.1 mg/mL. Tukey’s posthoc test confirmed that the antifungal activity of the sea cucumber extracts was comparable to the positive control Fluconazole (p>0.05). The present study revealed a potentially effective natural antifungal agent from sea cucumbers.