Minimum Fungicidal Concentration Research Articles

Unravelling the Antimicrobial, Antibiofilm, Suppressing Fibronectin Binding Protein A (fnba) and cna Virulence Genes, Anti-Inflammatory and Antioxidant Potential of Biosynthesized Solanum lycopersicum Silver Nanoparticles.

Background and Objectives: Urinary tract infections [UTIs] are considered the third most known risk of infection in human health around the world. There is increasing appreciation for the pathogenicity of Gram-positive and Gram-negative strains in UTIs, aside from fungal infection, as they have numerous virulence factors. Materials and Methods: In this study, fifty urine samples were collected from patients suffering from UTI. Among the isolates of UTI microbes, six isolates were described as MDR isolates after an antibiotic susceptibility test carried out using ten different antibiotics. An alternative treatment for microbial elimination involved the use of biosynthesized silver nanoparticles (AgNPs) derived from Solanum lycopersicum [S. cumin]. Results: The sizes and shapes of AgNPs were characterized through TEM imaging, which showed spherical particles in a size range of 35-80 nm, of which the average size was 53 nm. Additionally, the silver nanoparticles (AgNPs) demonstrated inhibitory activity against Staphylococcus aureus (OR648079), exhibiting a 31 mm zone of inhibition at a minimum inhibitory concentration (MIC) of 4 mg/mL and a minimum bactericidal concentration (MBC) of 8 mg/mL. This was followed by Aspergillus niger (OR648075), which showed a 30 mm inhibition zone at an MIC of 16 mg/mL and a minimum fungicidal concentration (MFC) of 32 mg/mL. Then, Enterococcus faecalis (OR648078), Klebsiella pneumoniae (OR648081), and Acinetobacter baumannii (OR648080) each displayed a 29 mm zone of inhibition at an MIC of 8 mg/mL and an MBC of 16 mg/mL. The least inhibition was observed against Candida auris (OR648076), with a 25 mm inhibition zone at an MIC of 16 mg/mL and an MFC of 32 mg/mL. Furthermore, AgNPs at different concentrations removed DPPH and H2O2 at an IC50 value of 13.54 μg/mL. Also, AgNPs at 3 mg/mL showed remarkable DNA fragmentation in all bacterial strains except Enterococcus faecalis. The phytochemical analysis showed the presence of different active organic components in the plant extract, which concluded that rutin was 88.3 mg/g, garlic acid was 70.4 mg/g, and tannic acid was 23.7 mg/g. Finally, AgNPs concentrations in the range of 3-6 mg/mL showed decreased expression of two of the fundamental genes necessary for biofilm formation within Staphylococcus aureus, fnbA (6 folds), and Cna (12.5 folds) when compared with the RecA gene, which decreased by one-fold when compared with the control sample. These two genes were submitted with NCBI accession numbers [OR682119] and [OR682118], respectively. Conclusions: The findings from this study indicate that biosynthesized AgNPs from Solanum lycopersicum exhibit promising antimicrobial and antioxidant properties against UTI pathogens, including strains resistant to multiple antibiotics. This suggests their potential as an effective alternative treatment for UTIs. Further research is warranted to fully understand the mechanisms of action and to explore the therapeutic applications of these nanoparticles in combating UTIs.

Oral Rinses: Some Kill and Some Cripple Candida albicans

Introduction: Oral healthcare professionals play a crucial role in guiding patients toward evidence-based choices among the many available oral rinses. In this study, we explored how specific oral rinse formulations affect the viability and modulate critical virulence traits of the opportunistic fungal pathogen Candida albicans. Materials and Methods: We assessed the effects of these oral rinses on the production of germ tube, production of phospholipase and hemolysin, as well as biofilm formation. Results: We found that oral rinses containing cetylpyridinium chloride (CPC) and chlorhexidine (CHX) showed the greatest fungicidal activity with the lowest minimum fungicidal concentrations (0.38% and 0.78%, respectively). Oral rinses based on zinc chloride and sodium fluoride with Miswak bark extract (MIS) or essential oils (EO) had much lower fungicidal activity (8–16 times lower) compared to CHX and CPC. However, they had a significantly greater impact on the virulence traits of C. albicans. They reduced germ tube production by 86–89% (vs. 42% for CHX and 29% for CPC), completely inhibited phospholipase and hemolysin production, and together with the CPC-based oral rinse, exerted the greatest reductions in biofilm formation across all tested concentrations. This was in contrast to both the controls and CHX, which had a minimal effect on biofilm formation. Conclusion: By inhibiting the virulence factors, the oral rinse can have a crippling effect on C. albicans, weakening this opportunistic pathogen and hindering its potential to cause infection.

In vitro activity of the anthelmintic drug niclosamide against Sporothrix spp. strains with distinct genetic and antifungal susceptibility backgrounds.

The drugs available to treat sporotrichosis, an important yet neglected fungal infection, are limited. Some Sporothrix spp. strains present reduced susceptibility to these antifungals. Furthermore, some patients may not be indicated to use these drugs, while others may not respond to the therapy. The anthelmintic drug niclosamide is fungicidal against the Sporothrix brasiliensis type strain. This study aimed to evaluate whether niclosamide also has antifungal activity against Sporothrix globosa, Sporothrix schenckii and other S. brasiliensis strains with distinct genotypes and antifungal susceptibility status. Minimal inhibitory and fungicidal concentrations (MIC and MFC, respectively) were determined using the microdilution method according to the CLSI protocol. The checkerboard method was employed to evaluate niclosamide synergism with drugs used in sporotrichosis treatment. Metabolic activity of the strains under niclosamide treatment was evaluated using the resazurin dye. Niclosamide was active against all S. brasiliensis strains (n = 17), but it was ineffective (MIC > 20µM) for some strains (n = 4) of other pathogenic Sporothrix species. Niclosamide MIC values for Sporothrix spp. were similar for mycelial and yeast-like forms of the strains (P = 0.6604). Niclosamide was fungicidal (MFC/MIC ratio ≤ 2) for most strains studied (89%). Niclosamide activity against S. brasiliensis is independent of the fungal genotype or non-wild-type phenotypes for amphotericin B, itraconazole, or terbinafine. These antifungal drugs presented indifferent interactions with niclosamide. Niclosamide has demonstrated potential for repurposing as a treatment for sporotrichosis, particularly in S. brasiliensis cases, instigating in vivo studies to validate the in vitro findings.

Unlocking the therapeutic potential of Carapa guianensis Aubl. (Meliaceae) leaves against Candida Sp.

Carapa guianensis Aubl., Meliaceae, is popularly known as andiroba, a plant species with wide therapeutic use, especially its fixed oil and leaves as a healing and anti-inflammatory agent. The work aimed to determine the antifungal and antioxidant activity of the extract and fractions of C. guianensis leaves. The leaves of C. guianensis collected in the municipality of São Luís, Maranhão, Brazil, were dried and pulverized. The powder was percolated with 70% ethanol, using a hydromodule of 1:12. The extract was developed with hexane and then partitioned (v/v) obtaining the following fractions: chloroform, ethyl acetate, and hydroethanol. The toxicological bioassay was carried out with Artemia salina and antifungal activity evaluated the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) with different species of the genus Candida. Toxicological tests showed progressive toxicity as partitioning was carried out. In the antifungal assay, the growth and death of C. albicans, C. glabrata, C. parapsilosis, and C. tropicalis were observed, demonstrating potential for use as an antifungal agent in pharmaceutical products.

In vitro Assessment of Antimicrobial potential of Siddha Polyherbal Formulation Tulasi oil against RTI pathogens

In Siddha medication, along with herbal formulations, some mineral formulations are used clinically to prevent or cure infectious diseases. The therapeutic values of some siddha formulations have been well documented earlier, but a huge number of them remain unexplored in terms of safety and efficacy. The aim of this study was to screen the antimicrobial potential of Siddha polyherbal formulation Tulasi oil (SPHTO) against respiratory tract infection (RTI) causing pathogens. The antimicrobial activity of SPHTO was examined by using the methods of agar well diffusion, Minimum inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC), Minimum Fungicidal Concentration (MFC) and Microbicidal action. Ciprofloxacin (bacteria) and Amphotericin B (fungi) were used as the positive control for this study. The potassium leakage was determined using a flame emission and atomic absorption spectroscopy. All tested concentrations gave notable antimicrobial activity against all tested pathogens. The 100mg/ml concentration showed the highest activity against E.coli followed by 200mg/ml against K. pnuemonia and C.neoformans when compared to 50 and 25mg/ml. For MIC, the highest activity was registered at 150mg/ml concentration against E. coli, while MBC/MFC was 200mg/ml concentration has shown the better activity against K. pneumonia and C. neoformans. Tulasi oil induced leakage of potassium ions from cells of E.coli and K.pneumonia suspended in glycil-glycine buffer solution and the leakage started within 10 minutes of contact of drugs with target cells. From this study, it may support the clinical claims of Tulasi oil to have potent antibacterial and antifungal activities and offer profound therapeutic benefits against respiratory tract infection.

Pharmacological potential of limonene against opportunistic fungi: Impact on Candida virulence

The present article aims to evaluate the antifungal and antivirulence effect of the phytoconstituent Limonene against Candida spp. Antifungal assays were performed, where the concentration capable of inhibiting 50 % of fungal growth, the growth inhibition curve, the minimum fungicidal concentration, the evaluation of the modifying effect with fluconazole, the inhibitory effect of the substances on the morphological transition of Candida spp. and the statistical analysis of the results were determined. With this study, it was seen that limonene demonstrated growth inhibition for the strains tested and when associated the natural compound with Fluconazole, there was potentiation of the effect of the drug, since the inhibition of growth by the combination occurred at lower concentrations against all strains tested, when compared to the drug alone, which inhibited growth at the highest concentration. In the test to determine the Minimum Fungicidal Concentration of the products tested alone and in combination, it was found that in the case of Candida strains, growth inhibition by limonene occurred at a concentration of 1024 μg/mL. For Fluconazole, growth impairment ranged from > 1024 μg/mL to 256 μg/mL for the strains. And when combined, limonene potentiated the action of FCZ, making fungal colonization unfeasible at concentrations below 1024 μg/mL. Regarding the morphological transition from yeast to hyphae, limonene was used at concentrations of 1024 μg/mL and 512 μg/mL, and it was found that, for CA and CK, the filaments were reduced in number and size at the highest concentration and against CT, the morphological transition from yeast to hyphae/pseudohyphae was totally inhibited, and if compared to the growth control, limonene was able to reduce fungal growth at concentrations greater than 512 μg/mL. This compound has antimicrobial activity described, due to its ability to interfere in the gene expression of the fungus, the limited therapeutic options and the recent emergence of multidrug-resistant Candida species represent a significant challenge for human medicine and highlight the need for new therapeutic approaches, and in this study a great potential of limonene was revealed in relation to the perspective of increasing the efficiency of commercial drug. This work can bring an important contribution to the scientific database, while emphasizing that in-depth studies and tests on the subject, in order to better investigate its effectiveness and mechanisms by which they exert their effects, are still necessary.

Antimicrobial effects of catnip (Nepeta cataria L.) essential oil against canine skin infection pathogens

Background and Aim: Catnip essential oils have antimicrobial effects against bacteria, yeast, and fungi; however, there is limited information regarding their antimicrobial activity against pathogens that cause canine skin infections. This study aimed to identify the phytochemical constituents of catnip essential oil and assay its antimicrobial activity against Staphylococcus pseudintermedius, Malassezia pachydermatis, Microsporum canis, Microsporum gypseum, Microsporum gallinae, and Trichophyton mentagrophytes. Materials and Methods: Catnip essential oil was extracted by hydrodistillation, and its chemical constituents were analyzed by gas chromatography–mass spectrometry (GC–MS). In vitro antimicrobial activity was investigated using broth microdilution and time-kill tests. To evaluate the effect of catnip essential oil on microbial morphology and cell membrane integrity, scanning electron microscopy (SEM) and leakage studies were conducted. Results: GC–MS analysis revealed that the principal components of catnip essential oil were cis- and trans-nepetalactone (57.09% of peak area), trans-, cis-nepetalactone (39.69% of peak area), trans-caryophyllene (1.88% of peak area), and caryophyllene oxide (1.34% of peak area). The minimum inhibitory concentration, minimum bactericidal concentration, and minimum fungicidal concentration values determined by broth microdilution ranged from 0.0625 mg/mL to 4.0 mg/ mL. Time-kill testing showed that the germicidal effects of catnip essential oil were time and concentration-dependent, respectively. Environmental SEM and cell leakage analysis indicated that catnip essential oil disrupted the integrity of cell membranes in the tested microorganisms. Conclusion: Catnip essential oil has potential as an alternative antimicrobial against a wide range of canine skin infection pathogens, including S. pseudintermedius, M. pachydermatis, Mi. canis, Mi. gypseum, Mi. gallinae, and T. mentagrophytes. Keywords: antimicrobial activity, canine dermatitis, catnip, Nepeta cataria.

Nitrogen-doped carbon quantum dots as a novel treatment for black fungal bone infections (Mucormycosis): in vitro and in vivo study

Most fungal bone and joint infections (arthritis) are caused by Mucormycosis (Mucor indicus). These infections may be difficult to treat and may lead to chronic bone disorders and disabilities, thus the use of new antifungal materials in bone disorders is vital, particularly in immunocompromised individuals, such as those who have contracted coronavirus disease 2019 (COVID-19). Herein, we reported for the first time the preparation of nitrogen-doped carbon quantum dots (N/CQDs) and a nitrogen-doped mesoporous carbon (N/MC) using a quick micro-wave preparation and hydrothermal approach. The structure and morphology were analysed using X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) and surface area analyser. Minimum inhibitory concentration (MIC), disc diffusion tests, minimum fungicidal concentration (MFC) and antifungal inhibitory percentages were measured to investigate the antifungal activity of N/CQDs and N/MC nanostructures. In addition to the in vivo antifungal activity in rats as determined by wound induction and infection, pathogen count and histological studies were also performed. According to in vitro and in vivo testing, both N/CQDs with small size and N/MC with porous structure had a significant antifungal impact on a variety of bone-infecting bacteria, including Mucor infection. In conclusion, the present investigation demonstrates that functional N/CQDs and N/MC are effective antifungal agents against a range of microbial pathogenic bone disorders in immunocompromised individuals, with stronger and superior fungicidal activity for N/CQDs than N/MC in vitro and in vivo studies.

Green synthesis and characterization of silver nanoparticles from Ducrosia flabellifolia Boiss. aqueous extract: Anti-quorum sensing screening and antimicrobial activities against ESKAPE pathogens.

Biosynthesis of silver nanoparticles using natural compounds derived from plant kingdom is currently used as safe and low-cost technique for nanoparticles synthesis with important abilities to inhibit multidrug resistant microorganisms (MDR). ESKAPE pathogens, especially MDR ones, are widely spread in hospital and intensive care units. In the present study, AgNPs using Ducrosia flabellifolia aqueous extract were synthesized using a reduction method. The green synthesized D. flabellifolia-AgNPs were characterized by UV-Vis spectrophotometer, Scanning electron microscopy (SEM), and X-ray diffraction assays. The tested D. flabellifolia aqueous extract was tested for its chemical composition using Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (LC-ESI-MS). Anti-quorum sensing and anti-ESKAPE potential of D. flabellifolia-AgNPs was also performed. Results from LC-ESI-MS technique revealed the identification of chlorogenic acid, protocatechuic acid, ferulic acid, caffeic acid, 2,5-dihydroxybenzoic acid, and gallic acid as main phytoconstituents. Indeed, the characterization of newly synthetized D. flabellifolia-AgNPs revealed spherical shape with mean particle size about 16.961±2.914 nm. Bio-reduction of silver was confirmed by the maximum surface plasmon resonance of D. flabellifolia-AgNPs at 430 nm. Newly synthetized D. flabellifolia-AgNPs were found to possess important anti-ESKAPE activity with low minimal inhibitory concentrations (MICs) ranging from 0.078 to 0.312 mg/mL, and low minimal bactericidal concentrations (MBCs) varying from 0.312 to 0.625 mg/mL. Moreover, D. flabellifolia-AgNPs were active against Candida utilis ATCC 9255, C. tropicalis ATCC 1362, and C. albicans ATCC 20402 with high mean diameter of growth inhibition at 5 mg/mL, low MICs, and minimal fungicidal concentrations values (MFCs). The newly synthetized D. flabellifolia-AgNPs were able to inhibit violacein production in Chromobacterium violaceum, pyocyanin in Pseudomonas aeruginosa starter strains. Hence, the newly synthesized silver nanoparticles using D. flabellifolia aqueous extract can be used as an effective alternative to combat ESKAPE microorganisms. These silver nanoparticles can attenuate virulence of Gram-negative bacteria by interfering with the quorum sensing system and inhibiting cell-to-cell communication.

In Vitro Antimycotic Activity and Structural Damage against Canine Malassezia pachydermatis Strains Caused by Mexican Stingless Bee Propolis.

This work describes the antimycotic activity of propolis from the stingless bees Scaptotrigona mexicana and Tetragonisca angustula, collected from two Mexican regions (Veracruz and Chiapas, respectively), against three clinical isolates and the reference strain ATCC 14522 of Malassezia pachydermatis, the causative agent of canine otitis. The chemical components of the ethanolic extracts of propolis were determined by gas chromatography coupled with mass spectrometry (GC-MS), and sesquiterpenes were the predominant compounds. The antimycotic activity was evaluated by plate microdilution. The induced changes in the yeasts were evaluated by fluorescence microscopy and staining with calcofluor white and propidium iodide. The minimum inhibitory concentration (MIC) was 7.11 mg/mL, and the minimum fungicidal concentration was 21.33 mg/mL for both extracts. The EPPs of Scaptotrigona mexicana and Tetragonisca angustula caused substantial damage to yeast morphology, where the propidium iodide staining of the yeasts treated with both EEPs revealed the penetration of this marker, which indicates the destruction of the cell wall and plasma membrane of the fungi. This result suggests that these types of propolis could be used as alternative treatments for canine external otitis. To the best of our knowledge, this seems to be the first scientific report that has demonstrated structural damage in Malassezia pachydermatis by Mexican stingless bee propolis.

Chemically engineered nano selective silver shapes: Novel synthesis and their potential activity as anti-candida agents

Oral candidiasis (OC) is an infection of oral mucosa caused by fungal overgrowth, especially Candida albicans due to its ability to form biofilms and the increased expression of virulence factors. In the present study, Ag Nano sphere & Ag Nano triangle were newly synthesized using Juniperus drupacea L leaves extract to combat the OC infections. Ag Nano sphere & Ag Nano triangle antifungal activity, mechanism of action against OC, and cytotoxic effect in L929 (ATCC CCL-1 NCTC) fibroblast cell monolayers were evaluated. The J. drupacea phytochemical analysis revealed the presence of three major compounds: α-pinene (34.27 %), thymol methylether (15.20 %) and camphor (9.01 %). The proposed synthesis approach includes the reduction of Ag+ by Juniperus drupacea L leaves extract to form Ag Nano spheres. One the other hand, the presence of polyvinylpyrrolidone (PVP) in the reaction mixture led to obtaining Ag Nano triangle shapes. The minimal fungicidal concentration (MFC) and minimal biofilm eradication concentration (MBEC) assays revealed the higher antifungal effect of Ag Nano sphere when compared to Ag Nano triangle. MFC of Ag Nano sphere and Nano triangle reached 62.5 and 125 µg/mL respectively. While MBECs were ranged between 4 and 32 and 16–64 µg/mL upon using Ag Nano sphere and Nano triangle respectively. The possible mechanism of action of the synthesized Ag Nano shapes was the attributed to the direct effect on the cell membrane especially after 6 h incubation. In vitro cytotoxic effect of the prepared Ag-Nano sphere on cell proliferation using L929 (ATCC CCL-1 NCTC) fibroblast cell monolayers suggested the Ag-Nano spheres are safe for possible biomedical application.

Marine Ascomycetes Extract Antifungal Susceptibility against Candida spp. Isolates from Oral Candidiasis HIV/AIDS Patient: An In Vitro Study.

The etiology of oral candidiasis (OC) was Candida albicans, C. krusei, C. dubliniensis, C. tropicalis that are frequently found in human immunodeficiency virus/ acquired immunodeficiency syndrome (HIV/AIDS) patients. Marine ascomycetes (MA) have been widely reported as an important producer of various antibiotic compounds. However, there is limited study of antifungal compounds from MA against Candida species. The aim of this study was to investigate the antifungal susceptibility of MA against Candida spp. isolates from OC HIV/AIDS patient. Trichoderma sp. is a sponge-associated fungus collected from Karimunjawa National Park, Central Java, Indonesia. The validation of C. albicans, C. krusei, C. dubliniensis, C. tropicalis. was done by ChromAgar. This study was true experimental with post-test only control group design; the sample was four replications for each group. Nystatin administration (K +), the golden standard antifungal drug, was used. The minimum fungicidal concentration (MFC), minimum inhibitory concentration (MIC), and diffusion zone methods were done. Analysis of variance difference test, and post-hoc Tukey's honest significant different were done to analyze the significant different between groups (p ≤ 0.05). The MFC and MIC of MA against C. albicans, C. krusei, C. dubliniensis, and C. tropicalis were found at 12.5%. In addition, the greatest diffusion zone of MA against C. albicans, C. krusei, C. dubliniensis, and C. tropicalis was found at 12.5%. There is no appreciable difference in antifungal activity between K + and 12.5% of MA extract (p ≥ 0.05). Concentration of 12.5% MA extract has antifungal susceptibility against Candida spp. isolates from OC HIV/AIDS patient.

Time-kill kinetic of nano-ZnO-loaded nanoliposomes against Aspergillus niger and Botrytis cinerea.

In vitro antimicrobial activity of nano-ZnO-loaded nanoliposomes at different levels of lecithin:nano-ZnO ratio (5:1, 15:1, and 25:1 w/w) against Aspergillus niger (IBRC-M 30095) and Botrytis cinerea (IBRC-M 30162) was evaluated. Nanoliposome formulations containing nano-ZnO were fabricated through thin-layer hydration sonication and heat methods. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of nano-ZnO-loaded nanoliposomes and free nano-ZnO against Aspergillus niger and Botrytis cinerea were determined. The time-kill experiments were performed for each isolate. Results showed that the encapsulation of nano-ZnO in nanoliposome systems significantly enhanced their antimicrobial activities by improving the penetration of ZnO nanoparticles the fungi cell membrane. In vitro antifungal activity of nano-ZnO-loaded nanoliposomes against Aspergillus niger and Botrytis cinerea was increased in thin-layer hydration sonication method compared with the heat method. The log phase for Aspergillus niger and Botrytis cinerea was around 70 h. Adding nano-ZnO-loaded nanoliposomes to the culture medium shortened the log phase for both Aspergillus niger and Botrytis cinerea. The highest antimicrobial activity of nanoliposomes was achieved using nanoliposomes containing the lecithin:nano-ZnO ratio of 25:1 (w/w) as compared to all samples. However, the length of the log phase growth cultures exposed to the nanoliposome formulations prepared by thin-layer hydration sonication method with the lecithin:nano-ZnO ratio of 25:1 (w/w) at MIC and MFC values was 60 and 40 h for both Aspergillus niger and Botrytis cinerea, respectively.

Phytochemical Characterization, Antimicrobial Activity, Pharmacokinetic,in silicoMolecular Docking and Interaction Analysis of Ajwa (Phoenix dactyliferaL.) Palm Date Seeds

BackgroundThere is growing evidence that antibiotic resistance is increasing. As a result, the efficacy of these antibiotics is gradually declining. This poses a serious health risk in terms of treatment strategies. Therefore, the scientific community is continuously putting all efforts into looking for novel and green therapeutics to overcome the problem of antibiotic resistance.ObjectivesThe aim of this study is to investigate the chemical composition of the methanolic extracts obtained from Phoenix dactylifera seeds, known agricultural waste, to identify new bioactive compounds and to examine the antibacterial and antifungal potentials.Materials and MethodsPhytochemical profiling was performed by the gas chromatography-mass spectrometry (GC-MS) technique. For the antibacterial activity of the methanolic extract, 12 microorganisms were selected, including eight bacteria ( Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Enterobacter cloacae, Enterococcus faecium, Staphylococcus hominis, Staphylococcus aureus, Staphylococcus epidermidis, and Klebsiella pneumoniae ATCC 27736) and four Candida species (C. albicans ATCC 10231, C. tropicalis ATCC 1362, C. utilis ATCC 9255, and C. guilliermondii ATCC 6260). The antimicrobial activity of the plant extract was supported by in silico molecular docking analysis.ResultsThe obtained results from the GC-MS technique revealed the presence of 51 compounds, of which 6-octadecenoic acid (Z) was identified as the major component. The tested extract exhibited the highest zone of inhibition against S. aureus (growth inhibition zone 14.66 ± 0.57 mm). In addition, minimal inhibitory concentration (MIC) and minimal bactericidal concentration values obtained against S. aureus were 1.4 and 5.6 mg/mL, respectively. Regarding the antifungal assay, C. albicans was the most sensitive one to the extract among all the tested Candida strains (14.33 mm) (MIC: 1.4 mg/mL, minimal fungicidal concentration (MFC): 11.2 mg/mL). The binding free energies and root mean square deviation values obtained from the interaction of phytochemicals with two different target receptors (1JIJ and 2QZW for TyrRS from S. aureus and aspartic proteinase from C. albicans) were also determined. Ergost-5-en-3-ol, (3beta) established hydrogen, electrostatic, alkyl, and pi-alkyl interactions with five different amino acid residues (Asp40, Asp80, Pro53, His50, and Phe54) located at the TyrRS receptor of S. aureus. On the other hand, hexadecanoic acid and methyl ester established hydrogen, electrostatic, pi-sigma, and alkyl interactions with five different amino acid residues (Thr222, Asp86, Asp218, Tyr84, and Ile123) located at the aspartic proteinase receptor of C. albicans. These interactions contributed to the stability of the complex and, hence, to the biological activity of the P. dactylifera compounds.ConclusionOur results highlighted that methanolic extract from date seeds can be a natural source of chemical compounds with promising antibacterial and antifungal activities, as demonstrated using both in vitro and in silico approaches.

Efficacy and Antifungal Mechanism of Rosemary Essential Oil against Colletotrichum gloeosporioides

The antifungal activity and mechanism of rosemary essential oil against Colletotrichum gloeosporioides, the walnut anthracnose pathogen, were investigated using scanning electron microscopy (SEM), index determination and transcriptome technique. The results showed that rosemary essential oil could inhibit the growth of C. gloeosporioides with minimum inhibitory (MIC) and fungicidal (MFC) concentrations of 15.625 μL/mL and 31.25 μL/mL, respectively. Scanning electron microscopy revealed that the mycelium morphology became shriveled, twisted, and severely deformed after being treated with rosemary essential oil. The activity of chitinase, which decomposes fungal cell wall components in C. gloeosporioides, increased. The ergosterol content in the plasma membrane decreased, while the cell contents including nucleic acids, soluble protein and soluble reducing sugar were released resulting in the extracellular electrical conductivity being changed. For metabolic activity, the enzymes succinate dehydrogenase (SDH), malate dehydrogenase (MDH), ATPase and ATP decreased, whereas phosphofructokinase (PFK) increased. Transcriptome sequencing results showed that the antifungal mechanism of rosemary essential oil involves the destruction of the cell wall and membrane, inhibition of genetic material synthesis, and cell division and differentiation. The results are helpful to understand the efficacy and antifungal mechanism of rosemary essential oil against C. gloeosporioides and provide a theoretical basis for the development of rosemary essential oil as a biological control agent.

Mechanistic insight into the membrane disrupting properties of thymol in Candida species

Candida infection is a major problem in immunocompromised patients and has become a significant public health concern in recent years due to high toxicity, multidrug resistance and unfavourable side effects of conventional antifungal drugs. This study aims to investigate the antifungal efficacy of thymol, a natural monoterpenoid, on the membrane integrity of Candida albicans, Candida glabrata and Candida tropicalis. Minimum inhibitory concentrations (MICs) of thymol were 32 µg/ml, 63 µg/ml, and 125 µg/ml for C. albicans, C. tropicalis and C. glabrata, respectively. The minimum fungicidal concentrations (MFC) were 64 µg/ml, 110 µg/ml and 240 µg/ml, respectively. At MIC, thymol treated cells showed complete suppression of cell growth (growth kinetics, disc diffusion), altered morphology (scanning electron microscopy), leakage of intracellular macromolecules (absorption at 260 nm), significant reduction in ergosterol levels and inhibition of plasma membrane H+-ATPase (Pma1) activity. The disruption of the fungal membranes by thymol was further confirmed by propidium iodide uptake using flow cytometry and confocal microscopy. In silico studies with membrane-bound proteins (lanosterol 14-α demethylase (LDM) and Pma1), showed good binding affinity of −6.3 kcal/mol and −6.7 kcal/mol, respectively, and formed hydrogen bonds with crucial amino acid residues at the active site of target proteins. With low MIC values, negligible host toxicity, desirable ADME properties, thymol has immense potential as a potent antifungal drug that can be used for developing mechanism-based drugs as it has specific binding affinity for specific target proteins. Studies using animal models are required for further validation.

Formulation and Development of a Herbal Antifungal Gel Containing Origanum vulgare and Syzygium aromaticum Essential Oils Against Oral Candida albicans.

Background Oral candidiasis is the most prevalent oral fungal infection, and existing antifungal agents have side effects such as drug intolerance, resistance, and toxicity. Herbal essential oils are emerging as an alternative therapeutic approach for treating fungal infections. Origanum vulgare (O. vulgare), commonly known as oregano, and Syzygium aromaticum (S. aromaticum), commonly known as clove, are known to have antifungal properties and are effective against fluconazole-resistant strains. A combination of essential oils has a synergistic effect and aids in achieving effective antifungal activity at sufficiently low concentrations, which could lead to reduced side effects and resistance. Aim of the study This study aimed to formulate and develop an herbal antifungal gel containing O. vulgareand S. aromaticum and evaluate its synergistic antifungal efficacy against oralCandida albicans (C. albicans). Methodology Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) determinations of O. vulgareand S. aromaticum essential oils were performed individually and in combination to assess the antifungal activity against C. albicans. Based on the obtained MIC and MFC of essential oils in combination, anherbal antifungal gel was formulated. Further, to determine the biocompatible nature of the gel, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. Results We found that a combination of O. vulgare and S. aromaticum essential oils showed antifungal activity at a lesser concentration, with a MIC of 0.19 μl/ml and MFC of 0.39μl/ml when compared to their individual concentrations. Based on our results, an antifungal herbal gel comprising a concentration of 0.6 μl/ml of both essential oils was developed to achieve synergistic antifungal activity against oral C. albicans. The MTTassay of the herbal gel did not show any cytotoxicity. Conclusion The novel herbal antifungal gel containing O. vulgareand S. aromaticum is biocompatible in nature and provides an alternative therapeutic approach for treating oral candidiasis.

Quantitative phytochemical, antioxidant and antimicrobial properties of the seeds of Lawsonia inermis L.

Lawsonia inermis, commonly known as Henna, is a plant that has been extensively utilized throughout the ages for its medicinal and cosmetic properties. This plant contains a variety of bioactive compounds and has attracted significant interest due to its potential antioxidant and antimicrobial activities. The aim of our study is to evaluate the antioxidant capacity and antibacterial activity of Lawsonia inermis seeds. L. inermis seed powder was extracted using hexane, dichloromethane, ethanol and water. The 4 fractions were quantified for polyphenols, flavonoids and tannins contents. The antioxidant capacity was assessed using 4 methods: 2,2?-Diphenyl- picrylhydrazyl hydrate (DPPH) , 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic) (ABTS), Ferric Reducing Antioxidant Power (FRAP), and Phosphomolybdate assay (PMA). Antimicrobial activity was determined by agar diffusion against Staphylococcus aureus, Bacillus cereus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The liquid dilution method was used to determine the minimum inhibitory concentration, minimum bactericidal concentration and minimum fungicidal concentration. The highest content of polyphenols and tannins was obtained in the ethanolic fraction, with values of 594.01±6.07 mg GAE/g dw and 242.4±1.6 mg CE/g dw respectively. The ethanolic fraction exhibited significant scavenging of free radicals (DPPH) with an IC50 of 3.06±0.01 µg/mL, lower than the IC50 of the standard quercetin (5.30±0.02 µg/mL). Moreover, the ethanolic fraction showed antimicrobial activity against S. aureus, B. cereus and E. faecalis strains, with zone of inhibition values of 15.5±0.7, 14.0±0.0 and 14.5±0.7 mm respectively. The minimum inhibitory concentration of the ethanolic fraction was found to be 1.563 mg/mL against both S. aureus and B. cereus strains, while the minimum bactericidal concentration was 3.125 mg/mL against B. cereus and 6.25 mg/mL against E. faecalis strains. L. inermis seeds exhibit significant antioxidant capacity and demonstrate antimicrobial activity. These findings indicate the potential of L. inermis seeds as a valuable source of bioactive compounds with antioxidant and antimicrobial properties.

Phytochemicals properties and antimicrobial activities of Carica papaya and Balanites aegyptiaca’ seeds aqueous infusion and decoction extracts

Carica papaya and Balanites aegyptiaca seeds extract have several uses, from food to traditional medicine. This study aims to determine their phytochemicals compound and appreciate their antimicrobial properties against clinical strains. Polyphenolic content was determined with standard methods while DPPH radical scavenging activity is used to appreciate the antioxidant properties, using ascorbic acid as standard. Antimicrobial activities were performed according to the inhibition diameters, using agar diffusion method. Minimum inhibitory concentrations and Minimum fungicidal concentrations were established based on broth dilution methods. The extraction yield is ranged between 6.3 and 20.3%. The results shown polyphenolic content varying from 1256.13±21.09 to 2315.81±53.52 µg GAE/100 mL for Carica papaya and from 303.98±0.93 to 948.06±3.23 µg GAE/100 mL for Balanites aegyptiaca while the antioxidant activity is ranged between 31.76±2.35 and 58.47±4.24 µg AEAC/100 mL for Carica papaya and from 29.80±0.32 to 78.44±2.81 µg AEAC/100 mL for Balanites aegyptiaca. Infusion’s extracts always have the highest values of polyphenols content and antioxidant activity than decoction extracts, be it papaya or balanites seeds. Neitheir flavonoids content nor antibacterial activity was found. The inhibition diameter varies from 14.5±0.5 to 40.5±0.5 mm for papaya seeds extracts and from 16.5±0.5 to 21.0±0.5 mm for balanites seeds extracts without a significative effect of mixture for both extracts. MICs vary from 11.71±0.1 to 75.0±0.0 mg. mL-1 for Carica papaya seeds extracts with 11.72±0.2 to 37.30±0.2 mg. mL-1 and 18.74±0.1 to 100±0.0 mg.mL-1 for Balanites aegyptiaca. Mixture extracts presented intermediaire values. Decoction extracts have lowest MICs values and infusion reveals itself to be the sweetable aqueous extraction techniques but the appropriate infusion time need to be highlight.
 Keywords: Carica papaya, Balanites aegyptiaca, Seeds extracts, Phytochemical, antifungal

Antifungal activity and potential mechanism of action of Huangqin decoction against Trichophyton rubrum.

Introduction. Trichophyton rubrum is a major causative agent of superficial dermatomycoses such as onychomycosis and tinea pedis. Huangqin decoction (HQD), as a classical traditional Chinese medicine formula, was found to inhibit the growth of common clinical dermatophytes such as T. rubrum in our previous drug susceptibility experiments.Hypothesis/Gap Statement. The antifungal activity and potential mechanism of HQD against T. rubrum have not yet been investigated.Aim. The aim of this study was to investigate the antifungal activity and explore the potential mechanism of action of HQD against T. rubrum.Methodology. The present study was performed to evaluate the antifungal activity of HQD against T. rubrum by determination of minimal inhibitory concentrations (MICs), minimal fungicidal concentrations (MFCs), mycelial growth, biomass, spore germination and structural damage, and explore its preliminary anti-dermatophyte mechanisms by sorbitol and ergosterol assay, HPLC-based ergosterol test, enzyme-linked immunosorbent assay and mitochondrial enzyme activity test.Results. HQD was able to inhibit the growth of T. rubrum significantly, with an MIC of 3.125 mg ml-1 and an MFC of 12.5 mg ml-1. It also significantly inhibited the hyphal growth, conidia germination and biomass growth of T. rubrum in a dose-dependent manner, and induced structural damage in different degrees for T. rubrum cells. HQD showed no effect on cell wall integrity, but was able to damage the cell membrane of T. rubrum by interfering with ergosterol biosynthesis, involving the reduction of squalene epoxidase (SE) and sterol 14α-demethylase P450 (CYP51) activities, and also affect the malate dehydrogenase (MDH), succinate dehydrogenase (SDH) and ATPase activities of mitochondria.Conclusion. These results revealed that HQD had significant anti-dermatophyte activity, which was associated with destroying the cell membrane and affecting the enzyme activities of mitochondria.