In vitro and in vivo antifungal activity of buparvaquone against Sporothrix brasiliensis

Sporotrichosis has become an important zoonosis in Brazil and Sporothrix brasiliensis is the primary species transmitted by cats. Improvement of animal treatment will help control and limit the spread and geographic expansion of sporotrichosis. Accordingly, buparvaquone, an antiprotozoal hydroxynaphthoquinone agent marketed as Butalex®, was evaluated in vitro and in vivo against feline-borne isolates of S. brasiliensis . Buparvaquone inhibited in vitro fungal growth at concentrations 4-fold lower than itraconazole (the first-choice antifungal used for sporotrichosis) and was 408 times more selective for S. brasiliensis than mammalian cells. Yeasts treated with a subinhibitory concentration of buparvaquone exhibited mitochondrial dysfunction, ROS and neutral lipid accumulation, and impaired plasma membranes. Also, scanning electron microscopy images revealed buparvaquone altered cell wall integrity and induced cell disruption. I n vivo experiments in a Galleria mellonella model revealed that buparvaquone (single dose of 5 mg/kg) is more effective than itraconazole against infections with S. brasiliensis yeasts. Combined, our results indicate that buparvaquone has a great in vitro and in vivo antifungal activity against S. brasiliensis , revealing the potential application of this drug as an alternative treatment for feline sporotrichosis.

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A Peptide Found in Human Serum, Derived from the C-Terminus of Albumin, Shows Antifungal Activity In Vitro and In Vivo

Microorganisms ◽

10.3390/microorganisms8101627 ◽

2020 ◽

Vol 8 (10) ◽

pp. 1627

Author(s):

Tecla Ciociola ◽

Pier Paolo Zanello ◽

Tiziana D’Adda ◽

Serena Galati ◽

Stefania Conti ◽

...

Keyword(s):

Antifungal Activity ◽

Human Serum ◽

Fungicidal Activity ◽

Galleria Mellonella ◽

Serum Proteins ◽

Morphological Alterations ◽

C Terminus ◽

Different Sources

The growing problem of antimicrobial resistance highlights the need for alternative strategies to combat infections. From this perspective, there is a considerable interest in natural molecules obtained from different sources, which are shown to be active against microorganisms, either alone or in association with conventional drugs. In this paper, peptides with the same sequence of fragments, found in human serum, derived from physiological proteins, were evaluated for their antifungal activity. A 13-residue peptide, representing the 597–609 fragment within the albumin C-terminus, was proved to exert a fungicidal activity in vitro against pathogenic yeasts and a therapeutic effect in vivo in the experimental model of candidal infection in Galleria mellonella. Studies by confocal microscopy and transmission and scanning electron microscopy demonstrated that the peptide penetrates and accumulates in Candida albicans cells, causing gross morphological alterations in cellular structure. These findings add albumin to the group of proteins, which already includes hemoglobin and antibodies, that could give rise to cryptic antimicrobial fragments, and could suggest their role in anti-infective homeostasis. The study of bioactive fragments from serum proteins could open interesting perspectives for the development of new antimicrobial molecules derived by natural sources.

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In Silico Predicted Antifungal Peptides: In Vitro and In Vivo Anti-Candida Activity

Journal of Fungi ◽

10.3390/jof7060439 ◽

2021 ◽

Vol 7 (6) ◽

pp. 439

Author(s):

Tecla Ciociola ◽

Walter Magliani ◽

Tiziano De Simone ◽

Thelma A. Pertinhez ◽

Stefania Conti ◽

...

Keyword(s):

In Silico ◽

Mammalian Cells ◽

Galleria Mellonella ◽

Ex Vivo ◽

Consensus Sequence ◽

In Silico Analysis ◽

Significant Activity ◽

Synthetic Antibody

It has been previously demonstrated that synthetic antibody-derived peptides could exert a significant activity in vitro, ex vivo, and/or in vivo against microorganisms and viruses, as well as immunomodulatory effects through the activation of immune cells. Based on the sequence of previously described antibody-derived peptides with recognized antifungal activity, an in silico analysis was conducted to identify novel antifungal candidates. The present study analyzed the candidacidal and structural properties of in silico designed peptides (ISDPs) derived by amino acid substitutions of the parent peptide KKVTMTCSAS. ISDPs proved to be more active in vitro than the parent peptide and all proved to be therapeutic in Galleria mellonella candidal infection, without showing toxic effects on mammalian cells. ISDPs were studied by circular dichroism spectroscopy, demonstrating different structural organization. These results allowed to validate a consensus sequence for the parent peptide KKVTMTCSAS that may be useful in the development of novel antimicrobial molecules.

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Synergistic Efficacy of Aedes aegypti Antimicrobial Peptide Cecropin A2 and Tetracycline against Pseudomonas aeruginosa

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00686-17 ◽

2017 ◽

Vol 61 (7) ◽

Cited By ~ 31

Author(s):

Zhaojun Zheng ◽

Nagendran Tharmalingam ◽

Qingzhong Liu ◽

Elamparithi Jayamani ◽

Wooseong Kim ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Aedes Aegypti ◽

Mammalian Cells ◽

Galleria Mellonella ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Synergistic Activity ◽

Content Type

ABSTRACT The increasing prevalence of antibiotic resistance has created an urgent need for alternative drugs with new mechanisms of action. Antimicrobial peptides (AMPs) are promising candidates that could address the spread of multidrug-resistant bacteria, either alone or in combination with conventional antibiotics. We studied the antimicrobial efficacy and bactericidal mechanism of cecropin A2, a 36-residue α-helical cationic peptide derived from Aedes aegypti cecropin A, focusing on the common pathogen Pseudomonas aeruginosa. The peptide showed little hemolytic activity and toxicity toward mammalian cells, and the MICs against most clinical P. aeruginosa isolates were 32 to 64 μg/ml, and its MICs versus other Gram-negative bacteria were 2 to 32 μg/ml. Importantly, cecropin A2 demonstrated synergistic activity against P. aeruginosa when combined with tetracycline, reducing the MICs of both agents by 8-fold. The combination was also effective in vivo in the P. aeruginosa/Galleria mellonella model (P < 0.001). We found that cecropin A2 bound to P. aeruginosa lipopolysaccharides, permeabilized the membrane, and interacted with the bacterial genomic DNA, thus facilitating the translocation of tetracycline into the cytoplasm. In summary, the combination of cecropin A2 and tetracycline demonstrated synergistic antibacterial activity against P. aeruginosa in vitro and in vivo, offering an alternative approach for the treatment of P. aeruginosa infections.

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Evaluation of Antifungal Activity of Chitosan Nanoparticles Against Fusarium Solani Phytopathogenic Fungi in in Vitro and in Vivo Assays

10.21203/rs.3.rs-362614/v1 ◽

2021 ◽

Author(s):

Pamela R. Avila ◽

Graciela Juez Castillo ◽

Carel E. Carvajal

Keyword(s):

Antifungal Activity ◽

Fusarium Solani ◽

Plant Root ◽

Chitosan Nanoparticles ◽

Fungal Growth ◽

Fungal Cell ◽

Tomato Root ◽

Root Cells

Abstract Fungal diseases are a current problem in agriculture causing significant losses in several crops whereby its prevention and treatment is of utmost importance. The Chitosan nanoparticles (ChNPs) were evaluated for their antimicrobial activity against the phytopathogen Fusarium solani. The chitosan concentration in nanoparticles that showed antifungal activity was 2.0 µg/mL. ChNPs showed to be a potential antifungal candidate with applications in phytosanitary control. Transmission electron microscopy (TEM) results showed damage to the fungal cell wall and membrane caused by the nanoparticles interaction with these structures affecting fungal growth and development in in vitro as in in vivo assay where microscopy demonstrated the internalization of nanoparticles aggregates within plant root cells cytoplasm up to 45 days. Therefore ChNPs nanoparticles could be an alternative method for diseases caused by Fusarium solani instead of chemical fungicides commonly used for treating tomato root rot.

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In vitro and in vivo activity of chelerythrine against Candida albicans and underlying mechanisms

Future Microbiology ◽

10.2217/fmb-2019-0178 ◽

2019 ◽

Vol 14 (18) ◽

pp. 1545-1557 ◽

Cited By ~ 4

Author(s):

Ying Gong ◽

Siwen Li ◽

Weixin Wang ◽

Yiman Li ◽

Wenli Ma ◽

...

Keyword(s):

Candida Albicans ◽

Antifungal Activity ◽

Calcium Concentration ◽

Galleria Mellonella ◽

Hyphal Growth ◽

Drug Transporter ◽

Antifungal Effect ◽

Underlying Mechanisms

Aim: To evaluate whether chelerythrine (CHT) exhibited antifungal activity against Candida albicans in vitro and in vivo and to explore the underlying mechanisms. Materials & methods: Broth microdilution assay and Galleria mellonella model were used to evaluate the antifungal effect in vitro and in vivo, respectively. Mechanism studies were investigated by morphogenesis observation, Fluo-3/AM, DCFH-DA and rhodamine6G assay, respectively. Results: CHT exhibited antifungal activity against C. albicans and preformed biofilms with minimum inhibitory concentrations ranged from 2 to 16 μg/ml. Besides, CHT protected G. mellonella larvae infected by C. albicans. Mechanisms studies revealed that CHT inhibited hyphal growth, increased intracellular calcium concentration, induced accumulation of reactive oxygen species and inhibited drug transporter activity. Conclusion: CHT exhibited antifungal activity against C. albicans.

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In Vitro and In Vivo Effect of Peptides Derived from 14-3-3 Paracoccidioides spp. Protein

Journal of Fungi ◽

10.3390/jof7010052 ◽

2021 ◽

Vol 7 (1) ◽

pp. 52

Author(s):

Liliana Scorzoni ◽

Ana Carolina Alves de Paula e Silva ◽

Haroldo Cesar de Oliveira ◽

Claudia Tavares dos Santos ◽

Junya de Lacorte Singulani ◽

...

Keyword(s):

Antifungal Activity ◽

Galleria Mellonella ◽

Therapeutic Potential ◽

Paracoccidioides Brasiliensis ◽

Therapeutic Vaccine ◽

Prolonged Treatment ◽

New Therapeutic Approaches ◽

Paracoccidioides Spp

Background: Paracoccidioidomycosis (PCM) is a chronic disease that causes sequelae and requires prolonged treatment; therefore, new therapeutic approaches are necessary. In view of this, three peptides from Paracoccidioides brasiliensis 14-3-3 protein were selected based on its immunogenicity and therapeutic potential. Methods: The in vitro antifungal activity and cytotoxicity of the 14-3-3 peptides were evaluated. The influence of the peptides in immunological and survival aspects was evaluated in vivo, using Galleria mellonella and the expression of antimicrobial peptide genes in Caenorhabditis elegans. Results: None of the peptides were toxic to HaCaT (skin keratinocyte), MRC-5 (lung fibroblast), and A549 (pneumocyte) cell lines, and only P1 exhibited antifungal activity against Paracoccidioides spp. The peptides could induce an immune response in G. mellonella. Moreover, the peptides caused a delay in the death of Paracoccidioides spp. infected larvae. Regarding C. elegans, the three peptides were able to increase the expression of the antimicrobial peptides. These peptides had essential effects on different aspects of Paracoccidioides spp. infection showing potential for a therapeutic vaccine. Future studies using mammalian methods are necessary to validate our findings.

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Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L.) Skeels leaves extract

Boletin Latinoamericano y del Caribe de Plantas Medicinales y Aromaticas ◽

10.37360/blacpma.21.20.5.40 ◽

2021 ◽

Vol 20 (5) ◽

pp. 536-555

Author(s):

Ernani Canuto Figueiredo Junior ◽

◽

Yuri Wanderley Cavalcanti ◽

Andressa Brito Lira ◽

Hilzeth de Luna Freire Pessoa ◽

...

Keyword(s):

Antioxidant Activity ◽

Antifungal Activity ◽

Galleria Mellonella ◽

In Vitro Assays ◽

Syzygium Cumini ◽

Candida Spp ◽

In Vivo Toxicity ◽

Phytochemical Composition

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125 μg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivo toxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

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Identification of Volatile Organic Compounds in Extremophilic Bacteria and Their Effective Use in Biocontrol of Postharvest Fungal Phytopathogens

Frontiers in Microbiology ◽

10.3389/fmicb.2021.773092 ◽

2021 ◽

Vol 12 ◽

Author(s):

Laura Toral ◽

Miguel Rodríguez ◽

Fernando Martínez-Checa ◽

Alfredo Montaño ◽

Amparo Cortés-Delgado ◽

...

Keyword(s):

Antifungal Activity ◽

Volatile Organic Compounds ◽

Organic Compounds ◽

Fruits And Vegetables ◽

Fungal Growth ◽

Postharvest Diseases ◽

Transmission Electron Microscopy Micrographs ◽

Volatile Organic

Phytopathogenic fungal growth in postharvest fruits and vegetables is responsible for 20–25% of production losses. Volatile organic compounds (VOCs) have been gaining importance in the food industry as a safe and ecofriendly alternative to pesticides for combating these phytopathogenic fungi. In this study, we analysed the ability of some VOCs produced by strains of the genera Bacillus, Peribacillus, Pseudomonas, Psychrobacillus and Staphylococcus to inhibit the growth of Alternaria alternata, Botrytis cinerea, Fusarium oxysporum, Fusarium solani, Monilinia fructicola, Monilinia laxa and Sclerotinia sclerotiorum, in vitro and in vivo. We analysed bacterial VOCs by using GC/MS and 87 volatile compounds were identified, in particular acetoin, acetic acid, 2,3-butanediol, isopentanol, dimethyl disulphide and isopentyl isobutanoate. In vitro growth inhibition assays and in vivo experiments using cherry fruits showed that the best producers of VOCs, Bacillus atrophaeus L193, Bacillus velezensis XT1 and Psychrobacillus vulpis Z8, exhibited the highest antifungal activity against B. cinerea, M. fructicola and M. laxa, which highlights the potential of these strains to control postharvest diseases. Transmission electron microscopy micrographs of bacterial VOC-treated fungi clearly showed antifungal activity which led to an intense degeneration of cellular components of mycelium and cell death.

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Synthesis, Biological Evaluation, and Structure–Activity Relationships of 4-Aminopiperidines as Novel Antifungal Agents Targeting Ergosterol Biosynthesis

Molecules ◽

10.3390/molecules26237208 ◽

2021 ◽

Vol 26 (23) ◽

pp. 7208

Author(s):

Jürgen Krauß ◽

Christoph Müller ◽

Monika Klimt ◽

Leandro Jorquera Valero ◽

José Francisco Martínez ◽

...

Keyword(s):

Antifungal Activity ◽

Galleria Mellonella ◽

Cholesterol Biosynthesis ◽

Biological Evaluation ◽

Ergosterol Biosynthesis ◽

In Vivo Model ◽

Candida Spp ◽

Mcf10a Cells

The aliphatic heterocycles piperidine and morpholine are core structures of well-known antifungals such as fenpropidin and fenpropimorph, commonly used as agrofungicides, and the related morpholine amorolfine is approved for the treatment of dermal mycoses in humans. Inspired by these lead structures, we describe here the synthesis and biological evaluation of 4-aminopiperidines as a novel chemotype of antifungals with remarkable antifungal activity. A library of more than 30 4-aminopiperidines was synthesized, starting from N-substituted 4-piperidone derivatives by reductive amination with appropriate amines using sodium triacetoxyborohydride. Antifungal activity was determined on the model strain Yarrowia lipolytica, and some compounds showed interesting growth-inhibiting activity. These compounds were tested on 20 clinically relevant fungal isolates (Aspergillus spp., Candida spp., Mucormycetes) by standardized microbroth dilution assays. Two of the six compounds, 1-benzyl-N-dodecylpiperidin-4-amine and N-dodecyl-1-phenethylpiperidin-4-amine, were identified as promising candidates for further development based on their in vitro antifungal activity against Candida spp. and Aspergillus spp. Antifungal activity was determined for 18 Aspergillus spp. and 19 Candida spp., and their impact on ergosterol and cholesterol biosynthesis was determined. Toxicity was determined on HL-60, HUVEC, and MCF10A cells, and in the alternative in vivo model Galleria mellonella. Analysis of sterol patterns after incubation gave valuable insights into the putative molecular mechanism of action, indicating inhibition of the enzymes sterol C14-reductase and sterol C8-isomerase in fungal ergosterol biosynthesis.

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Streptomyces rocheiACTA1551, an Indigenous Greek Isolate Studied as a Potential Biocontrol Agent againstFusarium oxysporumf.sp.lycopersici

BioMed Research International ◽

10.1155/2013/387230 ◽

2013 ◽

Vol 2013 ◽

pp. 1-10 ◽

Cited By ~ 24

Author(s):

Grammatiki S. Kanini ◽

Efstathios A. Katsifas ◽

Alexandros L. Savvides ◽

Amalia D. Karagouni

Keyword(s):

Antifungal Activity ◽

Biocontrol Agent ◽

Fungal Growth ◽

Medium Composition ◽

Phytopathogenic Fungus ◽

Antagonistic Action ◽

16S Rdna Sequence ◽

Potential Biocontrol Agent

Many studies have shown that several Greek ecosystems inhabit very interesting bacteria with biotechnological properties. ThereforeStreptomycesisolates from diverse Greek habitats were selected for their antifungal activity against the common phytopathogenic fungusFusarium oxysporum. The isolate encoded ACTA1551, member ofStreptomycesgenus, could strongly suppress the fungal growth when examined in antagonistic bioassaysin vitro. The isolate was found phylogenetically relative toStreptomyces rocheiafter analyzing its 16S rDNA sequence. The influence of different environmental conditions, such as medium composition, temperature, and pH on the expression of the antifungal activity was thoroughly examined.Streptomyces rocheiACTA1551 was able to protect tomato seeds fromF. oxysporuminfectionin vivowhile it was shown to promote the growth of tomato plants when the pathogen was absent. In an initial effort towards the elucidation of the biochemical and physiological nature of ACTA1551 antifungal activity, extracts from solid streptomycete cultures under antagonistic or/and not antagonistic conditions were concentrated and fractionated. The metabolites involved in the antagonistic action of the isolate showed to be more than one and produced independently of the presence of the pathogen. The above observations could support the application ofStreptomyces rocheiACTA1551 as biocontrol agent againstF. oxysporum.

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