RESUMEN
Wheat leaf blight caused by Bipolaris sorokiniana is a widespread fungal disease that poses a serious risk to wheat. Biological control without causing environmental pollution is one of the safest and most effective method to control plant diseases. The antagonistic bacterial strain HeN-7 (identified as Bacillus velezensis) was isolated from tobacco leaves cultivated in Henan province, China. The results of different concentrations of cell-free supernatant (CFS) from HeN-7 culture against B. sorokiniana mycelia showed that 20% HeN-7 CFS (v/v) reached the maximum inhibition rate of 96%. In the potted plants control assay, B. velezensis HeN-7 CFS exhibited remarkable biocontrol activity on the wheat infected with B. sorokiniana, the best pot control efficacy was 65% at 20% CFS. The research on the mechanism of action demonstrated that HeN-7 CFS induced the membrane lipid peroxidation in B. sorokiniana, leading to the disruption of cell membrane integrity and resulting in the leakage of cell contents; in addition, the intracellular mitochondrial membrane potential in mycelium dissipated and reactive oxygen species accumulated, thereby inhibiting the growth of B. sorokiniana. These results indicate that B. velezensis HeN-7 is a promising candidate as a biological control agent against Bipolaris sorokiniana infection.
Asunto(s)
Bacillus , Bipolaris , Nicotiana , Enfermedades de las Plantas , Hojas de la Planta , Bacillus/aislamiento & purificación , Bacillus/metabolismo , Bacillus/fisiología , Hojas de la Planta/microbiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Nicotiana/microbiología , Triticum/microbiología , Antifúngicos/farmacología , Antifúngicos/metabolismo , China , Especies Reactivas de Oxígeno/metabolismo , Micelio/crecimiento & desarrollo , AntibiosisRESUMEN
AIMS: Candida albicans is the most prevalent pathogenic fungus, exhibiting escalating multidrug resistance (MDR). Antimicrobial peptides (AMPs) represent promising candidates for addressing this issue. In this research, five antimicrobial peptides, ACP1 to ACP5 which named ACPs were studied as alternative fungicidal molecules. MAIN METHODS: CD assay was used to analyze the 2D structures, Absorbance method was used to test the antimicrobial activity, haemolytic activity, time-kill kinetics, biofilm inhibition and reduction activity, resistance induction activity and assessment against fluconazole-resistant C. albicans. SEM, TEM, CLSM, flow cytometer and FM were carried out to provide insight into the mechanisms of anti-Candida action. KEY FINDINGS: ACPs possessed an α-helical structure and strong anti-Candida activities, with minimum inhibitory concentrations (MICs) from 3.9 to 15.6 µg/mL. In addition, ACPs did not produce hemolysis at concentrations lower than 10 or 62 × MIC, indicating their low cytotoxicity. Fungicidal kinetics showed that they completely killed C. albicans within 8 h at 2 to 4 × MIC. Notably, ACPs were highly fungicidal against fluconazole-resistant C. albicans and showed low resistance. In addition, they were effective in inhibiting mycelium and biofilm formation. Fluorescence microscopy revealed that while fluconazole had minimal to no inhibitory effect on biofilm-forming cells, ACPs induced apoptosis in all of them. The research on mechanism of action revealed that ACPs disrupted the cell membranes, with ROS increasing and cellular mitochondrial membrane potential decreasing. SIGNIFICANCE: ACPs could be promising candidates for combating fluconazole-resistant C. albicans infections.
Asunto(s)
Antifúngicos , Péptidos Antimicrobianos , Biopelículas , Candida albicans , Fluconazol , Pruebas de Sensibilidad Microbiana , Candida albicans/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/química , Biopelículas/efectos de los fármacos , Péptidos Antimicrobianos/farmacología , Péptidos Antimicrobianos/química , Fluconazol/farmacología , Farmacorresistencia Fúngica/efectos de los fármacos , Hemólisis/efectos de los fármacos , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacosRESUMEN
Efficient and precise design of antimicrobial peptides (AMPs) is of great importance in the field of AMP development. Computing provides opportunities for peptide de novo design. In the present investigation, a new machine learning-based AMP prediction model, AP_Sin, was trained using 1160 AMP sequences and 1160 non-AMP sequences. The results showed that AP_Sin correctly classified 94.61% of AMPs on a comprehensive dataset, outperforming the mainstream and open-source models (Antimicrobial Peptide Scanner vr.2, iAMPpred and AMPlify) and being effective in identifying AMPs. In addition, a peptide sequence generator, AP_Gen, was devised based on the concept of recombining dominant amino acids and dipeptide compositions. After inputting the parameters of the 71 tridecapeptides from antimicrobial peptides database (APD3) into AP_Gen, a tridecapeptide bank consisting of de novo designed 17,496 tridecapeptide sequences were randomly generated, from which 2675 candidate AMP sequences were identified by AP_Sin. Chemical synthesis was performed on 180 randomly selected candidate AMP sequences, of which 18 showed high antimicrobial activities against a wide range of the tested pathogenic microorganisms, and 16 of which had a minimal inhibitory concentration of less than 10 µg/mL against at least one of the tested pathogenic microorganisms. The method established in this research accelerates the discovery of valuable candidate AMPs and provides a novel approach for de novo design of antimicrobial peptides.
Asunto(s)
Péptidos Antimicrobianos , Aprendizaje Automático , Pruebas de Sensibilidad Microbiana , Péptidos Antimicrobianos/farmacología , Péptidos Antimicrobianos/química , Diseño de Fármacos , Secuencia de AminoácidosRESUMEN
Trichophyton mentagrophytes is a zoophilic dermatophyte that can cause dermatophytosis in humans and animals. Antimicrobial peptides (AMPs) are considered as a promising agent to overcome the drug-resistance of T. mentagrophytes. Our findings suggest that cationic antimicrobial peptide (ACP5) not only possesses stronger activity against T. mentagrophytes than fluconazole, but also shows lower toxicity to L929 mouse fibroblast cells than terbinafine. Notably, its resistance development rate after resistance induction was lower than terbinafine. The present study aimed to evaluate the fungicidal mechanism of ACP5 in vitro and its potential to treat dermatophyte infections in vivo. ACP5 at 1 ×MIC completely inhibited T. mentagrophytes spore germination in vitro. ACP5 severely disrupts the mycelial morphology, leading to mycelial rupture. Mechanistically, ACP5 induces excessive ROS production, damaging the integrity of the cell membrane and decreasing the mitochondrial membrane potential, causing irreversible damage in T. mentagrophytes. Furthermore, 1% ACP5 showed similar efficacy to the commercially available drug 1% terbinafine in a guinea pig dermatophytosis model, and the complete eradication of T. mentagrophytes from the skin by ACP5 was verified by tissue section observation. These results indicate that ACP5 is a promising candidate for the development of new agent to combat dermatophyte resistance.
Asunto(s)
Arthrodermataceae , Tiña , Humanos , Ratones , Animales , Cobayas , Terbinafina/farmacología , Terbinafina/uso terapéutico , Trichophyton , Tiña/tratamiento farmacológico , Péptidos Antimicrobianos , Antifúngicos/farmacología , Fosfatasa Ácida Tartratorresistente/farmacologíaRESUMEN
Antimicrobial peptides and their analogues have become substitutes for antibiotics in recent years. The antimicrobial peptide analogue SAMP-A4-C8 (n-octanoic-VRLLRRRI) with high antimicrobial activity was found in our lab. We speculate that it may kill pathogens by some lethal mechanism of action. In the present investigation, the microbicidal activities of SAMP-A4-C8 and its mechanism of action were investigated. The results demonstrated that SAMP-A4-C8 had lethal activities against Staphylococcus aureus and Candida albicans by cell disruption. Based on its microbicidal activities, we believe that it is worth further research for its potential as drug candidate. The results showed that SAMP-A4-C8, with low propensity to induce the resistance of S. aureus and C. albicans, could kill the persister cells of S. aureus and C. albicans, exhibited biofilm forming inhibition activity and preformed biofilm eradication ability against S. aureus and C. albicans, and displayed therapeutic potential on pneumonia in S. aureus-infected mice by reducing lung inflammation. The present study provided a promising drug candidate in the war against multidrug resistance.
Asunto(s)
Antiinfecciosos , Infecciones Estafilocócicas , Animales , Ratones , Staphylococcus aureus , Péptidos Antimicrobianos , Antiinfecciosos/farmacología , Antibacterianos/farmacología , Candida albicans , Biopelículas , Pruebas de Sensibilidad MicrobianaRESUMEN
High-efficiency and low-toxic antimicrobial peptides (AMPs) are supposed to be the future candidates to solve the increasingly prominent problems of Candida albicans infection and drug resistance. Generally, introduction of hydrophobic moieties on AMPs resulted in analogues with remarkably increased activity against pathogens. CGA-N9, an antifungal peptide found in our lab, is a Candida-selective antimicrobial peptide capable of preferentially killing Candida spp. relative to benign microorganisms with low toxicities. We speculate that fatty acid modification could improve the anti-Candida activity of CGA-N9. In the present investigation, a set of CGA-N9 analogues with fatty acid conjugations at N-terminus were obtained. The biological activities of CGA-N9 analogues were determined. The results showed that the n-octanoic acid conjugation of CGA-N9 (CGA-N9-C8) was the optimal CGA-N9 analogue with the highest anti-Candida activity and biosafety; exhibited the strongest biofilm inhibition activity and biofilm eradication ability; and the highest stability against protease hydrolysis in serum. Furthermore, CGA-N9-C8 is less prone to develop resistance for C. albicans in reference with fluconazole; CGA-N9-C8 also exhibited Candidacidal activity to the planktonic cells and the persister cells of C. albicans; reduced C. albicans susceptibility in a systemic candidiasis mouse model. In conclusion, fatty acid modification is an effective method to enhance the antimicrobial activity of CGA-N9, and CGA-N9-C8 is a promising candidate to defend C. albicans infection and resolve C. albicans drug resistance.
Asunto(s)
Péptidos Antimicrobianos , Candida albicans , Animales , Ratones , Ácidos Grasos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Péptidos Catiónicos Antimicrobianos/uso terapéutico , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Antifúngicos/química , Cromogranina A/farmacología , Biopelículas , Pruebas de Sensibilidad MicrobianaRESUMEN
A new remediation method for petroleum-salt contaminated soil was proposed, in which wheat straw was applied to enhance salt leaching and meanwhile block salt upmovement along the soil capillary. It was shown that the existence of petroleum increased the surface hydrophobicity of soil and thus hindered the leaching process once the oil content was above 1.5% (mass fraction). The application of 5% (mass fraction) wheat straw into the soil increased the efficiency of salt leaching from 3% to 25%. The effectiveness of wheat straw layer in inhibiting the salt upmovement along the soil capillary was also proven. Field test of this method was carried out in an area of 6400 m2, in which wheat straw layer of 5 cm was distributed in the depth of 25 cm. After 50 days, 80% of the testing area showed a normal soil electronic conductivity (soil EC <5 mS x cm(-1)) in contrast to 17% before leaching process. The concentrations of Na+ and Cl- were decreased from 1642.5 mg x kg(-1) and 1 301.2 mg x kg(-1) to 499.3 mg x kg(-1) and 433.8 mg x kg(-1), respectively. The remediated land upon the implementation of wheat straw gave a 72% of the regular wheat production obtained from the normal farmland, while the control land without wheat straw gave 12%. These results demonstrated the effectiveness and the high potential of using wheat straw for the salt leaching in remediation of petroleum-salt contaminated soil.