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PLoS Neglected Tropical Diseases Sep 2023Filamentous fungi of the genus Madurella are the primary causative agents of mycetoma, a disease observed in tropical and subtropical regions. Since early diagnostics...
BACKGROUND
Filamentous fungi of the genus Madurella are the primary causative agents of mycetoma, a disease observed in tropical and subtropical regions. Since early diagnostics based on a morphological approach are difficult and have many shortcomings, a molecular diagnostic method suitable for rural settings is required. In this study, we developed the loop-mediated isothermal amplification (LAMP) method to present a foundational technique of the diagnosis of Madurella spp. (M. mycetomatis, M. pseudomycetomatis, M. tropicana, and M. fahalii), the common causative organisms of eumycetoma.
PRINCIPAL FINDINGS
We successfully designed a primer pair targeting the rDNAs of three Madurella spp. excluding M. fahalii, and detected up to 100 fg of genomic DNA extracted from isolates of M. mycetomatis and 1 pg of M. pseudomycetomatis and M. tropicana, within one hour. Second, a primer pair specific to M. mycetomatis, the most common causative species, or M. fahalii, a drug-resistant species, was constructed, and the detection limit of both primer pairs was 1 pg. The designed primers accurately distinguished 16 strains of the genus Madurella from various fungal species known to cause mycetomas.
CONCLUSION
In summary, we established the first model of a LAMP detection method that rapidly and sensitively detects and identifies Madurella isolates for clinical diagnostics. Moreover, the combined designed primer sets could identify mycetoma-causing strains simultaneously.
Topics: Mycetoma; Madurella; Nucleic Acid Amplification Techniques
PubMed: 37721946
DOI: 10.1371/journal.pntd.0011644 -
Medical Mycology Feb 2012Eumycetoma, a chronic granulomatous disease characterized by a subcutaneous mass, multiple sinuses and purulent discharge containing grains, remains difficult to...
Eumycetoma, a chronic granulomatous disease characterized by a subcutaneous mass, multiple sinuses and purulent discharge containing grains, remains difficult to diagnose and treat. Madurella mycetomatis is the most common causative agent of eumycetoma. Using a serum pool from patients with active mycetoma, we screened a M. mycetomatis-specific λgt11 cDNA library which was shown to contain 8% of cDNA inserts encoding proteins involved in glycolysis. Two of these enzymes, fructose-bisphosphate aldolase (FBA) and pyruvate kinase (PK), were produced in vitro and their antigenicity was studied with bead-based flow cytometry. It appeared that both FBA and PK IgG antibodies were present in eumycetoma patient sera. However, only FBA antibody levels were found to be significantly higher in eumycetoma patient sera when compared to healthy Sudanese controls. Furthermore, FBA and PK were also found to be expressed on the hyphae present in the mycetoma grain. In conclusion, this study presents two new antigenic proteins of M. mycetomatis next to the translationally controlled tumour protein (TCTP): the glycolytic enzymes FBA and PK. These antigens might be useful as vaccine-candidates in the prevention of mycetoma.
Topics: Antibodies, Fungal; Fructose-Bisphosphate Aldolase; Fungal Proteins; Histocytochemistry; Humans; Madurella; Male; Mycetoma; Phylogeny; Pyruvate Kinase; Recombinant Fusion Proteins; Spectrometry, Fluorescence; Statistics, Nonparametric; Sudan; Tumor Protein, Translationally-Controlled 1
PubMed: 21728753
DOI: 10.3109/13693786.2011.593005 -
Transactions of the Royal Society of... Mar 2011This prospective study aimed to determine the safety and efficacy of itraconazole for the treatment of patients with mycetoma due to Madurella mycetomatis. The study...
This prospective study aimed to determine the safety and efficacy of itraconazole for the treatment of patients with mycetoma due to Madurella mycetomatis. The study consisted of 13 patients with confirmed disease; all were treated with oral itraconazole in a dose of 400mg daily for three months after which the dose was reduced to 200mg daily for nine months. All patients showed good clinical response to 400mg itraconazole daily, but when the dose was reduced to 200mg daily, the clinical response was gradual and slow. Post-treatment surgical exploration showed that, in all patients, the lesions were well localized, encapsulated with thick capsule and they were easily removed surgically. In all these lesions, grains colonies were encountered and they were viable on culture. Post-operative biopsies showed no significant changes in the morphology of the grains. A constant finding was the presence of between 5-7 grains in a single cavity walled by fibrous tissue. The reaction surrounding the grains was a Type I tissue reaction characterized by a neutrophil zone around grains. Patients were followed up post-operatively for variable periods (range 18-36 months) and only one patient had recurrence. Initial pre-operative treatment with itraconazole may be recommended for eumycetoma patients to enhance lesions encapsulation and localization which can facilitate wide local excision to avoid unnecessary massive mutilating surgery and recurrence.
Topics: Adolescent; Adult; Antifungal Agents; Female; Humans; Itraconazole; Madurella; Male; Mycetoma; Prospective Studies; Sudan; Treatment Outcome; Young Adult
PubMed: 21247608
DOI: 10.1016/j.trstmh.2010.11.008 -
Microbes and Infection Jul 2007One of the hallmarks of eumycetoma is the formation of fungal grains, which are secreted by multiple sinuses in infected tissues. Madurella mycetomatis grains are black....
One of the hallmarks of eumycetoma is the formation of fungal grains, which are secreted by multiple sinuses in infected tissues. Madurella mycetomatis grains are black. This black colour was shown to be due to the presence of melanin. Melanin can be produced through various biochemical pathways. It appeared that M. mycetomatis melanisation could be blocked by inhibitors of the pyo- and dihydroxynaphthalene (DHN)-melanin pathways but not by inhibitors of the dihydroxyphenylalanine (L-DOPA)-melanin pathway. Melanin isolated from M. mycetomatis cells provides in vitro protection against the killing effects of the oxidant permanganate and several antifungals. When melanin was added to the culture medium, MICs were found to be 16-fold elevated in the case of itraconazole and 32-fold for ketoconazole. MICs for amphotericin B, fluconazole and voriconazole were not affected. Since itraconazole and ketoconazole are the main antifungal agents used to treat mycetoma, the clinical relevance of the in vitro rise in MIC should be studied further.
Topics: Antifungal Agents; Drug Resistance, Multiple, Fungal; Humans; Itraconazole; Ketoconazole; Madurella; Melanins; Mycetoma; Oxidants
PubMed: 17644456
DOI: 10.1016/j.micinf.2007.05.015 -
Molecules (Basel, Switzerland) Apr 2024In the search for new bioactive agents against the infectious pathogen responsible for the neglected tropical disease (NTD) mycetoma, we tested a collection of 27...
In the search for new bioactive agents against the infectious pathogen responsible for the neglected tropical disease (NTD) mycetoma, we tested a collection of 27 essential oils (EOs) in vitro against , the primary pathogen responsible for the fungal form of mycetoma, termed eumycetoma. Among this series, the EO of (Santalaceae), i.e., East Indian sandalwood oil, stood out prominently with the most potent inhibition in vitro. We, therefore, directed our research toward 15 EOs of species of different geographical origins, along with two samples of EOs from other plant species often commercialized as "sandalwood oils". Most of these EOs displayed similar strong activity against in vitro. All tested oils were thoroughly analyzed by GC-QTOF MS and most of their constituents were identified. Separation of the sandalwood oil into the fractions of sesquiterpene hydrocarbons and alcohols showed that its activity is associated with the sesquiterpene alcohols. The major constituents, the sesquiterpene alcohols ()-α- and ()-β-santalol were isolated from the oil by column chromatography on AgNO-coated silica. They were tested as isolated compounds against the fungus, and ()-α-santalol was about two times more active than the β-isomer.
Topics: Madurella; Plant Oils; Oils, Volatile; Mycetoma; Santalum; Sesquiterpenes; Antifungal Agents; Microbial Sensitivity Tests
PubMed: 38675665
DOI: 10.3390/molecules29081846 -
International Journal of Molecular... Jun 2024Eumycetoma is a neglected tropical disease (NTD) characterized by subcutaneous lesions and the formation of grains. Attempts to treat eumycetoma involve a combination of...
OBJECTIVES
Eumycetoma is a neglected tropical disease (NTD) characterized by subcutaneous lesions and the formation of grains. Attempts to treat eumycetoma involve a combination of antifungal treatment and surgery, although the outcome is frequently disappointing. Therefore, there is a need to identify novel antifungal drugs to treat eumycetoma. In this respect, Medicines for Malaria Venture (MMV) has assembled libraries of compounds for researchers to use in drug discovery research against NTD. Therefore, we screened two MMVOpen compound libraries to identify novel leads for eumycetoma.
METHODS
A total of 400 compounds from the COVID Box and the Global Health Priority Box were screened in vitro at 100 µM and 25 µM against the most common causative agents of eumycetoma, namely and and the resulting IC and MIC values were obtained. Compounds with an IC < 8 µM were identified for possible in vivo efficacy studies using an grain model in larvae.
RESULTS
Out of the 400 compounds, 22 were able to inhibit both and growth at 100 µM and 25 µM, with compounds MMV1593278, MMV020335, and MMV1804559 being selected for in vivo testing. Of these three, only the pyrazolopyrimidine derivative MMV1804559 was able to prolong the survival of -infected larvae. Furthermore, the grains in MMV1804559-treated larvae were significantly smaller compared to the PBS-treated group.
CONCLUSION
MMV1804559 shows promising in vitro and in vivo activity against .
Topics: Madurella; Mycetoma; Antifungal Agents; Animals; Microbial Sensitivity Tests; Larva; Humans
PubMed: 38892422
DOI: 10.3390/ijms25116227 -
The Pan African Medical Journal 2013
Topics: Diagnostic Imaging; Endemic Diseases; Foot Dermatoses; Humans; Male; Middle Aged; Morocco; Mycetoma
PubMed: 23504196
DOI: 10.11604/pamj.2013.14.24.2381 -
Journal of the American Academy of... Sep 1994Several subcutaneous and deep-seated mycoses are either observed more frequently in the tropical areas or are restricted to certain regions within the tropics. These... (Review)
Review
Several subcutaneous and deep-seated mycoses are either observed more frequently in the tropical areas or are restricted to certain regions within the tropics. These mycoses include sporotichosis, chromoblastomycosis, entomophthoromycosis, eumycetoma, lobomycosis, and paracoccidioidomycosis. In sporotrichosis and paracoccidioidomycosis, therapy often results in either complete resolution or marked improvement. For decades sporotrichosis has been treated successfully with potassium iodide, but recently the triazole compounds, especially itraconazole, have proved effective and free of major side effects. The usual therapy for paracoccidioidomycosis is sulfonamides or amphotericin B; the former requires prolonged treatment, whereas the latter causes a significant degree of toxicity. Various azole derivatives (ketoconazole, fluconazole, saperconazole, and itraconazole) allow shorter treatment courses, can be given orally, and are more effective. Presently, itraconazole is the drug of choice. Chromoblastomycosis is a difficult condition to treat, especially if it is caused by Fonsecaea pedrosoi. Several therapeutic approaches have been used, including heat, surgery, cryotherapy, thiabendazole, amphotericin B combined with flucytosine, and azole derivatives, but their success has been modest. A 65% response rate has been obtained with itraconazole given for periods of 6 to 19 months; in limited trials, saperconazole appears to be more effective and requires shorter treatment courses. Only a few patients with eumycetoma respond to therapy; 70% of patients with Madurella mycetomatis respond to prolonged treatment with ketoconazole. Griseofulvin has been tried in nonresponders with partial success. Limited data in patients with Fusarium species eumycetoma indicate good responses to itraconazole. Eumycetoma caused by Pseudallescheria boydii or Acremonium species has been refractory to therapy. Therapy of entomophthoromycosis is also difficult because the diagnosis is usually established late and not all patients respond to therapy; this situation applies to infection caused by either Basidiobolus haptosporus or Conidiobolus coronatus. Although there is no consensus, African physicians prefer to use potassium iodide or trimethoprim-sulfamethoxazole. Isolated reports indicate that the azole derivatives, including the triazoles, may be effective. As for lobomycosis, all attempts at medical treatment have failed. Surgery is successful only when the lesion is small and can be fully resected; repeated cryotherapy appears to be more successful.
Topics: Antifungal Agents; Blastomycosis; Chromoblastomycosis; Combined Modality Therapy; Drug Therapy, Combination; Humans; Mucormycosis; Mycetoma; Mycoses; Paracoccidioidomycosis; Physical Therapy Modalities; Potassium Iodide; Sporotrichosis; Tropical Climate
PubMed: 8077517
DOI: 10.1016/s0190-9622(08)81277-7 -
Tropical Medicine & International... Jun 2017To determine whether combination therapy would improve therapeutic outcome in eumycetoma caused by Madurella mycetomatis.
OBJECTIVE
To determine whether combination therapy would improve therapeutic outcome in eumycetoma caused by Madurella mycetomatis.
METHODS
Survival, colony-forming units (CFU), melanisation and histopathology in M. mycetomatis-infected Galleria mellonella larvae treated with amphotericin B, itraconazole, terbinafine or combinations thereof were determined.
RESULTS
Compared to larvae treated with 5% glucose, enhanced survival was obtained when M. mycetomatis-infected larvae were treated with amphotericin B, but not when they were treated with itraconazole or terbinafine. Combination therapy did not increase survival compared to 5% glucose-treated larvae, itraconazole-treated larvae or terbinafine-treated larvae. Compared to amphotericin B monotreatment, a significant decrease in survival was noted when this therapy was combined with either itraconazole or terbinafine. CFU, melanisation and histopathology did not differ between monotherapy, combination therapy or 5% glucose-treated larvae.
CONCLUSIONS
Combining different classes of antifungal agents did not enhance the survival of M. mycetomatis-infected G. mellonella larvae. Instead of improving the therapeutic outcome, combining either itraconazole or terbinafine with amphotericin B resulted in significantly lower survival rates of infected larvae than amphotericin B monotherapy. This experimental study does not provide support for the use of combined amphotericin B and itraconazole, combined itraconazole and terbinafine or combined terbinafine and amphotericin B and should be confirmed in other animal models.
Topics: Amphotericin B; Animals; Antifungal Agents; Disease Models, Animal; Drug Therapy, Combination; Itraconazole; Larva; Madurella; Moths; Mycetoma; Naphthalenes; Terbinafine
PubMed: 28342219
DOI: 10.1111/tmi.12871 -
Mycoses Jan 2024(1,3)-β-D-glucan is a panfungal biomarker secreted by many fungi, including Madurella mycetomatis, the main causative agent of eumycetoma. Previously we demonstrated...
INTRODUCTION
(1,3)-β-D-glucan is a panfungal biomarker secreted by many fungi, including Madurella mycetomatis, the main causative agent of eumycetoma. Previously we demonstrated that (1,3)-β-D-glucan was present in serum of patients with eumycetoma. However, the use of (1,3)-β-D-glucan to monitor treatment responses in patients with eumycetoma has not been evaluated.
MATERIALS AND METHODS
In this study, we measured (1,3)-β-D-glucan concentrations in serum with the WAKO (1,3)-β-D-glucan assay in 104 patients with eumycetoma treated with either 400 mg itraconazole daily, or 200 mg or 300 mg fosravuconazole weekly. Serial serum (1,3)-β-D-glucan concentrations were measured at seven different timepoints. Any correlation between initial and final (1,3)-β-D-glucan concentrations and clinical outcome was evaluated.
RESULTS
The concentration of (1,3)-β-D-glucan was obtained in a total of 654 serum samples. Before treatment, the average (1,3)-β-D-glucan concentration was 22.86 pg/mL. During the first 6 months of treatment, this concentration remained stable. (1,3)-β-D-glucan concentrations significantly dropped after surgery to 8.56 pg/mL. After treatment was stopped, there was clinical evidence of recurrence in 18 patients. Seven of these 18 patients had a (1,3)-β-D-glucan concentration above the 5.5 pg/mL cut-off value for positivity, while in the remaining 11 patients, (1,3)-β-D-glucan concentrations were below the cut-off value. This resulted in a sensitivity of 38.9% and specificity of 75.0%. A correlation between lesion size and (1,3)-β-D-glucan concentration was noted.
CONCLUSION
Although in general (1,3)-β-D-glucan concentrations can be measured in the serum of patients with eumycetoma during treatment, a sharp decrease in β-glucan concentration was only noted after surgery and not during or after antimicrobial treatment. (1,3)-β-D-glucan concentrations were not predictive for recurrence and seem to have no value in determining treatment response to azoles in patients with eumycetoma.
Topics: Humans; Madurella; Glucans; Azoles; Mycetoma; Proteoglycans
PubMed: 37872649
DOI: 10.1111/myc.13664