-
PLoS Neglected Tropical Diseases Jan 2020The genus Madurella comprising four species, M. fahalii, M. mycetomatis, M. pseudomycetomatis, and M. tropicana, represents the prevalent cause of eumycetoma worldwide....
The genus Madurella comprising four species, M. fahalii, M. mycetomatis, M. pseudomycetomatis, and M. tropicana, represents the prevalent cause of eumycetoma worldwide. The four species are phenotypically similar and cause an invariable clinical picture, but differ markedly in their susceptibility to antifungal drugs, and epidemiological pattern. Therefore, specific identification is required for optimal management of Madurella infection and to reveal proper epidemiology of the species. In this study, a novel multiplex real-time PCR targeting the four Madurella species was developed and standardized. Evaluation of the assay using reference strains of the target and non-target species resulted in 100% specificity, high analytical reproducibility (R2 values >0.99) and a lowest detection limit of 3 pg target DNA. The accuracy of the real-time PCR was further assessed using biopsies from eumycetoma suspected patients. Unlike culture and DNA sequencing as gold standard diagnostic methods, the real-time PCR yielded accurate diagnosis with specific identification of the causative species in three hours compared to one or two weeks required for culture. The novel method reduces turnaround time as well as labor intensity and high costs associated with current reference methods.
Topics: Biopsy; DNA, Fungal; Humans; Madurella; Mycetoma; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity
PubMed: 31940343
DOI: 10.1371/journal.pntd.0007845 -
Medical Mycology Nov 2023Mycetoma is a neglected tropical disease commonly caused by the fungus Madurella mycetomatis. Standard treatment consists of extensive treatment with itraconazole in...
Mycetoma is a neglected tropical disease commonly caused by the fungus Madurella mycetomatis. Standard treatment consists of extensive treatment with itraconazole in combination with surgical excision of the infected tissue, but has a low success rate. To improve treatment outcomes, novel treatment strategies are needed. Here, we determined the potential of manogepix, a novel antifungal agent that targets the GPI-anchor biosynthesis pathway by inhibition of the GWT1 enzyme. Manogepix was evaluated by determining the minimal inhibitory concentrations (MICs) according to the CLSI-based in vitro susceptibility assay for 22 M. mycetomatis strains and by in silico protein comparison of the target protein. The synergy between manogepix and itraconazole was determined using a checkerboard assay. The efficacy of clinically relevant dosages was assessed in an in vivo grain model in Galleria mellonella larvae. MICs for manogepix ranged from <0.008 to >8 mg/l and 16/22 M. mycetomatis strains had an MIC ≥4 mg/ml. Differences in MICs were not related to differences observed in the GWT1 protein sequence. For 70% of the tested isolates, synergism was found between manogepix and itraconazole in vitro. In vivo, enhanced survival was not observed upon admission of 8.6 mg/kg manogepix, nor in combination treatment with 5.7 mg/kg itraconazole. MICs of manogepix were high, but the in vitro antifungal activity of itraconazole was enhanced in combination therapy. However, no efficacy of manogepix was found in an in vivo grain model using clinically relevant dosages. Therefore, the therapeutic potential of manogepix in mycetoma caused by M. mycetomatis seems limited.
Topics: Animals; Itraconazole; Madurella; Mycetoma; Antifungal Agents
PubMed: 37960934
DOI: 10.1093/mmy/myad118 -
Acta Dermato-venereologica Mar 2015
Topics: Amputation, Surgical; Female; Foot Dermatoses; Humans; Madurella; Mycetoma; Pregnancy; Pregnancy Complications
PubMed: 25518866
DOI: 10.2340/00015555-2037 -
The Journal of Antimicrobial... Apr 2020Eumycetoma is currently treated with a combination of itraconazole therapy and surgery, with limited success. Recently, olorofim, the lead candidate of the orotomides, a...
OBJECTIVES
Eumycetoma is currently treated with a combination of itraconazole therapy and surgery, with limited success. Recently, olorofim, the lead candidate of the orotomides, a novel class of antifungal agents, entered a Phase II trial for the treatment of invasive fungal infections. Here we determined the activity of olorofim against Madurella mycetomatis, the main causative agent of eumycetoma.
METHODS
Activity of olorofim against M. mycetomatis was determined by in silico comparison of the target gene, dihydroorotate dehydrogenase (DHODH), and in vitro susceptibility testing. We also investigated the in vitro interaction between olorofim and itraconazole against M. mycetomatis.
RESULTS
M. mycetomatis and Aspergillus fumigatus share six out of seven predicted binding residues in their DHODH DNA sequence, predicting susceptibility to olorofim. Olorofim demonstrated excellent potency against M. mycetomatis in vivo with MICs ranging from 0.004 to 0.125 mg/L and an MIC90 of 0.063 mg/L. Olorofim MICs were mostly one dilution step lower than the itraconazole MICs. In vitro interaction studies demonstrated that olorofim and itraconazole work indifferently when combined.
CONCLUSIONS
We demonstrated olorofim has potent in vitro activity against M. mycetomatis and should be further evaluated in vivo as a treatment option for this disease.
Topics: Acetamides; Antifungal Agents; Humans; Madurella; Mycetoma; Piperazines; Pyrimidines; Pyrroles
PubMed: 31904836
DOI: 10.1093/jac/dkz529 -
Mycoses May 2023Mycetoma is a neglected tropical implantation disease caused by 70 different infectious agents. Identifying the causative organism to the species level is essential for...
OBJECTIVES
Mycetoma is a neglected tropical implantation disease caused by 70 different infectious agents. Identifying the causative organism to the species level is essential for appropriate patient management. Ultrasound, histopathology, culture and two species-specific PCRs are most the commonly used methods for species identification in endemic regions. The aim of this study was to compare the diagnostic performance of these commonly used assays using sequencing of barcoding genes as the gold standard.
METHODS
This descriptive cross-sectional study was conducted at the Mycetoma Research Centre, University of Khartoum, Sudan. It included 222 patients suspected of fungal mycetoma caused by Madurella mycetomatis.
RESULTS
154 (69.3%) were correctly identified by ultrasound, histology, culture and both species-specific PCRs. In 60 patients, at least one of the diagnostic tests failed to identify M. mycetomatis. Five patients had no evidence of eumycetoma, and for three, only the ultrasound was indicative of mycetoma. The two species-specific PCRs were the most sensitive and specific methods, followed by culture and histology. Ultrasound was the least specific as it only allowed differentiation between actinomycetoma and eumycetoma. The time to result was 9.38 minutes for ultrasound, 3.76 hours for PCR, 8.5 days for histopathology and 21 days for grain culturing.
CONCLUSION
Currently, PCR directly on DNA isolated from grains is the most rapid and reliable diagnostic tool to identify M. mycetomatis eumycetoma.
Topics: Humans; Mycetoma; Cross-Sectional Studies; Sudan; Polymerase Chain Reaction; Madurella; Diagnostic Tests, Routine
PubMed: 36583225
DOI: 10.1111/myc.13561 -
Mycoses Dec 2022Eumycetoma is a neglected tropical disease. It is a chronic inflammatory subcutaneous infection characterised by painless swellings which produce grains. It is currently...
BACKGROUND
Eumycetoma is a neglected tropical disease. It is a chronic inflammatory subcutaneous infection characterised by painless swellings which produce grains. It is currently treated with a combination of itraconazole and surgery. In an ongoing clinical study, the efficacy of fosravuconazole, the prodrug of ravuconazole, is being investigated. For both itraconazole and ravuconazole, no clinical breakpoints or epidemiological cut-off values (ECV) to guide treatment are currently available.
OBJECTIVE
To determine tentative ECVs for itraconazole and ravuconazole in Madurella mycetomatis, the main causative agent of eumycetoma.
MATERIALS AND METHODS
Minimal inhibitory concentrations (MICs) for itraconazole and ravuconazole were determined in 131 genetically diverse clinical M. mycetomatis isolates with the modified CLSI M38 broth microdilution method. The MIC distributions were established and used to determine ECVs with the ECOFFinder software. CYP51A sequences were sequenced to determine whether mutations occurred in this azole target gene, and comparisons were made between the different CYP51A variants and the MIC distributions.
RESULTS
The MICs ranged from 0.008 to 1 mg/L for itraconazole and from 0.002 to 0.125 mg/L for ravuconazole. The M. mycetomatis ECV for itraconazole was 1 mg/L and for ravuconazole 0.064 mg/L. In the wild-type population, two CYP51A variants were found for M. mycetomatis, which differed in one amino acid at position 499 (S499G). The MIC distributions for itraconazole and ravuconazole were similar between the two variants. No mutations linked to decreased susceptibility were found.
CONCLUSION
The proposed M. mycetomatis ECV for itraconazole is 1 mg/L and for ravuconazole 0.064 mg/L.
Topics: Humans; Madurella; Itraconazole; Mycetoma; Triazoles; Antifungal Agents
PubMed: 36005544
DOI: 10.1111/myc.13509 -
Medecine Tropicale Et Sante... Dec 2021The objective of this study was to describe the epidemiological and clinical aspects as well as the therapeutic methods of mycetomical lesions.
OBJECTIVE
The objective of this study was to describe the epidemiological and clinical aspects as well as the therapeutic methods of mycetomical lesions.
MATERIAL AND METHODS
This was a longitudinal retrospective study, which included all patients treated for mycetoma from January 2016 to December 2018 including two years of recruitment and one year of monitoring (2019). The study concerned 19 patients who were hospitalized and treated in the department of surgery.
RESULTS
Patients represented 2.3% of hospitalizations and consisted of 11 males and 8 females with an average age of 38 years with extremes of 15 - 70 years, and an average time between the onset of symptoms and presentation to the hospital of 10 years (range 1 - 40 years). Eight livestock breeders and seven farmers were concerned, 14 of whom have started the disease after trauma. The foot was involved in 13 patients. Twelve suffered from osteoarticular lesions. Black grains were present in 16 cases attributed to sp. We performed 12 amputations, six carcinological ablation to which specific local treatments were added (thin skin graft in two patients, fasciocutaneous flap in one patient and directed healing in the others) and local treatment in the last case.
CONCLUSION
Mycetoma should be discussed and diagnosed at an early stage in predisposed patients particularly in farmers and breeders. Prevention is necessary; it is based on wound disinfection and wearing safety shoes.
Topics: Adolescent; Adult; Animals; Child; Child, Preschool; Coleoptera; Female; Hospitals; Humans; Infant; Madurella; Male; Mali; Mycetoma; Retrospective Studies; Young Adult
PubMed: 35685855
DOI: 10.48327/mtsi.2021.170 -
PLoS Neglected Tropical Diseases Jul 2019Mycetoma is a persistent, progressive granulomatous inflammatory disease caused either by fungi or by bacteria. Characteristic of this disease is that the causative...
Mycetoma is a persistent, progressive granulomatous inflammatory disease caused either by fungi or by bacteria. Characteristic of this disease is that the causative agents organise themselves in macroscopic structures called grains. These grains are surrounded by a massive inflammatory reaction. The processes leading to this host tissue reaction and the immunophenotypic characteristics of the mycetoma granuloma are not known. Due to the massive immune reaction and the tissue remodeling involved, we hypothesised that the expression levels of interleukin-17 (IL-17) and matrix metalloprotease-9 (MMP-9) in the mycetoma granuloma formation were correlated to the severity of the disease and that this correlation was independent of the causative agent responsible for the granuloma reaction. To determine the expression of IL-17 and MMP-9 in mycetoma lesions, the present study was conducted at the Mycetoma Research Centre, Sudan. Surgical biopsies from 100 patients with confirmed mycetoma were obtained, and IL-17 and MMP-9 expression in the mycetoma granuloma were evaluated immunohistochemically. IL-17 was mainly expressed in Zones I and II, and far less in Zone III. MMP-9 was detected mainly in Zones II and III, and the least expression was in Zone I. MMP-9 was more highly expressed in Actinomadura pelletierii and Streptomyces somaliensis biopsies compared to Madurella mycetomatis biopsies. MMP-9 levels were directly proportional to the levels of IL-17 (p = 0.001). The only significant association between MMP9 and the patients' characteristics was the disease duration (p<0.001). There was an insignificant correlation between the IL-17 levels and the patients' demographic characteristics.
Topics: Actinobacteria; Actinomadura; Adolescent; Adult; Biopsy; Child; Collagen; Female; Gene Expression; Granuloma; Humans; Immunohistochemistry; Interleukin-17; Madurella; Male; Matrix Metalloproteinase 9; Middle Aged; Mycetoma; Qualitative Research; Severity of Illness Index; Streptomyces; Sudan; Young Adult
PubMed: 31295246
DOI: 10.1371/journal.pntd.0007351 -
Medical Mycology Feb 2022Eumycetoma is a neglected tropical disease, and Madurella mycetomatis, the most common causative agent of this disease forms black grains in hosts. Melanin was...
UNLABELLED
Eumycetoma is a neglected tropical disease, and Madurella mycetomatis, the most common causative agent of this disease forms black grains in hosts. Melanin was discovered to be one of the constituents in grains. Melanins are hydrophobic, macromolecular pigments formed by oxidative polymerisation of phenolic or indolic compounds. M. mycetomatis was previously known to produce DHN-melanin and pyomelanin in vitro. These melanin was also discovered to decrease M. mycetomatis's susceptibility to antifungals itraconazole and ketoconazole in vitro. These findings, however, have not been confirmed in vivo. To discover the melanin biosynthesis pathways used by M. mycetomatis in vivo and to determine if inhibiting melanin production would increase M. mycetomatis's susceptibility to itraconazole, inhibitors targeting DHN-, DOPA- and pyomelanin were used. Treatment with DHN-melanin inhibitors tricyclazole, carpropamid, fenoxanil and DOPA-melanin inhibitor glyphosate in M. mycetomatis infected Galleria mellonella larvae resulted in presence of non-melanized grains. Our finding suggested that M. mycetomatis is able to produce DOPA-melanin in vivo. Inhibiting DHN-melanin with carpropamid in combination with the antifungal itraconazole also significantly increased larvae survival. Our results suggested that combination treatment of antifungals and melanin inhibitors can be an alternative treatment strategy that can be further explored. Since the common black-grain eumycetoma causing agents uses similar melanin biosynthesis pathways, this strategy may be applied to them and other eumycetoma causative agents.
LAY SUMMARY
Melanin protects fungi from environmental stress and antifungals. We have discovered that Madurella mycetomatis produces DHN-, pyomelanin and DOPA-melanin in vivo. Inhibiting M. mycetomatis DHN-melanin biosynthesis increases therapeutic value of the antifungal itraconazole in vivo.
Topics: Animals; Antifungal Agents; Dihydroxyphenylalanine; Itraconazole; Madurella; Mycetoma
PubMed: 35064672
DOI: 10.1093/mmy/myac003 -
Studies in Mycology Jun 2019The genus is morphologically defined by having non-ostiolate ascomata with a thin peridium composed of , and smooth, single-celled, pigmented ascospores with one germ...
The genus is morphologically defined by having non-ostiolate ascomata with a thin peridium composed of , and smooth, single-celled, pigmented ascospores with one germ pore. is typified with that grows in close association with a hyphomycete which was traditionally identified as . Besides exhibiting the mycoparasitic nature, the majority of the described species are from soil, and some have economic and ecological importance. Unfortunately, no living type material of exists, hindering a proper understanding of the classification of . Therefore, was neotypified by material of a mycoparasite presenting the same ecology and morphology as described in the original description. We subsequently performed a multi-gene phylogenetic analyses (, , ITS and LSU) to resolve the phylogenetic relationships of the species currently recognised in . Our results demonstrate that is highly polyphyletic, being related to three family-level lineages in two orders. The redefined genus is restricted to its type species, , which belongs to the () and its host is demonstrated to be , one of the two species in the "" species complex. The new family is sister to the in the and accommodates the re-defined genera , and , with the last genus including two former species ( and ). This family also includes the genetic model species , which was combined in (as ). The remaining species fall in ten unrelated clades in the , leading to the proposal of nine new genera (, , , , , , , and ). The genus is transferred from () to based on its type species . is closely related to the human-pathogenic genus , and includes three thielavia-like species and one novel species. Three monotypic genera with a chaetomium-like morph ( and ) are introduced to better resolve the and the thielavia-like species in the family. and are closely related to and three newly introduced genera containing thielavia-like species; is closely related to the industrially important and thermophilic species (syn. ). This study shows that the thielavia-like morph is a homoplastic form that originates from several separate evolutionary events. Furthermore, our results provide new insights into the taxonomy of and the polyphyletic .
PubMed: 31824584
DOI: 10.1016/j.simyco.2019.08.002