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Journal of the European Academy of... Aug 2020At the dermatology service of the General Hospital of Mexico City, Mexico, two patients, father and son, with black-grain mycetoma were seen. The grains were isolated,...
BACKGROUND
At the dermatology service of the General Hospital of Mexico City, Mexico, two patients, father and son, with black-grain mycetoma were seen. The grains were isolated, and the cultured fungi were identified as Madurella mycetomatis based on morphology. Using the M. mycetomatis specific PCR, amplicons of a different size than that of the M. mycetomatis type strain were obtained.
OBJECTIVE
To determine the causative agent of the two black-grain mycetoma cases and develop non-culture-based diagnostic tools to identify them to the species level.
METHODS
The M. mycetomatis specific, the internal transcribed spacer (ITS) region, β-tubulin (BT) and ribosomal binding protein 2 (RBP2) PCRs were used to confirm the identity of the isolates. Genetic variation was established by amplification fragment length polymorphisms. To determine the antifungal susceptibility profile, the Sensititre™ YeastOne™ assay was used. To develop a species-specific PCR primers were designed on the sequenced PCR amplicon from the M. mycetomatis specific PCR.
RESULTS
By analyzing the ITS, BT and RBP2 regions the isolates were identified as Madurella pseudomycetomatis. The isolates from father and son were similar but not identical to M. pseudomycetomatis from Venezuela and one from an unknown origin. Madurella pseudomycetomatis isolates were inhibited by itraconazole, posaconazole and voriconazole but showed increased MIC values for amphotericin B and fluconazole. They were not inhibited by the echinocandins and five flucytosine. The two patients were treated with itraconazole resulting in cure for the father while the son was lost to follow-up. The species-specific PCR developed for M. pseudomyceotmatis was discriminative and specific.
CONCLUSION
Madurella pseudomycetomatis is genetically diverse with same susceptibility profile as M. mycetomatis and causes eumycetoma in Latin America. The M. pseudomycetomatis specific PCR can be used to identify this causative agent to the species level; however, this needs to be validated in an endemic setting.
Topics: DNA Primers; Humans; Madurella; Mexico; Mycetoma; Species Specificity
PubMed: 32233084
DOI: 10.1111/jdv.16402 -
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 -
BMC Dermatology Jan 2020Mycetoma is an uncommon chronic granulomatous infection of cutaneous and subcutaneous tissues that can be caused by filamentous bacteria (actinomycetoma) or fungi... (Review)
Review
BACKGROUND
Mycetoma is an uncommon chronic granulomatous infection of cutaneous and subcutaneous tissues that can be caused by filamentous bacteria (actinomycetoma) or fungi (eumycetoma). It is the prerogative of young men between the third and fourth decade and is transmitted through any trauma causing an inoculating point. The classic clinical triad associates a painless hard and swelling subcutaneous mass, multiple fistulas, and the pathognomonic discharge of grains. Although endemic in many tropical and subtropical countries, mycetoma can also be found in non-endemic areas as in Morocco, and causes then diagnosis problems leading to long lasting complications. Therefore, we should raise awareness of this neglected disease for an earlier management. Under medical treatment however, mycetoma has a slow healing and surgery is often needed, and relapses are possible.
CASE PRESENTATION
Herein we report a case of a 64 years old patient, with a history of eumycetoma occurring ten years ago treated with oral terbinafine coupled with surgery. A complete remission was seen after 2 years. He presented a relapse on the previous scar 6 months ago. There wasn't any bone involvement in the magnetic resonance imaging (MRI). The patient was put under oral terbinafine with a slow but positive outcome.
CONCLUSION
Through this case report, we perform a literature review and highlight the importance of increase awareness of mycetoma in clinical practice especially in non-endemic regions.
Topics: Antifungal Agents; Diagnosis, Differential; Foot; Foot Diseases; Humans; Madurella; Magnetic Resonance Imaging; Male; Middle Aged; Morocco; Mycetoma; Neglected Diseases; Tropical Medicine
PubMed: 31918687
DOI: 10.1186/s12895-019-0097-1 -
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 -
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 -
Medical Mycology Case Reports Dec 2019Mycetoma is a chronic-granulomatous disease characterized by the inflammation, swollen organ, draining sinuses containing blood, pus, and grains. We present a case of...
Mycetoma is a chronic-granulomatous disease characterized by the inflammation, swollen organ, draining sinuses containing blood, pus, and grains. We present a case of madura foot with novel etiologic agent Diagnosis was based on morphologic, physiologic, histipathologic and molecular methods. antifungal susceptibility tests revealed that MIC values for itraconazole, amphotericin B, and posaconazole were 0.0313 μg/ml, 0.0313 μg/ml, and 0.004 μg/ml, respectively. The patient was treated and recovered by itraconazole(400 mg/day) after prolonged course.
PubMed: 31667052
DOI: 10.1016/j.mmcr.2019.09.005 -
Epidemiology and Infection Oct 2019Mycetoma is a chronic granulomatous, suppurative and progressive inflammatory disease that usually involves the subcutaneous tissue and bones after traumatic inoculation...
Mycetoma is a chronic granulomatous, suppurative and progressive inflammatory disease that usually involves the subcutaneous tissue and bones after traumatic inoculation of the causative organism. In India, actinomycotic mycetoma is prevalent in south India, south-east Rajasthan and Chandigarh, while eumycetoma, which constitutes one third of the total cases, is mainly reported from north India and central Rajasthan. The objective was to determine the epidemiological profile and spectrum of eumycetoma from a tertiary care hospital in Delhi, North India. Thirty cases of eumycetoma were diagnosed by conventional methods of direct microscopy, culture and species-specific sequencing as per standard protocol. The spectrum of fungal pathogens included Exophiala jeanselmei, Madurella mycetomatis, Fusarium solani, Sarocladium kiliense, Acremonium blochii, Aspergillus nidulans, Fusarium incarnatum, Scedosporium apiospermum complex, Curvularia lunata and Medicopsis romeroi. Eumycetoma can be treated with antifungal therapy and needs to be combined with surgery. It has good prognosis if it is timely diagnosed and the correct species identified by culture for targeted therapy of these patients. Black moulds required prolonged therapy. Its low reporting and lack of familiarity may predispose patients to misdiagnosis and consequently delayed treatment. Hence health education and awareness campaign on the national and international level in the mycetoma belt is crucial.
Topics: Adolescent; Adult; Female; Humans; India; Male; Middle Aged; Mycetoma; Neglected Diseases; Prognosis; Retrospective Studies; Young Adult
PubMed: 31637988
DOI: 10.1017/S0950268819001766 -
Antimicrobial Agents and Chemotherapy Dec 2019The use of the Sensititre YeastOne YO10 alamarBlue assay for the susceptibility testing of was evaluated in isolates with and without pyomelanin secretion. Pyomelanin...
Pyomelanin Secretion in Madurella mycetomatis Interferes with Spectrophotometric Endpoint Reading Using the Sensititre YeastOne alamarBlue Assay but Not with Visual Endpoint Reading.
The use of the Sensititre YeastOne YO10 alamarBlue assay for the susceptibility testing of was evaluated in isolates with and without pyomelanin secretion. Pyomelanin secretion did not influence visual endpoint reading; however, it caused a shift in peak absorbance from 570 nm to 620 nm when read spectrophotometrically. Therefore, when choosing the method for endpoint reading, the presence of pyomelanin should be considered.
Topics: Azoles; Madurella; Melanins; Mycetoma
PubMed: 31611353
DOI: 10.1128/AAC.01532-19 -
PLoS Neglected Tropical Diseases Aug 2019Mycetoma is a devastating neglected tropical disease, caused by various fungal and bacterial pathogens. Correct diagnosis to the species level is mandatory for proper...
Mycetoma is a devastating neglected tropical disease, caused by various fungal and bacterial pathogens. Correct diagnosis to the species level is mandatory for proper treatment. In endemic areas, various diagnostic tests and techniques are in use to achieve that, and that includes grain culture, surgical biopsy histopathological examination, fine needle aspiration cytological (FNAC) examination and in certain centres molecular diagnosis such as PCR. In this retrospective study, the sensitivity, specificity and diagnostic accuracy of grain culture, surgical biopsy histopathological examination and FNAC to identify the mycetoma causative organisms were determined. The histopathological examination appeared to have better sensitivity and specificity. The histological examination results were correct in 714 (97.5%) out of 750 patients infected with Madurella mycetomatis, in 133 (93.6%) out of 142 patients infected with Streptomyces somaliensis, in 53 (74.6%) out of 71 patients infected with Actinomadura madurae and in 12 (75%) out of 16 patients infected with Actinomadura pelletierii. FNAC results were correct in 604 (80.5%) out of 750 patients with Madurella mycetomatis eumycetoma, in 50 (37.5%) out of 133 Streptomyces somaliensis patients, 43 (60.5%) out of 71 Actinomadura madurae patients and 11 (68.7%) out of 16 Actinomadura pelletierii. The mean time required to obtain the FNAC result was one day, and for the histopathological examinations results it was 3.5 days, and for grain it was a mean of 16 days. In conclusion, histopathological examination and FNAC are more practical techniques for rapid species identification than grain culture in many endemic regions.
Topics: Actinobacteria; Actinomadura; Adolescent; Adult; Aged; Biopsy; Child; Child, Preschool; Diagnostic Tests, Routine; Female; Humans; Madurella; Male; Middle Aged; Mycetoma; Pathology, Molecular; Polymerase Chain Reaction; Retrospective Studies; Sensitivity and Specificity; Streptomyces; Young Adult
PubMed: 31465459
DOI: 10.1371/journal.pntd.0007056 -
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