Abstract
Mycetoma is a tropical disease caused by several fungi and bacteria present in the soil. Fungal mycetoma and eumycetoma are especially challenging to treat; therefore, prevention, early diagnosis, and early treatment are important, but it is also necessary to understand the geographic distribution of these pathogenic fungi. In this study, we used DNA metabarcoding methodology to identify fungal species from soil samples. Soil sampling was implemented at seven villages in an endemic area of Sennar State in Sudan in 2019, and ten sampling sites were selected in each village according to land-use conditions. In total, 70 soil samples were collected from ground surfaces, and DNA in the soil was extracted with a combined method of alkaline DNA extraction and a commercial soil DNA extraction kit. The region for universal primers was selected to be the ribosomal internal transcribed spacer one region for metabarcoding. After the second PCR for DNA library preparation, the amplicon-based DNA analysis was performed using next-generation sequencing with two sets of universal primers. A total of twelve mycetoma-causative fungal species were identified, including the prime agent, Madurella mycetomatis, and additional pathogens, Falciformispora senegalensis and Falciformispora tompkinsii, in 53 soil samples. This study demonstrated that soil DNA metabarcoding can elucidate the presence of multiple mycetoma-causative fungi, which may contribute to accurate diagnosis for patient treatment and geographical mapping.
Highlights
Mycetoma is a chronic granulomatous and disabling inflammatory disease caused by specific groups of bacteria or fungi
The results suggest that multiple causative agents of fungal mycetoma are widespread regardless of the environment and can be a source of infection anywhere in an endemic area
Because the causative fungal pathogens exist in the soil in endemic areas, it is possible to determine the distribution of infection by analyzing collected soil samples from areas in or near-endemic areas to confirm their presence
Summary
Mycetoma is a chronic granulomatous and disabling inflammatory disease caused by specific groups of bacteria (actinomycetoma) or fungi (eumycetoma). It typically affects people living in poor, remote communities in tropical and subtropical regions within the so-called mycetoma belt, located between latitude 15 ̊S and 30 ̊N [1]. Most of the causative microorganisms inhabit the soil and invade the human body through minor unnoticed wounds on the skin, mainly in the foot and hand, and multiply to form multiple painless subcutaneous mass lesions that discharge seropurulent grains [1–4]. To understand the distributions of the causative fungi, it is necessary to comprehensively capture all fungi in an endemic area’s soil
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