Frequency of Actinomadura among Mycetoma Patients and its Relevance to Soil Actinomycetes in Sudan

Abstract

The present study aimed to determine the frequency of actinomadura among Sudanese patients suffering from mycetoma in endemic areas in Sudan and correlate these infections with soil actinomycetes isolated from these areas. One hundred clinical specimens were collected from mycetoma patients who attended the Mycetoma Research Center at soba teatching hospital as also as clinics in different areas in Sudan including Um-Rawaba Western Sudan, White Nile State and Wad Madani Hospital during the period from November 2008 to August 2010. Grains were collected from sinuses and incision biopsies in sterile containers. They were washed with normal saline, then sent to laboratory for culture and further investigations. Primary identification of the clinical isolates was performed based on the color of the grains, colony morphology and Grams.It stain revealed that 12% were actinomycetes among which 6 were identified as Actinomadura madurae, while the other were considered as eumycetoma (50%+88%). Further confirmation was done by subjecting the twelve isolated actinomycetes to conventional Polymerase Chain Reaction (PCR) and Real Time PCR (RT-PCR). The results showed that 6 (50%) were Streptomyces while the others (50%) were negative to stb1 gene. On the other hand, one hundred soil samples were collected from different geographical areas including Um-Rawaba Northen Kordofan, Western Sudan, White Nile State and Khartoum State. Soil samples were collected in clear dry containers, and sent tothe laboratory for culture and further investigations. Serial dilution from the soil were performed and inoculated on Tryptic Soy Agar (TSA) and Actinomycetes Isolation Agar (AIA) media, and incubated aerobically at 37oC. Soil isolates were identified by colony morphology, Gram Stain, modified Zihel Neelson stain, biochemical tests and mycolic acids profile. The results showed that 20% of the isolates were Gram positive branched bacilli and week acid fast bacilli with distinguished growth on TSA and AIA. Among these 20 isolates 3 (15%) were Streptomyces species, while the other 17 (85%) were Nocardia and Nocardia like. In conclusion that, actinomadura exist with high frequency among Sudanese patients with mycetoma. Since a number of bacterial agents which are known to cause actinomadura have been isolated from soil in target areas, the study suggests that soil may represent a source of the infection.