Abstract
Human rhinosporidial tissue was used as the source of the various developmental stages of Rhinosporidium seeberi - endospores with electron dense bodies, juvenile, and immature sporangia. After homogenisation in phosphate buffered saline (PBS) and removal of tissue fragments by centrifugation, the rhinosporidial bodies were isolated on centrifuged Percoll columns with gradients of densities or on triple-layered columns of varying density. The separated bands, after repeated washing in PBS gave bodies free from human tissue as shown on Leishman and PAS staining and indirect immunofluorescence with rabbit and human patients' anti-rhinosporidial sera. Sonicates of these bodies were tested on agarose gel for precipitation with antisera, and on SDS-PAG electrophoresis and Coomassie Blue staining. Percoll columns were shown to be capable of isolating these stages of R. seeberi, free from human tissue and contaminating bacteria.
Similar content being viewed by others
References
Rao SN. A technique for harvesting intact sporangia from Rhinosporidium seeberi. Ind J Exp Biol 1966; 4: 189–190.
Grover Shobha, Rhinosporidium seeberi: A preliminary study of the morphology and life cycle. Sabouraudia 1970; 7: 249–251.
Kannan-Kutty M, Gomez JB. The ultrastructure and life history of Rhinosporidium seeberi. South East Asian J Trop Med Public Health 1971; 2: 9–16.
Arseculeratne SN, Ajello L. Rhinosporidium seeberi. In Leslie Collier, Albert Balows and Max Sussman, eds. Topley and Wilson's Microbiology and Microbial Infections. London: Edward Arnold., Vol. 4, Medical Mycology, 1998: 595–615.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Atapattu, D.N., Arseculeratne, S.N., Rajapakse, R.P.V.J. et al. Purification of the Endospores and Sporangia of Rhinosporidium Seeberi on Percoll Columns. Mycopathologia 145, 113–119 (1999). https://doi.org/10.1023/A:1007086732491
Issue Date:
DOI: https://doi.org/10.1023/A:1007086732491