Isolate of Encephalitozoon intestinalis from stools of a Colombian patient with AIDS
Keywords:
Acquired immunodeficiency sindrome, microsporidiosis, patient isolate, diarrhea, cell cultures, feces
Abstract
Introduction. Microsporidia are obligate intracellular parasites that are recognized as important opportunistic pathogens of immunocompromised and transplanted patients. Enterocytozoon bieneusi and, less frequently, Encephalitozoon intestinalis are the most prevalent species in humans; both of them are associated with enteric infections. Cell cultures have been useful in the study of microsporidia biology. In Colombia, however, no isolates of microsporidia from patients with AIDS have been obtained. Objective. A cell culture of intestinal microsporidia was established from stools of positive patients in order to isolate a native strain. Materials and methods. Stool from a single AIDS patient was concentrated with the water-ether technique, and the sediment was treated with a mixture of antibiotics and antifungal agents for 18 hours at 37o C. Vero cells were cultivated in 24-well plates with Gibco® RPMI medium supplemented with 10% bovine fetal serum and antibiotics. The culture was subsequently inoculated with previously concentrated spores. The medium was changed every second day and the presence of spores was evaluated with the Quick Hot Gram chromotrope stain. Results. Two weeks post-infection, microsporidial spores were identified with characteristic morphology and staining properties. PCR results showed that Encephalitozoon intestinalis was the isolated species. Conclusions. A cell culture of microsporidia was established from a stool sample. This protocol is important to isolate and maintain additional native Colombian strains and it will contribute to biochemical, immunological and epidemiological studies of the currently established strain.Downloads
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References
1. Sprague VV, Becnel JJ. Note on the name-author-date combination for the taxon Microsporidies Balbiani, 1882, when ranked as Phylum. J Invertebr Pathol. 1998;71:91-4.
2. Franzen C, Müller A. Microsporidiosis: human diseases and diagnosis. Microbes Infect. 2001;3:389-400.
3. Mathis A, Weber R, Deplazes P. Zoonotic potential of the microsporidia. Clin Microbiol Rev. 2005;18:423-45.
4. Franzen C, Müller A. Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia. Clin Microbiol Rev. 1999;12:243-85.
5. Wasson K, Peper RL. Mammalian microsporidiosis. Vet Pathol. 2000;37:113-28.
6. Didier S. Microsporidiosis: An emerging and opportunistic infection in humans and animals. Acta Trop. 2005;94:61-76.
7. Kotler DP, Orenstein JM. Prevalence of intestinal microsporidiosis in HIV-infected individuals referred for gastroenterological evaluation. Am J Gastroenterol. 1994;8:1998-2002.
8. Gamboa Domínguez A, Bencosme Vinas C, Kato Maeda M. Microsporidiasis in AIDS patients with chronic diarrhea. Experiences at the National Institute of Nutrition "Salvador Zubriran". Rev Gastroenterol Mex. 1999;64: 70-4.
9. Brasil P, de Lima DB, de Paiva DD, Lobo MS, Sodre FC, Silva SP, et al. Clinical and diagnostic aspects of intestinal microsporidiosis in HIV-infected patients with chronic diarrhea in Rio de Janeiro, Brazil. Rev Inst Med Trop Sao Paulo. 2000;42:299-304.
10. Flórez AC, García DA, Moncada LI, Beltrán M. Prevalencia de microsporidios y otros parásitos intestinales en pacientes con infección por VIH, Bogotá, 2001. Biomédica. 2003;23:274-82.
11. Botero J, Montoya MN, Agudelo S, Vanegas AL, Diaz A, Bornay FJ, et al. Frecuencia de microsporidiosis intestinal en pacientes VIH positivos determinada mediante las técnicas de quick-hot gram chromotropo y PCR. Biomédica. 2004;24:375-84.
12. Visvesvara GS. Microsporidia: in vitro cultivation of microsporidia of clinical importance. Clin Microbiol Rev. 2002;15:401-13.
13. Del Aguila C, Croppo GP, Moura H, da silva AJ, Leitch GJ, Moss DM, et al. Ultrastructure, immunofluorescence, western blot, and PCR analysis of eight isolates of Encephalitozoon (septata) intestinalis established in culture from sputum and urine samples and duodenal aspirates of five patients with AIDS. J Clin Microbiol. 1998;36:1201-8.
14. van Gool T, Canning EU, Gilis H, van den Bergh Weerman MA, Eeftinck Schattenkerk JK, Dankert J. Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method. Parasitology. 1994;109:281-9.
15. Visvesvara GS, da Silva AJ, Croppo GP, Pieniazek NJ, Leitch GJ, Ferguson D, et al. In vitro culture and serologic and molecular identification of Septata intestinalis isolated from urine of a patient with AIDS. J Clin Microbiol. 1995;33:930-6.
16. da Silva AJ, Schwartz DA, Visvesvara GS, de Moura H, Slemenda SB, Pieniazek NJ. Sensitive PCR diagnosis of infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol. 1996;34:986-7.
2. Franzen C, Müller A. Microsporidiosis: human diseases and diagnosis. Microbes Infect. 2001;3:389-400.
3. Mathis A, Weber R, Deplazes P. Zoonotic potential of the microsporidia. Clin Microbiol Rev. 2005;18:423-45.
4. Franzen C, Müller A. Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia. Clin Microbiol Rev. 1999;12:243-85.
5. Wasson K, Peper RL. Mammalian microsporidiosis. Vet Pathol. 2000;37:113-28.
6. Didier S. Microsporidiosis: An emerging and opportunistic infection in humans and animals. Acta Trop. 2005;94:61-76.
7. Kotler DP, Orenstein JM. Prevalence of intestinal microsporidiosis in HIV-infected individuals referred for gastroenterological evaluation. Am J Gastroenterol. 1994;8:1998-2002.
8. Gamboa Domínguez A, Bencosme Vinas C, Kato Maeda M. Microsporidiasis in AIDS patients with chronic diarrhea. Experiences at the National Institute of Nutrition "Salvador Zubriran". Rev Gastroenterol Mex. 1999;64: 70-4.
9. Brasil P, de Lima DB, de Paiva DD, Lobo MS, Sodre FC, Silva SP, et al. Clinical and diagnostic aspects of intestinal microsporidiosis in HIV-infected patients with chronic diarrhea in Rio de Janeiro, Brazil. Rev Inst Med Trop Sao Paulo. 2000;42:299-304.
10. Flórez AC, García DA, Moncada LI, Beltrán M. Prevalencia de microsporidios y otros parásitos intestinales en pacientes con infección por VIH, Bogotá, 2001. Biomédica. 2003;23:274-82.
11. Botero J, Montoya MN, Agudelo S, Vanegas AL, Diaz A, Bornay FJ, et al. Frecuencia de microsporidiosis intestinal en pacientes VIH positivos determinada mediante las técnicas de quick-hot gram chromotropo y PCR. Biomédica. 2004;24:375-84.
12. Visvesvara GS. Microsporidia: in vitro cultivation of microsporidia of clinical importance. Clin Microbiol Rev. 2002;15:401-13.
13. Del Aguila C, Croppo GP, Moura H, da silva AJ, Leitch GJ, Moss DM, et al. Ultrastructure, immunofluorescence, western blot, and PCR analysis of eight isolates of Encephalitozoon (septata) intestinalis established in culture from sputum and urine samples and duodenal aspirates of five patients with AIDS. J Clin Microbiol. 1998;36:1201-8.
14. van Gool T, Canning EU, Gilis H, van den Bergh Weerman MA, Eeftinck Schattenkerk JK, Dankert J. Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method. Parasitology. 1994;109:281-9.
15. Visvesvara GS, da Silva AJ, Croppo GP, Pieniazek NJ, Leitch GJ, Ferguson D, et al. In vitro culture and serologic and molecular identification of Septata intestinalis isolated from urine of a patient with AIDS. J Clin Microbiol. 1995;33:930-6.
16. da Silva AJ, Schwartz DA, Visvesvara GS, de Moura H, Slemenda SB, Pieniazek NJ. Sensitive PCR diagnosis of infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol. 1996;34:986-7.
How to Cite
1.
Galván AL, Bedoya K, Montoya MN, Botero J. Isolate of Encephalitozoon intestinalis from stools of a Colombian patient with AIDS. biomedica [Internet]. 2008 Sep. 1 [cited 2024 May 12];28(3):441-7. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/83
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Published
2008-09-01
Issue
Section
Technical note
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