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[First isolate of Encephalitozoon intestinalis from stools of a Colombian patient with AIDS]
Katherine Bedoya1, Martha Nelly Montoya, Jorge Botero
1Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales - GIEPI, Corporación para el Estudio de las Patologías Tropicales - CAEPT, Universidad de Antioquia, Medellín, Colombia.
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 37 degrees 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.
Insights
Researchers successfully established a microsporidia cell culture from an AIDS patient
Area of Science:
- Parasitology
- Infectious Diseases
- Cell Biology
Background:
- Microsporidia are opportunistic intracellular parasites causing enteric infections, particularly in immunocompromised individuals.
- Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common human microsporidia species.
- Establishing microsporidia cell cultures is crucial for studying their biology, but native Colombian strains were previously unobtained.
Purpose of the Study:
- To establish a cell culture of intestinal microsporidia from a patient in Colombia.
- To isolate a native strain of microsporidia for further research.
Main Methods:
- Concentration of microsporidian spores from an AIDS patient's stool using the water-ether technique.
- Treatment of concentrated spores with antibiotics and antifungals.
- Inoculation of Vero cells with treated spores and monitoring via Quick Hot Gram chromotrope stain.
Main Results:
- Microsporidial spores with characteristic morphology and staining properties were identified two weeks post-infection.
- Polymerase Chain Reaction (PCR) confirmed the isolated species as Encephalitozoon intestinalis.
Conclusions:
- A viable cell culture of microsporidia was successfully established from a clinical stool sample.
- This protocol enables the isolation and maintenance of native Colombian microsporidia strains.
- The established strain will facilitate future biochemical, immunological, and epidemiological studies.
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