Produção Nacional
Culture and Propagation of Microsporidia
2014; Linguagem: Inglês
10.1128/9781555818227.ch11
AutoresGovinda S. Visvesvara, Hércules Moura, Gordon J. Leitch, David A. Schwartz,
Tópico(s)Insect symbiosis and bacterial influences
ResumoIsolation in culture should always be attempted even when a presumptive diagnosis has been made. This is particularly important for the purpose of establishing a bank of isolates to be used for antigenic, molecular, and biochemical analyses. The first attempt to culture microsporidia was made in 1937 by Trager, who was partially successful in establishing Nosema bombycis infection of a cell culture developed from the ovarian tube lining cells of the silkworm (Bombyx mori). Cultures of Encephalitozoon cuniculi were initiated in several different ways: by adding infected tissue explants to cultured cells, by allowing infected cells in explanted cells to grow, by allowing germination of spores in the presence of cells and thereafter infecting cells, and by scraping infected cells from infected cultures and adding them to fresh cell cultures. The first microsporidian isolated from a human, Vittaforma corneae (Nosema corneum) was cultured from a corneal biopsy that was shipped to the laboratory overnight in Hanks' balanced salt solution (HBSS). Only a few attempts have been made to isolate microsporidia into culture from feces because enteric bacteria and yeast usually overgrow the rich culture medium that is used, which impedes isolation of the fastidious microsporidia, especially Enterocytozoon bieneusi. Inoculated cell cultures should be examined frequently with an inverted microscope preferably equipped with phase-contrast or differential interference-contrast optics. One great advantage of obtaining spores from in vitro cultures is the absence of bacterial and fungal contaminants.
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