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Cultivation and partial characterization of spiroplasmas in cell cultures

Cultivation and partial characterization of spiroplasmas in cell cultures

Infection and Immunity 35(1): 296-304

Spiroplasmas were propagated in the Drosophila melanogaster cell line Dm-1. Spiroplasma citri and unidentified strains (corn stunt organism, 277F [tick isolate], powder puff, BNR-1, honey bee and OBMG) grew to 108-109 colony forming units per ml and could be passaged. Cytopathic effect (CPE) varied with the infecting spiroplasma. The honey bee isolate killed DM-1 within 2-4 days and produced CPE in 4 mammalian cells tested. At 25.degree. C, suckling mouse cataract agent produced no CPE in Dm-1 cells. Dm-1 cells did not support growth of the spiroplasmal sex ratio organism. Spiroplasmas could be detected in the cell cultures by agar inoculation, dark-field microscopy, scanning electron microscopy and DNA fluorescent staining. The uridine phosphorylase test showed significant levels of conversion of [14C]uridine to [14C]uracil for all but some plant isolates: S. citri, corn stunt organism, lettuce, cactus and powder puff strains. These are the 1st mycoplasmas to lack the enzyme. Primary isolations of corn stunt organism from infected corn plants were made in Dm-1 and I-XII cultures. The course of corn stunt organism infection of Dm-1 was monitored for 3 passages. The use of agarose and Dienes staining of the colonies improved growth and colony counting of corn stunt organism. The number of viable infected Dm-1 cells decreased from 1.2 .times. 107 at passage 1 to 7.0 .times. 106 at passage 2 and 3 .times. 105 at passage 3.

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Accession: 001059691

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PMID: 6797950

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