Incidence and Detection of Pleuropneumonia-Like Organisms in Cell Cultures by Fluorescent Antibody and Cultural Procedures

Abstract

B3ARILE, MICHAEL F. (National Institutes of Health, Bethesda, AMd.), WX ALTER F. iMALIZIA, AND DONALD B. RIGGS. Incidence and detection of pleuropneumonia-like organisms in cell cultures by fluorescent antibody and cultural procedures. J. Bacteriol. 84:130-136. 1962-A total of 102 tissue-cell cultures from 17 separate laboratories was examined for pleuropneumonia-like organisms (PPLO) by the fluorescent antibody and cultural procedures. PPLO were isolated from 48 of the 49 tissue-cell cultures found positive for PPLO by the fluorescent antibody l)rocedure, and results of the two procedures agreed in 101 of the 102 (99%) cases. PPLO were isolated from none of 10 primary-cell cultures prepared from six animal species and from 48 of 92 (52%) continuous-cell cultures prepared from eight animal species. Cells grown in media containing antibiotics were more frequently contaminated with PPLO (72%) than cells grown in antibiotic-free media (7 %). Cultures (91%) from tissue-cultureproducing laboratories and cultures (76%) used for propagation of microorganisms were contaminated with PPLO, although none used for tissueculture metabolic studies was contaminated. In addition, our findings support the view that PPLO contamination of cell cultures is probably owing to bacterial contaminants which revert to L forms in the presence of antibiotics. Continuous tissue-cell cultures are frequently contaminated with pleuropneunmonia-like organisms (PPLO) or L forms of bacteria or both

The serological differentiation ofMycoplasmastrains (pleuropneumonia-like organisms) from various sources

“…The results of this investigation as well as those of Collier (1957), Corriell et al (1960) and Bailey et al (1961) establish that a large proportion of PPLO from tissue cultures are M. hominis type 1. Barile, Malizia & Riggs (1962), using a fluorescent antibody technique, found that 48 strains from tissue cultures were antigenically related, but did not identify the serotype. One of these, H.Ep2, proved in this present survey to be M. hominis type 1-further evidence of the widespread contamination of cell lines by this species.…”

Potential Use of Surface-active Agents for Controlling Mycoplasma Contamination in Animal Cell Cultures

“…Mycoplasma species was first reported by Robinson et al (14) and has since been found to be a relatively common occurrence (2,3,6,15). The effects of mycoplasmata contaminants in cell cultures are numerous and depend on the species involved and the type of cell infected.…”

Respiratory infection of lambs with Mycoplasma ovipneumoniae

“…The indirect immunofluorescent technique (IIP) of Barile et al (1962) and Del Guidice et al 1967 Control slides were treated with normal horse serum or PBS in place of specific antiserum. All isolates of M. ovipneumoniae had "typical" colony morphology, i.e., centerless and lacy type colonies when grown on FA, while all other mycoplasmas isolated had "fried egg" type colony morphology (Figs 1 and 2).…”