Comparison of various atmospheric conditions for isolation and subcultivation of Mycoplasma hyorhinis from cell cultures
- 38 Downloads
The efficiency of aerobic incubation was compared with incubation under various oxygen and carbon dioxide conditions for the isolation and subcultivation of three strains of Mycoplasma hyorhinis from VERO-cell cultures and subcultivation of three laboratory strains.
Under anaerobic conditions with a low oxidation-reduction potential (at or below-115 mV) as obtained in jars, with catalysts, containing mixtures of 5%–10% CO2 in H2, very poor or no growth of any of the six M. hyorhinis strains was observed. When traces of oxygen were present (that is, under conditions with higher oxidation-reduction potentials, e.g. when omitting the catalyst in the above gas mixtures or in 5% CO2+95% N2) isolation from cell cultures was successful in most tests, but subcultivation of these primary isolates was seldom possible under these semi-anaerobic conditions. However, in most cases these primary isolates could be subcultivated aerobically, although aerobic conditions were unsatisfactory for isolation in about half of the experiments.
Isolation of M. hyorhinis was optimal in 5% O2+95% N2, under which condition the isolates could also always be subcultivated. Isolation failed occasionally when 5% O2+5% CO2+90% N2 was used, thus indicating that 5% CO2 was slightly inhibitory. 5% CO2 in air and 10% CO2 either in air, H2 or N2 were also inadequate for isolation from cell cultures.
In contrast to the findings with these cell culture-adapted M. hyorhinis strains, the laboratory strains could be subcultivated easily under all conditions tested except those with an oxidation-reduction potential at or below-115 mV; 100% CO2 was inhibitory for all 6 strains.
Our findings may partly explain why M. hyorhinis is often considered “noncultivable” on artificial media once adapted to cell cultures. The findings emphasize the need to employ also a micro-aerophilic condition (5% O2 in 95% N2) in the examination of cell cultures for mycoplasma.
KeywordsCell Culture Carbon Dioxide Anaerobic Condition Aerobic Condition Atmospheric Condition
Unable to display preview. Download preview PDF.
- Barile, M. F. 1979. Mycoplasma-tissue cell interactions. p. 425–474. In J. G. Tully and R. F. Whitcomb (eds), The mycoplasmas. Vol. 2 — Academic Press, New York, San Francisco and London.Google Scholar
- Barile, M. F. and Schimke, R. T. 1963. A rapid chemical method for detecting PPLO contamination of tissue cell cultures. — Proc. Soc. Exp. Biol. Med. 114: 676–679.Google Scholar
- Butler, M. and Leach, R. H. 1964. A mycoplasma which induces acidity and cytopathic effect in tissue culture. — J. Gen. Microbiol. 34: 285–294.Google Scholar
- Central Veterinary Laboratory, Weybridge 1977. Specifications for the production and control of avian live virus vaccines, 2nd edition. Ministry of Agriculture, Fisheries and Food, appendix III, 22.Google Scholar
- Del Guidice, R. A., Gardella, R. A. and Hopps, H. E. 1980. Cultivation of formerly non-cultivable strains of Mycoplasma hyorhinis. — Curr. Microbiol. 4: 75–80.Google Scholar
- Dinter, Z. and Taylor-Robinson, D. 1969. Susceptibility and resistance of various strains of Mycoplasma hyorhinis to antisera, polymyxins and low pH values. — J. Gen. Microbiol. 57: 263–272.Google Scholar
- Herderschêe, D. 1963. An improved medium for the cultivation of the Eaton agent. — Antonie van Leeuwenhoek 29: 154–156.Google Scholar
- Hers, J. F. Ph. and Masurel, M. 1967. Infection with Mycoplasma pneumoniae in civilians in the Netherlands. — Ann. N. Y. Acad. Sci. 143: 447–460.Google Scholar
- Holdeman, L. V., Cato, E. P., Moore (eds). 1977. Anaerobe laboratory manual, 4th edition. — Anaerobe Laboratory Virginia Polytechnic Institute and State University, Blacksburg.Google Scholar
- Hopps, H. E., Meyer, B. C., Barile, M. F. and Del Guidice, R. A. 1973. Problems concerning “non-cultivable” contaminants in tissue cultures. — Ann. N. Y. Acad. Sci. 225: 265–276.Google Scholar
- McGarrity, G. J. and Coriell, L. L. 1973. Detection of anaerobic mycoplasmas in cell cultures. — In Vitro 9: 17–18.Google Scholar
- Morgan, J. F., Mortan, H. J. and Parker, R. 1950. Nutrition of animal cells in tissue culture. I. Initial studies on a synthetic medium. — Proc. Soc. Exp. Biol. Med. 73: 1–8.Google Scholar
- Morton, H. E., Smith, P. F. and Leberman, P. R., 1951. Investigation of the cultivation of pleuropneumonia-like organisms from man. Am. J. Syph., Gonorrhea vener. Dis. 35: 361–369.Google Scholar
- Polak-Vogelzang, A. A., Reijgers, R. and Hekkens, F. E. N. 1980. Isolation of Mycoplasma hyorhinis and Mycoplasma fermentans from cell cultures. — J. Biol. Standard. 8: 243–254. 8: 243–254.Google Scholar
- Seip, W. F. and Evans, G. L. 1980. Atmospheric analysis and redox potentials of culture media in the Gas Pak system. — J. Clin. Microbiol. 11: 226–233.Google Scholar