Preservation Evaluation of Mycoplasma fermentans in Different Conditions

Author(s): Rivera A., Rodriguez-Preval N., Giono-Cerezo S., Sanchez-Hernandez J. A.

Different species of mycoplasmas are considered pathogenic in humans, in addition to being characterized by their difficult cultivation in the laboratory and demanding nutritional requirements, their preservation is complicated for future research based on clinical isolates. The objective of this work was to evaluate the viability of Mycoplasma fermentans preserving it with different cryopreservatives and temperatures. The preservation evaluation of Mycoplasma fermentans was performed with SP4 broth supplemented with 40% fetal bovine serum (FBS) and 10% dimethylsulfoxide (DMSO), 40% gelatin (GEL) and 10% DMSO, 40% glycerol (GLI) and 10% DMSO, and supplement free SP4 broth as a control. 10 mL of each of the SP4 broths supplemented with 10 mL of Mycoplasma fermentans culture were mixed at a concentration of 1X106 CFU/mL, stored at -20oC and -80oC for twelve months. Every two months, it was thawed and its viability was evaluated on SP4 agar and quantification of the CFU/mL. Trials with fetal bovine serum, gelatin and glycerol at -20oC and -80oC showed efficiency in keeping Mycoplasma fermentans viable during the twelve months. Glycerol being the cryopreservative agent that favored the greatest viability, showing significant difference (P <0.05), with respect to the control group. Supplementation of SP4 broth with glycerol showed the best viability for Mycoplasma fermentans.

Preservation, Mycoplasma fermentans, Fetal Bovine Serum, Glycerol, Dimethylsulfoxide

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