Thallium acetate

All laboratory manipulations with Mycoplasma were carried out under BSL2 conditions (Mwirigi et al., 2016 (link)). Mycoplasma mycoides subsp. capri, Mycoplasma mycoides subsp. mycoides and Mycoplasma capricolum subsp capricolum (Table 2, Table 3) were cultured in Pleuropneumonia Like-Organism (PPLO) broth (Difco™ PPLO Broth) media prepared as follows: 21 g of PPLO was dissolved in 700 ml of distilled water and autoclaved for 15 min at 121 °C. The mixture was cooled in a water bath to 55 °C and supplemented with phenol red (Carl Roth GmbH) to a final concentration of 3%, 200 ml horse serum (Sigma), 0.25% of glucose (Carl Roth GmbH), 0.15% of penicillin G (Carl Roth GmbH) and 0.25% of thallium acetate (Carl Roth GmbH). A forty-eight well plate was used to grow Mycoplasma strains where they were incubated at 37 °C for a minimum period of seven days. Growth of Mycoplasma cells was determined by color change from red to yellow (Stemke and Robertson, 1982 (link)), as a result of pH change due to the growth of Mycoplasma strains. Stock cultures of both Mmc, Mmm and Mcc liquid cultures were grown to a density of approximately 1–3×10⁶ cells per ml as measured by the colony forming units method (Stemke and Robertson, 1982 (link)) and cryopreserved at −80 °C.

All laboratory manipulations with Mycoplasma were carried out under BSL2 conditions (Mwirigi et al., 2016 (link)). Two Mycoplasma mycoides subsp. capri (211/94 and 95010), two Mycoplasma mycoides subsp. Mycoides (Afadé and Gladysdale), and one Mycoplasma capricolum subsp capricolum (6443-90) (Tables 2, 3) were cultured in Pleuropneumonia Like-Organism (PPLO) broth (Difco^TM PPLO Broth) media prepared as follows: 21 g of PPLO was dissolved in 700 ml of distilled water and autoclaved for 15 min at 121°C. The mixture was cooled in a water bath to 55°C and supplemented with phenol red (Carl Roth GmbH) to a final concentration of 3%, 200 ml horse serum (Sigma), 0.25% of glucose (Carl Roth GmbH), 0.15% of penicillin G (Carl Roth GmbH) and 0.25% of thallium acetate (Carl Roth GmbH). A 48 well-plate was used to grow Mycoplasma strains where they were incubated at 37°C for a minimum period of seven days. Growth of Mycoplasma cells was determined by color change from red to yellow (Stemke and Robertson, 1982 (link)), as a result of pH change due to the growth. Stock cultures of Mmc, Mmm, and Mcc were grown to a density of approximately 1–3 10⁶ cells per ml as measured by both the colony forming units (CFU) method (Stemke and Robertson, 1982 (link)) and the use of FCM (Assunção et al., 2006 (link)) after cryopreservation in a freezer at -80°C for further antimycoplasmal activity tests.