Culture of Neospora caninum in the presence of a Mycoplasma Removal Agent results in the selection of a mutant population of tachyzoites

Abstract

S U M M A R YMycoplasmas are common contaminants of eukaryotic cells grown in tissue culture. A commercially available Mycoplasma Removal Agent (MRA) was therefore assessed for its effect on tachyzoites of Neospora caninum, in order to determine its suitability for further use in parasite cell cultures. Analyses of tachyzoite and excreted-secreted proteins and antigens by SDS-PAGE and Western blotting show that MRA treatment results in the rapid selection of a mutant population that differs from the control and parental lines in its protein and antigen content. The treatment of N. caninum cultures with MRA is therefore not recommended for the eradication of Mycoplasma.

Safe, Effective, and Inexpensive Clearance of Mycoplasma Contamination from Cultures of Apicomplexan Parasites with Sparfloxacin

“…This poor selectivity is due to their effect on the parasite apicoplast, an essential chloroplast-derived organelle that retains translation and DNA replication machinery of cyanobacterial origin. The lone validated exception is Mycoplasma removal agent (MRA), which contains an undisclosed fluoroquinolone and has successfully been used to clear Mycoplasma from cultures of apicomplexan parasites, including P. falciparum ( 6 ), Toxoplasma gondii , and Neospora caninum ( 15 ). The use of MRA has some downsides: clearance of Mycoplasma with MRA can take 1 to 3 weeks at the recommended concentration of 0.5 μg/mL, and some Mycoplasma strains have shown resistance to clearance with MRA ( 12 , 13 ).…”

Safe, effective, and inexpensive clearance of mycoplasma contamination from cultures of apicomplexan parasites with Sparfloxacin

“…This poor selectivity is due the effect of these antibiotics on the parasite apicoplast, an essential chloroplast-derived organelle that retains translation and DNA-replication machinery of cyanobacterial origin. The lone validated exception is Mycoplasma Removal Agent (MRA), which contains an undisclosed fluoroquinolone and has successfully been used to clear mycoplasma from cultures of apicomplexan parasites, including P. falciparum 6 , Toxoplasma gondii , and Neospora caninum 15 . At the recommended concentration of 0.5 μg/mL clearance of mycoplasma with MRA can take 1-3 weeks, is relatively expensive at $270 per liter of culture medium, and some mycoplasma strains have shown resistance to clearance with MRA 12,13 .…”