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

TitleSafe, Effective, and Inexpensive Clearance of Mycoplasma Contamination from Cultures of Apicomplexan Parasites with Sparfloxacin.
Publication TypeJournal Article
Year of Publication2022
AuthorsMalave-Ramos DR, Kennedy K, Key MN, Dou Z, Kafsack BFC
JournalMicrobiol Spectr
Volume10
Issue5
Paginatione0349722
Date Published2022 Oct 26
ISSN2165-0497
KeywordsAnimals, Anti-Bacterial Agents, Fluoroquinolones, Mycoplasma, Parasites
Abstract

Most commercial products cannot be used for clearance of Mycoplasma contamination from cultures of apicomplexan parasites due to the parasites' dependence on the apicoplast, an essential organelle with DNA replication and translation machinery of cyanobacterial origin. The lone exception, mycoplasma removal agent (MRA), is relatively expensive, and some mycoplasma strains have shown resistance to clearance with MRA. Here, we report that the fluoroquinolone antibiotic sparfloxacin is a safe, effective, and inexpensive alternative for treatment of mycoplasma contamination in cultures of apicomplexan parasites. Sparfloxacin cleared both MRA-sensitive and MRA-resistant mycoplasma species from P. falciparum cultures at 1 and 4 μg/mL, respectively. We show that cultures of three different apicomplexan parasites can be maintained at concentrations of sparfloxacin required to clear mycoplasma without resulting in substantial deleterious effects on parasite growth. We also describe an alternative low-cost, in-house PCR assay for detecting mycoplasma. These findings will be useful to laboratories maintaining apicomplexan parasites in vitro, especially in low-resource environments, where the high cost of commercial products creates an economic barrier for detecting and eliminating mycoplasma from culture. IMPORTANCE These findings will be useful to laboratories maintaining apicomplexan parasites in vitro, especially in low-resource environments, where the high cost of commercial products creates an economic barrier for detecting and eliminating Mycoplasma from culture.

DOI10.1128/spectrum.03497-22
Alternate JournalMicrobiol Spectr
PubMed ID36190416
PubMed Central IDPMC9603166
Grant ListR01 AI141965 / AI / NIAID NIH HHS / United States
R01 AI143707 / AI / NIAID NIH HHS / United States
R21 AI166436 / AI / NIAID NIH HHS / United States

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