EC Microbiology

Research Article Volume 19 Issue 4 - 2023

Antibiotic Treatment for Mycoplasma Contamination in Plasmodium falciparum Cultures

Suman Tamang and Vineeta Singh*

Parasite and Host Biology Group, ICMR-National Institute of Malaria Research (NIMR), Dwarka Sector 8, New Delhi, India

*Corresponding Author: Vineeta Singh, Parasite and Host Biology Group, ICMR-National Institute of Malaria Research (NIMR), Dwarka Sector 8, New Delhi, India.
Received: January 24, 2023; Published: March 27, 2023

Mycoplasmas, the simplest and smallest living prokaryotes are the most common contaminants that pose a serious impediment in the long-term maintenance of Plasmodium falciparum (Pf) cultures. The comparative efficacy of two antibiotics - Mycoplasma Removal Agent (MRA) and ciprofloxacin at different concentrations for eliminating Mycoplasma contamination in Pf cultures was determined. Mycoplasma contaminated and uncontaminated Pf cultures were maintained for 12 days and Giemsa-stained thin smears were prepared daily to monitor parasite growth. The presence of Mycoplasma was ascertained daily by 16S rRNA PCR kit. We found that MRA successfully eliminated Mycoplasma from contaminated culture at both 0.5 (7-day treatment) and 1 µg/mL (5-day treatment) concentrations, without affecting parasite growth but treatment with three different concentrations of ciprofloxacin (5, 10 and 20 µg/mL) showed an anti-plasmodial activity limiting its usefulness in Pf cultures.

Keywords: Plasmodium falciparum; Mycoplasma Removal Agent (MRA); Ciprofloxacin; Mycoplasma; Culture Contamination

  1. Olarerin-George AO and Hogenesch JB. “Assessing the prevalence of mycoplasma contamination in cell culture via a survey of NCBI’s RNA-seq archive”. Nucleic Acids Research 43 (2015): 2535-2542.
  2. Rottem S and Barile MF. “Beware of mycoplasmas”. Trends in Biotechnology 11 (1993): 143-151.
  3. Armstrong SE., et al. “The scope of mycoplasma contamination within the biopharmaceutical industry”. Biologicals 38 (2010): 211-213.
  4. Rottem S., et al. “Contamination of Tissue Cultures by Mycoplasmas”. Biomedical Tissue Culture (2012).
  5. Chernova OA., et al. “Mycoplasmas and Their Antibiotic Resistance: The Problems and Prospects in Controlling Infections”. Acta Naturae 8 (2016): 24.
  6. Gautier-Bouchardon Av. “Antimicrobial Resistance in Mycoplasma spp”. Microbiology Spectrum (2018): 6.
  7. Visser IJR., et al. “Failure of antibiotics gentamycin, tylosin, lincomycin and spectinomycin to eliminate Mycoplasma bovis in artificially infected frozen bovine semen”. Theriogenology 51 (1999): 689-697.
  8. Schultz KK., et al. “Optimization of an antibiotic sensitivity assay for Mycoplasma hyosynoviae and susceptibility profiles of field isolates from 1997 to 2011”. Veterinary Microbiology 158 (2012): 104-108.
  9. Drexler HG and Uphoff CC. “Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention”. Cytotechnology 39 (2002): 75-90.
  10. Rowe JA., et al. “Implications of mycoplasma contamination in Plasmodium falciparum cultures and methods for its detection and eradication”. Molecular and Biochemical Parasitology (1998): 92.
  11. Rottem S. “Interaction of mycoplasmas with host cells”. Physiological Reviews 83 (2003): 417-432.
  12. Turrini F., et al. “Mycoplasma contamination of Plasmodium cultures - a case of parasite parasitism”. Parasitology Today 13 (1997): 367-368.
  13. Singh S., et al. “Treatment and control of mycoplasma contamination in Plasmodium falciparum culture”. Parasitology Research 104 (2008): 181-184.
  14. Agarwal P., et al. “Management of mycoplasma contamination in in vitro culture of Plasmodium falciparum without antibiotic treatment - a preliminary report”. Microbiological Research 164 (2013): 211-215.
  15. Stanisic DI., et al. “Development of cultured Plasmodium falciparum blood-stage malaria cell banks for early phase in vivo clinical trial assessment of anti-malaria drugs and vaccines”. Malaria Journal (2015): 14.
  16. Pall Corporation. Mycoplasma Reduction - Laboratory | Pall Corporation n.d (2022).
  17. Rawadi G and Dussurget O. “Advances in PCR-based detection of mycoplasmas contaminating cell cultures”. PCR Techniques and Applications 4 (1995): 199-208.
  18. Uphoff CC and Drexler HG. “Eradication of Mycoplasma contaminations from cell cultures”. Current Protocols in Molecular Biology (2014): 106.
  19. Geary TG and Jensen JB. “Effects of Antibiotics on Plasmodium Falciparum in Vitro”. The American Journal of Tropical Medicine and Hygiene 32 (1983): 221-225.
  20. Pradines B., et al. “In Vitro Activities of Antibiotics against Plasmodium falciparum Are Inhibited by Iron”. Antimicrobial Agents and Chemotherapy 45 (2001): 1746.
  21. Gaillard T., et al. “Antibiotics in malaria therapy: which antibiotics except tetracyclines and macrolides may be used against malaria?” Malaria Journal 15 (2016): 1-10.
  22. Uphoff CC., et al. “Elimination of mycoplasma from leukemia-lymphoma cell lines using antibiotics”. Leukemia 16 (2002): 284-288.
  23. Romorini L., et al. “Effect of antibiotics against Mycoplasma on human embryonic stem cells undifferentiated status, pluripotency, cell viability and growth”. PLoS One (2013): 8.
  24. Taylor-Robinson D and Bébéar C. “Antibiotic susceptibilities of mycoplasmas and treatment of mycoplasmal infections”. Journal of Antimicrobial Chemotherapy 40 (1997): 622-630.
  25. Drexler HG., et al. “Treatment of mycoplasma contamination in a large panel of cell cultures”. In Vitro Cellular and Developmental Biology 30A (1994): 344-347.
  26. Trager W and Jensen JB. “Human malaria parasites in continuous culture”. Science 193 (1976): 673-675.
  27. World Health Organization. Malaria parasite counting. Malaria Microscopy Standard Operating Procedure (2016).
  28. HiMedia-EZdetect TM. “Animal Cell Culture | Mycoplasma Detection and Elimination Platform | EZdetect TM Mycoplasma Detection Kits n.d (2016).
  29. Harasawa R., et al. “Detection and tentative identification of dominant mycoplasma species in cell cultures by restriction analysis of the 16S-23S rRNA intergenic spacer regions”. Microbiological Research 144 (1993): 489-493.
  30. Sasaki T., et al. “Inhibition of growth of mammalian cell cultures by extracts of arginine-utilizing mycoplasmas”. In Vitro 20 (1984): 369-375.
  31. Miller CJ., et al. “Mycoplasma infection significantly alters microarray gene expression profiles”. Biotechniques 35 (2003): 812-814.
  32. Somasundaram C., et al. “Use of ciprofloxacin and BM-Cyclin in mycoplasma decontamination”. In Vitro Cellular and Developmental Biology 28A (1992): 708-710.
  33. Schmitt K., et al. “A safe and efficient method for elimination of cell culture mycoplasmas using ciprofloxacin”. The Journal of Immunological Methods 109 (1988): 17-25.
  34. Mowles JM. “The use of ciprofloxacin for the elimination of mycoplasma from naturally infected cell lines”. Cytotechnology 1 (1988): 355-358.
  35. Mahmoudi N., et al. “In vitro activities of 25 quinolones and fluoroquinolones against liver and blood stage Plasmodium spp”. Antimicrobial Agents and Chemotherapy 47 (2003): 2636-2639.
  36. Bové JM. “Molecular features of mollicutes”. Clinical Infectious Diseases1 (1993): S10-S31.
  37. Nikfarjam L and Farzaneh P. “Prevention and Detection of Mycoplasma Contamination in Cell Culture”. Cell Journal 13 (2012): 203.
  38. Razin S., et al. “Adhesion of mycoplasmas to eukaryotic cells”. Ciba Foundation Symposium 80 (1981): 98-118.
  39. Peterson BW., et al. “Bacterial Cell Surface Damage Due to Centrifugal Compaction”. Applied and Environmental Microbiology 78 (2012): 120.
  40. Kiss F., et al. “The effect of centrifugation at various g force levels on rheological properties of rat, dog, pig and human red blood cells”. Clinical Hemorheology and Microcirculation 62 (2016): 215-227.
  41. Malave-Ramos DR., et al. “Safe, effective, and inexpensive clearance of mycoplasma contamination from cultures of apicomplexan parasites with Sparfloxacin”. Bio Rxiv (2022): 505931.

Suman Tamang and Vineeta Singh. Antibiotic Treatment for Mycoplasma Contamination in Plasmodium falciparum Cultures. EC Microbiology  19.4 (2023): 58-67.