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Home / Equine Research Bank / Parasitol Res / Continuous in vitro cultivation of erythrocytic stages of Ba...
Parasitology research1995; 81(4); 355-358; doi: 10.1007/BF00931544

Continuous in vitro cultivation of erythrocytic stages of Babesia equi.

Abstract: The protozoan parasite Babesia equi, a causative agent of equine piroplasmosis, was continuously cultivated in horse erythrocytes. The parasites were isolated from a carrier horse at a time when no parasite was detected in a thin blood smear. The culture medium consisted of modified medium 199 supplemented with 40% non-heat-inactivated horse serum in a humidified atmosphere containing 5% CO2, 2% O2, and 93% N2 at 37 degrees C. Parasites were detected after 2 days in culture. When the percentage of parasitized erythrocytes (PPE) reached 1%, the cultures were transferred into a humidified atmosphere of 5% CO2 in air. After 7 days the cultures were split at a ratio of 1:2, and after another 5 days they were split at a ratio of 1:4. From them on, cultures were split at a ratio of 1:4 routinely at 2-day intervals. The PPE ranged between 10% and 25%. Supplementation with hypoxanthine was essential for the initiation and propagation of cultures. In established cultures, hypoxanthine could be replaced by equimolar concentrations of adenosine or guanosine. Parasites from cultures could be cryopreserved and resuscitated. Cultures were maintained for more than 300 days.
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Publication Date: 1995-01-01 PubMed ID: 7624296DOI: 10.1007/BF00931544Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses an approach for in vitro continuous cultivation of Babesia equi, a protozoan parasite causing equine piroplasmosis, in horse erythrocytes.

Research Objectives and Methods

  • The researchers aimed to find a reliable method to cultivate Babesia equi, a parasite responsible for equine piroplasmosis, in a lab setting.
  • Babesia equi was isolated from a carrier horse at a time no parasites were visually present in a thin blood smear.
  • The culture medium used in the experiment consisted of modified medium 199 supplemented with 40% non-heat-inactivated horse serum.
  • The ambient conditions for the cultivation environment included a humidified atmosphere containing 5% CO2, 2% O2, and 93% N2, and maintained at a temperature of 37 degrees Celsius.

Cultivation Process and Result

  • The researchers detected parasites in the culture after two days.
  • Once the percentage of parasitized erythrocytes (PPE) reached 1%, the cultures were moved to a humidified atmosphere of 5% CO2 in the air.
  • The cultures then went through a process of splitting at given interval: 7 days with a ratio of 1:2 and after another 5 days at a ratio of 1:4.
  • From this point, the cultures were divided regularly every two days at a ratio of 1:4.
  • The PPE was noted to fluctuate between 10% and 25% during this process.

Key Observations and Conclusion

  • Addition of hypoxanthine was found to be crucial for initiating and maintaining the cultures.
  • In original cultures, hypoxanthine could be replaced by equivalent concentrations of adenosine or guanosine, which are other purine derivatives commonly used in cell cultures.
  • The parasites developed in these cultures could be cryopreserved (preserved by freezing at very low temperatures) for future use.
  • These cultures remained viable and were maintained for over 300 days, providing a consistent in vitro method for growing the Babesia equi parasite.

Cite This Article

APA
Zweygarth E, Just MC, de Waal DT. (1995). Continuous in vitro cultivation of erythrocytic stages of Babesia equi. Parasitol Res, 81(4), 355-358. https://doi.org/10.1007/BF00931544

Publication

Parasitology research
ISSN: 0932-0113
NlmUniqueID: 8703571
Country: Germany
Language: English
Volume: 81
Issue: 4
Pages: 355-358

Researcher Affiliations

Zweygarth, E
  • Onderstepoort Veterinary Institute, South Africa.
Just, M C
    de Waal, D T

      MeSH Terms

      • Animals
      • Babesia / drug effects
      • Babesia / growth & development
      • Babesia / isolation & purification
      • Babesiosis / parasitology
      • Carrier State
      • Cells, Cultured
      • Cryopreservation
      • Culture Media
      • Erythrocytes / parasitology
      • Horse Diseases / parasitology
      • Horses / parasitology
      • Hypoxanthine
      • Hypoxanthines / pharmacology
      • Parasitology / methods

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      Citations

      This article has been cited 12 times.
      1. Allred DR. Integration of DNA Repair, Antigenic Variation, Cytoadhesion, and Chance in Babesia Survival: A Perspective. Front Cell Infect Microbiol 2022;12:869696.
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        doi: 10.3389/fphys.2018.01696pubmed: 30568595google scholar: lookup
      3. Silva MG, Villarino NF, Knowles DP, Suarez CE. Assessment of Draxxin(®) (tulathromycin) as an inhibitor of in vitro growth of Babesia bovis, Babesia bigemina and Theileria equi. Int J Parasitol Drugs Drug Resist 2018 Aug;8(2):265-270.
        doi: 10.1016/j.ijpddr.2018.04.004pubmed: 29689532google scholar: lookup
      4. Zweygarth E, Josemans AI. L-cysteine replaces microaerophilous culture conditions for the in vitro initiation of Theileria equi. Parasitol Res 2014 Jan;113(1):433-5.
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        doi: 10.1038/s41598-024-54327-ypubmed: 38351295google scholar: lookup
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