Babesia equi erythrocytic stage continuously cultured in an enriched medium.
Abstract: Babesia equi was continuously cultured through 90 passages in an enriched chemically defined basal medium (HL-1) supplemented with 20% fetal bovine serum and serum replacement factors, including lipid-rich bovine serum albumin, bovine insulin, and human transferrin. Cryopreservation and subsequent recovery of B. equi were easily achieved. Inoculation of a splenectomized and an intact horse with cultured infected erythrocytes resulted in parasitemias and B. equi in vitro reisolation from both animals. In vitro forms of the parasite resembled in vivo forms. After establishment, parasitemias of 10-15% were commonly observed.
Publication Date: 1994-04-01 PubMed ID: 8158466
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The study presents the successful and continuous culture of the Babesia equi parasite, a cause of equine babesiosis, in a chemically enriched medium over 90 passages, also demonstrating the parasite can be cryopreserved and recovered, as well as infecting horses under experimental conditions.
Objective
The objective of this research was to create a sustainable and continuous culture of the Babesia equi parasite, a disease-causing micro-organism in horses, in a specially enriched medium. An added goal was to assess the possibility of cryopreservation and recovery of the parasite, and to observe its behavior when introduced into both a horse with a removed spleen and one with an intact spleen.
Methodology
- The parasites were cultured in a chemically defined enriched medium, known as HL-1. This medium was supplemented with 20% fetal bovine serum and other serum replacement factors. The replacements included lipid-rich bovine serum albumin, bovine insulin, and human transferrin.
- The process of culture was carried out continuously through 90 passages, which refers to 90 generations of the parasites.
- The researchers also conducted a procedure of cryopreservation, a process where organisms are preserved by cooling to very low temperatures, and then attempted to recover the parasites from this state.
- Two horses, one with a removed spleen (splenectomized) and one with an intact spleen, were inoculated with the cultured parasites. Following inoculation, the team observed parasitemias, or the presence of parasites in the blood,
- The team completed an in vitro reisolation of B. equi from both of these infected animals.
Results & Conclusions
- The study succeeded in its main aim of developing a continuous culture of Babesia equi in an enriched medium, with the culture flowing through 90 passages.
- The team was also successful in cryopreserving the parasites and subsequently recovering them, demonstrating the practicality of long-term storage of the parasites.
- After inoculation, both the splenectomized and the intact horse showed signs of parasitemias, and B. equi could be reisolated from them, illustrating the viability of these parasites after culture and their potential infectivity.
- The in vitro forms of the parasite were found to be similar to in vivo forms, indicating the cultured parasites maintained traits equivalent to naturally-occurring parasites.
- Following establishment, parasitemias of 10-15% were commonly observed, signifying a significant presence of these parasites in the blood post-infection.
Cite This Article
APA
Holman PJ, Chieves L, Frerichs WM, Olson D, Wagner GG.
(1994).
Babesia equi erythrocytic stage continuously cultured in an enriched medium.
J Parasitol, 80(2), 232-236.
Publication
Researcher Affiliations
- Department of Veterinary Pathobiology, Texas A&M University, College Station 77843-4467.
MeSH Terms
- Animals
- Babesia / growth & development
- Babesiosis / blood
- Babesiosis / parasitology
- Culture Media
- Erythrocytes / parasitology
- Horse Diseases / blood
- Horse Diseases / parasitology
- Horses
Citations
This article has been cited 13 times.- Li D, Wang L, Guan X, Wang S, Liu Q, Chen F, Zheng Y, He L, Zhao J. Establishment of Continuous In Vitro Culture of Babesia gibsoni by Using VP-SFM Medium with Low-Concentration Serum.. Microbiol Spectr 2023 Jun 15;11(3):e0025823.
- Jiang W, Wang S, Li D, Zhang Y, Luo W, Zhao J, He L. Continuous In Vitro Culture of Babesia duncani in a Serum-Free Medium.. Cells 2023 Feb 2;12(3).
- Grimsley M, Hicks J, Zeineldin M, Murphy G, Sigafoose T. Complete Genome Sequence of Theileria equi NVSL354.. Microbiol Resour Announc 2023 Feb 16;12(2):e0080922.
- McCormack KA, Alhaboubi A, Pollard DA, Fuller L, Holman PJ. In vitro cultivation of Babesia duncani (Apicomplexa: Babesiidae), a zoonotic hemoprotozoan, using infected blood from Syrian hamsters (Mesocricetus auratus).. Parasitol Res 2019 Aug;118(8):2409-2417.
- Hines SA, Ramsay JD, Kappmeyer LS, Lau AO, Ojo KK, Van Voorhis WC, Knowles DP, Mealey RH. Theileria equi isolates vary in susceptibility to imidocarb dipropionate but demonstrate uniform in vitro susceptibility to a bumped kinase inhibitor.. Parasit Vectors 2015 Jan 20;8:33.
- 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.
- Adaszek Ł, Winiarczyk S. In vitro cultivation of Babesia canis canis parasites isolated from dogs in Poland.. Parasitol Res 2011 May;108(5):1303-7.
- Holman PJ, Spencer AM, Droleskey RE, Goethert HK, Telford SR 3rd. In vitro cultivation of a zoonotic Babesia sp. isolated from eastern cottontail rabbits (Sylvilagus floridanus) on Nantucket Island, Massachusetts.. J Clin Microbiol 2005 Aug;43(8):3995-4001.
- Holman PJ, Bendele KG, Schoelkopf L, Jones-Witthuhn RL, Jones SO. Ribosomal RNA analysis of Babesia odocoilei isolates from farmed reindeer (Rangifer tarandus tarandus) and elk (Cervus elaphus canadensis) in Wisconsin.. Parasitol Res 2003 Nov;91(5):378-83.
- Schuster FL. Cultivation of Babesia and Babesia-like blood parasites: agents of an emerging zoonotic disease.. Clin Microbiol Rev 2002 Jul;15(3):365-73.
- Samuel T, Böse R. The 18 kDa antigen of Theileria equi is a specific but less abundant protein also expressed by parasites cultured in vitro.. Vet Res Commun 2001 Apr;25(3):169-78.
- Holman PJ, Hietala SK, Kayashima LR, Olson D, Waghela SD, Wagner GG. Case report: field-acquired subclinical Babesia equi infection confirmed by in vitro culture.. J Clin Microbiol 1997 Feb;35(2):474-6.
- Zweygarth E, Just MC, de Waal DT. Continuous in vitro cultivation of erythrocytic stages of Babesia equi.. Parasitol Res 1995;81(4):355-8.
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