Abstract
In this study, we report Babesia bigemina proliferation in culture medium free of components of animal origin supplemented with a lipid mixture. Babesia bigemina continuously proliferated in VP-SFM with a higher percent parasitized erythrocyte as compare to using other animal component-free culture media. Compared with Advanced DMEM/F12 (ADMEM/F12), VP-SFM had a similar percent parasitized erythrocyte (PPE). Supplementation of VP-SF with a lipid acid mixture improved B. bigemina proliferation in vitro culture, with a maximum PPE of 11.3%. Growth of B. bigemina in a perfusion bioreactor using VP-SFM medium supplemented with lipid mixture resulted in a PPE above 28%. In conclusion, we demonstrated that B. bigemina proliferated in an animal component-free medium supplemented with the fatty acid mixture. This innovation to B. bigemina in vitro culture method presented herein is an important source of biological material for live vaccine production and understanding the mechanisms and molecules involved in parasite attachment and invasion of bovine erythrocytes.
Highlights
Bovine babesiosis is a tick-borne disease caused by Babesia bigemina and Babesia bovis
Discussionbetween VP-SFM + lipid mixture and ADMEM/F12 control medium. (B) Giemsa-stained blood smear showing B. bigemina parasitized erythrocytes. This is the first report of the continuous in vitro growth of B. bigemina in a medium free of components of animal origin supplemented with a lipid mixture
We showed that VP-SFM combined with a lipid mixture achieved high efficiency of B. bigemina proliferation in a perfusion bioreactor reaching a maximum percentage of parasitized erythrocytes (PPE) of 29.63%
Summary
Bovine babesiosis is a tick-borne disease caused by Babesia bigemina and Babesia bovis. This disease affects bovines in tropical and subtropical areas and creates a negative economic impact on the cattle industry [1]. The success of Babesia in vitro growth in erythrocytes using the microaerophilic stationary phase system (MASP) facilitated the study of gene regulation necessary for parasite development, drug efficiency evaluation, and Babesia parasite attenuation for vaccines [4,5,6,7]. More than 100 species of Babesia have been isolated. Few Babesia species have been successfully cultivated in vitro system [10]. The primary concern is that to maintain continuous in vitro Babesia growth is the requirement of high sera concentrations in the growth medium
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