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Viability assays of intra-erythrocytic organisms using fluorescent dyes

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Fletcher, T. I., Wigg, J. L., Rolls, P. J. and de Vos, A. J. (2009) Viability assays of intra-erythrocytic organisms using fluorescent dyes. Veterinary Parasitology, 163 (1-2). pp. 144-147. ISSN 0304-4017

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Article Link: https://doi.org/10.1016/j.vetpar.2009.03.029


Three intra-erythrocytic tick fever organisms of cattle (Babesia bovis, Babesia bigemina and Anaplasma centrale) were subjected to a range of stressors, including heat, storage over time, specific chemotherapy and cryopreservation. Various stains, both alone and in combination, were used in an attempt to assess viability of these organisms before and after the stressors were applied. Carboxyfluorescein diacetate succinimidyl ester (CFSE) stained live Babesia spp. very well while fluorescein diacetate (FDA) stained A. centrale successfully. Propidium iodide (PI) and ethidium-homodimer-1 (Eth-D) were used as counter stains to identify dead organisms. Stain combinations allowed differentiation between living and dead Babesia organisms after exposure to heat and after chemotherapy. PI and Eth-D as counter stains were of little value after deglycerolisation of cryopreserved organisms. Possible reasons for this limited success in determining death or viability of tick fever organisms after some treatments include the impermeability of red blood cells to PI and Eth-D counter stains or the loss of live and/or dead organisms during sample processing.

Item Type:Article
Keywords:Viability assay; Fluorescent staining; Babesia bigemina; Babesia bovis; Anaplasma centrale; CFSE; FDA; Propidium iodide; Eth-D
Subjects:Veterinary medicine > Veterinary epidemiology. Epizootiology
Veterinary medicine > Veterinary parasitology
Veterinary medicine > Diseases of special classes of animals > Cattle
Live Archive:21 Feb 2024 02:27
Last Modified:21 Feb 2024 02:27

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