Parasitology Research

, Volume 115, Issue 5, pp 2023–2034 | Cite as

Metabolic signatures of Besnoitia besnoiti-infected endothelial host cells and blockage of key metabolic pathways indicate high glycolytic and glutaminolytic needs of the parasite

  • A. Taubert
  • C. Hermosilla
  • L. M. R. Silva
  • A. Wieck
  • K. Failing
  • S. Mazurek
Original Paper
First Online:
Received:
Accepted:

Abstract

Besnoitia besnoiti is an obligate intracellular and emerging coccidian parasite of cattle with a significant economic impact on cattle industry. During acute infection, fast-proliferating tachyzoites are continuously formed mainly in endothelial host cells of infected animals. Given that offspring formation is a highly energy and cell building block demanding process, the parasite needs to exploit host cellular metabolism to meet its metabolic demands. Here, we analyzed the metabolic signatures of B. besnoiti-infected endothelial host cells and aimed to influence parasite proliferation by inhibitors of specific metabolic pathways. The following inhibitors were tested: fluoro 2-deoxy-d-glucose and 2-deoxy-d-glucose (FDG, DG; inhibitors of glycolysis), 6-diazo-5-oxo-l-norleucin (DON; inhibitor of glutaminolysis), dichloroacetate (DCA; inhibitor of pyruvate dehydrogenase kinase which favorites channeling of glucose carbons into the TCA cycle) and adenosine-monophosphate (AMP; inhibitor of ribose 5-P synthesis). Overall, B. besnoiti infections of bovine endothelial cells induced a significant and infection rate-dependent increase of glucose, lactate, glutamine, glutamate, pyruvate, alanine, and serine conversion rates which together indicate a parasite-triggered up-regulation of glycolysis and glutaminolysis. Thus, addition of DON, FDG, and DG into the cultivation medium of B. besnoiti infected endothelial cells led to a dose-dependent inhibition of parasite replication (4 μM DON, 99.5 % inhibition; 2 mM FDG, 99.1 % inhibition; 2 mM DG, 93 % inhibition; and 8 mM DCA, 71.9 % inhibition). In contrast, AMP had no significant effects on total tachyzoite production up to a concentration of 20 mM. Together, these data may open new strategies for the development of therapeutics for B. besnoiti infections.

Keywords

Coccidia Metabolic signature Glycolysis Glutaminolysis DON 2-Deoxy-d-glucose 
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Notes

Acknowledgments

The authors thank A. Wehrend (Clinic for Obstetrics, Gynecology and Andrology of Large and Small Animals, Justus Liebig University, Giessen, Germany) for the continuous supply of bovine umbilical cords. We also acknowledge the outstanding work of B. Hofmann in all cell culture experiments.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • A. Taubert
    • 1
  • C. Hermosilla
    • 1
  • L. M. R. Silva
    • 1
  • A. Wieck
    • 2
  • K. Failing
    • 3
  • S. Mazurek
    • 2
  1. 1.Institute of Parasitology, Biomedical Research Center SeltersbergJustus Liebig University GiessenGiessenGermany
  2. 2.Institute of Veterinary Physiology and BiochemistryJustus Liebig University GiessenGiessenGermany
  3. 3.Unit for Biomathematics and Data ProcessingJustus Liebig University GiessenGiessenGermany

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