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In vitro effects of arylimidamides against Besnoitia besnoiti infection in Vero cells

Published online by Cambridge University Press:  24 February 2011

H. C. E. CORTES ,
N. MULLER ,
D. BOYKIN ,
C. E. STEPHENS  and
A. HEMPHILL
H. C. E. CORTES
Affiliation:
Laboratório de Parasitologia Victor Caeiro, Núcleo da Mitra, ICAAM, Universidade de Évora, Apartado 94, 7000-554 Évora, Portugal
N. MULLER
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
D. BOYKIN
Affiliation:
Department of Chemistry, Georgia State University, PO Box 4098, Atlanta, Georgia 30302-4098, USA
C. E. STEPHENS
Affiliation:
Department of Chemistry and Physics, Augusta State University, Augusta, Georgia 30904-2200, USA
A. HEMPHILL*
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
*
*Corresponding author:Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland. Tel: +41 31 631 2474. Fax: +41 31 631 2477. E-mail: hemphill@ipa.unibe
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Summary

The in vitro effects of 4 arylimidamides (DB811, DB786, DB750 and DB766) against the proliferative tachyzoite stage of the apicomplexan parasite Besnoitia besnoiti were investigated. These four compounds had been shown earlier to exhibit in vitro activities in the nanomolar range against the related apicomplexans Neospora caninum and Toxoplasma gondii. Real-time-PCR was used to assess B. besnoiti intracellular proliferation in vitro. Preliminary assessment by light microscopy identified DB811 and DB750 as the most promising compounds, while DB786 and DB766 were much less effective. Three-day-growth assays and quantitative real-time PCR was used for IC50 determination of DB811 (0·079 μM) and DB750 (0·56 μM). Complete growth inhibition was observed at 1·6 μM for DB 811 and 1·7 μM for DB750. However, when infected cultures were treated for 14 days, proliferation of parasites occurred again in cultures treated with DB750 from day 4 onwards, while the proliferation of DB811-treated tachyzoites remained inhibited. Electron microscopy of B. besnoiti-infected fibroblast cultures fixed and processed at different time-points following the initiation of drug treatments revealed that DB811 exerted a much higher degree of ultrastructural alterations compared to DB750. These results show that arylimidamides such as DB811 could potentially become an important addition to the anti-parasitic arsenal for food animal production, especially in cattle.

Keywords

arylimidamideBesnoitia besnoitiin vitro drug treatmentbesnoitiosis
Type
Research Article
Information
Parasitology , Volume 138 , Issue 5 , April 2011 , pp. 583 - 592
Copyright
Copyright © Cambridge University Press 2011

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