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In vitro efficacy of nitro- and bromo-thiazolyl-salicylamide compounds (thiazolides) against Besnoitia besnoiti infection in Vero cells

Published online by Cambridge University Press:  19 February 2007

H. C. E. CORTES ,
N. MUELLER ,
M. ESPOSITO ,
A. LEITÃO ,
A. NAGULESWARAN  and
A. HEMPHILL
H. C. E. CORTES*
Affiliation:
Laboratório de Parasitologia Victor Caeiro, Núcleo da Mitra, ICAM, Universidade de Évora, Apartado 94, 7000-554 Évora, Portugal
N. MUELLER
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
M. ESPOSITO
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
A. LEITÃO
Affiliation:
Instituto de Investigação Científica Tropical, CIISA, Faculdade de Medicina Veterinária, Av. da Universidade Técnica, 1300-447 Lisboa, Portugal
A. NAGULESWARAN
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
A. HEMPHILL*
Affiliation:
Institute of Parasitology, University of Berne, Längass-Strasse 122, CH-3012 Berne, Switzerland
*
*Corresponding authors: Laboratório de Parasitologia Victor Caeiro, Núcleo da Mitra, ICAM, Universidade de Évora, Apartado 94, 7000-554 Évora, Portugal. Tel: 00351 266760860. Fax: 00351 266 760912. E-mail: hcec@uevora.pt; Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012 Berne, Switzerland. Tel: +41 31 6312384. Fax: +41 31 6312477. E-mail: hemphill@ipa.unibe.ch
*Corresponding authors: Laboratório de Parasitologia Victor Caeiro, Núcleo da Mitra, ICAM, Universidade de Évora, Apartado 94, 7000-554 Évora, Portugal. Tel: 00351 266760860. Fax: 00351 266 760912. E-mail: hcec@uevora.pt; Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012 Berne, Switzerland. Tel: +41 31 6312384. Fax: +41 31 6312477. E-mail: hemphill@ipa.unibe.ch
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Summary

Nitazoxanide (NTZ) and its deacetylated metabolite tizoxanide (TIZ) exhibit considerable in vitro activity against Besnoitia besnoiti tachyzoites grown in Vero cells. Real-time-PCR was used to assess B. besnoiti tachyzoite adhesion, invasion, and intracellular proliferation in vitro. A number of NTZ-derivatives, including Rm4822 and Rm4803, were generated, in which the thiazole-ring-associated nitro-group was replaced by a bromo-moiety. We here show that replacement of the nitro-group on the thiazole ring with a bromo (as it occurs in Rm4822) does not impair the efficacy of the drug, but methylation of the salicylate ring at the ortho-position in a bromo-derivative (Rm4803) results in complete abrogation of the antiparasitic activity. Treatment of extracellular B. besnoiti tachyzoites with NTZ has an inhibitory effect on host cell invasion, while treatments with TIZ, Rm4822 do not. TEM demonstrates that the effects of Rm4822 treatment upon the parasites are similar to the damage induced by NTZ. This includes increased vacuolization of the parasite cytoplasm, and loss of the structural integrity of the parasitophorous vacuole and its membrane. Thus, Rm4822, due to the absence of a potentially mutagenic nitro-group, may represent an important potential addition to the anti-parasitic arsenal for food animal production, especially in cattle.

Keywords

thiazolidesBesnoitia besnoitiin vitro drug treatmentbesnoitiosis
Type
Research Article
Information
Parasitology , Volume 134 , Issue 7 , July 2007 , pp. 975 - 985
Copyright
Copyright © Cambridge University Press 2007

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References

REFERENCES

Coombs, G. H. and Muller, S. (2002). Recent advances in the search for new anticoccidial drugs. International Journal for Parasitology 32, 497–508.CrossRefGoogle Scholar
Cortes, H., Ferreira, M. L., Silva, J. F., Vidal, R., Serra, P. and Caeiro, V. (2003). Contribuição para o estudo da besnoitiose bovina em Portugal. Revista Portuguesa de Ciências Veterinárias 98, 43–46.Google Scholar
Cortes, H., Leitão, A., Vidal, R., Vila-Vicosa, M. J., Ferreira, M. L., Caeiro, V. and Hjerpe, C. A. (2005). Besnoitiosis in bulls in Portugal. The Veterinary Record 157, 262–264.CrossRefGoogle ScholarPubMed
Cortes, H. C., Reis, Y., Waap, H., Vidal, R., Soares, H., Marques, I., Pereira, I., Fazendeiro, I., Ferreira, M. L., Caeiro, V., Shkap, V., Hemphill, A. and Leitao, A. (2006). Isolation of Besnoitia besnoiti from infected cattle in Portugal. Veterinary Parasitology 141, 226–233.CrossRefGoogle ScholarPubMed
Darius, A. K., Mehlhorn, H. and Heydorn, A. O. (2004). Effects of toltrazuril and ponazuril on the fine structure and multiplication of tachyzoites of the NC-1 strain of Neospora caninum (a synonym of Hammondia heydorni) in cell cultures. Parasitology Research 92, 453–458.CrossRefGoogle ScholarPubMed
Dubey, J. P. (1999). Recent advances in Neospora and neosporosis. Veterinary Parasitology 84, 349–367.CrossRefGoogle ScholarPubMed
Esposito, M., Stettler, R., Moores, S. L., Pidathala, C., Muller, N., Stachulski, A., Berry, N. G., Rossignol, J. F. and Hemphill, A. (2005). In vitro efficacies of nitazoxanide and other thiazolides against Neospora caninum tachyzoites reveal antiparasitic activity independent of the nitro group. Antimicrobial Agents and Chemotherapy 49, 3715–3723.CrossRefGoogle ScholarPubMed
Esposito, M., Mueller, N. and Hemphill, A. (2007). Structure-activity relationships from in vitro efficacies of the thiazolide series against the intracellular apicomplexan protozoan Neospora caninum. International Journal for Parasitology 37, 183–190. Epub ahead of print (doi:10.1016/j.ijpara.2006.10.009)CrossRefGoogle ScholarPubMed
Fox, L. M. and Saravolatz, L. D. (2005). Nitazoxanide: a new thiazolide antiparasitic agent. Clinical and Infectious Diseases 40, 1173–1180.CrossRefGoogle ScholarPubMed
Hemphill, A. and Croft, S. L. (1997). Electron microscopy in parasitology. In Analytical Parasitology (ed. Rogan, M. T.), pp. 227–268. Springer Verlag, Heidelberg.CrossRefGoogle Scholar
Hemphill, A., Mueller, J. and Esposito, M. (2006 a). Nitazoxanide, a broad-spectrum thiazolide anti-infective agent for the treatment of gastrointestinal infections. Expert Opinion on Pharmacotherapy 7, 953–964.CrossRefGoogle ScholarPubMed
Hemphill, A., Vonlaufen, N. and Naguleswaran, A. (2006 b). Cellular and immunological basis of the host-parasite relationship during infection with Neospora caninum. Parasitology 133, 261–278.CrossRefGoogle ScholarPubMed
Juste, R. A., Cuervo, L. A., Marco, J. C. and Oregui, L. M. (1990). La besnoitiosis bovina: desconocida en España? Medicina Veterinária 7, 613–618.Google Scholar
Mitchell, S. M., Zajac, A. M., Davis, W. L., Kennedy, T. J. and Lindsay, D. S. (2005). The effects of ponazuril on development of apicomplexans in vitro. Journal of Eukaryotic Microbiology 52, 231–235.CrossRefGoogle ScholarPubMed
Mordue, D., Desai, N., Dustin, M. and Sibley, D. L. (1999). Invasion by Toxoplasma gondii establishes a moving junction that selectively excludes host cell plasma membrane proteins on the basis of their membrane anchoring. The Journal of Experimental Medicine, 190, 1783–1792.CrossRefGoogle ScholarPubMed
Muller, J., Ruhle, G., Muller, N., Rossignol, J. F. and Hemphill, A. (2006). In vitro effects of thiazolides on Giardia lamblia WB clone C6 cultured axenically and in coculture with Caco2 cells. Antimicrobial Agents and Chemotherapy 50, 162–170.CrossRefGoogle ScholarPubMed
Naguleswaran, A., Muller, N. and Hemphill, A. (2003). Neospora caninum and Toxoplasma gondii: a novel adhesion/invasion assay reveals distinct differences in tachyzoite-host cell interactions. Experimental Parasitology 104, 149–158.CrossRefGoogle ScholarPubMed
Pankuch, G. A. and Appelbaum, P. C. (2006). Activities of tizoxanide and nitazoxanide compared to those of five other thiazolides and three other agents against anaerobic species. Antimicrobial Agents and Chemotherapy 50, 1112–1117.CrossRefGoogle ScholarPubMed
Pols, J. W. (1960). Studies on bovine besnoitiosis with special reference to the aetiology. The Onderstepoort Journal of Veterinary Research 28, 265–356.Google Scholar
Schwab, J. C., Beckers, C. J. and Joiner, K. A. (1994). The parasitophorous vacuole membrane surrounding intracellular Toxoplasma gondii functions as a molecular sieve. Proceedings of the National Academy of Sciences, USA 91, 509–513.CrossRefGoogle ScholarPubMed
Shkap, V., Pipano, E. and Ungar-Waron, H. (1987). Besnoitia besnoiti: chemotherapeutic trials in vivo and in vitro. Revue d'Élevage et de Médecine Vétérinaire des Pays Tropicaux 40, 259–264.Google ScholarPubMed
Sisson, G., Goodwin, A., Raudonikiene, A., Hughes, N. J., Mukhopadhyay, A. K., Berg, D. E. and Hoffman, P. S. (2002). Enzymes associated with reductive activation and action of nitazoxanide, nitrofurans, and metronidazole in Helicobacter pylori. Antimicrobial Agents and Chemotherapy 46, 2116–2123.CrossRefGoogle ScholarPubMed