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Enhancement of the drug susceptibility of a triclabendazole-resistant isolate of Fasciola hepatica using the metabolic inhibitor ketoconazole

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Abstract

A study has been carried out to investigate whether the action of triclabendazole (TCBZ) is altered by using the metabolic inhibitor, ketoconazole (KTZ) to inhibit the cytochrome P450 (CYP 450) system within Fasciola hepatica. The Oberon TCBZ-resistant and Cullompton TCBZ-susceptible isolates were used for these experiments. The CYP 450 enzyme system was inhibited by a 2 h pre-incubation in KTZ (40 µM). Flukes were then incubated for a further 22 h in NCTC medium containing either KTZ; KTZ + nicotinamide adenine dinucleotide phosphate (NADPH; 1 nM); KTZ + NADPH + TCBZ (15 μg/ml); or KTZ + NADPH + triclabendazole sulphoxide (TCBZ.SO;15 μg/ml). Morphological changes resulting from drug treatment and following metabolic inhibition were assessed using scanning electron microscopy. After treatment with either TCBZ or TCBZ.SO alone, there was greater disruption to the TCBZ-susceptible isolate than the TCBZ-resistant isolate. However, co-incubation with KTZ and TCBZ/TCBZ.SO led to more severe surface changes to the TCBZ-resistant isolate than with each drug on its own, with greater swelling and blebbing of the tegument and even the loss of the apical plasma membrane in places. With the Cullompton isolate, there was limited potentiation of drug action in combination with KTZ, and only with TCBZ.SO. The results support the concept of altered drug metabolism within TCBZ-resistant isolates and indicate that this process may play a role in the development of drug resistance.

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Acknowledgements

This investigation was supported by a DARDNI Postgraduate Studentship to Catherine Devine. It was also partially supported by a grant from the European Union (DELIVER grant, no. FOOD-CT-200X-023025) and by a BBSRC/Defra grant (C00082X/1).

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Authors and Affiliations

  1. Parasite Therapeutics Group, School of Biological Sciences, Medical Biology Centre, The Queen’s University of Belfast, 97 Lisburn Road, Belfast, BT9 7BL, Northern Ireland, UK

    Catherine Devine, Gerard P. Brennan, Alan Trudgett, Elizabeth Hoey & Ian Fairweather

  2. Laboratorio de Farmacología, Departamento de Fisiopatología, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Campus Universitario (UNCPBA), 7000, Tandil, Argentina

    Carlos E. Lanusse & Luis I. Alvarez

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  1. Catherine Devine
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  2. Gerard P. Brennan
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  3. Carlos E. Lanusse
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  4. Luis I. Alvarez
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  5. Alan Trudgett
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  6. Elizabeth Hoey
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  7. Ian Fairweather
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Correspondence to Ian Fairweather.

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Devine, C., Brennan, G.P., Lanusse, C.E. et al. Enhancement of the drug susceptibility of a triclabendazole-resistant isolate of Fasciola hepatica using the metabolic inhibitor ketoconazole. Parasitol Res 107, 337–353 (2010). https://doi.org/10.1007/s00436-010-1866-2

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  • DOI: https://doi.org/10.1007/s00436-010-1866-2

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