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Morphological response of triclabendazole-susceptible and triclabendazole-resistant isolates of Fasciola hepatica to treatment in vitro with nitroxynil (Trodax)

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Abstract

Adult liver flukes belonging to three isolates of differing sensitivity to triclabendazole were incubated for 24 h in vitro in nitroxynil at a concentration of 100 μg/ml. Fine structural changes to the tegument, sub-tegumental region and gut were assessed by transmission electron microscopy. Similar changes were observed in all three isolates. In the tegumental syncytium, the basal infoldings and mitochondria were swollen, and there was an accumulation and accelerated release of secretory bodies at the apex. The crystalline core of the spines was disrupted, and the tegumental covering sloughed off. Mitochondria in the tegumental cells were also swollen, the Golgi complexes were affected and reduced numbers of T1 secretory bodies were evident in the T1-type of tegumental cell. In the sub-tegumental region, large spaces were present between cells and tissues, indicative of severe internal flooding. Swelling of mitochondria and cisternae of the granular endoplasmic reticulum was seen in the gastrodermal cells, which contained few secretory bodies. The extent of disruption varied between the isolates: the triclabendazole-resistant Sligo isolate was the most severely affected, while the Fairhurst triclabendazole-susceptible isolate was the least affected. In all three isolates, the tegument was more severely affected than the gut.

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References

  • Alvarez LI, Solana HD, Mottier ML, Virkel GL, Fairweather I, Lanusse CE (2005) Altered drug influx/efflux and enhanced metabolic activity in triclabendazole-resistant liver flukes. Parasitology 131:501–510

    Article  PubMed  CAS  Google Scholar 

  • Álvarez-Sánchez MA, Mainar-Jaime RC, Pérez-García J, Rojo-Vázquez FA (2006) Resistance of Fasciola hepatica to triclabendazole and albendazole in sheep. Vet Rec 159:424–425

    PubMed  Google Scholar 

  • Alvinerie M, Perez C, Sutra J-F, Floc’h R, Bayle R, Galtier P (1991a) Disposition of [14C]-nitroxynil in sheep. Acta Vet Scand 87:401–403

    Google Scholar 

  • Alvinerie M, Floc’h R, Galtier P (1991b) Plasma protein binding of nitroxynil in several species. J Vet Pharmacol Ther 14:170–173

    Article  PubMed  CAS  Google Scholar 

  • Boray JC (1990) Drug resistance in Fasciola hepatica. In: Boray JC, Martin PJ, Roush RT (eds) Resistance of parasites to antiparasitic drugs. MSD AGVET, Rahway, NJ, pp 51–60

    Google Scholar 

  • Boray JC (1994) Disease of domestic animals caused by flukes. Food and Agricultural Organization of the United Nations, Rome, pp 49

    Google Scholar 

  • Boray JC, Crowfoot PD, Strong MB, Allison JR, Schellenbaum M, Von Orelli M, Sarasin G (1983) Treatment of immature and mature Fasciola hepatica infections in sheep with triclabendazole. Vet Rec 113:315–317

    PubMed  CAS  Google Scholar 

  • Borgsteede FHM, Moll L, Vellema P, Gaasenbeek CPH (2005) Lack of reversion in triclabendazole-resistant Fasciola hepatica. Vet Rec 156:350–351

    PubMed  CAS  Google Scholar 

  • Caffrey CR (2007) Chemotherapy of schistosomiasis: present and future. Curr Opin Chem Biol 11:433–439

    Article  PubMed  CAS  Google Scholar 

  • Coles GC, Stafford KA (2001) The activity of oxyclozanide, nitroxynil, clorsulon and albendazole against adult triclabendazole-resistant Fasciola hepatica. Vet Rec 148:732–724

    Google Scholar 

  • Coles GC, Rhodes AC, Stafford KA (2000) The activity of closantel against adult triclabendazole-resistant Fasciola hepatica. Vet Rec 146:504

    PubMed  CAS  Google Scholar 

  • Dawes B (1966) Experimental fascioliasis: some effects on Fasciola hepatica of treatment of rat hosts with bithionol (Actamer). Helminthologia 7:297–307

    Google Scholar 

  • Fairweather I (2005) Triclabendazole: new skills to unravel an old(ish) enigma. J Helminthol 79:227–234

    Article  PubMed  CAS  Google Scholar 

  • Fairweather I, Boray JC (1999) Fasciolicides: efficacy, actions, resistance and its Management. Vet J 158:1–32

    Article  Google Scholar 

  • Fairweather I, Holmes SD, Threadgold LT (1984) Fasciola hepatica: motility responses to fasciolicides in vitro. Exp Parasitol 57:209–224

    Article  PubMed  CAS  Google Scholar 

  • Fairweather I, Anderson HR, Threadgold LT (1986) Fasciola hepatica: tegumental changes induced in vitro by the deacetylated (amine) metabolite of diamphenethide. Exp Parasitol 62:336–348

    Article  PubMed  CAS  Google Scholar 

  • Hernāndez-Campos A, Ibarra-Velarde F, Vera-Montenegro Y, Rivera-Fernandez N, Castillo R (2002) Synthesis and fasciolicidal activity of 5-chloro-2methylthio-6-(1-naphthyloxy)-1H-benzimidazole. Chem Pharm Bull 50:649–652

    Article  PubMed  Google Scholar 

  • Keiser J, Utzinger J (2007a) Artemisinins and synthetic trioxolanes in the treatment of helminth infections. Curr Opin Infect Dis 20:605–612

    PubMed  CAS  Google Scholar 

  • Keiser J, Utzinger J (2007b) Food-borne trematodiasis: current chemotherapy and advances with artemisinins and synthetic trioxolanes. Trends Parasitol 23:555–562

    Article  PubMed  CAS  Google Scholar 

  • Keiser J, Utzinger J, Vennerstrom JL, Dong Y, Brennan GP, Fairweather I (2007) Activity of artemether and OZ78 against triclabendazole-resistant Fasciola hepatica. Trans R Soc Trop Med Hyg 101:1219–1222

    Article  PubMed  CAS  Google Scholar 

  • Lloyd GM (1986) Energy metabolism and its regulation in the adult liver fluke Fasciola hepatica. Parasitology 93:217–248

    PubMed  CAS  Google Scholar 

  • McConville M, Brennan GP, McCoy M, Castillo R, Hernández-Campos A, Ibarra F, Fairweather I (2006) Adult triclabendazole-resistant Fasciola hepatica: surface and subsurface tegumental responses to in vitro treatment with the sulphoxide metabolite of the experimental fasciolicide compound alpha. Parasitology 133:195–208

    Article  PubMed  CAS  Google Scholar 

  • McConville M, Brennan GP, Flanagan A, Edgar HWJ, McCoy M, Castillo R, Hernández-Campos A, Fairweather I (2008) Surface and internal tegumental changes in juvenile Fasciola hepatica following treatment in vivo with the experimental fasciolicide, compound alpha. Vet Parasitol 153:52–64

    Article  PubMed  CAS  Google Scholar 

  • McCoy MA, Fairweather I, Brennan GP, Kenny JM, Forbes AB (2005) The efficacy of nitroxynil and triclabendazole administered synchronously against juvenile triclabendazole-resistant Fasciola hepatica in sheep. Res Vet Sci 78:33

    Article  CAS  Google Scholar 

  • McKinstry B (2008) Ultrastructural changes observed in Fasciola hepatica following treatment with nitroxynil and triclabendazole, alone and in combination. PhD Thesis, The Queen’s University of Belfast, UK

  • McKinstry B, Fairweather I, Brennan GP, Forbes AB (2003) Fasciola hepatica: tegumental surface alterations following treatment in vivo and in vitro with nitroxynil (Trodax). Parasitol Res 91:251–263

    Article  PubMed  CAS  Google Scholar 

  • McKinstry B, Brennan GP, Halferty L, Forbes AB, Fairweather I (2007) Ultrastructural changes induced in the tegument and gut of Fasciola hepatica following in vivo and in vitro drug treatment with nitroxynil (Trodax). Parasitol Res 101:929–941

    Article  PubMed  CAS  Google Scholar 

  • Meaney M, Fairweather I, Brennan GP, Forbes AB (2004) Transmission electron microscope study of the ultrastructural changes induced in the tegument and gut of Fasciola hepatica following in vivo drug treatment with clorsulon. Parasitol Res 92:232–241

    Article  PubMed  CAS  Google Scholar 

  • Meaney M, Haughey S, Brennan GP, Fairweather I (2005a) A scanning electron microscopy study on the route of entry of clorsulon into the liver fluke, Fasciola hepatica. Parasitol Res 95:117–128

    Article  PubMed  CAS  Google Scholar 

  • Meaney M, Haughey S, Brennan GP, Fairweather I (2005b) Ultrastructural observations on oral ingestion and trans-tegumental uptake of clorsulon by the liver fluke, Fasciola hepatica. Parasitol Res 95:201–212

    Article  PubMed  CAS  Google Scholar 

  • Meaney M, Allister J, McKinstry B, McLauglin K, Brennan GP, Forbes AB, Fairweather I (2006) Fasciola hepatica: morphological effects of a combination of triclabendazole and clorsulon against mature fluke. Parasitol Res 99:609–621

    Article  PubMed  CAS  Google Scholar 

  • Meaney M, Allister J, McKinstry B, McLaughlin K, Brennan GP, Forbes AB, Fairweather I (2007) Fasciola hepatica: ultrastructural effects of a combination of triclabendazole and clorsulon against mature fluke. Parasitol Res 100:1091–1104

    Article  PubMed  CAS  Google Scholar 

  • Moll L, Gaasenbeek CP, Vellema P, Borgsteede FH (2000) Resistance of Fasciola hepatica against triclabendazole in cattle and sheep in the Netherlands. Vet Parasitol 91:153–158

    Article  PubMed  CAS  Google Scholar 

  • Mottier L, Alvarez L, Fairweather I, Lanusse C (2006a) Resistance-induced changes in triclabendazole transport in Fasciola hepatica: ivermectin reversal effect. J Parasitol 92:1355–1360

    Article  PubMed  CAS  Google Scholar 

  • Mottier L, Alvarez L, Ceballos L, Lanusse C (2006b) Drug transport mechanisms in helminth parasites: passive diffusion of benzimidazole anthelmintics. Exp Parasitol 113:49–57

    Article  PubMed  CAS  Google Scholar 

  • Overend DJ, Bowen FL (1995) Resistance of Fasciola hepatica to triclabendazole. Aust Vet J 72:275–276

    Article  PubMed  CAS  Google Scholar 

  • Parnell MJ (1970) Determination of nitroxynil residues in sheep and calves. Pestic Sci 1:138–143

    Article  CAS  Google Scholar 

  • Robinson G, Threadgold LT (1975) Electron microscope studies of Fasciola hepatica. XIII. The fine structure of the gastrodermis. Exp Parasitol 37:20–36

    Article  PubMed  CAS  Google Scholar 

  • Robinson MW, Trudgett A, Hoey EM, Fairweather I (2002) Triclabendazole-resistant Fasciola hepatica: β-tubulin and response to in vitro treatment with triclabendazole. Parasitology 124:325–338

    Article  PubMed  CAS  Google Scholar 

  • Robinson MW, Lawson J, Trudgett A, Hoey EM, Fairweather I (2004) The comparative metabolism of triclabendazole sulphoxide by triclabendazole-susceptible and triclabendazole-resistant Fasciola hepatica. Parasitol Res 92:205–210

    Article  PubMed  Google Scholar 

  • Ryan LA, Hoey E, Trudgett A, Fairweather I, Fuchs, Robinson MW, Chambers E, Timson DJ, Ryan E, Feltwell T, Ivens A, Bentley G, Johnston D (2008) Fasciola hepatica expresses multiple α- and β-tubulin isotypes. Mol Biochem Parasitol 159:73–78

    Article  PubMed  CAS  Google Scholar 

  • Skuce PJ, Fairweather I (1990) The effect of the hydrogen ionophore closantel upon the pharmacology and ultrastructure of the adult liver fluke Fasciola hepatica. Parasitol Res 76:241–250

    Article  PubMed  CAS  Google Scholar 

  • Threadgold LT (1963) The tegument and associated structures of Fasciola hepatica. Quart J Microsc Soc 104:505–512

    Google Scholar 

  • Threadgold LT (1967) Electron-microscope studies of Fasciola hepatica. III. Further observations on the tegument and associated structures. Parasitology 57:633–637

    Article  PubMed  CAS  Google Scholar 

  • Tielens AGM, den Heuvel JM, den Bergh SG (1984) The energy metabolism of Fasciola hepatica during its development in the final host. Mol Biochem Parasitol 13:301–307

    Article  PubMed  CAS  Google Scholar 

  • Toner E, McConvery F, Brennan GP, Meaney M, Fairweather I (2008) A scanning electron microscope study on the route of entry of triclabendazole into the liver fluke, Fasciola hepatica. Parasitology (in press)

  • Utzinger J, Shu-Hua X, Tanner M, Keiser J (2007) Atremisinins for schistosomiasis and beyond. Curr Opin Investig Drugs 8:105–116

    PubMed  CAS  Google Scholar 

  • Vera Montenegro Y, Ibarra Velarde F, Liébano Hernàndez E, Quiroz Romero H, Castillo Bocanegra R, Hernández Campos A, Ochoa Galvan P (2004) Efficacy of an experimental faasciloicide against immature and mature Fasciola hepatica in artificially infected calves. Parasitol Res 92:211–214

    Article  PubMed  CAS  Google Scholar 

  • Verheyen A, Vanparijs O, Lauwers H, Thienpont D (1980) The influence of closantel administration to sheep on the ultrastructure of the adult liver fluke, Fasciola hepatica L. In: Van den Bossche H (ed) The host-invader interplay. Elsevier, Amsterdam, pp 705–708

    Google Scholar 

  • Walker SM, McKinstry B, Boray JC, Brennan GP, Trudgett A, Hoey EM, Fletcher H, Fairweather I (2004) Response of two isolates of Fasciola hepatica to treatment with triclabendazole in vivo and in vitro. Parasitol Res 94:427–438

    Article  PubMed  CAS  Google Scholar 

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Acknowledgement

This investigation was supported by a DARDNI Postgraduate Studentship to Bryan McKinstry.

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Authors and Affiliations

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

    B. McKinstry, L. Halferty, G.P. Brennan & I. Fairweather

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  1. B. McKinstry
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  2. L. Halferty
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  3. G.P. Brennan
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  4. I. Fairweather
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Correspondence to I. Fairweather.

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McKinstry, B., Halferty, L., Brennan, G. et al. Morphological response of triclabendazole-susceptible and triclabendazole-resistant isolates of Fasciola hepatica to treatment in vitro with nitroxynil (Trodax). Parasitol Res 104, 645–655 (2009). https://doi.org/10.1007/s00436-008-1241-8

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  • DOI: https://doi.org/10.1007/s00436-008-1241-8

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