Hostname: page-component-745bb68f8f-hvd4g Total loading time: 0 Render date: 2025-01-11T07:51:46.781Z Has data issue: false hasContentIssue false
  • English
  • Français

Adult triclabendazole-resistant Fasciola hepatica: surface and subsurface tegumental responses to in vitro treatment with the sulphoxide metabolite of the experimental fasciolicide compound alpha

Published online by Cambridge University Press:  02 May 2006

M. McCONVILLE ,
G. P. BRENNAN ,
M. McCOY ,
R. CASTILLO ,
A. HERNANDEZ-CAMPOS ,
F. IBARRA  and
I. FAIRWEATHER
M. McCONVILLE
Affiliation:
Parasite Proteomics and Therapeutics Research Group, Queens University Belfast
G. P. BRENNAN
Affiliation:
Parasite Proteomics and Therapeutics Research Group, Queens University Belfast
M. McCOY
Affiliation:
Veterinary Sciences Division, Department of Agriculture and Rural Development for Northern Ireland (DARDNI), Belfast
R. CASTILLO
Affiliation:
Facultads de Quimica y Medicina Veterinaria, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City
A. HERNANDEZ-CAMPOS
Affiliation:
Facultads de Quimica y Medicina Veterinaria, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City
F. IBARRA
Affiliation:
Facultads de Quimica y Medicina Veterinaria, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City
I. FAIRWEATHER
Affiliation:
Parasite Proteomics and Therapeutics Research Group, Queens University Belfast
Get access Rights & Permissions [Opens in a new window]

Abstract

Mature Fasciola hepatica of the triclabendazole-resistant Sligo isolate were incubated in vitro with 10 μg/ml of the sulphoxide metabolite of compound alpha [5-chloro-2-methylthio-6-(1-naphthyloxy)-H-benzimidazole]; the metabolite will be referred to as alpha.SO. Changes resulting from drug treatment were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and tubulin immunocytochemistry (ICC). SEM revealed that disruption to the tegumental surface mainly took the form of swelling and blebbing. Extensive spine loss occurred on the ventral surface of the oral cone, and sloughing of the tegument was observed along the lateral margins of the fluke. Examination of sections from the anterior mid-body region at the TEM level revealed that treatment with alpha.SO led to swelling of the basal infolds and mitochondria within the tegumental syncytium; also, accumulations of secretory bodies beneath the apical plasma membrane. The tegumental cell bodies contained swollen mitochondria and cisternae of granular endoplasmic reticulum, but few Golgi complexes were observed. An increase in T2 secretory bodies was observed, whilst in the T1 tegumental cells, the T1 secretory bodies had decreased in number. Immunocytochemical (ICC) studies showed that incubation with alpha.SO, ABZ.SO and TCBZ.SO did not cause significant changes to the distribution of tubulin within the tegumental syncytium of the Sligo isolate. In contrast, alpha.SO, ABZ.SO and TCBZ.SO caused severe disruption to tubulin organization within the syncytial layer of the TCBZ-susceptible Cullompton isolate. The EM results confirm that compound alpha is a fasciolicide capable of disrupting the tegument of mature TCBZ-resistant F. hepatica; however, this was not accompanied by any change in tubulin immunoreactivity.

Keywords

Fasciola hepaticatriclabendazole resistancealpha.SOimmunocytochemistrytegumenttubulin
Type
Research Article
Information
Parasitology , Volume 133 , Issue 2 , August 2006 , pp. 195 - 208
Copyright
2006 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Alvarez, L., Solana, H., Mottier, L., Virkel, G., Fairweather, I. and Lanusse, C. ( 2005). Altered drug influx/efflux and enhanced metabolic activity in triclabendazole-resistant liver flukes. Parasitology 131, 501–510.CrossRefGoogle Scholar
Anderson, H. R. and Fairweather, I. ( 1995). Fasciola hepatica: ultrastructural changes to the tegument of juvenile flukes following incubation in vitro with the deacetylated (amine) metabolite of diamphenethide. International Journal for Parasitology 25, 319–333.CrossRefGoogle Scholar
Boray, J. C., Crowfoot, P. D., Strong, M. B., Allison, J. R., Schellenbaum, M., Von Orelli, M. and Sarasin, G. ( 1983). Treatment of immature and mature Fasciola hepatica infections in sheep with triclabendazole. Veterinary Record 113, 315–317.CrossRefGoogle Scholar
Buchanan, J. F., Fairweather, I., Brennan, G., Trudgett, A. and Hoey, E. M. ( 2003). Fasciola hepatica: surface and internal tegumental changes induced by treatment in vitro with the sulphoxide metabolite of albendazole (‘Valbazen’). Parasitology 126, 141–153.CrossRefGoogle Scholar
Coles, G. C. and Stafford, K. A. ( 2001). Activity of oxyclozanide, nitroxynil, clorsulon and albendazole against adult triclabendazole-resistant Fasciola hepatica. Veterinary Record 148, 723–724.CrossRefGoogle Scholar
Coles, G. C., Rhodes, A. C. and Stafford, K. A. ( 2000). Activity of closantel against adult triclabendazole-resistant Fasciola hepatica. Veterinary Record 146, 504.CrossRefGoogle Scholar
Fairweather, I. ( 2005). Triclabendazole: new skills to unravel an old(ish) enigma. Journal of Helminthology 79, 227–234.CrossRefGoogle Scholar
Fairweather, I., Anderson, H. R. and Threadgold, L. T. ( 1986). Fasciola hepatica: tegumental changes induced in vitro by the deacetylated (amine) metabolite of diamphenethide. Experimental Parasitology 62, 336–348.CrossRefGoogle Scholar
Fairweather, I., Anderson, H. R. and Baldwin, T. M. A. ( 1987). Fasciola hepatica: tegumental surface alterations following treatment in vitro with the deacetylated (amine) metabolite of diamphenethide. Parasitology Research 73, 99–106.CrossRefGoogle Scholar
Hernández-Campos, A., Ibarra-Velarde, F., Vera-Montenegro, Y., Rivera-Fernández, N. and Castillo, R. ( 2002). Synthesis and fasciolicidal activity of 5-chloro-2-methylthio-6-(1-naphthyloxy)-1 H-benzimidazole. Chemical and Pharmaceutical Bulletin 50, 649–652.CrossRefGoogle Scholar
Ibarra, F., Vera, Y., Quiroz, H., Cantó, J., Castillo, R., Hernández, A. and Ochoa, P. ( 2004). Determination of the effective dose of an experimental fasciolicide in naturally and experimentally infected cattle. Veterinary Parasitology 120, 65–74.CrossRefGoogle Scholar
Lacey, E. ( 1988). The role of the cytoskeletal protein, tubulin, in the mode of action and mechanism of drug resistance to benzimidazoles. International Journal for Parasitology 18, 885–936.CrossRefGoogle Scholar
Mas-Coma, S., Bargues, M. D. and Valero, M. A. ( 2005) Fascioliasis and other plant-borne trematode zoonoses. International Journal for Parasitology 35, 1255–1278.CrossRefGoogle Scholar
McCoy, M. A., Fairweather, I., Brennan, G. P., Kenny, J. M., Ellison, S. and Forbes, A. F. ( 2005). The efficacy of nitroxynil and triclabendazole administered synchronously against juvenile triclabendazole-resistant Fasciola hepatica in sheep. Research in Veterinary Science 78 (Suppl. A), 33.Google Scholar
McKinstry, B., Fairweather, I., Brennan, G. P. and Forbes, A. B. ( 2003). Fasciola hepatica: tegumental surface alterations following treatment in vivo and in vitro with nitroxynil (Trodax). Parasitology Research 91, 251–263.CrossRefGoogle Scholar
Meaney, M., Fairweather, I., Brennan, G. and Forbes, A. B. ( 2004). Transmission electron microscope study of the ultrastructural changes induced in the tegument and gut of Fasciola hepatica following treatment with clorsulon. Parasitology Research 92, 232–241.CrossRefGoogle Scholar
Meaney, M., Haughey, S., Brennan, G. P. and Fairweather, I. ( 2005 a). A scanning electron microscope study on the route of entry of clorsulon into the liver fluke, Fasciola hepatica. Parasitology Research 95, 117–128and 96, 189–198.Google Scholar
Meaney, M., Haughey, S., Brennan, G. P. and Fairweather, I. ( 2005 b). Ultrastructural observations on oral ingestion and trans-tegumental uptake of clorsulon by the liver fluke, Fasciola hepatica. Parasitology Research 95, 201–212.Google Scholar
Meaney, M., Fairweather, I., Brennan, G. P., Ramasamy, P. and Subramanian, P. B. ( 2002). Fasciola gigantica: tegumental surface alterations following treatment in vitro with the sulphoxide metabolite of triclabendazole. Parasitology Research 88, 315–325.CrossRefGoogle Scholar
Meaney, M., Fairweather, I., Brennan, G. P., McDowell, L. S. L. and Forbes, A. B. ( 2003). Fasciola hepatica: effects of the fasciolicide clorsulon in vitro and in vivo on the tegumental surface, and a comparison of the effects on young- and old-mature flukes. Parasitology Research 91, 238–250.CrossRefGoogle Scholar
Moll, L., Gaasenbeek, C. P. H., Vellema, P. and Borgsteede, F. H. M. ( 2000). Resistance of Fasciola hepatica against triclabendazole in cattle and sheep in the Netherlands. Veterinary Parasitology 91, 153–158.CrossRefGoogle Scholar
Overend, D. J. and Bowen, F. L. ( 1995). Resistance of Fasciola hepatica to triclabendazole. Australian Veterinary Journal 72, 275–276.CrossRefGoogle Scholar
Rivera, N., Ibarra, F., Zepeda, A., Fortoul, T., Cantó, G., Hernández, A. and Castillo, R. ( 2005). The effect of the 5-chloro-2-methylthio-6-(1-naphtyloxy)-H-benzimidazole on the tegument of immature Fasciola hepatica in their natural host. Parasitology Research 95, 379–383.CrossRefGoogle Scholar
Rivera, N., Ibarra, F., Zepeda, A., Fortoul, T., Hernández, A., Castillo, R. and Cantó, G. ( 2004). Tegumental surface changes in adult Fasciola hepatica following treatment in vitro and in vivo with an experimental fasciolicide. Parasitology Research 93, 283–286.CrossRefGoogle Scholar
Rivero, L. M., Jung, H., Castillo, R. and Hernández Campos, A. ( 1998). High-performance liquid chromatographic assay for a new fasciolicide agent, α-BIOF10, in biological fluids. Journal of Chromatography B 712, 237–241.CrossRefGoogle Scholar
Robinson, M. W., Trudgett, A., Hoey, E. M. and Fairweather, I. ( 2002). Triclabendazole-resistant Fasciola hepatica: β-tubulin and response to in vitro treatment with triclabendazole. Parasitology 124, 325–338.CrossRefGoogle Scholar
Robinson, M. W., Trudgett, A., Hoey, E. M. and Fairweather, I. ( 2003). The effect of the microtubule inhibitor tubulozole-C on the tegument of triclabendazole-susceptible and triclabendazole-resistant Fasciola hepatica. Parasitology Research 91, 117–129.CrossRefGoogle Scholar
Robinson, M. W., Lawson, J., Trudgett, A., Hoey, E. M. and Fairweather, I. ( 2004). The comparative metabolism of triclabendazole sulphoxide by triclabendazole-susceptible and triclabendazole-resistant Fasciola hepatica. Parasitology Research 92, 205–210.CrossRefGoogle Scholar
Rogan, M. T. and Threadgold, L. T. ( 1984). Fasciola hepatica: tegumental changes as a result of lectin binding. Experimental Parasitology 57, 248–260.CrossRefGoogle Scholar
Skuce, P. J. and Fairweather, I. ( 1990). The effect of the hydrogen ionophore closantel upon the pharmacology and ultrastrucure of the adult liver fluke Fasciola hepatica. Parasitology Research 76, 241–250.CrossRefGoogle Scholar
Skuce, P. J., Anderson, H. R. and Fairweather, I. ( 1987). The interaction between the deacetylated (amine) metabolite of diamphenethide (DAMD) and cytochemically demonstrable Na+/K+-ATPase activity in the tegument of Fasciola hepatica. Parasitology Research 74, 161–167.CrossRefGoogle Scholar
Smeal, M. G. and Hall, C. A. ( 1983). The activity of triclabendazole against immature and adult Fasciola hepatica infections in sheep. Australian Veterinary Journal 60, 329–331.CrossRefGoogle Scholar
Stitt, A. W. and Fairweather, I. ( 1993 a). Fasciola hepatica: tegumental surface changes in adult and juvenile flukes following treatment in vitro with the sulphoxide metabolite of triclabendazole (Fasinex). Parasitology Research 79, 529–536.Google Scholar
Stitt, A. W. and Fairweather, I. ( 1993 b). Fasciola hepatica: the effect of the microtubule inhibitors colchicine and tubulozole-C on the ultrastructure of the adult fluke. Parasitology 107, 297–309.Google Scholar
Stitt, A. W. and Fairweather, I. ( 1994). The effect of the sulphoxide metabolite of triclabendazole (‘Fasinex’) on the tegument of mature and immature stages of the liver fluke, Fasciola hepatica. Parasitology 108, 555–567.CrossRefGoogle Scholar
Turner, K., Armour, J. and Richards, R. J. ( 1984). Anthelmintic efficacy of triclabendazole against Fasciola hepatica in sheep. Veterinary Record 114, 41–42.CrossRefGoogle Scholar
Vera Montenegro, Y., Ibarra Velarde, F., Liébano Hernández, E., Quiroz Romero, H., Castillo Bocanegra, R., Hernández Campos, A. and Ochoa Galván, P. ( 2004). Efficacy of an experimental fasciolicide against immature and mature Fasciola hepatica in artificially infected calves. Parasitology Research 92, 211–214.CrossRefGoogle Scholar
Walker, S. M., Mckinstry, B., Boray, J. C., Brennan, G. P., Trudgett, A., Hoey, E. M., Fletcher, H. and Fairweather, I. ( 2004). Response of two isolates of Fasciola hepatica to treatment with triclabendazole in vivo and in vitro. Parasitology Research 94, 427–438.CrossRefGoogle Scholar
Wolstenholme, A. J., Fairweather, I., Prichard, R., von Samson-Himmelstjerna, G. and Sangster, N. C. ( 2004). Drug resistance in veterinary helminths. Trends in Parasitology 20, 469–476.CrossRefGoogle Scholar