The experiments described here were designed to characterize the status of susceptibility/resistance to albendazole (ABZ) and triclabendazole (TCBZ) of a Fasciola hepatica isolate (named CEDIVE isolate) recovered from infected sheep (Gualeguay, Argentina) and maintained under laboratory conditions. Two separate clinical efficacy experiments were performed on sheep artificially infected with the CEDIVE isolate. Experiment 1: sheep were randomly distributed either in an untreated control group or in an ABZ (7.5 mg/kg) treated group (n = 4 each). Additionally, the systemic exposure of ABZ metabolites was assessed in those ABZ-treated infected animals. In Experiment 2, an untreated control group and a TCBZ (10 mg/kg) treated group was included (n = 4 each). The fluckicidal efficacy of ABZ and TCBZ was assessed by the comparison of the number of flukes recovered from untreated and treated sheep at 15 days post-treatment. The efficacy against the CEDIVE isolate of F. hepatica was 29% (ABZ) and 100% (TCBZ). The plasma drug exposure (expressed as AUC and C-max) observed in the ABZ treated animals (Experiment 1) was in agreement with data obtained in the previous studies, which indicate that the low ABZ efficacy was not related to the quality of the pharmaceutical product and/or to a low systemic availability of the active drug/metabolite. The results reported here clearly show that the CEDIVE isolate of F. hepatica behaves as resistant to ABZ and susceptible to TCBZ. (C) 2012 Elsevier B.V. All rights reserved.

The main goal of the current work was to develop and validate anin vitrofluke egg hatch test, as a method for the detection of albendazole (ABZ) resistance in the liver fluke,Fasciola hepatica. Fluke eggs (200/ml,n= 5) from six different isolates were used in the current experimental work. They were obtained from different geographical locations and named Cullompton (UK), CEDIVE (Chascomus, Argentina), INTA-Bariloche (Bariloche, Argentina), Rubino (Uruguay), Cajamarca (Perú) and Río Chico (Catamarca, Argentina). The fluke eggs were incubated (25°C) for a 12-h period in the presence of either ABZ or its sulphoxide metabolite (ABZ.SO) (5, 0.5 or 0.05 nmol/ml). Untreated eggs were incubated as a control. Incubated eggs (with or without drug present) were kept in darkness at 25°C for 15 days. Afterwards, the trematode eggs were exposed to daylight over a 2-h period. Hatched and unhatched eggs were evaluated using an optical microscope, and the ovicidal activity was assessed for each fluke isolate. A very low ovicidal activity ( ≤ 13.4%) was observed in the ABZ-resistant CEDIVE isolate for both ABZ and ABZ.SO. Conversely, in the INTA-Bariloche and Río Chico isolates, which are suspected to be susceptible to ABZ, ovicidal activities ≥ 70.3% were observed after incubation with ABZ at the lowest concentration tested (0.05 nmol/ml). This finding correlates with that previously described for the ABZ-susceptible Cullompton. Finally, the Cajamarca and Rubino isolates behaved as ABZ resistant, since no ovicidal activity was observed after eggs were incubated with ABZ at 0.5 nmol/ml. Considering the specific results obtained for each isolate under assessment, the egg hatch test described here may be a suitable method for detection of ABZ resistance inF. hepatica.

Flubendazole (FLBZ) is a poor water solubility broad-spectrum BZD methylcarbamate anthelmintic compound. Cyclodextrins (CDs) are usually used to increase aqueous solubility of poor hydrosoluble compounds. The comparative in vitro aqueous solubility of FLBZ and other BZD anthelmintics in the presence of hydroxypropyl-β-cyclodextrin (HPβCD) was evaluated in the current work. Additionally, the comparative pharmacokinetic behaviour of FLBZ (and its metabolites) administered by the intraruminal (i.r.) or intraabomasal (i.a.) routes to sheep as either an aqueous CDs-based solution or a conventional carboximethylcellulose (CMC) suspension was assessed. Drug solubility studies involving albendazole, mebendazole, oxfendazole and FLBZ were performed in an aqueous solution (pH 1.2 or 7.4) with or without HPβCD (10%, w/v). The pharmacokinetic study involved two experiments. Experiment 1: In a crossover study, sheep received either a FLBZ-CDs solution (n = 3) or a FLBZ-CMC suspension (n = 3) by the i.r. route (3.8 mg/kg). The treatment Groups were reversed after a 21-days washout period. Experiment 2: sheep (n = 4) were treated by the i.a. route with the FLBZ-CDs solution (3.8 mg/kg). Plasma and abomasal fluid samples were collected between 0 and 72 h post-treatment. Samples were analysed by HPLC.