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Fasciolosis caused by Fasciola spp. is considered the most important helminth infection of ruminants in tropical countries. Anthelmintic resistance has become a global concern. This study compared the efficacy of the commonly used anthelmintics, determined the toxicity level and any indication of resistance. Thirty two water buffaloes naturally-infected with Fasciola spp. were used to determine the efficacy of triclabendazole (TBZ), albendazole (ABZ), and bromofenofos (BRO) using Fecal Egg Count Reduction Test (FECRT). To test the toxicity of the drugs given, serum glutamic-pyruvic transaminase (SGPT) was evaluated before and within one week after treatment. One dose administration of ABZ registered an efficacy of 79.17%, 73.33% for TBZ and 70.83% for BRO. Efficacy in two dosetreatment group was 83.33% for both BRO and ABZ, and 90.00% for TBZ. Two dose-treatment was effective for TBZ (90%), ineffective for BRO and ABZ. SGPT levels were not significantly different between pre-treatment and posttreatment across all treatments. Giving one or two doses of anthelmintics, at one month interval, does not increase the efficacy of the three drugs tested. The study also implies that anthelmintic resistance may have developed in the animals.
The aim of this study was to clone new secretory antigen of Fasciola hepatica and to predict its availability for immune system. The new cathepsin L - FhPcW1 (F. hepatico cysteine proteinase Warsaw 1), GenBank accession: EF407948, cDNA was cloned from adult F. hepatica flukes using RACE-PCR method. FhPcW1 is encoded by a 1,066 bp mRNA with a predicted open reading frame (ORF) of 326 amino acids (predicted pl = 5.41, m.w. = 37.137 kDa). Performed bioinformatic analysis included alignments of the nucleotide and amino acid sequences, the ExPASy (Expert Protein Analysis System) proteomics server of the Swiss Institute of Bioinformatics and National Center for Biotechnology Information. Performed analyses allowed to suppose that FhPcW1 is a secreted protein, which contains signal peptide, serine, threonine, tyrosine phosphorylation sites and four tyrosine sulfation sites, and does not contain glycosylation sites. The ORF corresponding to FhPcW1 exhibited strong similarity to previously cloned cathepsins L from the F. hepatico as well as F. gigantica. Predicted biochemical characteristics fits to the described before F. hepatica cathepsin Ls. Moreover, three dimensional model and MHC types ligation strength prediction were performed. Analysis of MHC type I and II peptide binding suggests that FhPcW1 may have significant immunogenic potential. The potential HLA II epitopes are situated at the outer surface of this protein. Thus, these epitopes seems to be available for immune response, especially for antibodies. This result may show that FhPcW1 seems to be a promising antigen for vaccination against F. hepatica.
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