SCHISTOSOMICIDAL ACTIVITY OF THE CRUDE EXTRACT OF ARTOCARPUS LAKOOCHA.

Puag-Haad is a traditional anthelmintic drug used to treat taeniasis in Thailand and Lao PDR. It is derived from the aqueous extract of the plant Artocarpus lakoocha. We investigated the in vitro anthelmintic properties of Puag-Haad against Schistosoma mansoni. Adult worms were incubated in M-199 medium containing 250, 500 and 750 µg/ml of Puag-Haad or praziquantel (PZQ) at a concentration of 175 µg/ml for 3, 6, 12 and 24 hours. The relative motility (RM value), survival index (SI) and tegument alterations seen under scanning electron microscope were assessed at each incubation time. The results showed the crude extract of A. lakoocha at a concentration of 250 µg/ml was more effective in causing damage than PZQ at a concentration of 175 µg/ml using RM and SI values. The major target organ affected by Puag-Haad was the tegument. The damage was greater at higher concentrations of the crude extract. It is likely tetrahydroxystilbene (THS), the main compound in Puag-Haad, caused the damage. THS could be a future candidate as a schistosomal drug. Further studies are needed to explore its mechanism, efficiency and safety in vivo.

Schistosoma mekongi cathepsin B and its use in the development of an immunodiagnosis.

Schistosomiasis mekongi is one of the most important human parasitic diseases caused by Schistosoma mekongi in South-east Asia. The endemic area is the Mekong River sub-region from Laos to Cambodia. This parasite also infects dogs and pigs which are its alternative host species. Currently, the lack of reliable rapid diagnosis makes it difficult to monitor the infection and spreading of the disease. In this study, we screened the antigens of the parasite with sera of infected mice using Western blotting and identified proteins of interest with LC-MS/MS to obtain potential candidate proteins for diagnostic development. This assay yielded 2 immunoreactive bands at molecular masses of 31 and 22kDa. The 31kDa protein was the major band identified as cathepsin B, and its gene was cloned to obtain a full cDNA sequence (SmekCatB). The cDNA consisted of 1123bp and its longest reading frame encoded for 342 amino acids with some putative post translation modifications. The recombinant SmekCatB (rSmekCatB) with hexahistidine tag at the C-terminus was expressed in Escherichia coli and purified by Ni-NTA resin under denaturing conditions. The rSmekCatB reacted with sera of S. mekongi-infected mice. Indirect ELISA using rSmekCatB as the antigen to detect mouse antibodies, revealed a sensitivity of 91.67% for schistosomiasis mekongi and the specificity of 100%. Our data suggested that SmekCatB is one of the most promising parasitic antigens that could be used for the diagnosis of S. mekongi infection.