Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro.

Fasciolosis, a parasitic disease caused by the trematode Fasciola hepatica underreported is expanding both in human and animal population, throughout the world. The constant use of synthetic drugs to treat this condition has led to the natural selection of resistant strains of the parasite. Hence, there is a growing focus on the potential anti-helminthic properties of medicinal plants and phytopharmaceuticals. The current study assessed the potential anti-fasciolicide action of Momordica charantia leaf extracts and fractions on the eggs of F. hepatica parasites. The lyophilized crude extract (CE) of M. charantia leaves and its sub-fractions, obtained from liquid-liquid partitioning with organic solvents, were analysed by High Performance Liquid Chromatography (HPLC), suspended in 1% DMSO and used in in vitro tests. Quadruplicates of 50F. hepatica eggs were incubated at 23°C with M. charantia leaf CE in different concentrations. After 12days no larvae were formed in eggs incubated with CE concentrations above 12.5mg/mL. Eggs incubated with CE sub-fractions at concentrations of 1000, 100, 10, 1, 0.1, 0.01µg/mL affected embryonic development, with n-butanol presenting the strongest inhibition of miracidia formation. In contrast, on the 12th day, 90% of the miracidia hatched in the control experiments using 0.03% DMSO whereas embryogenesis was completely abolished with any concentration of albendazole sulphoxide ABZ(SO). Chemical analysis of the CE and sub-fractions revealed a prominent presence of flavonoids. HPLC-MS confirmed Quercetin to be one of the main flavonoids present in the CE and the n-butanol subfraction. This is the first study to analyse the potential anti-fasciolicide action of M. charantia leaf CE and subfractions.

Proteomic analysis of embryonic Fasciola hepatica: characterization and antigenic potential of a developmentally regulated heat shock protein.

Fasciola hepatica is responsible for human disease and economic livestock loss on a global scale. We report the first post-genomic investigation of cellular proteins expressed by embryonic F. hepatica via two-dimensional electrophoresis, image analysis and tandem mass spectrometry. Antioxidant proteins and protein chaperones are prominently expressed by embryonic F. hepatica. Molecular differences between the egg and other characterized F. hepatica lifecycle stages were noted. Furthermore, proteins expressed within liver fluke eggs differ to those isolated from the well-characterized eggs of the human blood flatworm Schistosoma mansoni were revealed. Plasticity in expression of major proteins, particularly a prominently expressed 65kDa protein cluster was seen between natural populations of embryonating F. hepatica eggs suggesting that liver fluke embryogenisis is a plastic process. Immunoblotting revealed that the abundant 65kDa protein cluster is recognised by infection sera from three F. hepatica challenged host species. Mass spectrometry and BLAST analyses demonstrated that the 65kDa antigen shows homology to egg antigens of other flatworm parasites, and is represented in a F. hepatica EST database constructed from adult fluke transcripts. EST clones encoding the egg antigen were re-sequenced, predicting two forms of the protein. Four clones predict a 312 aa polypeptide, three clones encode a putative 110 amino acid extension at the N-terminus which may be involved in protein secretion, although this extension was not expressed by natively extracted proteins. Consistent expression of alpha crystallin domains confirmed the protein to be a member of the alpha crystallin containing small heat shock protein (AC/sHSP) superfamily. AC/sHSPs are ubiquitous in nature, however, this is the first time a member of this protein superfamily has been described from F. hepatica. The antigenic AC/sHSP was named Fh-HSP35alpha based on predictions of molecular weight. Production of recombinant Fh-HSP35alpha reveals considerable mass discrepancy between native and recombinant proteins, although descriptions of other characterized flatworm AC/sHSPs, suggest that the native form is a dimer. Immunoblot analyses confirm that the recombinant protein is recognised by F. hepatica challenged hosts, but does not react with sera from non-infected animals. We discuss the potential of recombinant Fh-HSP35alpha as an egg-based diagnostic marker for liver fluke infection.

Comparative studies on the productivity of Fasciola gigantica and F. hepatica sporocysts in Lymnaea tomentosa that died after a cercarial shedding or without emission.

A histology study was performed on Fasciola gigantica- or F. hepatica-infected Lymnaea tomentosa that died after a cercarial shedding or without emission to compare the parasite productivity of each trematode. Degenerated rediae increased in number throughout the experiment. Their number rose rapidly after day 79 in snails that died after shedding in the F. gigantica group; they increased more slowly in snails that died without shedding. In the F. hepatica group the number rose after day 63 in the former snails and after day 79 in the latter. The contents of normal rediae evolved inversely. The number of morulae, for example, dropped rapidly after day 79 in the F. gigantica group. In the F. hepatica group it dropped after day 63 in snails that died after shedding and decreased after day 79 in the other dead snails. Free and degenerated cercariae were more numerous in snails that died without emission than in those that died at shedding. They rapidly increased in number after day 77. The numbers of rediae of F. gigantica were substantially greater than those of F. hepatica. In each group considered separately, it was likewise higher in snails that died without shedding than in those that died after the shedding of cercariae.

[Helminth eggs: trace elements of neolithic and paleolithic parasitosis on French sites]. / Les oeufs d'Helminthes: éléments traces des parasitoses néolithique et paléolithique en sites français.

On the neolithic site of Chalain (Jura, France), the analysis of human coprolites has revealed the presence of many well preserved eggs of Helminths: eggs of Trichuris spP., Capillaria spp., Fasciola hepatica, Diphyllobothrium spP. In the paleolithic picturial sanctuary of the Grande Grotte at Arcy-sur-Cure (Yonne, France), eggs of Ascaris spP. have been discovered. The presence of these parasits open a new way of search about the knowledge of ancient populations.