SUMMARYFascioliasis is an emerging/re-emerging vector-borne disease with the widest known distribution. Approximately 17 million people are infected around the world, being the Andean region the most affected area. There is an important necessity to develop sensitive and specific diagnostic tools to treat patients early and to avoid complications. In this paper we evaluated the immune response of infected humans against two antigenic preparations: the total soluble extract (FhTSE) and the adult worm vomit (FhAWV) in order to identify antigenic fractions specific for Fasciola hepatica. Both preparations were processed by SDS-PAGE and Western blot with human sera with fascioliasis (F), other parasitosis and healthy individuals. In the immunoblot of FhTSE, sera F recognised 16 bands with MW between eight and 110 kDa, from which those of 8, 9, 10, 38, 45 and 57 kDa were specific. In the preparation FhAWV, sera F recognised nine bands with MW from eight to 85 kDa, from which those of 8, 12, 15 and 24 kDa were specific. Some bands of cross-reaction were evident with sera from patients with other parasitoses, more frequent with the FhTSE. Bands within the MW mentioned, particularly that of eight kDa, have been shown to be specific by others, and deserve additional characterisation for their potential use in immunodiagnosis.

Schistosoma mansoni is a parasitic trematode of the portal-mesenteric veins with a closed-end intestine. Adult worms regurgitate their intestinal content after digestion, together with constituents of the lining gut. Some of these molecules circulate in the blood and are antigenic. We obtain a "vomit" preparation and preliminary evaluate its biochemical composition and antigenic capacity. The "vomit" preparation was obtained after changes in temperature and solutions of incubation of adult worms between 4 and 37 degrees C. Supernatant was assayed for protein, carbohydrate concentration and enzymatic activities associated to the intestine and to the worm tegument. The antigenicity of the product was evaluated using Western blot (WB) analysis against sera of experimentally infected mice, before and after drug cure, sera from people infected with S. mansoni and from individuals infected with other parasitoses. More carbohydrate than protein was detected in the preparations. Cysteine proteinase (CP), N-acetyl-beta-D: -glucosaminidase and alkaline phosphatase activities were detected. The latter enzyme activity is a marker of the tegument, suggesting that in spite of careful conditions used to avoid the presence of tegumental material, manipulation of the worms always resulted in the release of tegumental molecules. Cationic exchange chromatography was useful to separate various components of this "vomit" preparation, particularly enzymes responsible for CP activity. Two highly immunogenic and specific duplets were observed in the WB analysis, 31/32- and 38/40-kDa components, the former probably referring to the intestinal CPs Sm31/Sm32. None of the two duplets disappeared after successful chemotherapy during the time of evaluation in mice or humans.

A recently described hyperendemic onchocerciasis area, located in the Unturán Mountains (between the Siapa and Orinoco basins) of southern Venezuela was studied using a cocktail of 3 low molecular weight onchocercal recombinant antigens (OvMBP/10, OvMBP/11, and OvMBP/29). The resulting seroepidemiological data were compared with those from a hypoendemic community (Altamira) situated in the northern coastal mountain range. Parasitological (skin biopsy) and serological (enzyme-linked immunosorbent assay, ELISA) methods for the specific diagnosis of Onchocerca volvulus in these 2 very different endemic areas were, respectively, 88% and 96% sensitive in Unturán, and 57% and 91% sensitive in Altamira. The mean microfilarial load, the mean optical density (OD), and the seropositivity rates all increased significantly with age in both communities. The serological variables (mean OD and prevalence of anti-O. volvulus antibodies) were both significantly higher in Unturán than in Altamira for children and young adults (aged < 25 years), although above this age no differences between communities were detected. Seroprevalence had already reached 50% in the under 15 year-olds examined at Unturán but was just 5% at Altamira for the same age-class. The prevalence of specific antibodies (mainly a marker of exposure to risk of infection) exceeded 85% in the remaining age-categories at the hyperendemic area. This is in agreement with the high community microfilarial load recorded in Unturán (> 20 mf/mg) and the presence of sclerosing keratitis and hanging groin, suggesting that onchocerciasis is a public health problem in this community. The ELISA test used here, based on a cocktail of 3 low molecular weight onchocercal recombinant antigens, appears, therefore, to constitute a practical tool for the description of endemicity levels in remote areas, particularly given the fact that finger-prick blood samples are routinely taken from children in the Upper Orinoco region for surveys of malaria incidence. Such studies could aid in defining the true extent of the Amazon focus (still unknown) and providing priority indicators for the selection of communities where onchocerciasis control programmes should be implemented.

A new region with human onchocerciasis is reported in the Unturán mountains, South Venezuela, affecting Yanomamö populations not surveyed in previous studies conducted in the Venezuelan-Brazilian border area. Its distribution probably extends towards the Upper Toototobi endemic area in Brazil. The age-standardized prevalence of Onchocerca volvulus microfilariae (mf) (67%), the prevalence of infection in those aged > or = 20 years (86%), and the community microfilarial load (CMFL) (24 mf/mg), are consistent with hyperendemic transmission. Both prevalence and mean intensity increased monotonically with age without reaching a plateau, the highest values being recorded in the > or = 45 years age class (respectively, 95% and 42 [geometric mean of Williams] or 172 [arithmetic mean] mf/mg). The degree of parasite overdispersion (measured by the variance/mean ratio) also increased with host age. The CMFL value, the presence of sclerosing keratitis, hanging groin, and severe skin lesions, indicated that the infection poses an important public health problem in the region.

Hydatid disease in tropical areas poses a serious diagnostic problem due to the high frequence of cross-reactivity with other endemic helminthic infections. The enzyme-linked-immunosorbent assay (ELISA) and the double diffusion arc 5 showed respectively a sensitivity of 73% and 57% and a specificity of 84-95% and 100%. However, the specificity of ELISA was greatly increased by using ovine serum and phosphorylcholine in the diluent buffer. The hydatic antigen obtained from ovine cyst fluid showed three main protein bands of 64, 58 and 30 KDa using SDS PAGE and immunoblotting. Sera from patients with onchocerciasis, cysticercosis, toxocariasis and Strongyloides infection cross-reacted with the 64 and 58 KDa bands by immunoblotting. However, none of the analyzed sera recognized the 30 KDa band, that seems to be specific in this assay. The immunoblotting showed a sensitivity of 80% and a specificity of 100% when used to recognize the 30 KDa band.