Occurrence of spermic diploid and aspermic triploid forms of Fasciola in Vietnam and their molecular characterization based on nuclear and mitochondrial DNA.

Fasciola spp. found in Asian countries are diversified in nature, and they should therefore be characterized by spermatogenesis, ploidy and genetic differentiation as well as morphology. The present study showed that spermic diploid and aspermic triploid forms of Fasciola occurred in Vietnam. The spermic diploid specimens were accurately identified as F. gigantica, while the aspermic triploids could not be identified on the basis of their morphology by the ratio of body length and width and DNA sequences of nuclear ribosomal ITS1 and mitochondrial NDI and COI genes. The molecular data also indicated that Vietnamese aspermic triploids might be hybrids and/or their offspring between Fasciola hepatica and F. gigantica, because they showed the ITS1-Fh/Fg haplotype, which had chimeric sequences of the two species. Furthermore, the aspermic triploids seem to have originated in countries other than Vietnam and to have rapidly spread to that country with infected animals.

Hepatic lesions caused by migrating larvae of Ascaris suum in chickens.

Group A consisted of chickens infected with a single dose of Ascaris suum and group B of chickens infected with two successive doses. At days 1, 3, 7, 14 and 21 after the first or second infection dose, six chickens from each group were sacrificed. In both groups, larvae were recovered from the livers on days 1, 3, and 7 and lungs on days 3 and 7. No larvae were detected in chickens on day 14. Clear white lesions were noticed only on the livers from chickens of group B at day 7 but had disappeared at day 14. A comparison with group B showed mild histological changes that developed relative to the livers from group A.

Recombinant Ascaris 16-Kilodalton protein-induced protection against Ascaris suum larval migration after intranasal vaccination in pigs.

We recently cloned a protective antigen that is commonly expressed in Ascaris species that infect humans and pigs. We evaluated the vaccinal effects of this 16-kilodalton protein (As16) in pigs, the natural host of Ascaris suum, by intranasal immunization. Pigs that received Escherichia coli-expressed recombinant As16 (rAs16) coupled with cholera toxin (CT) had significantly elevated levels of rAs16-specific serum immunoglobulin G (IgG) and mucosal-associated IgA antibodies. rAs16 evoked a type II immune response characterized by elevated levels of interleukin-4 and -10 in the culture supernatants of peripheral blood mononuclear cells of the vaccinated pigs. An increased level of rAs16-specific serum IgG1 was also detected. Pigs vaccinated with rAs16-CT were protected from migration of A. suum larvae through the lungs, as indicated by a 58% reduction in the recovery of lung-stage third-stage larvae (L3), compared with that in nonvaccinated controls. Purified immunoglobulin from rAs16-CT-vaccinated pigs inhibited survival of infective L3 and interrupted the molting of lung-stage L3. Immunofluorescence studies revealed that this immunoglobulin bound to the digestive tracts of L3, suggesting that it might inactivate functions of the gut tissues of Ascaris species. We conclude that rAs16 is a promising mucosal vaccine candidate for pig and human ascariasis.

Ingestion of Fasciola gigantica metacercariae by the intermediate host snail, Lymnaea ollula, and infectivity of discharged metacercariae.

The rate of ingestion of Fasciola normal metacercariae (NMc) encysted on plants by Lymnaea ollula was examined, and the infectivity of the ingested metacercariae (IMc) in the feces of the host snail to mice was studied. As a result of ingestion by snails, the metacercarial outer cyst disappeared in about 50% of IMc in feces. There was no significant difference in the liver juvenile recovery at autopsy between mice inoculated with NMc and IMc kinds of metacercariae. Compared with NMc, the number of IMc could more easily be counted, because the separation of IMc from fecal contents under a microscope was not laborious.

Lung-stage protein profile and antigenic relationship between Ascaris lumbricoides and Ascaris suum.

The protein profile and antigenic properties of lung-stage larvae of Ascaris lumbricoides and A. suum were studied using 2-dimensional electrophoresis and immunoblot analysis, respectively. The protein profiles of the 2 parasites were identical except for the presence of only 1 major protein spot specific for each. There was a complete cross-reactivity between the 2 parasites at the immunological level, and no specific antigen was recognized using specific antibody raised against the 2 parasites in rabbits.

Species-specific proteins identified in Ascaris lumbricoides and Ascaris suum using two-dimensional electrophoresis.

The protein profile of adult female Ascaris lumbricoides and Ascaris suum originating from humans and pigs, respectively, was studied using two-dimensional polyacrylamide gel electrophoresis. Six different major protein spots specific for A. lumbricoides were identified irrespective of their geographical origin and no major specific spot was encountered in A. suum. No major differences in the protein profiles between the extract by phosphate-buffered saline and urea were encountered for either Ascaris species. It is therefore possible to use 2D-PAGE as a tool for discriminating the closely related Ascaris species from humans and pigs.