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Seasonality of the nematode Gnathostoma turgidum in Virginia opossums (Didelphis virginiana) in the wild has been reported; however, the mechanisms involved in deworming are unknown. We monitored the parasitologic and biologic changes in four Virginia opossums naturally infected with G. turgidum by coproparasitologic examination and abdominal ultrasonography. Eggs became detectable in the feces of opossums in May, peaked in July and August, and suddenly decreased in October. Adults of G. turgidum were expelled in the feces mainly in September. Ultrasonography of the liver showed slight damage during May. Lesions in the stomach appeared in April and persisted until September. The abnormalities of the liver and stomach were resolved in November. These data suggest that G. turgidum is likely expelled as a result of host immunologic mechanisms, although termination of a natural life span cannot be definitively excluded.

The low-molecular weight proteins (LMWPs) of Taenia solium metacestode (TsM) constituted pertinent serodiagnostic antigens for cysticercosis. We established a novel single-step purification of the LMWPs from TsM cyst fluid (CF). When the CF was precipitated with trichloroacetic acid/acetone mixture at the final concentrations of 5 and 50%, most LMWPs (ranging 7-38kDa) remained in the supernatant fraction. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses revealed that the LMWPs consisted mainly with the previously reported subunit proteins of the 120- and 150-kDa antigen complexes. Serum samples from neurocysticercosis (NC) and other helminthic infections, as well as those from healthy controls were tested by both immunoblotting and micro-ELISA. In 135 cases of active stage NC patients, 132 cases (97.7%) showed positive reactions. Serum samples from other helminthic diseases (n=125) and healthy controls (n=100) exhibited no positive reactions except for cystic echinococcosis, of which 12% (3/25 cases) exhibited low levels of cross-reactivity. The LMWPs from different geographical regions (Korea and Mexico) showed diagnostic sensitivity and specificity of 97.7% and 98.7% against active stage NC. Our single-step separation method for the LMWPs provided excellent performance with easy applicability and high reproducibility, which has a great benefit for preparation of potent antigen in endemic areas., (Copyright © 2010 Royal Society of Tropical Medicine and Hygiene.)

Gnathostoma turgidum is a nematode that parasitizes the stomach of opossums, Didelphis virginiana. Despite its wide distribution in the Americas, its natural life cycle is poorly understood. Recently, we found an endemic area for G. turgidum infection in Sinaloa, Mexico (Diaz-Camacho et al., 2009). Based on sporadic surveys for several years, the prevalence was apparently high in summer and extremely low in winter. To confirm that this is really a seasonal variance, we conducted a longitudinal survey on G. turgidum infection in opossums from November 2007 to November 2008. The results showed amazing seasonal changes in the prevalence, with synchronized migration and maturation of worms in opossums. Between February and March, many juvenile worms, with occasional AL3, were found in the liver, but no worms were found in the stomach. Mature adult worms began to appear in the stomach around April and rapidly increased in number toward July, when all worms resided in the stomach. Then, the worms disappeared almost completely by November. These results suggest that G. turgidum is an annual parasite of the opossum, D. virginiana, in Mexico.

Gnathostomosis, caused by Gnathostoma binucleatum, is a serious public health issue in Mexico. Although 2 other Gnathostoma spp., G. turgidum and G. lamothei, have been found in wild animals, their natural life cycle or their relation to human disease remains unclear. While we were conducting an epidemiological survey on Gnathostoma spp. in Sinaloa State, Mexico, we found an endemic area for G. turgidum in common opossums, Didelphis virginiana, located in Tecualilla, Sinaloa. The species identification was carried out by morphological and molecular biological methods. This is the first record of an endemic area for G. turgidum infection in opossums, D. virginiana, in the Americas.

Gnathostomosis is a typical fish-borne zoonotic parasitosis and is currently a serious public health issue in Mexico. Among several Gnathostoma species present in wild animals in Mexico, Gnathostoma binucleatum is the only proven species responsible for human diseases, and the advanced third stage larvae (AL3) of G. binucleatum have been found in over 20 species of fish in this country. In Sinaloa State, two fish species, Dormitator latifrons and Eleotris picta, were heavily contaminated with G. binucleatum AL3. When we analyzed the relationship between the size of the fish and the density of infection with G. binucleatum AL3, the distribution patterns of AL3 were markedly different between these two fish species. Apparent size-dependent accumulation was observed in E. picta but not in D. latifrons, suggesting that E. picta is a paratenic host whereas D. latifrons is a second intermediate host.

The antigenic similarities of adult-worm extracts of Gnathostoma spinigerum, G. hispidum and G. doloresi, all of which are important food-borne parasites causing larva migrans in humans, were evaluated. The 40 sera used came from gnathostomiasis cases in Mexico, where G. binucleatum is endemic, or in Japan, where G. doloresi predominates. When used as the fixed antigens in microplate-ELISA, the adult-worm extracts from the three different species of Gnathostoma were found to have equal binding capacity to the Gnathostoma-specific IgG antibodies in the sera of the Mexican and Japanese patients. The correlation coefficients for the optical densities seen in the ELISA, between any two of the three Gnathostoma extracts, were all >0.900. The dose-response curves produced when four sera were tested, in the microplate-ELISA, against the three different Gnathostoma extracts were nearly identical, indicating that the antigens in each of the extracts had similar avidity. Furthermore, the results of competitive-inhibition ELISA indicated that the antigenic specificities of the three extracts were almost identical. An antigen of 40 kDa, which SDS-PAGE and immunoblot analysis revealed to be present in all three extracts, was recognized by the sera from the gnathostomiasis cases. When the sera were investigated by dot-blot ELISA, they also gave similar results whichever extract was used as the antigen source. It appears that, in the serodiagnosis of gnathostomiasis by microplate- or dot-ELISA, each of the three adult-worm extracts would be equally useful, regardless of the causative species of Gnathostoma.

Gnathostomosis is an emerging public health problem in Sinaloa, Mexico, where an increasing number of human cases have been diagnosed since 1989. The present study was carried out to determine the presence of the parasite in other natural hosts from the area. Birds, fish, opossums and raccoons were captured from local dams and lagoons. The flesh from bird and fish specimens was ground and examined under a 100 W light bulb. Larvae were processed for light and electron microscopy. A total of 368 advanced stage 3 (AL3) larvae were found in 300 ichthyophagous birds, with Egretta alba exhibiting the highest infection rate. A total of 4,156 fish were examined, of which six species were infected with AL3 larvae: Arius guatemalensis (blue sea catfish), Dormitator latifrons (Pacific fat sleeper), Gobiomorus sp. (fat sleeper), Oreochromis sp. (Nile tilapia), Cichlasoma beani (Sinaloan cichlid or green guapote) and Eleotris picta (spotted sleeper). Twenty larvae from birds were used to infect domestic cats and dogs. Young adult worms were recovered from the stomach of a cat with a 17 day infection and from a dog with a 35 day infection. Larvae exhibited four rows of hooklets on the head bulb, whereas the young adults had nine rows of hooklets. The cuticular spines of adult worms along the body evolved from single-pointed, bi- or trifurcated spines. Nuclei were counted in intestinal cells examined in serial sections of larvae recovered from a great heron and a fish, in which a mean of 1.6 nuclei/cell was found, corresponding to data published for Gnathostoma binucleatum. Although the external morphology of both larvae and adults are in agreement with previous descriptions of Gnathostoma spinigerum, the results indicate that natural host infections in Sinaloa may be caused by either G. spinigerum or G. binucleatum.

Gnathostomiasis is an important food-borne parasitic zoonosis that is endemic mainly in Asian countries where some people prefer to eat raw freshwater fish. In North America, the first recorded case of gnathostomiasis was in Mexico in 1970, and the numbers of gnathostomiasis patients in Mexico seems to be increasing dramatically with time. However, the epidemiology of this disease in Mexico has never been described in detail. Here we review the current status of gnathostomiasis in Mexico.