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PubMed1990

[Antigenic recognition of rats infected with Gnathostoma hispidum: comparison of antigens derived from the advanced third stage larvae of Gnathostoma hispidum and Gnathostoma doloresi].

To increase understanding of immune response of host against G. hispidum, antigenic recognition of rodent gnathostomiasis were identified by the immunoblot method after PAGE of the advanced third stage larvae of G. hispidum and G. doloresi. The molecular weight of the major antigenic bands of G. hispidum extracts were 35 kDa and 16 kDa, and of G. doloresi were 48 kDa, 37 kDa and 35 kDa. Of these bands, the 35 kDa antigen of G. hispidum and G. doloresi was recognized with the sera of rats infected with G. hispidum 5 weeks after inoculation. Lecting staining with endoglycosidase-H or hydrolytic degradation on nitrocellulose strips revealed that the 35 kDa band of G. hispidum possessed the mannose-rich oligosaccharides. On the other hand, the band of G. doloresi had the hybrid-type glycopeptides. The larval extracts and excretory-secretory materials of G. hispidum were not antigenitically identical.

Animals↗
PubMed2003

Evaluation of the antigenic similarities of adult-worm extracts from three Gnathostoma species, using sera from Mexican and Japanese patients with Gnathostoma infections.

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.

Animals↗
PubMed2000

ITS-2 rDNA sequencing of Gnathostoma species (Nematoda) and elucidation of the species causing human gnathostomiasis in the Americas.

From several gnathostome species the complete internal transcribed spacer ITS-2 ribosomal DNA (rDNA) repeat sequence and a fragment of the 5.8S rDNA were obtained by direct polymerase chain reaction cycle-sequencing and silver-staining methods. The size of the complete ITS-1 sequence in agarose gel electrophoresis was also obtained. The ITS-2 enabled the differentiation of Gnathostoma spinigerum from Thailand and Gnathostoma binucleatum from Mexico and Ecuador and confirmed the validity of the latter. Gnathostoma turgidum, Gnathostoma sp. I (=Gnathostoma procyonis sensu Almeyda-Artigas et al., 1994), and Gnathostoma sp. II (=G. turgidum sensu Foster, 1939 pro parte), all from Mexico, proved to be independent species, but Gnathostoma sp. III, also from Mexico, could not be differentiated from G. turgidum. In Mexico and Ecuador, gnathostomes involved in human infection and that had been classified as G. spinigerum belong to G. binucleatum. The 5.8S rDNA sequences of the 6 Gnathostoma species studied were identical. The results of the ITS-1 agreed with those results of ITS-2.

Animals↗
PubMed2005

A new species of Gnathostoma (Nematoda: Gnathostomatidae) in Procyon lotor hernandezii from Mexico.

Gnathostoma lamothei n. sp., inhabiting the stomach of Procyon lotor hernandezii Wagler, 1831, in Tlacotalpan, Veracruz State, and Rio Sapo, Oaxaca, Mexico, is described. This new species differs from all other congeners by having the posterior half of the body surface covered by rows of tiny round bosses instead of spines, or lacking ornamentations. Sequences of the ITS2 of the ribosomal DNA of G. lamothei n. sp. are compared with sequences of other species of the genus recorded in Mexico; they show a wide divergence (<50%) with Gnathostoma binucleatum Almeyda-Artigas, 1991, and Gnathostoma turgidum Stossich, 1902, and high similarity with Gnathostoma sp. I sequence (99.2%). On the basis of morphometric traits and sequences, previous records of Gnathostoma sp. I (=Gnathostoma procyonis of Almeyda-Artigas et al., 1994, not Chandler, 1942, and Gnathostoma neoprocyonis nomen nudem) in Mexico are referred to as the new species.

Animals↗
PubMed2004

Gnathostoma infection in fish caught for local consumption in Nakhon Nayok Province, Thailand. II. Deasonal variation in swamp eels.

From August 2000 to August 2001, 1844 swamp eels (Monopterus albus) were purchased from several local markets in Nakhon Nayok Province, Thailand, and examined for the presence of Gnathostoma advanced third-stage larvae. The overall prevalence was 30.1% and the mean number of larvae/eel (infection intensity) was 10.0. The highest infection rate (44.1%) was found in August 2000 and the lowest (10.7%) in March 2001. The greatest mean number of larvae/eel (75.1) was found in August 2000, whereas the fewest (2.3) was in July 2001. It is suggested that the prevalence and intensity of infection decreased within two months after the end of the rainy season and started to rise again about two months after the next rainy season began. A total of 5,532 Gnathostoma larvae were recovered from 555 infected eels, with a maximum number of 698 larvae/eel. The highest rates of Gnathostoma infection according to eel body length and weight were 87.5% in the group 91-100 cm, and 100% in groups of 901-1100 g, respectively. There were significant correlations between eel body lengths and infection rates, body lengths and infection intensities; eel body weights were also significantly correlated with infection rates and infection intensities. It was noted that the longer/ heavier the eels were, the higher would be the infection rates and the greater the infection intensities. Tissue distributions of Gnathostoma larvae in the livers and muscles of swamp eels were as follows: 43.0% of the total number of larvae were found in the muscles and 57.0% were in the liver; 29.7, 51.7, and 18.6% were in the anterior, middle, and posterior parts, respectively; 35.1% were in the dorsal part, while 64.9% were in the ventral part; 9.0, 18.7, 7.4, 20.6, 33.1, and 11.2% were in the anterodorsal, mediodorsal, posterodorsal, anteroventral, medioventral and posteroventral parts, respectively. Of the 5,532 Gnathostoma larvae examined, 1101 (19.9%) were found to possess morphological variants or abnormal cephalic hooklets. The most common unusual feature was that there were few to numerous extra rudimentary hooklets below row 4 and between the 4 rows of hooklets (7.6%), the presence of a fifth row of hooklets (3.5%), abnormal hooklets in any of the 4 rows of hooklets (5.2%), spiral arrangement of the 4 rows of hooklets (1.8%), and larvae having only 3 rows of hooklets (0.3%).

Animals↗
PubMed2004

A note on the correlation between rainfall and the prevalence of Gnathostoma spp. infective stage larvae in swamp eels in Thailand.

Gnathostoma spinigerum is the major causative agent of human gnathostomiasis, a parasitic zoonosis with a great variety of clinical manifestations. Generally, humans are infected by consumption of third-stage larvae (L3) of G. spinigerum in infected hosts in the form of partially cooked or uncooked food. Surveys of the contamination of Gnathostoma spp. L3 in swamp eels are useful for prevention and control of diseases and have been continuously performed in Thailand. The author performed a retrospective study on 33 previous cross-sectional surveys with geographical data and the prevalence of Gnathostoma spp. L3 that covered 12 provinces in Thailand. The relation between rainfall (derived from the geographical data) and the prevalence of Gnathostomo spp. L3 in swamp eels (derived from the overall infection rate of Gnathostoma spp. L3) was investigated. The least-square equation plot rainfall (y) versus prevalence (x) is y= 9.68x + 1,035.12 (r = 0.83; p < 0.01). A significant correlation was discerned between rainfall and the prevalence of eel infection but not for the season of the survey. Similar to the previous study, the prevalence of eel infection may depend on rainfall rather than season. However, this study focused on only 33 cross-sectional surveys in Thailand; further similar study in other countries to assess the correlation between rainfall and the prevalence of infection is required to substantiate this conclusion.

Animals↗
PubMed1995

A new type of advanced third-stage larvae of the genus Gnathostoma in freshwater eels, Fluta alba, from Nakhon Nayok, central Thailand.

Five advanced third-stage larvae of a newly identified type of genus Gnathostoma were collected from freshwater eels, Fluta alba, which were purchased at a market in Nakhon Nayok, central Thailand. The most remarkable characteristic of the newly identified larvae was the larger body size compared with any other larva of Gnathostoma spp. They were also distinguishable from other species by the shape of their hooklets, which branched in a complex manner at the base: this had not been previously observed in any other larval Gnathostoma. The newly described larvae had an average number of 44.5, 45.0, 49.0 and 55.1 hooklets on the head-bulb from the first to the fourth rows, respectively, which were comparable to those of larval G. spinigerum. However, the average number of nuclei in each intestinal cell was 2.21 and fewer than those of the larvae of G. spinigerum. These results suggest that the new type of larvae belong to either G. vietnamicum, G. malaysiae, or constitute a new species of the genus Gnathostoma.

Animals↗
PubMed1986

Enzyme-linked immunosorbent assay for detection of antibody to Gnathostoma antigen in patients with intermittent cutaneous migratory swelling.

A sensitive enzyme-linked immunosorbent assay is described for the detection of immunoglobulin G antibody to Gnathostoma antigen in the sera of patients with intermittent cutaneous migratory swelling who were suspected of being infected by the tissue nematode Gnathostoma spinigerum. The antigen used was a crude somatic aqueous extract of the third larval stage obtained from naturally infected eels. The 1:320 dilution of all sera from 46 patients gave enzyme-linked immunosorbent assay values above the mean (+2 standard deviations) of normal healthy controls, which was arbitrarily used as a cutoff point. The sera from patients suspected of being infected by another tissue nematode, Angiostrongylus cantonensis, as well as from patients with intestinal roundworm infections, also reacted weakly in the test system used in this study. Nonetheless, when the assay was carried out using both Gnathostoma and Angiostrongylus antigens simultaneously, it appeared to be a reliable laboratory test that can be used to support a clinical diagnosis of gnathostomiasis in patients with intermittent cutaneous migratory swelling.

Antibodies↗
PubMed1998

The first discovery of larval Gnathostoma hispidum (Nematoda: Gnathostomidae) from a snake host, Agkistrodon brevicaudus.

The present study was performed to observe the infection status of several kind of animals with indigenous Gnathostoma in Korea, and morphological characteristics of gnathostome larvae detected from pit-viper, Agkistrodon brevicaudus, for the species identification. To know the existence of Gnathostoma in Korea, 3,450 loaches, 24 bullfrogs, several kinds of snakes, i.e., 55 Elaphe rufodorsata, 2 Dinodon rufozonatum rufozonatum, 62 Rhabdophis tigrinus tigrinus and 87 Agkistrodon spp., and 438 cats were examined. A total of 21 larval gnathostomes was detected from 12 pit-vipers, A. brevicaudus. They were 2,233 x 0.343 mm in average size and covered with about 210 transverse rows of minute cuticular spines. Their characteristic head bulbs were provided with 4 rows of hooklets of which average numbers in each row were 36.8, 39.0, 41.7 and 44.3, posteriorly. In the cross sections of midgut level, the intestinal wall consisted of a single layer of 19-25 elongate epithelial cells with a single nucleus. SEM observation of the larvae revealed unique features of head bulb, cuticular spines on transverse striations and a cervical papilla. On the basis of above morphological characteristics, they were identified as the advanced third-stage larvae of Gnathostoma hispidum. It was first confirmed that the pit-viper, Agkistrodon brevicaudus is the snake intermediate host of G. hispidum.

Agkistrodon↗
PubMed1983

Eosinophilic meningoradiculomyelitis caused by Gnathostoma spinigerum. A case report.

A 51-year-old man had excruciating pains in the left arm and chest approximately four weeks after ingestion of live loaches. Eosinophilia, eosinophilic pleocytosis in the CSF, and a high serum IgE level were noted. Skin tests and antigen-antibody reactions were positive for Gnathostoma infection. His clinical signs and symptoms ameliorated with symptomatic treatment within six months. Only 34 cases of gnathostomiasis involving the CNS have been reported in the English literature, and ours is the first Japanese case, to the best of our knowledge, of eosinophilic meningoradiculomyelitis caused by Gnathostoma spinigerum.

Eosinophilia↗
PubMed2002

Morphology of Gnathostoma spp. isolated from natural hosts in Sinaloa, Mexico.

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.

Animals↗
PubMed1994

Intraocular Gnathostoma spinigerum. Clinicopathologic study of two cases with review of literature.

BACKGROUND: Live intraocular nematode is a rare occurrence that is mostly reported in Southeast Asian countries. Common nematodes that are seen live in the eye are microfilaria, Gnathostoma, and Angiostrongylus. Approximately 12 cases of intraocular gnathostomiasis have been reported in the literature. METHOD: Two cases of intraocular gnathostoma, removed by vitrectomy in the first case and by paracentesis in the second case, are reported. Morphologic study of the parasites in wet preparation was performed under dissecting microscope and fixed in Karnovosky's fixative. Light microscopic and scanning electron microscopic studies were also performed. RESULTS: The first patient had anterior uveitis, multiple iris holes, and dense vitreous haze with fibrous proliferation over the optic disc. On resolution of the vitreous haze, a live worm was seen in the vitreous cavity. The second patient had anterior uveitis with secondary glaucoma, multiple iris holes, mild vitritis, and focal subretinal haemorrhage with subretinal tracts. Four days later a live worm was seen in the anterior chamber and removed. Microscopic study of the parasites from both patients revealed typical head bulb with four circumferential rows of hooklets, and fine cuticular spines were seen on the surface of the body. CONCLUSIONS: Iris holes, uveitis, and subretinal haemorrhage with subretinal tract can be characteristic features of intraocular gnathostomiasis. Identification of this parasite can be made by typical features, which can be identified on light and scanning electron microscopic study.

Adult↗
PubMed1988

Eosinophilic meningitis and radiculomyelitis in Thailand, caused by CNS invasion of Gnathostoma spinigerum and Angiostrongylus cantonensis.

During the 6 year period from January 1980 to December 1985 44 patients with infection of the central nervous system by Gnathostoma spinigerum or Angiostrongylus cantonensis were admitted to the Division of Neurology, Ramathibodi Hospital, Bangkok, Thailand. In 16 patients the diagnosis could be confirmed serologically by means of ELISA techniques. In gnathostomiasis encephalitis, myelitis, radiculitis and subarachnoid haemorrhage formed the majority of clinical syndromes. Intracerebral haematoma and transitory obstructive hydrocephalus are described in this report as being caused by Gnathostoma spinigerum infection for the first time. In angiostronglyus infections the clinical syndrome of meningitis was predominant, but one patient, whose angiostrongyliasis was proved serologically, also showed bilateral paresis of abducens nerve. The main laboratory finding was eosinophilic pleocytosis in the CSF (greater than 10%) which in patients originating or returning from South-East-Asia, particularly Thailand, is highly suggestive of these parasitic infections. Increasing transcontinental travel, influx of refugees and those seeking asylum as well as importation of food from South East Asian countries demand greater awareness of these parasitic infections even in Central Europe.

Adult↗
PubMed2003

Characterization of monoclonal antibodies against Gnathostoma nipponicum.

Monoclonal antibodies (mAbs) were produced against the proteins of advanced third-stage larvae (AdL3) of Gnathostoma nipponicum. Six mAbs (Gn2C3, Gn2H3, Gn4C3, Gn4E9, GnSH1, and Gn10B7) were obtained as determined by enzyme-linked immunosorbent assay (ELISA). Gn4E9 and GnSH1 seemed to be genus-specific, as they did not cross-react with Anisakis sp., Dirofilaria immitis, Gongylonema pulchrum, Toxocara canis, Trichinella sp., Trichuris vulpis, Metagonimus sp., or Spirometra erinaceieuropaei by ELISA. Immunohistochemistry showed that Gn2C3, Gn4E9, and Gn5H1 reacted strongly with the central esophagus; Gn2H3 reacted with cuticle,muscle, intestine, and the cervical sac; and Gn4C3 and Gn10B7 reacted with cuticle, muscle, esophagus, intestine, and the cervical sac of AdL3. In Western blotting analysis, Gn2C3, Gn4E9, and Gn5H1 reacted to 60-, 53-, 46-, and 41-kDa proteins; Gn4C3 reacted to the AdL3 protein of G. nipponicum (>42 kDa). Moreover, proteins purified using a mAb Gn4E9 immunoprecipitation method (sizes 60-, 53-, 46-, and 41-kDa) were used as antigens in ELISAs. A significant difference (P < 0.01) was shown between mouse sera infected with G. nipponicum and sera infected with Trichnella sp. or not infected. These results provide a rationale for evaluating esophageal proteins for the development of diagnostic methods for detecting G. nipponicum or Gnathostoma sp. infections.

Animals↗
PubMed2005

Confirmation of Gnathostoma binucleatum Almeyda-Artigas, 1991, advanced third-stage larvae in Tres Palos Lagoon, Mexico, by morphological and molecular data.

Advanced third-stage larvae of Gnathostoma sp. corresponding to 6 morphotypes, distinguished on the basis of the shape of the cephalic hooklets, were collected from the muscle tissue of 5 species of freshwater fish (i.e., Dormitator latifrons, Eleotris picta, Gobiomorus maculatus, Ariopsis guatemalensis, and Cichlasoma trimaculatum) in Tres Palos Lagoon, Guerrero, Mexico. Principal components analysis of 4 morphological characters cluster all samples in a single taxonomic group. A minimum amount of variation was observed among ITS2 sequences of 3 morphotypes and Gnathostoma binucleatum obtained from GenBank (0-0.84%). The observed variation among morphotypes 1, 2, and 3 is the result of intraspecific variability of G. binucleatum supported by morphology and DNA. Morphotypes 4, 5, and 6 belong to the same taxon on the basis of morphology of the hooklets only. For an accurate morphological diagnosis of the causative agent of gnathostomiasis, it is necessary to develop similar studies with other species of the genus.

Animals↗
PubMed1996

Surface ultrastructures of the third-stage larvae of Gnathostoma hispidum.

A scanning electron microscopic study was performed to observe the surface ultrastructures of the third-stage larvae of Gnathostoma hispidum. The early third-stage larvae (EL3) were collected from the viscera of Chinese loaches by the artificial digestion method. The advanced third-stage larvae (ADL3) were recovered from mice experimentally infected with EL3. Both larval worms were fixed with 2.5% glutaraldehyde, dehydrated in graded alcohol, dryed in critical point dryer, and coated with gold. The specimens were observed with a SEM (DS-130C). On the head bulb of both larval stage, the mouth had a pair of lateral lips of equal size and of half moon shape. Each lip had a couple of labial papillae and a small amphid located between the two papillae. The hooklets on the head bulb had single-pointed tips and curved posteriorly. The cuticular spines of EL3 were larger and more densely distributed in the anterior area (about 1.8 micron in length) and gradually decreased in size and number posteriorly. The cuticular spines in the anterior area of ADL3 were sharp-pointed and about 4.5 micron in length, and those in the middle area were about 1.75 micron. The velvety cuticular folds and dot-like cuticular spines were distributed in the posterior area. A cervical papilla was located between the 7th and 8th transverse striations. A dome-like body papilla was located at the posterior 1/4 of body. An ellipsoidal excretory pore was located between the 17th and 18th striations. From the above results, it is suggested that the characteristic SEM findings obtained from this study may be helpful on the species identification of larval Gnathostoma.

Animals↗
PubMed1995

A case report of colonic ileus due to eosinophilic nodular lesions caused by Gnathostoma doloresi infection.

Gnathostomiasis is primarily a disease of the skin characterized as creeping eruption or mobile erythema. However, larval Gnathostoma sometimes migrate into an unexpected site to elicit serious illness. Here we describe a case of colonic ileus caused by Gnathostoma doloresi. The patient was a 57-year-old man living in Miyazaki Prefecture, Japan, which is known as an area endemic for this parasite. One week after having eaten a few slices of the flesh of a snake (Agkistrodon halys), he developed severe abdominal pain. An abdominal radiograph revealed multiple gas-fluid levels with a distended bowel of an inverted U shape. A barium enema revealed a tumor in the ascending colon near the hepatic flexure that was surgically removed by simple colonic resection. An oblique section of a parasite surrounded by massive infiltration of eosinophils was found by postoperative histopathologic examination. The entire body of the advanced third-stage larva of G. doloresi was dissected from a specimen-embedded paraffin block.

Animals↗
PubMed2000

Gnathostomiasis possibly caused by Gnathostoma malaysiae.

Gnathostomiasis is rarely reported in travelers, although the disease remains a major public health problem in Southeast Asia. A creeping eruption and Quincke's edema (slowly migrating erythema with pruritus) appeared in two Japanese men who had eaten raw freshwater shrimp in Myanmar. A Gnathostoma larva was found in subcutaneous tissue from one of the men. Four species causing human gnathostomiasis, G. hispidum, G. doloresi, G. nipponicum and G. spinigerum, can be distinguished based on the number of nuclei in intestinal epithelial cells of infected larvae, in cross-section. In G. hispidum, only a single large nucleus is found. Morphologically, our larva was initially identified as G. hispidum. However, since the number of epithelial cells was greater and the body width was larger than those of a "large-type" 3rd-stage larva of G. hispidum, the larva was then identified as a 3rd-stage larva of G. malaysiae, Miyazaki and Dun, 1965, as reported by Setasuban et al, (1991). Since no human cases caused by this species of Gnathostoma have previously been encountered, this appears to be the first report of gnathostomiasis due to G. malaysiae.

Animals↗