Larval Gnathostoma spinigerum Detected in Asian Swamp Eels, Monopterus albus, Purchased from a Local Market in Yangon, Myanmar

The present study was performed to determine the infection status of swamp eels with Gnathostoma sp. larvae in Myanmar. We purchased total 37 Asian swamp eels, Monopterus albus, from a local market in Yangon in June and December 2013 and 2014. All collected eels were transferred with ice to our laboratory and each of them was examined by the artificial digestion technique. A total of 401 larval gnathostomes (1-96 larvae/eel) were detected in 33 (89.2%) swamp eels. Most of the larvae (n=383; 95.5%) were found in the muscle. The remaining 18 larvae were detected in the viscera. The advanced third-stage larvae (AdL3) were 2.3-4.4 mm long and 0.25-0.425 mm wide. The characteristic head bulb (0.093 x 0.221 mm in average size) with 4 rows of hooklets, muscular long esophagus (1.025 mm), and 2 pairs of cervical sacs (0.574 mm) were observed by light microscopy. The average number of hooklets in the 1st, 2nd, 3rd, and 4th rows was 41, 45, 48, and 51, respectively. As scanning electron microscopic findings, the characteristic 4-5 rows of hooklets on the head bulb, a cervical papilla, tegumental spines regularly arranged in the transverse striations, and an anus were well observed. Based on these morphological characters, they were identified as the AdL3 of Gnathostoma spinigerum. By the present study, it has been confirmed for the first time that Asian swamp eels, M. albus, from Yangon, Myanmar are heavily infected with G. spinigerum larvae.

The Jeju Weasel, Mustela sibilica quelpartis, A New Definitive Host for Gnathostoma nipponicum Yamaguti, 1941

Adult gnathostomes were discovered in the stomach of the Jeju weasel, Mustela sibilica quelpartis, road-killed in Jeju-do (Province). Their morphological characters were examined to identify the species. Total 50 gnathostome adults were collected from 6 out of 10 weasels examined. In infected weasels, 4-6 worms were grouped and embedded in each granulomatous gastric tumor, except 1 weasel. Male worms were 25.0x1.4 mm in average size, and had a tail with pedunculate papillae, a spicule, and minute tegumental spines. Females were 40.0x2.5 mm in average size, and had a tail without tegumental spines. Pointed and posteriorly curved hooklets were arranged in 8-10 rows on the head bulb. Tegumental spines were distributed from behind the head bulb to the middle portion of the body. The spines were different in size and shape by the distribution level of the body surface. Fertilized eggs were 65.5x38.9 microm in average size, and had a mucoid plug at 1 pole. These gnathostomes from Jeju weasels were identified as Gnathostoma nipponicum Yamaguti, 1941. By the present study, it was confirmed for the first time that G. nipponicum is distributed in Jeju-do, the Republic of Korea, and the Jeju weasel, M. sibilica quelpartis, plays a crucial role for its definitive host.

Various morphologic features of Gnathostoma spinigerum in histologic sections: report of 3 cases with reference to topographic study of the reference worm.

Gnathostomiasis is common in Southeast Asian countries and can be found sporadically in other parts of the world mainly due to human migration. The definitive diagnosis can be given either by identification of the parasite isolated from the patient or through histologic section of the lesion. It is therefore important for pathologists to be familiar with the morphology of parasitic larvae which varies according to the levels of section-cutting so that the diagnosis will not be misled. We present three cases of gnathostomiasis with different features of parasitic morphology and compare these with the reference adult worm.

A survey of infective larvae of Gnathostoma in eels sold in Ho Chi Minh City.

To investigate the distribution of Gnathostoma spp in Ho Chi Minh City (HCM city), 1,081 eels were purchased from a local market twice a month from March 1998 to February 1999. Infective larvae of Gnathostoma spp detected from the flesh and liver of eels by the press preparation technique were examined and identified. Three hundred and fifty advanced third-stage larvae were recovered from liver, none from the flesh. The average rate of infection was 0.11; a high rate of infection was found from August to November and a low rate of infection from February to May. The average number of larvae/eel was 2.9; the greatest number of larvae/eel was in January whereas the lowest was in March and April. There was a marked decrease in both prevalence and intensity of infection from February to May, followed by a rise from June. The finding suggests that in HCM city, the infection rate abruptly decreases soon after the end of the rainy season and starts to rise when the rain comes and reaches its peak at the end of the rainy season. All recovered larvae were identified as G. spinigerum.

Three cases of human gnathostomiasis caused by Gnathostoma hispidum, with particular reference to the identification of parasitic larvae.

Human gnathostomiasis is well known to be caused by Gnathostoma spinigerum, G. hispidum, G. doloresi and G. nipponicum in Japan. In the present reported cases, skin biopsies were performed on three Japanese patients. As a result, cross sections of the larvae were clearly observed in the biopsied specimens. All cross sectioned larvae had an intestinal canal, which consisted from 25-35 cells. A large nucleus was observed at the center of each intestinal cell. The morphological features completely correlated with the characteristics of larval G. hispidum. Based on these findings, all three human cases were thus determined to be confirmed cases of gnathostomiasis caused by G. hispidum.

Gnathostome infection in swamp eels, Fluta alba, in central Thailand.

To investigate the distribution of gnathostome worms in central Thailand, the infective larvae of Gnathostoma spp were examined from the flesh and liver of swamp eels, Fluta alba. Seven hundred and eighty-eight eels were purchased from markets in 11 provinces; Ang Thong (30), Ayutthaya (36), Chachoengsao (30), Lop Buri (30), Nakhon Nayok (437), Pathum Thani (30), Prachin Buri (48), Ratchaburi (53), Saraburi (30), Samut Prakan (30) and Suphan Buri (34). The highest rate of gnathostome infection was observed in swamp eels from Nakhon Nayok (68.7%). The infection rates in Ayutthaya, Ang Thong, Prachin Buri, Ratchaburi, Saraburi and Lop Buri were 33.3%, 26.7%, 25.0%, 18.9%, 13.3% and 10.0% respectively. Gnathostome larvae were not found in swamp eels from Chachoengsao, Pathum Thani, Samut Prakan and Suphan Buri. Among the 9,573 larvae recovered, almost all were the advanced third stage larvae of G. spinigerum, except one larva from Nakhon Nayok and two larvae from Ratchaburi which were identified as the advanced third stage larvae of G. vietnamicum and G. hispidum respectively. This study is the first report of swamp eels as natural intermediate hosts of G. vietnamicum and G. hispidum.

Seasonal variation in the intensity of Gnathostoma larvae in swamp eels (Fluta alba) sold in a local market in Bangkok.

The viscera of swamp eels were obtained from a local market in Bangkok twice a month from June 1996 to May 1997. The livers were separated, weighed and counted. Gnathostome larvae were recovered from the livers by the digestion technic, examined, identified, and counted. A total of 12,278 Gnathostoma larvae were obtained from 18,561.1 g (15,264 pieces) of eel livers. The overall average number of larvae/g liver and the overall average number of larvae/liver are 0.91 and 0.94, respectively. The greatest number of larvae/g liver (on average) was in December (high levels of infection during the months of October to December) whereas the lowest was in April (lowest levels of infection during the months of March to April). Thus there was a marked decrease in the average number of larvae/g liver during January to April, which then started to rise in May. This finding suggests that the level of infection abruptly decreases soon after the completion of the rainy season, starts to rise when the rain has come, and reaches its peak when the amount of rainfall is highest. More than 99% of the total gnathostome larvae recovered were identified to be G. spinigerum, and 25.4% of the entire larvae recovered bore variant or abnormal cephalic hooklets. The most common unusual feature was that there were extra rudimentary hooklets above row one, below row four and in between the four rows of hooklets which comprised 21.4%. In addition, the body size and the number of cephalic hooklets of G. spinigerum are also discussed.

Morphological variation and abnormality of cephalic hooklets of Gnathostoma spinigerum hepatic stage larvae from laboratory infected mice.

One thousand advanced third-stage larvae of Gnathostoma spinigerum from laboratory infected mice, two to five weeks after being fed with infected cyclops, were examined specifically for the morphology of their cephalic hooklets. Among these, only the 15-day old (early hepatic-stage) larvae and the 30-day old (late hepatic-stage) larvae were measured for the size of their body and hooklets. The average body size of the 15-day old and 30-day old larvae were 3.4 +/- 0.4 x 0.4 +/- 0.04 mm and 4.9 +/- 0.4 x 0.5 +/- 0.04 mm, respectively. The average size of the hooklets from rows one to four of the 15-day old larvae was 14.6 +/- 1.7 x 6.8 +/- 0.6 microm, 15.6 +/- 2.0 x 7.2 +/- 0.5 microm, 16.0 +/- 1.8 x 7.4 +/- 0.6 microm and 15.9 +/- 1.9 x 7.3 +/- 0.6 microm, respectively. Those of the 30-day old larvae were 15.1 +/- 1.7 x 7.1 +/- 0.6 microm, 16.3 +/- 1.6 x 7.5 +/- 0.7 microm, 16.5 +/- 1.7 x 7.8 +/- 0.6 microm and 16.3 +/- 1.7 x 7.6 +/- 0.8 microm, respectively. The average number of cephalic hooklets from rows one to four of the two- to five-week old larvae were 42.8 +/- 2.6, 45.3 +/- 2.8, 46.9 +/- 2.8 and 50.2 +/- 2.9, respectively. Several types of morphological variation and abnormality of the cephalic hooklets were observed. The most common ones were extra rudimentary hooklets below row four or within the four rows of hooklets (10.8%), the present of a fifth row of hooklets (1.9%), abnormal hooklets in only row four (1.2%), lobed or branched hooklets (0.5%), spiral arrangement of the four rows of hooklets (0.4%), and fragmented hooklets (0.4%).

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.

Anatomical localization of Gnathostoma spinigerum larval antigens by an indirect fulorescent antibody test.

Indirect fluorescent antibody test (IFAT) was performed on sections of Gnathostoma spinigerum advanced third-stage larva with gnathostomiasis, angiostrongyliasis, trichinosis, strongyloidiasis and cysticercosis sera. Positive fluorescence was observed with the first three sera. Fluorescence was associated with the anterior part of the esophagus, surface of the cuticle and cytoplasmic granules of the intestine. Absorption of sera with gnathostome antigen did not elicit fluorescence. The results suggest that substances secreted from the esophagus and intestine constitute antigens in excretory-secretory products of the larva.

A brief report on Gnathostoma spinigerum specimens obtained from human cases.

A total of 23 gnathostome specimens different stages from the Hospital for Tropical Diseases, other general hospitals and medical clinics in Bangkok metropolitan area and central Thailand were identified as G. spinigerum. The various stages were as follows: Five advanced third-stage larvae (22.0%) of various sizes; 2.20-3.50 mm X 0.40-0.63 mm were removed from the skin, mucous membrane and the eyeball, of which the smallest measured 2.20 X 0.40 mm. Fourteen immature worms of both sexes were from the skin and mucous membrane, one spontaneously voided in the urine. The sizes varied from 4.63-9.35 mm X 0.60- 1.05 mm (8 male worms), and 3.83-16.25 mm. X 0.83-1.0 mm (6 female worms). All these immature worms had 8 rows of complete cephalic-hooklets with the exception of two with 6 and 7 rows of complete cephalic-hooklets. Four mature males size ranged from 9.9-12.5 mm X 1.0-1.25 mm. The smallest was removed from the abdominal skin of a newly born 7-day old baby evidently the case was prenatally transmitted.

Gnathostoma malaysiae Miyazaki and Dunn, 1965 from Rattus surifer in Thailand.

Gnathostoma malaysiae Miyazaki and Dunn, 1965 was found in the stomach wall of Rattus surifer, captured in Phuket Island and Khao Yai National Park of Thailand. This is the first to be recorded in Thailand and the second discovery after the first description of Miyazaki and Dunn (1965). Pathological findings of infected animals were also described. Some new morphological descriptions were added to the original.

Another case of human gnathostomiasis in Indonesia.

A male Gnathostoma spinigerum was removed from the abdominal wall of a 29-year-old woman, who was in the habit of eating "naniura", a dish containing raw carp (Cyprinus carpio). After the removal of the worm she still had pain in her chest and noticed intermittent swelling of her right lower arm. Probably the patient still had one or more Gnathostoma worms migrating in her body.

Gnathostoma spinigerum Owen, 1836 (Nematoda: Gnathostomidae) from a civet cat, Prionodon linsang Hardwick, with reference to its dietary habits.

The recovery of six adult Gnathostoma spinigerum Owen, 1936 from a civet cat, Prionodon linsang Hardwick, constitutes the second reported record of this parasite and the first authenticated case of adult worms found in a wild animal from Malaysia. The food habits of the infected P. linsang as an important link in the transmission of G. spinigerum in the intermediate and definitive hosts together with the probable distribution of this parasite are discussed.