Survey of Gymnophalloides seoi Metacercariae in Natural and Cultured Oysters from Several Western Coastal Areas, Korea

Gymnophalloides seoi (Digenea: Gymnophallidae) is a human intestinal trematode contracted by eating raw oysters (Crassostrea gigas) in the Republic of Korea (=Korea). It has been known to be highly endemic in Aphae Island, Shinan-gun, Jeollanam-do (Province). However, recent epidemiological status of G. seoi has not been reported since the 1990s. In this study, we investigated the prevalence of G. seoi metacercariae in natural and cultured oysters collected from 3 islands and 2 coastal areas in western parts of Korea. The oysters were examined using the artificial digestion method followed by stereomicroscopy. The overall positive rate of G. seoi metacercariae in natural oysters was 66.0% (99/150), and the oysters collected from Yubu Island showed the highest infection rate (74.0%). However, the metacercarial density per oyster was relatively low (1.5–2.4 per oyster). By contrast, no metacercaria was found in cultured oysters purchased from 2 coastal areas in Chungcheongnam-do. Thus, we could confirm that natural oysters produced from 3 western coastal islands are infected with G. seoi metacercariae, whereas cultured oysters purchased from 2 coastal areas were free from infection.

Increased Intestinal Epithelial Cell Turnover and Intestinal Motility in Gymnophalloides seoi-Infected C57BL/6 Mice

The changing patterns of goblet cell hyperplasia, intestinal epithelial cell turnover, and intestinal motility were studied in ICR and C57BL/6 mice infected with Gymnophalloides seoi (Digenea: Gymnophallidae). Whereas ICR mice retained G. seoi worms until day 7 post-infection (PI), C57BL/6 mice showed a rapid worm expulsion within day 3 PI. Immunosuppression with Depo-Medrol significantly delayed the worm expulsion in C57BL/6 mice. Goblet cell counts were increased in both strains of mice, peaking at day 1 PI in C57BL/6 mice and slowly increasing until day 7 PI in ICR mice. In C57BL/6 mice infected with G. seoi, newly proliferating intestinal epithelial cells were remarkably increased in the crypt, and the increase was the highest at day 1 PI. However, in ICR mice, newly proliferating intestinal epithelial cells increased slowly from day 1 to day 7 PI. Intestinal motility was increased in G. seoi-infected mice, and its chronological pattern was highly correlated with the worm load in both strains of mice. Meanwhile, immunosuppression of C57BL/6 mice abrogated the goblet cell proliferation, reduced the epithelial cell proliferation, and suppressed the intestinal motility. Goblet cell hyperplasia, increased intestinal epithelial cell turnover, and increased intestinal motility should be important mucosal defense mechanisms in G. seoi-infected C57BL/6 mice.

STAT6 Expression and IL-13 Production in Association with Goblet Cell Hyperplasia and Worm Expulsion of Gymnophalloides seoi from C57BL/6 Mice

In intestinal helminth infections, Th2 immune respones are generally associated with mucin secretion for worm expulsion from the host intestine. In particular, IL-4 and IL-13 are the important cytokines related with intestinal mucus production via STAT6 signalling in nematode infections. However, this perspective has never been studied in Gymnophalloides seoi infection. The present study aimed to observe the STAT6 signalling and cytokine responses in C57BL/6 mice, a mouse strain resistant to infection with this trematode. The results showed that worm expulsion occurred actively during days 1-2 post-infection (PI), when goblet cells began to proliferate in the small intestine. The STAT6 gene expression in the mouse spleen became remarkable from day 2 PI. Moreover, G. seoi infection induced a significant increase of IL-13 from day 4 PI in the spleen of infected mice. Our results suggested that goblet cell hyperplasia and worm expulsion in G. seoi-infected mice should be induced by STAT6 signalling, in which IL-13 may be involved as a dominant triggering cytokine.

Intestinal Helminth Infections in Feral Cats and a Raccoon Dog on Aphaedo Island, Shinan-gun, with a Special Note on Gymnophalloides seoi Infection in Cats

Four feral cats and a raccoon dog purchased from a local collector on Aphaedo Island, Shinan-gun, where human Gymnophalloides seoi infections are known to be prevalent, were examined for their intestinal helminth parasites. From 2 of 4 cats, a total of 310 adult G. seoi specimens were recovered. Other helminths detected in cats included Heterophyes nocens (1,527 specimens), Pygidiopsis summa (131), Stictodora fuscata (4), Acanthotrema felis (2), Spirometra erinacei (15), toxocarids (4), and a hookworm (1). A raccoon dog was found to be infected with a species of echinostome (55), hookworms (7), toxocarids (3), P. summa (3), and S. erinacei (1). No G. seoi was found in the raccoon dog. The results indicate that feral cats and raccoon dogs on Aphaedo are natural definitive hosts for intestinal trematodes and cestodes, including G. seoi, H. nocens, and S. erinacei. It has been first confirmed that cats, a mammalian species other than humans, play the role of a natural definitive host for G. seoi on Aphaedo Island.

Transition of Parasitic Diseases in Korea

The transition of important parasitic diseases in Korea is briefly reviewed in this article. Soiltransmitted helminthiases, such as ascariasis, trichuriasis, and hookworm infections, which had been prevalent all over the country, decreased remarkably, largely owing to the national control activities by means of mass examination-mass treatment schemes for school students. Paragonimiasis has shown significant reduction in its incidence, but clonorchiasis still remains as an important health-threatening trematode infection. Intestinal trematodiases, including metagonimiasis, heterophyidiases, and echinostomiases are prevalent, and new species, such as Gymonphalloides seoi and Neodiplostomum seoulense, have been documented as new human parasitic diseases. Tapeworm infections including larval cestodiases are also decreasing. Among the protozoan infections, amebiasis and malaria showed an apparent decreasing trend, but in recent years, vivax malaria has become a re-emerging disease. Brugian filariasis, which was prevalent in southern parts of the Korean peninsula, has been completely eliminated nowadays. In terms of the prevalence and distribution, clonorchiasis, enterobiasis, and vivax malaria are currently the major parasitoses of public health importance in Korea.

A new endemic focus of Gymnophalloides seoi infection on Aphae Island, Shinan-gun, Jeollanam-do

A new endemic focus of Gymnophalloides seoi infection has been discovered on Aphae Island (Shinangun, Jeollanam-do), Republic of Korea. This area, which is referred to as Bokyong-ri, is a small seashore village located in the northern portion of the island. Fecal samples were collected from a total of 57 residents and examined by the Kato-Katz and formalin-ether sedimentation techniques. Helminth eggs were detected in 37 samples (64.9%); 33 samples were positive for G. seoi eggs (57.9%), 4 for Pygidiopsis summa (7.0%), 13 for other heterophyids (22.8%), 1 for Clonorchis sinensis (1.7%), and 1 for Trichuris trichiura (1.7%). Women (70.4%) revealed higher rates of G. seoi infection than did men (46.7%), and individuals older than 50 years of age also evidenced higher rates of G. seoi infection than in other age groups (P<0.05). In worm collection from 13 G. seoi egg positive cases, G. seoi (total 86,558 specimens), Heterophyes nocens (278), Stictodora sp. (10), Heterophyopsis continua (3), P. summa (3), and unidentified flukes (42) were collected. Oysters, the source of G. seoi infection, were collected from an area proximal to the village and 50 were examined for metacercariae; 47 (94%) were found infected and the observed metacercarial density was 9.5+/-8.9 per oyster. The results of this study indicate that Bokyong-ri is a new endemic area of G. seoi infection, with high rates of infection in humans and oysters.

Prevalence of Gymnophalloides seoi infection in coastal villages of Haenam-gun and Yeongam-gun, Republic of Korea

One coastal village in Haenam-gun and two in Yeongam-gun, Jeollanam-do were surveyed for intestinal parasite infections by fecal examination. The egg positive rates of Gymnophalloides seoi were high, 24.1% (14/58) in Haenam-gun and 9.3% (11/118) in Yeongam-gun. The egg positive rates of heterophyids, including Heterophyes nocens, and of Clonorchis sinensis were 10.3% and 6.9% in Haenam-gun, and 14.4% and 8.5% in Yeongam-gun, respectively. After praziquantel treatment and purgation, a total of 37,761 fluke specimens were recovered from 17 patients; 11 in Haenam-gun and 6 in Yeongam-gun. Gymnophalloides seoi was the most commonly recovered species, with 37,489 specimens in total (2,205 per person). Other recovered flukes included Heterophyes nocens, Stictodora fuscata, Heterophyopsis continua, Pygidiopsis summa, and undetermined species. These results indicate that the areas surveyed are new endemic foci of G. seoi.

A case of colonic lymphoid tissue invasion by Gymnophalloides seoi in a Korean man

A 65-year old Korean man, living in Mokpo-city, Jeollanam-do, Republic of Korea, visited a local clinic complaining of right upper quadrant pain and indigestion. At colonoscopy, he was diagnosed as having a carcinoma of the ascending colon, and thus, a palliative right hemicolectomy was performed. Subsequently, an adult fluke of Gymnophalloides seoi was incidentally found in a surgical pathology specimen of the lymph node around the colon. The worm was found to have invaded gut lymphoid tissue, with characteristic morphologies of a large oral sucker, a small ventral sucker, and a ventral pit surrounded by strong muscle fibers. This is the first reported case of mucosal tissue invasion by G. seoi in the human intestinal tract.