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Experimental and epidemiological studies on the life cycle of Echinostoma hortense Asada, 1926 (Trematoda:Echinostomatidae)
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Original Article
Korean J Parasitol. 1986 Dec;24(2):121-136. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1986.24.2.121
Copyright © 1986 by The Korean Society for Parasitology
Experimental and epidemiological studies on the life cycle of Echinostoma hortense Asada, 1926 (Trematoda:Echinostomatidae)
Yung Kyum Ahn and Yong Suk Ryang
Department of Parasitology, Yonsei University Wonju College of Medicine, Korea.
Department of Medical Technology, Yonsei University College of Health Science, Korea.
Abstract

Recently there have been some reports on human infections of Echinostoma hortense in Korea. It was found that a few species of freshwater fishes were playing the role of the second intermediate host of E. hortense. However, molluscan intermediate host has not been identified yet in Korea. The present study aimed to establish the life cycle of E. hortense in laboratory. Experimental studies such as egg production from the rat, development of the eggs in vitro, exposure of miracidia to freshwater snails, shedding pattern of cercariae from infected snails, morphology of cercariae, cercarial infection to the second intermediate host and infection of metacercariae to the difinitive hosts were done. In addition, epidemiological surveys on the infection status in inhabitants and house rats, and on the natural infection of larval echinostomes in the snails and fishes were carried out along the South Hangang-river. The results obtained were as follows: The eggs deposited from adults in physiological saline were cultivated at room temperature (20-24C). The miracidia were firstly observed on 8 days after cultivation, and 85.5 per cent of the eggs contained the mature miracidia on 11 days after cultivation. More than 90 per cent formed the miracidia when cultivated at temperature 22-27C. Hatching of the miracidia began on 12 days after cultivation and continued for a week. The size of the miracidia was 103.0 × 51.4 µm in average. The motility of miracidia were active up to 8 hours after shedding, but they were all dead within 10 hours after shedding. A freshwater snail, Radix auricularia coreana was cultivated in aquaria. A hatched F1 snails from the egg masses were exposed to 20 miracidia respectively. Escape of cercariae started on 15 days after infection. Radix auricularia coreana was experimentally identified as the first intermediate host of E. hortense in Korea. Cercarial shedding started on 15-20 days after infection by snail, continued for about 10 days (8.8 days in average). Infected snails were dead within 32 days after the miracidial infection. About 1,335 cercariae (328-1,994) per snail were shed in its life, and 119 cercariae in average per snail per day were shed. The cercariae were motile for more than 24 hours, and then squirming at the bottom until death. The body and tail sizes of cercariae were 356 × 186 µm and 510 × 68 µm in average, respectively. The rediae parasitized in the snail hosts were found mainly around the pericardial regions, and their size was 1,575 × 258 µm in average. The numbers of developing cercariae in a mature redia were 14 in average (7-20 in range). The numbers of rediae in a snail were 102 in average on 15 days after miracidial infection and 221 in average on 28 days. Three uninfected Misgurnus anguillicaudatus, less than 6.5 cm long were used in for the cercaria1 infection. They were all exposed with 755 cercariae, and examined at 5-day intervals starting from 10 days after infection. All the fihes were infected with metacercariae of E. hortense and a total of 275 was found infected (36.4 per cent). The metacercariae were fed to rats and the adult worms were obtained on 15 days after infection. The infected rats began to deposit the eggs on 11 days after infection. The number of eggs deposited per day per worm (EPD/worm) was 400-500 on 3 weeks after infection and was increased to 1,000-1,500 on 4 to 17 weeks, then decreased to 800 on 2l weeks after infection. A total of 745 stool specimens collected from 576 male and 169 female residents of 8 different villages along South Hangang basin was examined. Out of 745 specimens, the eggs of Echinostoma sp. were found in 2 cases (0.3 per cent). Of 34 house rats one showed egg-positive (2.9 per cent). Total 971 Radix auricularia coreana collected from 7 sampling stations were examined for shedding of cercariae. Three snails (0.3 per cent) shed the cercariae of E. hortense. A total of 119 out of 542 freshwater fishes (22.0 per cent) had the metacercariae of E. hortense. The fishes parasitized with the metacercariae were 4 out of 14 examined species. The infection rate of 4 species were 34.1 per cent (106 out of 311) in Misgurnus anguillicaudatus, 30.4 per cent 7 out of 23) in Misgurnus mizolepis, 4.3 per cent (2 out of 46) in Moroco oxycephalus and 22.2 per cent (4 out of 18) in Odontobutis obscura interrupta. In summarizing the above results, the first intermediate host of E. hortense was found as Radix auricularia coreana in Korea. Also it took about 46 days for the shortest completion of a life cycle of E. hortense in summer; that is, 10 days for miracidial development in eggs, 15 days for cercarial development in the snail, about 10 days for metacercarial development in the second intermediate hosts, 11 days for the maturation as the adults in the definitive hosts. The natural infection rates of E. hortense in the intermediate hosts were relatively high but those in the definitive hosts were low in the middle areas of South Hangang basin.

Figures


Fig. 1
Surveyed areas in this study.


Fig. 2
Number of shedding cercariae per day of Echinostoma hortense from experimentally infected snails, Radix auricularia coreana.


Fig. 3
Egg laying capacity (E. P. D./worm) of Echinostoma hortense in rat.


PLATE I
Fig. 1. Adult of E. hortense from the experimentally infected rat, 3 weeks old (OS, oral sucker; CS, cirrus sac; U, uterus; O, ovary; VT, vitellaria), stained by carmine(scale: 1mm).

Figs. 2,3. Matured eggs, operculated, yellowish brown with wrinkled thickening of shell at non-operculated end, and showing the miracidium in egg shell (scale: 100/µm).

Fig. 4.Radix auricularia coreana, the snail of the first intermediate host of E. hortense.

Fig. 5. Miracidium. Hatching start on 10th day of cultivation (scale: 50µm).

Fig. 6. Snail eggs laid on the stem of water plant in the aquaria.

Fig. 7. Veliger stage of the snail, grew 10 days (scale: 2mm).

Fig. 8. Matured Radix snails, reared about 50 days in laboratory (length: 5~7mm).



PLATE II
Fig. 9. Redia, average 1,575/µm long and contain many cercariae and germ balls (scale: 400/µm).

Fig. 10. An equipment for shedding cercariae from the snail in a day.

Fig. 11. Motile cercaria under a cover slip, showing lustrous excretory granules, (scale: 300µm).

Fig. 12,13. Metacercarial cysts subspherical or oval, average 157×l30µm, collar spines and excretory granules conspicuous. Metacercariae occurred in 'the intestinal muscle (Fig. 12) and gills (Fig. 13) of loach under experimental condition (scale: 200µm).


Tables


Table 1
Maturity and hatch of Echinostoma hortense eggs in tap water


Table 2
Measurements of shedding cercariae from snails, Radix auricularia coreana which were experimentally infected with miracidia of Echinostoma hortense (unit: µm)


Table 3
Shedding pattern of cercariae of Echinostoma hortense from experimentally infected snails, Radix auricularia coreana


Table 4
Measurements of redia in snails, Radix auricularia coreana experimentally infected with miracidia of Echinostoma hortense (unit: µm)


Table 5
Location and density of metacercariae in loaches experimentally infected with cercariae of Echinostoma hortense


Table 6
Egg laying capacity (E. P. D./worm) of Echinostoma hortense in rat


Table 7
Prevalence of Echinostoma infection in the final hosts (inhabitants and domestic rats) and detection rate of larvae (shedding cercaria & metacercaria) in the snail and fishes caught (or purchased) along the south Hangang

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