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 x 51.4 micrometer 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 x 186 micrometer and 510 x 68 micrometer in average, respectively. The rediae parasitized in the snail hosts were found mainly around the pericardial regions, and their size was 1,575 x 258 micrometer 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.
parasitology-helminth-trematoda ; Echinostoma hortense ; epidemiology ; biology ; Misgurnus anguillicaudatus ; Misgurnus mizolepis ; Moroco oxycephalus ; Odontobutis obscura interrupta ; Radix auricularia

Freshwater snail is an important biological group in aquatic ecosystem and an intermediate host of many parasites. Intestinal flora plays an important role in animal energy metabolism and resistance to pathogens. We analyzed the intestinal microbiota diversity of Radix auricularia (RA) and Planorbella trivolvis (PL) by 16S rRNA high-throughput sequencing. At the phylum level, RA had 23 phyla, including Proteobacteria (33.63%), Cyanobacteria (15.33%), Chloroflexi (13.95%), and Actinomycetes (12.99%). PL had 13 phyla, including Proteobacteria (54.88%), Bacteroidetes (28.49%), and Actinomycetes (7.65%). At the genus level, there were 445 genera in RA, including Pleurocapsa, Thiodictyon, Leptotrichia, and Nocardioides. There were 238 genera in PL, including Cloacibacterium, OM60NOR5_clade, Pseudomonas, and Rhodobacter. Ninety-three genera were the common core flora of the two snail species (all the samples were present), and 27 genera had an abundance greater than 0.5%. The structure of intestinal microbiota was significantly different between the two groups (P=0.027). We performed the functional prediction of intestinal microbiota using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), and the results show that the KEGG functional composition of the intestinal flora of the two snails was similar, and the abundance of the amino acid metabolism, carbohydrate metabolism and membrane transport were large. In summary, the intestinal microbiota of the two snails was high in diversity and significantly different, but there were a large number of common core flora.
Animals ; Auricularia ; Ecosystem ; Fresh Water ; Gastrointestinal Microbiome/genetics* ; High-Throughput Nucleotide Sequencing ; Phylogeny ; RNA, Ribosomal, 16S/genetics* ; Snails

The Jinyang Lake is dammed up Nam River, and surrounded by Jinju city and four counties; Jinyang, Sanchong, Hadong, and Sachon in Kyongsang-Nam-Do, Korea. The area around this man-made lake have been known as an endemic focus of clonorchiasis in Korea. The present study was first aimed to know the distribution of freshwater mollusks including medically important snails, and larval trematodes shed from Parafossarulus manchouricus and Semisulcospira libertina. In addtion to above studies, water analyses in each snail habitat were carried out in order to figure out a part of their environmental factors. This malaco-ecological survey was done at the six areas around upper, middle and lower parts of the lake for 4 months, August-November, 1983. Total nine species of freshwater mollusks were collected throughout the study: 4 species of gastropods; Semisulcospira libertina, Cipangopaludina chinensis, Parafossarulus manchouricus and Radix auricularia, and 5 species of bivalves; Unio douglasiae, Anodonta woodiana, Lamprotula gottschei, Corbicula fluminea and Limnoperma lucustris. Out of nine species of freshwater mollusks, three species of gastropods; S. libertina, P. manchouricus and R. auricularia were medically important in terms of the transmission of digenetic trematodes to humans. P. manchouricus and R. auricularia were mainly collected from the shallow ponds and the irrigation channels with the muddy basin, but S. libertina and the bivalves were only collected from the stream of Nam river where the gravels and rocks were dominant. The levels of dissolved oxygen(D.O.) and biochemical oxygen demand (B.O.D.(5)) of the water specimens sampled from the study areas ranged from 6.0 to 9.6 ppm and from 0.4 to 1.6 ppm respectively. As a result, it is considered that water system around the Jinyang Lake might be relatively clean without any heavy pollution of aquatic microorganisms and organic materials during the period of this study. On the other hand, eight metalic constituents from the water samples were also assayed, and all metalic ions detected were remarkably low below the legal criteria. However, calcium ion in the water samples from the habitats of P. manchouricus was considerably higher than others. Infection rates of digenetic trematodes in the snails were 6.9 percent in P. manchouricus and 4.8 poercent in S. libertina, respectively. P. manchouricus snails harboring with the cercariae of Clonorchis sinensis were only 0.14 percent among the snails examined and other trematode cercariae except cercaria of C. sinensis were; furcocercus cercariae, cercaria of Loxogenes liberum type I and II. S. libertina snails parasitized with the cercariae of Metagonimus yokogawai were 1.5 percent out of the snails examined and no cercaria of Paragonimus westermani was found in S. libertina snails in the present study. Digenetic trematode cercariae other then M. yokogawai in S. libertina snails were: Cercaria yoshidae (B type), Cercaria cristata, Cercaria innominatum, Cercaria of Centrocestus formosanus and Cercaria nipponensis.
parasitology-helminth-trematoda ; Clonorchis sinensis ; Loxogenes liberum ; Metagonimus yokogawai ; Centrocestus formosanus ; Cercaria cristata ; Cercaria innominatum ; Cercaria nipponensis ; epidemiology ; Parafossarulus manchouricus ; Semisulcospira libertina ; Radix auricularia