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Cercaria of Nanophyetus Japonensis from the Freshwater Snail, Semisulcospira Libertina, in Japan and Its Experimental Infection

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Cercaria of Nanophyetus Japonensis from the Freshwater Snail, Semisulcospira Libertina, in Japan and Its Experimental Infection [Jpn. J. Parasitol., Vol. 34, No. 2, 41-53, April, 1985] Cercaria of Nanophyetus japonensis from the Freshwater Snail, Semisulcospira libertina, in Japan and Its Experimental Infection Susumu SAITO (Received for publication ; July 2, 1984) Key words : Nanophyetus japonensis, cercaria, metacercaria, adult, experimental exposure, morphology worms were obtained from rats, the final Introduction hosts, 12 days postadministration of the met The freshwater snail, Semisulcospira liber acercariae. From the morphological features tina, is widely distributed in the rivers, of the metacercariae and the adult worms, ponds and lakes of Japan. The search for this parasite was identified as Nanophyetus cercariae parasitizing the snail has been car japonensis. The present paper will demons ried out by many parasitologists, because the trate the above-mentioned results in detail. snail is known as a first intermediate host of Materials and Methods important human trematodes, such as Parag- onimus westermani, Metagonimus yokoga- 1. Detection of cercariae wai, etc. The author started a survey of Each freshwater snail, Semisulcospira liber the snail in 1975 in the Tohoku District, and tina, was put into a 4 cm wide petri dish con found on October 5, 1976 a microcercous xi- taining dicloried tap water. The cercariae phidiocercaria from snails collected in the began to come out from the snails within a drainage way at Tsunagi-Onsen, Morioka few minutes. The snails were left in the City, Iwate Prefecture. This cercaria was dishes for 2 days and the cercariae which morphologically nearly identical with the came out into the water were observed by dis cercaria of Nanophyetus salmincola harboring section microscope. Negative snails were in the freshwater snail, Oxytrema silicula, crushed in the water and re-examined for from the Pacific Coast of North America cercaria. (Sinitsin, 1930; Simms et al., 1931) or the 2. Experimental exposure of freshwater cercaria of N. schikhobalowi harboring in the fishes to cercariae freshwater snails, Semisulcospira laevigata Two species of freshwater fishes, the brook and S. cancella, from the Far East of the trout, Oncorhynchus masou, and the goldfish, USSR (Filimonova, 1965). Therefore, the Carassius carassius auratus, were used for ex second intermediate hosts, the brook trout, perimental exposure. The trout were 18-19 Oncorhynchus masou, and the goldfish, Car- cm in body length and were bred for 7 months assius carassius auratus, were experimentally at Yasuka Fish Farm, Shizukuishi Town, Iwate exposed to the cercaria. The metacercariae Prefecture. Fifteen of them were exposed for were recovered from the hosts 16 to 45 days 30 minutes to cercariae obtained from 5 posi postexposure to the cercariae. The adult tive crushed snails in a plastic container, 26 cm long, 38 cm wide, and 23 cm high with Department of Parasitology, Yamagata Univer water 5 cm in depth. The trout were bred for sity School of Medicine, Yamagata City 990- 45 days in a concrete ditch, 10 m long and 23, Japan. 1 m wide, that was divided into two parts with 42 wire netting. The exposed trout were left at Results the lower part of the stream, and the control trout at the upper part. Mountain water con 1. Geographical distribution of the cercaria tinuously flowed through the ditch. of Nanophyetus japonensis in Semisulcospira Colorless goldfish about 5 cm in body length libertina were obtained for the experimental exposure The cercariae of N. japonensis were detect to cercariae. Three to 5 goldfish and 2-3 ed in the freshwater snail, Semisulcospira li cercarial positive crushed snails were placed bertina, collected in Iwate, Akita and Yama- together for 1 to 2 hours in a plastic contain gata Prefectures (Table 1). In Iwate Prefec er, 21 cm long, 9 cm wide, and 11 cm high ture the snails were collected from the drain with water 3 cm in depth, and then the fish age way of the hot-springs in Tsunagi-Onsen, were bred in a plastic container, 33 cm long, Morioka City, 8 times from October 5, 1976 18 cm wide, and 23 cm high. to September 11, 1977. The cercariae were The water was continuously aerated to detected in the snails every time except on create many chances of contact between the December 15, 1976. In Akita Prefecture, only cercariae and fishes during the exposure one out of 102 snails collected on September period. 21, 1982 were positive with cercariae. From 3. Experimental infection in rats 1975 to 1983, the snails collected from 16 areas Metacercariae encysted in the fish were li of the Shonai Plains and 12 areas of the inland berated from the tissue of the fish at 37 °C by in Yamagata Prefecture were examined. placing in artificial gastric juice consisting of Snails positive with the cercariae of N. 100jHg/ml pepsin and 0.08 N HC1 for 1 to 2 japonensis were found in 2 rivers in the hours. The obtained metacercariae were ad inland. In the upper stream of the Sukawa ministrated to rats. Adult flukes were recov River, Kaminoyama City, 6 out of 94 snails ered from the small intestines of rats one to collected on November 27, 1977, 2 out of 87 two weeks postadministration. snails on May 3, 1978 and 6 out of 76 snails on 4. Procedure for the observation of flukes May 28, 1978 were positive with cercariae. For the morphological observations of cer- In the Daimon River, Yamagata City, the caria and metacercaria, especially of the ex snails were collected 6 times during the period cretory system, the materials were put into a from June 7, 1978 to July 18, 1983, and only half diluted physiological saline solution and 2 out of 80 snails collected on July 18, 1983 were pressed by a cover glass till they became were positive with cercariae. very thin, flat and transparent. These materi 2. Morphology of rediae and cercariae als were preservable for observation for a long Rediae and cercariae were obtained by time when the margin of the cover glass was crushing the snail. Numerous rediae and cer sealed with vaseline. cariae harbored in the mid-gut of the snail. The measurements of redia and cercaria Mature cercariae came out from the snails were carried out with materials fixed in 10 % when they were left in dicloried tap water for hot formalin and slightly pressed under the one day. cover glass. The observations of excysted Redia (Fig. 1-A, Photo. 1-A) : The rediae metacercariae and adults were mainly carried were found among the cercariae. They great out with materials fixed in Schaudinn's ly varied in size, the length being 200//m to solution under cover glass pressure, stained 1945 /im and the width 50 fim to 444 //ra. with Haidenhain's iron-hematoxylin or borax The shape of the redia was generally cylindri carmine, and mounted in balsam. All cal, but the smaller ones tapered from the an measurements were recorded in /mi with the terior end to a point at the posterior end. A aid of a camera lucida. spherical pharynx at the anterior end was 42- 75 X 46-70 //m, and led into a sac-like or cy lindrical gut which reached from one third to ( 2 ) 43 Table 1 Geographical distribution of the cercaria, Nanophyctus japonensis, from the snail host, Semisulcospira liberti?iay in Tohoku District, Japan Locality Date No. exam. No. infect. 0 > Iwate Tsunagi-Onsen, Morioka 5 Oct. 1976 237 1 0.4 10 tt tt 140 2 1.4 29 tt tt 460 3 0.7 15 Dec. tt 147 0 0 5 May 1977 188 2 1.1 20 Jul. tt 311 3 1.0 28 tt tt 650 5 0.8 11 Sept. tt 72 1 1.4 Akita Nishiki-Mura 5 Oct. 1976 55 0 0 1 Sept. 1982 107 0 0 21 tt tt 102 1 1.0 13 ft 1983 130 0 0 31 Aug. 1975 Other 2 areas 665 0 0 8 Aug. 1983 Yamagata Inland areas 7 Jun. 1978 20 0 0 Daimon River, Yamagata 19 Oct. tt 150 0 0 11 Nov. tt 300 0 0 8 Jul. 1979 266 0 0 7 Nov. 1982 262 0 0 18 Jul. 1983 80 2 2.5 Sukawa River, Kaminoyama 27 Nov. 1977 94 6 6.4 3 May 1978 87 2 2.3 28 ti tt 76 6 7.9 Other 10 areas 3 May 1978 1439 0 0 8 Aug. 1983 Shonai Plain 16 areas 9 Jul. 1975 2793 0 0 17 Aug. 1976 two thirds of the length of the redia. The minute backward directed spines and long birth pore was located on the body surface sensory hairs. They were distributed abound- near the side of the pharynx. The redia antly on the anterior part of the body. The contained numerous cercariae in various stag oral sucker was subglobular in shape, 46-75 es of development. fim long by 46-68 fiva wide, accompanied by Cercaria (Table 2, Fig. 1-B, Photo. 1-B) : a conspicuous 13-17 fim long stylet, which had The cercaria was microcercous xiphidiocer- a round base and sharply pointed tip. The carial in type. The body was oval in shape mouth, surrounded by the oral sucker, was and the size of the material fixed with 10 % followed by the pharynx, but the esophagus hot formalin was 229-387//m long by 105-141 and intestine could not be traced. The nerve (im wide. The body surface, cuticle, was commissure ran transversely through the some transparent, and was covered with many what posterior part of the pharynx. The ( 3 ) 44 c D Fig. 1 45 G Photo. 1 46 Table 2 Measurements of various stages of Nanophyetus japonensis in Japan (in Cercaria Metacercaria Adult Body length 229-387(305)* 253-460(342) 560-690(640) Body width 105-141(125) 207-298(247) 245-340(311) Oral sucker length 46- 75( 54) 83-103( 95) 113-150(138) Oral sucker width 46- 68( 57) 103-130(115) 128-170(155) Acetabulum length 44- 56( 50) 63- 83( 69) 108-125(119) Acetabulum width 47- 58( 53) 83-112( 96) 103-138(129) Pharynx length 18- 26( 22) 30- 35( 31) 34- 45( 38) Pharynx width 14- 22( 18) 25- 30( 27) 34- 45( 40) Ovary length 38 ( 38) 50- 75( 66) Ovary width 25- 38( 28) 23- 50( 36) Right testis length 69- 90( 75) 105-140(122) Right testis width 48- 63( 54) 53- 63( 58) Left testis length 60-100( 78) 110-135(125) Left testis width 43- 73( 52) 63- 78( 67) Seminal vesicle length 96-135(113) Seminal vesicle width 71-126( 99) Tail length 15- 19( 18) Tail width 12- 17( 14) Stylet length 13- 17( 15) Stylet width 4- 5( 5) Cyst length 181-345(236) Cyst width 165-338(226) Excretory bladder length 70-125(108) Excretory bladder width 138-233(192) range (average) Fig.
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