Parvatrema chaii n. sp. (Digenea: Gymnophallidae) from mice experimentally infected with metacercariae collected from surf-clam, Mactra veneriformis

Parvatrema chaii n. sp. (Digenea: Gymnophallidae) is described using the worms recovered from experimentally infected mice in Korea. The metacercariae were collected from surf-clams, Mactra veneriformis, from a tidal flat in Sochon-gun, Chungchongnam-do. The metacercariae were elliptical (0.262 x 0.132 mm), and the genital pore had an anterior arch of 16-17 sensory papillae in scanning electron microscopic view. Adult worms were ovoid to foliate (0.275-0.303 by 0.140-0.150 mm), and their characteristic features included the presence of lateral lips, short esophagus, genital pore located some distance anterior to the ventral sucker, club-shaped seminal vesicle, a compact to slightly lobed vitellarium, elliptical eggs (0.018-0.020 by 0.010-0.013 mm), and absence of the ventral pit. This gymnophallid is classified as a member of the genus Parvatrema because of the location of the wide genital pore some distance from the ventral sucker, and the absence of the ventral pit. It differs from previously reported Parvatrema species, including the type species, P. borinquenae. In particular, the morphologies of the vitellarium and the genital pore with an anterior arch of 16-17 sensory papillae are unique features. Therefore, we propose it as a new species, Parvatrema chaii n. sp. (Digenea: Gymnophallidae).     

Identification of Acanthotrema felis (Digenea: Heterophyidae) metacercariae encysted in the brackish water fish Acanthogobius flavimanus

The metacercariae of Acanthotrema felis Sohn et al., 2003 (Digenea: Heterophyidae) were discovered in a species of the brackish water fish, Acanthogobius flavimanus, in the Republic of Korea. They were experimentally fed to kittens, and adult flukes were harvested 7 days later. The adults were morphologically characterized by the presence of a bipartite seminal vesicle, the ventral sucker associated with a ventrogenital sac enclosing 3 sclerites (2 long and pointed, and 1 short and thumb-like), and an unarmed gonotyl. The adult flukes were identified as A. felis Sohn et al., 2003, and the brackish water fish A. flavimanus has been verified as one of its second intermediate hosts.     

Discovery of Maritrema jebuensis n. sp. (Digenea: Microphallidae) from the Asian Shore Crab, Hemigrapsus sanguineus, in Korea

Maritrema spp. (Digenea: Microphallidae) are parasites of birds, but have not been found in the Republic of Korea. In this study, metacercariae of Maritrema sp. were discovered in the Asian shore crab, Hemigrapsus sanguineus, caught in the mud-flats of Jebu-do, Hwasung-gun, Gyeonggi-do, and the adult flukes were confirmed by experimental infection into mice. Based on the symmetric ribbon-like vitellarium, adult flukes of Maritrema sp. were identified, but did not belong to previously described species in terms of the following morphologic characteristics: ceca reaching to the lateral wall at the anterior border of the ovary; ventral sucker larger than oral sucker; a prominent metraterm; and vitellarium forming a complete ring. Hence, we named this microphallid M. jebuensis n. sp. after the island where the second intermediate hosts were collected. From this study, it has been shown that Maritrema sp. is distributed in Korea and transmitted by the Asian shore crab, H. sanguineus.     

Mactra veneriformis, an intertidal clam, as a new second intermediate host for Acanthoparyphium marilae (Digenea: Echinostomatidae)

Metacercariae of Acanthoparyphium marilae Yamaguti, 1934 (Digenea: Echinostomatidae) were discovered in an intertidal clam, Mactra veneriformis, in a southwestern coastal area of the Republic of Korea. A total of 128 metacercariae were detected from 10 clams examined. They were round, 320 microm in average diameter, with 23 collar spines. They were fed experimentally to chicks, and 10 days later adult flukes were obtained. The adults were morphologically characterized by the head collar with a single row of 23 dorsally uninterrupted spines, without special end group spines, a round ventral sucker, 2 round and tandem testes, and vitellaria extending at lateral fields from the posterior extremity not beyond the middle level of the posterior testis. The most characteristic feature of this species was the limited distribution of vitellaria, which differs from Acanthoparyphium tyosenense Yamaguti, 1939, the metacercariae of which are encysted in the same mollusk species. This is the first report in which the metacercariae of this species were detected, and the intertidal bivalve, M. veneriformis, has been identified as a second intermediate host for A. marilae.     

Massaliatrema misgurni n. sp. (Trematoda: Heterophyidae) whose metacercariae encyst in loaches (Misgurnus anguillicaudatus)

Hitherto unknown metacercariae were found encysted in loaches (Misgurnus anguillicaudatus) from China. They were experimentally fed to golden hamsters, and gravid adults were recovered 1 week post-infection from their small intestines. A new species, Massaliatrema misgurni n. sp. (Heterophyidae), is described from the adults. This new species is different from M. gyrinicola Dollfus and Timon-David, 1960, in having a smaller acetabulum/oral sucker ratio, less branched vitellaria widely entering the intercecal anteroacetabular area and an almost median seminal receptacle; and from M. yamashitai Kamiya and Ohbayashi, 1975, in having a larger acetabulum/oral sucker ratio, with the seminal vesicle situated in the uterine loop and the vitellaria entering the intercecal anteroacetabular area. This report is the first record of M. anguillicaudatus as a second intermediate host of the genus Massaliatrema Dollfus and Timon-David, 1960.     

Bartolius pierrei n. g., n. sp. (Digenea: Gymnophallidae) from the Península Valdés, Argentina

Bartolius pierrei n. g., n. sp. (Digenea: Gymnophallidae) is described from metacercariae and naturally and cultivated obtained adults from southern Argentina. The second intermediate host is Darina solenoides (King) (Bivalvia: Mactridae) and the definitive host is Larus dominicanus Lichtenstein (Aves: Laridae). The diagnostic characters are as follows: Body small, oval. Oral sucker without lateral projections, twice size of ventral sucker (except in young metacercariae). Caeca short (in adults), without dorsal diverticula. Ventral sucker in posterior third of body. Ventral pit absent. Seminal vesicle bipartite. Ovary post-testicular. Vitelline glands paired, compact, close to ventral sucker. Uterus in fore- and hindbody. Genital atrium tubular. Genital pore inconspicuous, close to anterior margin of ventral sucker. Excretory vesicle Y-shaped with very short stem. Excretory formula: 2[(2+2)+(2+2)]=16. Bartolius is distinguished from other genera of the Gymnophallidae by the post-testicular position of the ovary.     

Holostephanus metorchis (Digenea: Cyathocotylidae) from chicks experimentally infected with metacercariae from a fish, Pseudorasbora parva, in the Republic of Korea

Holostephanus metorchis (Digenea: Cyathocotylidae) is a parasite of birds, transmitted by freshwater fishes. H. metorchis adults were recovered from chicks experimentally infected with metacercariae collected from freshwater fishes, Pseudorasbora parva. The metacercariae were oval, surrounded with thick fibrous capsules. In adult flukes, the holdfast organ occupied the ventral concavity, and the anterior testis did not reach the level of the ventral sucker. Based on these morphological characteristics, these flukes were identified as H. metorchis.     

Redescription and surface ultrastructure of Pygidiopsis macrostomum (Digenea: Heterophyidae)

Pygidiopsis macrostomum Travassos, 1928, a poorly known species originally described from a single specimen from Rattus norvegicus (Erxleben, 1777) in Brazil, is redescribed on the basis of metacercariae from the mesenteries of naturally infected guppies Poecilia vivipara Bloch and Schneider, 1801 (Poeciliidae), and adults obtained from an experimental infection of hamsters. Pygidiopsis macrostomum is characterized by the absence of oral spines, vitellaria extending forward to ventral sucker, uterus reaching pharyngeal level, X-shaped excretory vesicle, and an oral sucker/acetabulum ratio of 1:0.8. The surface ultrastructure shows that the tegument of the metacercaria does not strongly differ from that of adults. The brush-shaped spines of P. macrostomum are similar to those reported for Pygidiopsis summa and Pygidiopsis ardeae, but no differences in spine shape were observed throughout the body.     

Rhipidocotyle genovi n. sp. (Digenea: Bucephalidae) from the intestine ofGaidropsarus mediterraneus (L.) (Gadiformes: Gadidae) from the Black Sea

Rhipidocotyle genovi n. sp. is described from the fishGaidropsarus mediterraneus (L.) (Gadiformes: Gadidae) in the Black Sea. Morphological features characteristic of this species include: an anterior sucker with seven prominent muscular appendages; a pharynx rather posterior in position; ovary and testes relatively anterior in position, usually anterior to the pharynx; a long (in relation to body length) sucker, cirrus-sac and excretory vesicle; the presence of a pre-oral lip; and contiguous vitelline fields which form an arch. The new species is distinguished fromR. galeata, R. lamberti andBucephalus marinum. This is the first member of the genus whose adults are recovered from Black Sea marine fishes. Metacercariae, which encyst in large numbers in the musculature and fins of the blenniid fishesBlennius tentacularis, B. sanguinolentus, B pavo, B. sphinx andB. zvonimiri and exhibit a similar morphology to the adult, are described.     

Parvatrema duboisi (Digenea: Gymnophallidae) Life Cycle Stages in Manila Clams,Ruditapes philippinarum,from Aphae-do (Island), Shinan-gun,Korea

Life cycle stages, including daughter sporocysts, cercariae, and metacercariae, of Parvatrema duboisi (Dollfus, 1923) Bartoli, 1974 (Digenea: Gymnophallidae) have been found in the Manila clam Ruditapes philippinarum from Aphae-do (Island), Shinan-gun, Jeollanam-do, Korea. The daughter sporocysts were elongated sac-like and 307–570 (av. 395) μm long and 101–213 (av. 157) μm wide. Most of the daughter sporocysts contained 15–20 furcocercous cercariae each. The cercariae measured 112–146 (av. 134) μm in total length and 35–46 (av. 40) μm in width, with 69–92 (av. 85) μm long body and 39–54 (av. 49) μm long tail. The metacercariae were 210–250 (av. 231) μm in length and 170–195 (av. 185) μm in width, and characterized by having a large oral sucker, genital pore some distance anterior to the ventral sucker, no ventral pit, and 1 compact or slightly lobed vitellarium, strongly suggesting P. duboisi. The metacercariae were experimentally infected to ICR mice, and adults were recovered at day 7 post-infection. The adult flukes were morphologically similar to the metacercariae except in the presence of up to 20 eggs in the uterus. The daughter sporocysts and metacercariae were molecularly (ITS1–5.8S rDNA-ITS2) analyzed to confirm the species, and the results showed 99.8–99.9% identity with P. duboisi reported from Kyushu, Japan and Gochang, Korea. These results confirmed the presence of various life cycle stages of P. duboisi in the Manila clam, R. philippinarum, playing the role of the first as well as the second intermediate host, on Aphae-do (Island), Shinan-gun, Korea.     

A new cercaria and metacercaria of Acanthoparyphium (Echinostomatidae) found in an intertidal snail Zeacumantus subcarinatus (Batillaridae) from New Zealand

A new 23-collar-spined cercaria and metacercaria are described from intertidal molluscs of the coast of New Zealand. The new cercaria found emerging from the mud snails Zeacumantus subcarinatus (Sowerby) (Prosobranchia: Batillariidae) is characterized mainly by the number and arrangement of the cephalic glands, the size of the suckers, and the size and number of the collar spines. The cercaria encysts in the cockle Austrovenus stutchburyi (Wood) (Bivalvia: Veneridae) which lives in sympatry with the first intermediate host throughout New Zealand. Laboratory infections of cockles by cercariae from naturally infected snails resulted in metacercariae identical to those found in naturally infected cockles. The main features of the cercaria and metacercaria are the presence of a reniform collar with 23 spines, two pairs of small cephalic glands at the oral sucker level, another two pairs of much longer ones posterior to the pharynx, and the excretory vesicle Y-shaped with the main collecting canals extending to the anterior level of the ventral sucker. Each of the main collecting canals had 10-11 pairs of bilateral diverticula between the anterior edge of the ventral sucker and anterior body end. A brief discussion of its possible life cycle and ecology is also provided.     

Survey of Zoonotic Trematode Metacercariae in Fish from Water systems of Geum-gang (River) in Republic of Korea

The infection status of zoonotic trematode metacercariae (ZTM) was surveyed in freshwater fishes from the water systems of Geum-gang (River) in the Republic of Korea (Korea). A total of 1,161 freshwater fishes from 6 local sites of Geum-gang were examined with the artificial digestion method for 4 years (2012–2015). Clonorchis sinensis metacercariae were detected in 122 (37.2%) out of 328 fishes in the positive fish species from 4 surveyed areas, and their mean intensity was 43 per fish infected. Metagonimus spp. metacercariae were found in 432 (51.7%) out of 835 fishes in the positive fish species from all 6 surveyed areas, and their mean intensity was 30 per fish infected. Centrocestus armatus metacercariae were detected in 285 (75.0%) out of 380 fishes in the positive fish species from 6 surveyed areas, and their mean intensity was 2,100 per fish infected. Echinostoma spp. metacercariae were found in 56 (19.7%) out of 284 fishes in the positive fish species from 5 surveyed areas, and their mean intensity was 10 per fish infected. Clinostomum complanatum metacercariae were detected in 98 (57.3%) out of 171 fishes in the positive fish species from only 2 surveyed areas, and their mean intensity was 11 per fish infected. Conclusively, the endemicity of ZTM is not so high in fishes from water systems of Geum-gang in Korea although it is more or less different by fish species, surveyed areas and ZTM species.     

Taxonomic status, redescription, and surface ultrastructure of Ascocotyle (Phagicola) pindoramensis n. comb. (Digenea: Heterophyidae)

Pygidiopsis pindoramensis Travassos, 1928, is redescribed from the holotype and specimens obtained from experimental infections of hamsters with metacercariae of the naturally infected poeciliids Poecilia vivipara and Phalloptychus januarius from Rio de Janeiro, Brazil. Ascocotyle (Phagicola) pindoramensis (Travassos, 1928) n. comb. is proposed, based on the presence of a solid, tapering muscular posterior prolongation of the oral sucker and Ascocotyle (Phagicola) mollienisicola (Sogandares-Bernal and Bridgman, 1960) is synonymized with A. (P.) pindoramensis because of the identical morphology and measurements of adults and metacercariae, similar spectrum of fish intermediate hosts (poeciliids), and the same site of infection of the metacercariae. Trematodes designated as Pygidiopsis pindoramensis, previously reported from Argentina and Mexico, represent another species of Pygidiopsis Looss, 1907, because they possess circumoral spines and their oral sucker is devoid of the posterior muscular prolongation. Data on the surface morphology of metacercariae and adults of A. (P.) pindoramensis are inferred from scanning electron microscopy observations.     

Surface ultrastructure of Metagonimus takahashii metacercariae and adults

A scanning electron microscopic study was performed on the surface ultrastructure of metacercariae and adults of Metagonimus takahashii. Metacercariae were collected from the scale of crucian carp (Carassius auratus), and adult flukes were harvested 1-4 weeks after infection to rats. In excysted metacercariae, the oral sucker had type I (numerous) and type II (seven in total) sensory papillae. Tegumental spines were dense and digitated into 5-7 points on the surface anterior to the ventral sucker, but became sparse and less digitated posteriorly toward the end of the body. In adults, seven type II sensory papillae were characteristically arranged around the lip of the oral sucker, and on the inner side of the lip four small and two large type I sensory papillae were symmetrically seen on each side (12 in total). Tegumental spines on anterior two-thirds of the body, were digitated with 9-12 tips ventrally and 8-13 tips dorsally. Sperms entering into the Laurer's canal were observed. The results show that the surface ultrastructure of M. takahashii is generally similar to those of M. yokogawai and M. miyatai except for the digitation of tegumental spines.     

Two species of goby, Boleophthalmus pectinirostris and Scartelaos sp., as the new second intermediate hosts of heterophyid fluke in Korea

A survey was performed in order to determine the infection status of the metacercariae of heterophyid fluke in two goby species, Boleophthalmus pectinirostris and Scartelaos sp., collected from Gangjin-gun, and Shinangun, Sooncheon-shi, Jeollanam-do, Republic of Korea. A total of three metacercariae of Heterophyopsis continua was found in only one B. pectinirostris (10.0%) from Gangjin-gun. Heterophyes nocens metacercariae were detected in 24 B. pectinirostris (96.0%) and 14 Scartelaos sp. (63.6%) from Shinan-gun. Heterophyopsis continua metacercariae were found in 11 B. pectinirostris (44.0%) and 21 Scartelaos sp. (95.5%) from Shinan-gun. Stictodora fuscata metacercariae were detected in 18 B. pectinirostris (72.0%) from Shinan-gun. No metacercariae were detected in 20 B. pectinirostris from Sooncheon-shi. From the above results, this study is the first to prove that B. pectinirostris and Scartelaos sp. serve as the second intermediate hosts of some heterophyid flukes in Korea.     

Cercaria caribbea LVIII Cable, 1963 (Digenea: Cyathocotylidae) in the Republic of Korea and its surface ultrastructure

Cercaria caribbea LVIII Cable, 1963 (Digenea: Cyathocotylidae) was detected from a brackish water gastropod species (Cerithideopsilla cingulata) in a coatal area of Shinan-gun, Jeollanam-do (Province), the Republic of Korea, and its surface ultrastructure was studied using a scanning electron microscope. The cercariae were found freely swimming or enveloped within daughter sporocysts when the snail host was mechanically broken. They were morphologically characterized by a linguiform and ventrally concave body, a long and bifurcated tail, and the presence of a holdfast (=tribocytic) organ posterior to the ventral sucker. On the whole ventral and dorsal surfaces, peg-like tegumental spines were densely distributed. Around the oral sucker, several sensory papillae, each with a short cilium, were distributed, and on the tail, sensory papillae, each with an extensively long cilium, were observed. This is the first record describing a cyathocotylid cercaria from a brackish water gastropod in the Republic of Korea.     

Tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis]

The tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis (Trematoda: Echinostomatidae) was observed by scanning electron microscopy. Three-day (juvenile) and 16-day (adult) worms were harvested from rats (Sprague-Dawley) experimentally fed the metacercariae from the laboratory-infected fresh water snail, Hippeutis cantori. The worms were fixed with 2.5% glutaraldehyde, processed routinely, and observed by an ISI Korea DS-130 scanning electron microscope. The 3-day old juvenile worms were elongated and ventrally curved, with their ventral sucker near the anterior two-fifths of the body. The head crown was bearing 37-38 collar spines arranged in a zigzag pattern. The lips of the oral and ventral suckers had 8 and 5 type II sensory papillae respectively, and between the spines, a few type III papillae were observed. Tongue or spade-shape spines were distributed anteriorly to the ventral sucker, whereas peg-like spines were distributed posteriorly and became sparse toward the posterior body. The spines of the dorsal surface were similar to those of the ventral surface. The 16-day old adults were leaf-like, and their oral and ventral suckers were located very closely. Aspinous head crown, oral and ventral suckers had type II and type III sensory papillae, and numerous type I papillae were distributed on the tegument anterior to the ventral sucker. Scale-like spines, with broad base and round tip, were distributed densely on the tegument anterior to the ventral sucker but they became sparse posteriorly. At the dorsal surface, spines were observed at times only at the anterior body. The results showed that the tegument of E. cinetorchis is similar to that of other echinostomes, but differs in the number and arrangement of collar spines, shape and distribution of tegumenal spines, and type and distribution of sensory papillae.     

Acanthotrema felis n. sp. (Digenea: Heterophyidae) from the small intestine of stray cats in the Republic of Korea

Acanthotrema felis n. sp. (Digenea: Heterophyidae) is described from adult flukes recovered in the small intestine of stray cats caught in the southwestern part of the Republic of Korea. The worms were characterized by minute body size, the presence of a bipartite seminal vesicle, and a ventral sucker associated with a ventrogenital sac, which enclosed 3 sclerites (2 long and pointed, and I short and thumblike), and an unarmed gonotyl. They were assigned to Acanthotrema Travassos, 1928, which currently contains 4 species. The new species resembles A. acanthotrema, the type species, because both have 3 sclerites on the ventrogenital sac. However, all 3 sclerites in the new species are armed with minute spines on their base, whereas only 2 sclerites in A. acanthotrema have minute spines near their tips. Three species of Stictodora Looss, 1899, namely,Stictodora tridactyla Martin and Kuntz, 1955, S. cursitans Kinsella and Heard, 1974, and S. tanayensis Velasquez, 1973, are transferred to Acanthotrema, and the new species is distinguished from them particularly in terms of the number and shape of its sclerites. The generic diagnosis of Acanthotrema is emended.     

A new neodiplostomid (Digenea) from the intestine of chicks infected with metacercariae from the grass snake, Rhabdophis tigrina

Three species of neodiplostomula are known to inhabit the European grass snake, Rhabdophis tigrina, in the Republic of Korea: Pharyngostomum cordatum (large-sized neodiplostomula), an intestinal trematode of cats; Neodiplostomum seoulense (small-sized neodiplostomula), an intestinal trematode of humans and rodents; and Neodiplostomum leei (small-sized neodiplostomula), which migrates to the livers of rodents and is an intestinal trematode of birds. The present study describes a fourth species, Neodiplostomum (Conodiplostomum) boryongense n. sp. (Digenea: Neodiplostomidae), based on adult flukes recovered from the small intestines of chicks experimentally infected with small-sized neodiplostomula from the grass snake. The new species differs from 13 previously known species. It also differs from N. seoulense in its larger body size, severely bilobed testes, and smaller genital atrium, and from N. leei in its larger body size, smaller ventral sucker, presence of a genital cone, and vitelline follicles distributed chiefly in the forebody. The new species does not migrate to the livers of rodents nor does it develop to adulthood in the rodent intestines. However, the neodiplostomula of the new species are indistinguishable from those of the other 2 species. Results show that at least 4 species of neodiplostomula inhabit the grass snake in the Republic of Korea.     

Blood flukes (Digenea: Aporocotylidae) of stingrays (Myliobatiformes: Dasyatidae): Orchispirium heterovitellatum from Himantura imbricata in the Bay of Bengal and a new genus and species of Aporocotylidae from Dasyatis sabina in the northern Gulf of Mexico

We redescribe Orchispirium heterovitellatum based on the holotype and 3 original voucher specimens collected from the mesenteric blood vessels of scaly whiprays Himantura imbricata (Bloch and Schneider, 1801) (as Dasyatis imbricatus) captured in the western Bay of Bengal off Waltair, India. We emend the diagnosis of Orchispirium to include anterior sucker present, testis looping, cirrus sac enveloping large internal seminal vesicle, oviducal seminal receptacle present, and metraterm short and thin-walled. We describe Myliobaticola richardheardi n. gen., n. sp. based on live observations, light microscopy, and scanning electron microscopy of adult specimens collected from between the cardiac trabeculae of Atlantic stingrays Dasyatis sabina (Lesueur, 1824) captured in Mississippi Sound (type locality), Mississippi, and Apalachicola Bay, Florida. The new species has a minute, aspinous body lacking lateral tubercles; an aspinous and eversible anterior sucker lacking a peduncle; a posterior esophageal swelling; an inverse U-shaped intestine; smooth ceca terminating in the anterior body half; a looping testis lacking lobes; a cirrus sac enveloping a large internal seminal vesicle; a medial and primarily post-testicular ovary; an oviducal seminal receptacle; a postgonadal uterus flanking the internal seminal vesicle; a short and thin-walled metraterm; and a common genital pore. It lacks a pharynx and Laurer's canal. No other named aporocotylids infect a member of cohort Batoidea or have the combination of an aspinous body, an aspinous anterior sucker, a posterior esophageal swelling, an inverse U-shaped intestine, a looping testis, a cirrus sac enveloping a large internal seminal vesicle, and a common genital pore; these observations indicate that O. heterovitellatum and M. richardheardi are closely related. The discovery of a second species representing a second genus of Aporocotylidae in diamond stingrays (Dasyatidae) suggests that Batoidea is an undersampled host group for aporocotylid infections.