Comparative morphology of minute intestinal fluke eggs that can occur in human stools in the republic of Korea

The egg morphology of minute intestinal flukes (MIF) that can occur as human infections in the Republic of Korea, i.e., Metagonimus yokogawai, M. miyatai, M. takahashii, Heterophyes nocens, Heterophyopsis continua, Stellantchasmus falcatus, Stictodora fuscata, Pygidiopsis summa, and Gymnophalloides seoi, was studied in comparison with Clonorchis sinensis. The adult worms were obtained from residents of endemic areas, and their intrauterine eggs were studied and measured using light microscopy; the length, width, length-width ratio (LWR), and Faust-Meleney index (FMI). Several specimens were processed for scanning electron microscopy (SEM), and before gold-coating, the uterine portion of each fluke was etched with a sharp pin in order to expose the eggs. The MIF eggs were ovoid, pyriform, or elliptical with a size range of 21-35×12-21 μm. S. fuscata eggs revealed the highest FMI (largest in the area) and lowest LWR, whereas P. summa eggs showed the lowest FMI and medium LWR. SEM revealed that G. seoi and S. fuscata had remarkably clean shell surface lacking the muskmelon-like structure which is prominent in C. sinensis eggs. In Metagonimus spp., H. continua, H. nocens, and S. falcatus eggs, minute surface ridges were recognizable though less prominent compared with C. sinensis. On the surface of P. summa eggs, thread-like curly structures were characteristically seen. The results revealed that important differential keys for MIF eggs include the length, width, area (FMI), shape of the eggs, and the extent of the muskmelon-like structure or ridges on their shell surface and operculum.     

Prevalence of Metagonimus metacercariae in sweetfish, Plecoglossus altivelis, from eastern and southern coastal areas in Korea

The present study was performed to determine the current infecion status of Metagonimus yokogawai metacercariae in sweetfish, Plecoglossus altivelis, collected from several streams in eastern and southern coastal areas of Korea. The sweetfish collected were artificially digested with pepsin-HCl solution and examined under a stereomicroscope in August and September, 2007. Out of 145 sweetfish collected from 10 streams in eastern coasts (Gangwon-do and Gye-ongsangbuk-do), 88 (60.7%) were infected with Metagonimus metacercariae. The average metacercarial density was 61 per infected fish. Among 141 sweetfish collected from 10 streams in southern coasts (Gyeongsangnam-do, Ulsan Metropolitan city, and Jeollanam-do), 140 (99.3%) were infected with Metagonimus metacercariae, and their average density was 949 per infected fish. The present study confirmed that M. yokogawai metacercariae are still prevalent in the sweetfish from several eastern and southern coastal localities. The prevalence and metacercarial density were much higher in the sweetfish from streams of southern coasts than in those of eastern coasts. Therefore, attention should be paid to this small fluke infection, and consumption of raw sweetfish naturally produced in these areas should be prohibited.     

High endemicity of Metagonimus yokogawai infection among residents of Samchok-shi, Kangwon-do

A small-scale epidemiological survey was undertaken during 1997-1998 on the residents along the Osib-chon (Stream), Samchok-shi (City), Kangwon-do (Province), to evaluate the status of Metagonimus yokogawai infection. A total of 165 fecal samples was collected and examined by cellophane thick smear and formalin-ether sedimentation techniques. The egg positive rate of M. yokogawai was 29.7%, showing a remarkable difference between males (46.6%) and females (16.3%). To obtain the adult flukes of M. yokogawai, 11 egg positive persons were treated with praziquantel and purged with magnesium sulfate. A total of 242,119 adult flukes (average 22,010 per person, 367-119,650 in range) was collected from diarrheic stools, all of which were identified as M. yokogawai. The results show that M. yokogawai is still highly endemic in this area.     

Decrease of Metagonimus yokogawai Endemicity along the Tamjin River Basin

The Tamjin River which flows from Jangheung-gun via Gangjin-gun to the South Sea was reported to be a highly endemic area of Metagonimus yokogawai infection in 1977 and 1985. However, there were no recent studies demonstrating how much change occurred in the endemicity, in terms of prevalence and worm burden, of metagonimiasis in this river basin. Thus, a small-scale epidemiological survey was carried out on some residents along the Tamjin River basin in order to determine the current status of M. yokogawai infection. A total of 48 fecal samples were collected and examined by the Kato-Katz thick smear and formalin-ether sedimentation techniques. The egg positive rate of all helminths was 50.0%, and that of M. yokogawai was 37.5%, followed by C. sinensis 22.9% and G. seoi 4.2%. To obtain the adult flukes of M. yokogawai, 6 egg positive cases were treated with praziquantel 10 mg/kg in a single dose and purged with magnesium sulfate. A total of 5,225 adult flukes (average 871 specimens per person) of M. yokogawai were collected from their diarrheic stools. Compared with the data reported in 1977 and 1985, the individual worm burdens appeared to have decreased remarkably, although the prevalence did not decrease at all. It is suggested that the endemicity of M. yokogawai infection along the Tamjin River has been reduced. To confirm this suggestion, the status of infection in snail and fish intermediate hosts should be investigated.     

Surface ultrastructure of Metagonimus miyatai metacercariae and adults

A scanning electron microscopic study was performed to observe surface ultrastructures of excysted metacercariae and adults of Metagonimus miyatai. Metacercariae were collected from the scale of the pale chub (Zacco platypus), and adult flukes were harvested 1-4 weeks after infection to rats. In excysted metacercariae, the oral sucker was devoid of tegumental spines and had type I and type II sensory papillae. Anteriorly to the ventral sucker, spines were dense and digitated into 5-7 points, whereas near the posterior end of the body spines were sparse and digitated into 2-3 points. In one-week adults, 7 type II sensory papillae were arranged around the lip of the oral sucker, and at inner side of the lip one pair of small and two pairs of large type 1 sensory papillae were seen on each side. The distribution of tegumental spines was similar to that of metacercariae, but they were more differentiated with 9-11 pointed tips. In two- to four-week old adults, the surface ultrastructure was nearly the same as in one-week old adults, however, sperms were frequently seen entering into the Laurer''s canal. Conclusively, the surface ultrastructure of M. miyatai was generally similar to that of M. yokogawai, however, differentiation of tegumental spines and distribution of sensory papillae around the oral sucker were different between the two species, which may be of taxonomic significance.     

Four Human Cases of Acanthotrema felis (Digenea: Heterophyidae) Infection in Korea

Acanthotrema felis is an intestinal trematode of cats originally reported from the Republic of Korea. Only 1 human case infected with a single adult worm has been previously recorded. In the present study, we report 4 human cases infected with a total of 10 worms recovered after anthelmintic treatment and purging. All 4 patients reside in coastal areas of Jeollanam-do, Korea, and have consumed brackish water fish including the gobies, Acanthogobius flavimanus. The worms averaged 0.47 mm in length and 0.27 mm in width, and had 3 sclerites on the ventrogenital sac; 1 was short and thumb-like, another was long and blunt-ended, and the 3rd was long and broad-tipped. They were identified as A. felis Sohn, Han, & Chai, 2003. Surveys on coastal areas to detect further human cases infected with A. felis are required.     

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.     

Metagonimus yokogawai: a 100-kDa Somatic Antigen Commonly Reacting with Other Trematodes

This study was undertaken to characterize the properties of a 100 kDa somatic antigen from Metagonimus yokogawai. Monoclonal antibodies (mAbs) were produced against this 100 kDa antigen, and their immunoreactivity was assessed by western blot analysis with patients'' sera. The mAbs against the 100 kDa antigen commonly reacted with various kinds of trematode antigens, including intestinal (Gymnophalloides seoi), lung (Paragonimus westermani), and liver flukes (Clonorchis sinensis and Fasciola hepatica). However, this mAb showed no cross-reactions with other helminth parasites, including nematodes and cestodes. To determine the topographic distribution of the 100 kDa antigen in worm sections, indirect immunoperoxidase staining was performed. A strong positive reaction was observed in the tegumental and subtegumental layers of adult M. yokogawai and C. sinensis. The results showed that the 100 kDa somatic protein of M. yokogawai is a common antigen which recognizes a target epitope present over the tegumental layer of different trematode species.     

Heterophyid metacercarial infections in brackish water fishes from Jinju-man (Bay), Kyongsangnam-do, Korea

Heterophyid metacercarial infections in brackish water fishes, i.e., perch, shad, mullet, redlip mullet, and goby, of Jinju-man (Bay), Kyongsangnam-do, Korea, were investigated using a digestion technique. Among 45 perch (Lateolabrax japonicus), the metacercariae of Heterophyopsis continua were found in 55.6% (18.5 metacercariae per fish), Stictodora spp. in 28.9% (3.6), and Metagonimus takahashii in 6.7% (17.0). The metacercariae of H. continua were detected in 23 (65.7%) of 35 shad (Konosirus punctatus). Among 15 mullet (Mugil cephalus), the metacercariae of Pygidiopsis summa were found in 100% (105.9 metacercariae per fish), Heterophyes nocens in 40.0% (8.5), H. continua in 13.3%, and Stictodora spp. in 6.7%. Among 12 redlip mullet (Chelon haematocheilus), the metacercariae of P. summa were detected in 91.7% (1,299 metacercariae per fish), H. nocens in 16.7%, and Stictodora spp. in 16.7%. Among 35 gobies (Acanthogobius flavimanus), the metacercariae of Stictodora spp. were found in 82.9% (44.5 metacercariae per fish), and H. continua in 25.7% (2.8). The adult flukes recovered from a cat experimentally infected with metacercariae from perches were compatible with M. takahashii. The results confirm that the estuarine fish in Jinju-man (Bay) are heavily infected with heterophyid metacercariae. Furthermore, it was found for the first time that the perch acts as a second intermediate host for M. takahashii.     

Surface ultrastructure of Pygidiopsis summa (Digenea: Heterophyidae) adult flukes

A scanning electron microscopic study was performed on the surface ultrastructure of Pygidiopsis summa (Digenea: Heterophyidae) adults. Metacercariae were collected from gills and muscles of mullets (Mugil cephalus) caught in a known endemic area, and adult flukes were harvested from dogs after 8 weeks of experimental infection. The worm was calabash form with its posterior part broader than the anterior part. Tegumental spines were densely distributed over the body surface, except on the suckers and genital apparatus, and around the excretory pore. Well differentiated spines were observed on the anterior half of the body, with 14-16 tips ventrally, and 19-20 tips dorsally. On the oral sucker, three pairs of type I sensory papillae (uni-ciliated knob-like swellings) and one pair of type II sensory papillae (aciliated round-swellings) were observed on the anterior and posterior parts of the lip, respectively. On the lip of the ventral sucker, one pair of type II sensory papillae was distributed only on its posterior part. Sperms were seen emerging from or entering into the genital apparatus. The results showed that the surface ultrastructure of P. summa was unique among the heterophyid trematodes, especially in digitation of tegumental spines and in distribution of sensory papillae on oral and ventral suckers.     

Discovery of Parvatrema duboisi and Parvatrema homoeotecnum (Digenea: Gymnophallidae) from Migratory Birds in Korea

Adult worms of Parvatrema spp. (Digenea: Gymnophallidae) were found in the intestines of 2 species of migratory birds, i.e., a great knot, Calidris tenuirostris, and 2 Mongolian plovers, Charadrius mongolus, in the coastal area of Gunsan-si, Jeollabuk-do in October 2009. The recovered Parvatrema worms were 79 in total number and composed of 2 species. The worms from a great knot were 289 µm in length with the oral and ventral sucker ratio of 2 : 1. They had a single vitellarium, and their intrauterine eggs were 25.0 × 17.5 µm in size. These findings were compatible with P. duboisi (Dollfus, 1923) Bartoli, 1974 (syn. P. timondavidi Bartoli, 1963). The worms recovered from the Mongolian plovers were smaller in length than P. duboisi and had 2 vitellaria. The oral and ventral sucker ratio was 2.5 : 1, and the eggs were 17.5 × 8.8 µm in size. These worms were assigned to be P. homoeotecnum James, 1964. This is the first report on the natural final hosts of Parvatrema spp. in Korea.     

A Case of Chaunocephalosis by Chaunocephalus ferox (Digenea: Echinostomatidae) in an Oriental White Stork,Ciconia boyciana,in Korea

We intended to describe a case of chaunocephalosis and morphological characteristics of its causative agent, Chaunocephalus ferox, recovered from an oriental white stork, Ciconia boyciana, in the Republic of Korea. An oriental white stork was referred to the Wildlife Center of Chungbuk in Korea in February 2014 for severe depression with cachexia and it died the next day. At necropsy, the stomach was severely expanded and 7 thick-walled nodules were observed in the upper part of the intestine. Although the stomach was filled with full of foreign materials, the intestine was almost empty. The nodules were globular and total 9 flukes were recovered. They were 8,030–8,091 μm in length and 3,318–3,333 μm in maximum width. Because the flukes had bulbous forebody with short narrow subcylindrical hindbody, 27 collar spines, and vitelline follicles not reaching to the posterior end, the specimens were identified as being C. ferox. The cyst formation induced thickening of the intestinal wall with narrowing of the lumen that could have contributed to the gastric impaction to the death of the host. This is the first described case of chaunocephalosis and its causative agent C. ferox found from an oriental white stork in Korea.     

Discovery of Endocotyle incana and Spelotrema pseudogonotyla (Digenea: Microphallidae) from Scolopacid Migratory Birds in Korea

Migratory birds have been suspected as playing a central role in transmission of various trematodes, but few surveys have been undertaken in the Republic of Korea. In the present study, we describe new fauna of microphallid flukes in Korea. Two species of microphallids were found in the intestines of 2 migratory bird species, namely the red necked stints (Calidris ruficollis) and the terek sandpiper (Xenus cinereus), in a coastal area of Gunsan-si, Jeollabuk-do. The microphallids recovered from the latter were 451 μm in length and 217 μm in width, and the eggs were very small, 13 μm by 8 μm. They had the characteristic extracecal vitellaria intruding into the median posttesticular area, belonging to Endocotyle incana. Another microphallids were discovered from both migratory birds, 417 μm in length and 249 μm in width. The cirrus was projecting into the genital atrium in form of male papilla, and bounded by a muscular flap, termed pseudogonotyl. They were identified as Spelotrema pseudogonotyla in consideration of the distinctive male papilla and pseudogonotyl. Besides these flukes, Maritrema obstipum and Gynaecotyla squatarolae also were recovered. This paper is in fact the first report on the presence of Spelotrema and Endocotyle in Korea.     

Surveys of Gynaecotyla squatarolae and Microphallus koreana (Digenea: Microphallidae) Metacercariae in Two Species of Estuarine Crabs in Western Coastal Areas, Korea

To figure out the geographical distribution of Gynaecotyla squatarolae and Microphallus koreana metacercariae in Korea, shore crabs of southwestern coastal areas were examined. Eight coastal areas in Inchon-si (A), Gyeonggi-do (B), Chungcheongnam-do (C, D, and E), Jeollabuk-do (F), and Jeollanam-do (G and H) were selected, and 2 kinds of crabs, Macrophthalmus dilatatus and/or Macrophthalmus japonicus, were caught. After transportation to the laboratory, 15 crabs per each group were grouped and ground in a mortar and pestle, and examined for microphallid metacercariae. In M. dilatatus, G. squatarolae metacercariae were recovered from 3 (C, E, and H) out of 6 regions, but M. koreana metacercariae were not recovered. In the case of M. japonicus, G. squatarolae metacercariae were recovered from 6 (B, D, E, F, G, and H) of 7 areas surveyed, and M. koreana matacercariae were detected from 5 regions (A, B, D, F, and H). These results indicate that the life cycle of G. squatarolae is maintained in the western coastal areas using M. dilatatus and M. japonicus as intermediate hosts, while that of M. koreana is maintained only using M. japonicus.     

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.     

Austropeplea ollula (Pulmonata: Lymnaeidae): a new molluscan intermediate host of a human intestinal fluke, Echinostoma cinetorchis (Trematoda: Echinostomatidae) in Korea

Three freshwater snail species of the family Lymnaeidae have been reported from Korea, Radix auricularia coreana, Austropeplea ollula and Fossaria truncatula. Out of 3 lymnaeid snail species, A. ollula was naturally infected with the Echinostoma cinetorchis cercariae (infection rate = 0.7%). In the experiments with the laboratory-bred snails, F. truncatula as well as A. ollula was also susceptible to the E. cinetorchis miracidia with infection rates of 25% and 40%, respectively. All of three lymnaeid snail species exposed to the E. cinetorchis cercariae were infected with the E. cinetorchis metacercariae. It is evident that A. ollula acts as the first molluscan intermediate host of E. cinetorchis in Korea, and F. truncatula may be a possible candidate for the first intermediate host of this intestinal fluke. Also, three lymnaeid snail species targeted were experimentally infected with E. cinetorchis metacercariae.     

Then there were five: a reexamination of the ant genus Paratrechina (Hymenoptera,Formicidae)

The ant genus Paratrechina is reexamined based on the discovery of two new species from Madagascar (P. ankaranasp. n. and P. antsingysp. n.). Paratrechina kohli, a species known from central Africa, is transferred to Paratrechina from Prenolepis based on a new morphological interpretation of the genus and an updated morphological diagnosis of the genus is provided. This means that other than the widespread P. longicornis, whose origins remain uncertain, all Paratrechina are restricted either to the Afrotropical or Malagasy regions. It would also appear that of the five Paratrechina species now known, three are from dry forest habitats. With this reexamination of the genus, the possible origins of P. longicornis are discussed. A key to the genera of the Prenolepis genus-group is provided, as is a key to the workers of Paratrechina. In addition, we also designate a lectotype for Paratrechina kohli.     

Centrocestus formosanus (Digenea: Heterophyidae) encysted in the freshwater fish, Puntius brevis, from Lao PDR

The metacercariae of Centrocestus formosanus, a minute intestinal trematode of mammals and birds, were detected in the freshwater fish, Puntius brevis, from Vientiane Municipality, Lao PDR. The metacercariae were experimentally fed to mice, and adult flukes were recovered in their small intestines 7 days later. The adult flukes were morphologically characterized by having 32 (rarely 34) circumoral spines arranged in 2 alternative rows, a large bipartite seminal vesicle, an oval-shaped ovary, and an X-shaped excretory bladder. Based on these characters, the adults were identified as Centrocestus formosanus (Nishigori, 1924). The taxonomic significance of C. formosanus, in relation to a closely related species, C. caninus (Leiper, 1913), is briefly discussed. It has been first verified by adult worm recovery that C. formosanus is prevalent in Vientiane areas of Lao PDR, taking the freshwater fish, P. brevis, as a second intermediate host.