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.     

Corbicula fluminea (Bivalvia: Corbiculidae): a possible second molluscan intermediate host of Echinostoma cinetorchis (Trematoda: Echinostomatidae) in Korea.

More than 1,500 clams of Corbicula fluminea, the most favorable food source of freshwater bivalves in Korea, were collected from 5 localities to examine cercarial and metacercarial infection with Echinostoma cinetorchis. Although 3 clams infected with suspicious E. cinetorchis metacercariae out of 200 specimens collected at Kangjin, Chollanam-do were detected, no cercarial and metacercarial infections with E. cinetorchis were observed in field-collected Corbicula specimens. In the susceptibility experiments with laboratory-reared clams, those infected with miracidia of E. cinetorchis did not release their cercariae up to 60 days after infection. To confirm the identity of second intermediate host of E. cinetorchis experimentally, a total of 30 clams were exposed to the cercariae from Segmentina hemisphaerula that had been infected with miracidia of E. cinetorchis. The clams were susceptible to cercariae of E. cinetorchis with an infection rate of 93.3%. Metacercariae from clams taken more than 7 days after cercarial exposure were fed to rats (S/D strain), and adult worms of E. cinetorchis, characterized by 37-38 collar spines on the head crown, were recovered from the ileocecal regions. This is the first report of C. fluminea as a possible second intermediate host of E. cinetorchis.     

Host specificity of Pisidium coreanum (Bivalvia: Sphaeriidae) to larval infection with a human intestinal fluke Echinostoma cinetorchis (Trematoda: Echinostomatidae) in Korea

The fingernail clam, Pisidium coreanum, has been traditionally consumed raw as a so-called drug therapy by patients with bone fractures in Korea. The present study was designed to determine the possible occurrence and, if present, the prevalence of Echinostoma cinetorchis in P. coreanum collected at a local site, and to determine the susceptibility of the clams in the laboratory to infection with miracidia and cercariae of E. cinetorchis. No cercariae or metacercariae of E. cinetorchis were observed in field-collected P. coreanum clams. In susceptibility experiments with laboratory-reared clams, individuals exposed to miracidia of E. cinetorchis did not release cercariae by 20 days after exposure; necropsy of exposed clams failed to show development of any sporocysts or rediae. To confirm the possibility of these clams serving as an experimental second intermediate host of E. cinetorchis, 20 of them were exposed to E. cinetorchis cercariae from experimentally infected Segmentina hemisphaerula that had been previously exposed to miracidia of E. cinetorchis; all exposed clams became infected. Metacercariae from clams at 14 days postinfection were fed to rats, and adult worms were recovered from the ileocecal regions. This is the first report of P. coreanum serving as second intermediate host of E. cinetorchis.     

Segmentina hemisphaerula: a new molluscan intermediate host for Echinostoma cinetorchis in Korea.

Three species of Planorbidae have been reported from Korea, e.g., Gyraulus convexiusculus, Hippeutis (Helicorbis) cantori, and Segmentina (Polypylis) hemisphaerula. Of these, only H. cantori was reported as the first and second intermediate host for Echinostoma cinetorchis, an important human intestinal parasite in Korea. Segmentina hemisphaerula has also been found to be an intermediate host. In field-collected planorbids, only S. hemisphaerula was found shedding echinostome cercariae and infected with metacercariae of E. cinetorchis, whereas no G. convexiusculus and H. cantori were found to be infected. In experiments with laboratory-bred snails, G. convexiusculus and S. hemisphaerula were susceptible to infection by miracidia of E. cinetorchis, but H. cantori could not be infected. Tadpoles of Rana nigromaculata and laboratory-bred snails of the 3 planorbid species were exposed to E. cinetorchis cercariae shed from field-collected S. hemisphaerula. All tadpoles, S. hemisphaerula, and G. convexiusculus became infected, but no H. cantori were infected. Metacercariae from tadpoles, S. hemisphaerula, and G. convexiusculus were fed to rats per os, and eggs of E. cinetorchis were detected in the rat feces 1 wk later. The rats were killed, and adult E. cinetorchis were recovered from the small intestines. This is the first report of G. convexiusculus as a potential first and second intermediate host and of S. hemisphaerula as a new first and second intermediate host for E. cinetorchis in Korea.     

Experimental life history of Echinostoma cinetorchis

The life history of Echinostoma cinetorchis was completed in the laboratory using Hippeutis cantori as the first and second intermediate host. The incubation for maturation of eggs and hatching of miracidia took 24 days at 28 degrees C. On the 66th day after miracidial challenge, 16 snails were crushed and examined for the presence of E. cinetorchis larvae. The metacercariae were detected in all of the snails, and from three of them were found the rediae and cercariae. The morphological characteristics of the larvae and the experimentally obtained adults were compatible with that of E. cinetorchis.     

Host specificity of Austropeplea ollula (Gastropoda: Lymnaeidae) to miracidial infection with a human intestinal fluke Neodiplostomum seoulense (Trematoda: Diplostomatidae) in Korea

Three species of freshwater pulmonate snails of Lymnaeidae have been reported from Korea, Radix auricularia coreana, Austropeplea ollula, and Fossaria truncatula. Of these 3 species, only A. ollula was naturally infected with Neodiplostomum seoulense cercariae (prevalence, 0.3%). In experiments with the laboratory-reared snails, A. ollula was found to be susceptible to the N. seoulense miracidia (prevalence, 10%). This is the first report of A. ollula as the first molluscan intermediate host for N. seoulense in Korea.     

Cercarial shedding of Echinostoma cinetorchis and experimental infection of the cercariae to several kinds of snails

The development of Echinostoma cinetorchis in several snail species reared in laboratory aquaria was observed. The eggs from adult flukes collected from the intestine of rats were cultivated to miracidia, and exposed to Hippeutis sp. snails. Observations were made for cercarial shedding from the exposed snails. The cercariae shed from the snails were again exposed to several species of fresh water snails in order to observe metacercarial formation in the snails and their infectivity to final hosts. The results obtained in this study were as follows: 1. Twenty miracidia were exposed to each snail of Hippeutis sp. About 58.3% of the above snails (7 out of 12) were dead before shedding the cercariae, and the remainder shed the cercariae for a period of 7 to 9 days before death. 2. Cercarial shedding from the infected snails started from the 25th day after the exposure to miracidia, and the total number of cercariae shed per snail was 684 in average (range; 482-904). 3. The size of rediae developed in the infected Hippeutis sp. snails was 1,242 x 214 microns in average, and the number of rediae per snail was 350 in average (range; 120-510). 4. About 40 to 50 cercariae shed from the Hippeutis sp. snails were each exposed to several species of snails reared in the laboratory. The metacercarial formation was confirmed by dissecting the infected snails, 12 to 16 days after the infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

The role of light and gravity in the experimental transmission of Echinostoma caproni (Digenea: Echinostomatidae) cercariae to the second intermediate host, Biomphalaria glabrata (Gastropoda: Pulmonata)

Trematode cercariae inhabit predictable environments and respond to trigger cues with genetically fixed releaser responses when foraging for the upstream host. The effect of light and gravity on the transmission of Echinostoma caproni cercariae to Biomphalaria glabrata was investigated experimentally. Transmission chambers were constructed of clear polyvinyl chloride pipe. Snails were constrained within the chamber to prevent movement, while permitting the cercariae to swim freely. A trial consisted of 2 infected B. glabrata shedding E. caproni cercariae placed at the center of the chamber, with 5 uninfected B. glabrata placed 10 cm on either side (or above and below) of the shedding snails as sentinels. There was no significant difference in the prevalence of infection sentinel snails in either experiment (light vs. dark or top vs. bottom); however, mean intensity was significantly higher in sentinel snails in the dark portion of the chamber (42.5 vs. 10.4; P = 0.001) and the top of the transmission chamber (66.1 vs. 38.0; P = 0.0003). There was a high correlation between the number of metacercariae collected from sentinel snails and the total number of infective units (metacercariae + unsuccessful cercariae): r = 0.992 (light vs. dark) and r = 0.957 (top vs. bottom), respectively, at cercariae densities estimated from 22 to 3,304/L. The results suggest that cercariae of E. caproni exhibit negative photo- and geotaxis in searching for a second intermediate host. Stereotypical releaser responses to environmental trigger cues (light and gravity) allow E. caproni cercariae to exploit flexible strategies for completing the life cycle consistent with the broad range second intermediate and definitive hosts used by E. caproni cercariae and adults, respectively.     

Encystment and metacercariae development of Echinostoma cinetorchis cercariae in an in vitro culture system

For some species of 37-collar-spined Echinostoma, their cercariae successfully encyst and develop to metacercariae in vitro. In our study, we cultured Echinostoma cinetorchis cercariae in 12 different media to study the formation of metacercariae. Locke's solution, medium 199, and RPMI 1640 were used as media for culture. RPMI 1640 produced the highest encystment and normal metacercariae development. The osmolality of the media was related to their ability to encyst and develop. The 0.5 X media induced higher encystment and normal metacercaria formation than the 1x media. The addition of fetal bovine serum to RPMI 1640 increased the level of encystment and normal metacercariae development. In the mixture of 0.5 x RPMI 1640 and 10% fetal bovine serum, encystment was highest, at 96.0%, and the development ratio of normal metacercariae was also the highest, at 91.5%, after 48 hr of cultivation. In a viability test, 7 day- and 14 day-cultured metacercariae were successfully matured to adult worms in experimentally infected rats. These results showed that E. cinetorchis cercariae could be cultured to viable metacercariae in an in vitro culture system and that 0.5 x RPMI 1640 plus 10% fetal bovine serum was the most useful medium for cultivation. This culture system can be adapted for additional studies on the E. cinetorchis life cycle, especially to supply large numbers of metacercariae for other studies on this echinostome.     

Larval trematodes (Digenea) of the great pond snail, Lymnaea stagnalis (L.), (Gastropoda, Pulmonata) in Central Europe: a survey of species and key to their identification

A survey of cercariae and metacercariae (Trematoda, Digenea) from the great pond snail (Lymnaea stagnalis) in Central Europe (Austria, Czech Republic, South-East Germany, Poland and Slovak Republic) is presented, based on a study of 3,628 snails examined from 1998 to 2005. A total of 953 (26.3%) L. stagnalis were infected with 24 trematode species comprising 19 species of cercariae and 11 species of metacercariae (six species occurred both as cercarie and metacercarie) of eight families. The dominant cercariae were those of Opisthioglyphe ranae (159 hosts infected), Plagiorchis elegans (141) (both family Plagiorchiidae) and Echinoparyphium aconiatum (153) (Echinostomatidae); 14 double infections were found. The most frequent metacercariae were those of Neoglyphe locellus (71) (Omphalometridae), E. aconiatum (66), Echinostoma sp. (59) and Moliniella anceps (48) (Echinostomatidae). In the previous studies carried out in Central Europe, a very similar spectrum of nine trematode families of 22 cercariae determined to species level and 43 types of cercariae reported under generic or provisional names, which can be in many cases conspecific with the previous taxa, were found. A simple key to identification of cercariae and metacercariae, together with their illustrations, is provided.     

Occurrence of a diploid type and a new first intermediate host of a human lung fluke, Paragonimus westermani, in Korea

The biology, chromosome number, and karyotype of a lung fluke, Paragonimus westermani (Kerbert, 1878) collected in Haenam, Haenam-gun Chollanam-do, Korea were analyzed. We compared the size of metacercariae from Haenam with those taken from a crayfish collected at Youngam, Youngam-gun, Chollanam-do, Korea. The mean length of P. westermani metacercariae from Haenam was 300.3 microm and that from Youngam was 362.0 microm. Adult worms were recovered from the lungs of experimentally infected dogs. The mean egg sizes obtained from adult flukes were 72.1 x 46.8 microm from Haenam and 93.5 x 54.2 microm from Youngam. Semisulcospira tegulata collected in the Youngam area were found to be infected with cercariae of P. westermani, one of the snail-borne human lung fluke trematodes in Korea. Of 4218 snails studied, 5 (0.12%) harbored P. westrermani larvae. This is the first report of S. tegulata serving as the initial intermediate host of P. westermani. The chromosome numbers of P. westermani from Haenam and Youngam were 2n = 22 and 3n = 33. The diploid type of P. westermani has not been previously reported in Korea.     

Acanthoparyphium tyosenense (Digenea: Echinostomatidae): experimental confirmation of the cercaria and its complete life history in Korea

Cercaria yamagutii Ito, 1957, was found in the marine mesogastropods Lunatia fortuni and Glossaulax didyma from the tidelands of Simpo located at the estuary of the Mankyoung River, which runs to the western coast of Korea. Metacercariae were found in a marine bivalve Mactra veneriformis after being infected with C. yamagutii experimentally. When a sea gull, Larus crassiostris, was fed with the metacercariae collected from the infected M. veneriformis, adult worms were recovered 10 days later. It was confirmed that the parasites collected from L. crassiostris were Acanthoparyphium tyosenense Yamaguti, 1939. From the results of this life cycle study, it was determined that the first intermediate hosts of A. tyosenense are L. fortuni and G. didyma. The second intermediate and final hosts are M. veneriformis and L. crassiostris, respectively. Mactra veneriformis was experimentally infected with C. yamagutii isolated from L. fortuni and G. didyma by maintaining them in a water tank for 30 min at about 20 C. The cercariae entered M. veneriformis through their incurrent siphons. Five hours after infection, the cercariae tails began to separate from the bodies, and the cercariae formed cysts. Mature cysts were formed 340 hr (14 days) after infection and identified as the metacercariae of A. tyosenense. The prevalence of A. tyosenense metacercariae was 99.5% in naturally infected M. veneriformis. This is the first report of C. yamagutii as the cercaria of A. tyosenense, and the complete life cycle of A. tyosenense was established in Korea.     

Larval trematodes (Digenea) of planorbid snails (Gastropoda: Pulmonata) in Central Europe: a survey of species and key to their identification

A survey of the larval stages (cercariae and metacercariae) of trematodes (Digenea) found in planorbid snails in Central Europe (Austria, Czech Republic, south-east Germany, Hungary, Poland and the Slovak Republic) is presented based on a study of 7,628 snails of 12 species examined between 1998-2006. A total of 34 trematode larval stages, comprising cercariae of 28 species and metacercariae of seven species (one species occurred both as cercaria and metacercaria) of nine families were found in 898 (11.5%) snails of eight species. The dominant cercariae were those belonging to the Rubenstrema exasperatum (Rudolphi, 1819)/Neoglyphe locellus (Kossack, 1910) species complex, Tylodelphys excavata (Rudolphi, 1803) and Echinostoma spiniferum (La Valette, 1855) sensu Nasincová (1992), all from Planorbarius corneus (Linnaeus). Almost the same spectrum of cercariae of the families Echinostomatidae, Plagiorchiidae and Omphalometridae was found in the present study as in previous reports; however, a considerably lower spectrum of cercariae of the families Diplostomidae and Strigeidae was recorded. The most frequent metacercariae were those of Echinoparyphium aconiatum Dietz, 1909, Neoglyphe locellus and Moliniella anceps (Molin, 1859), all occurring mainly in P. corneus. The most heavily infected snail species was P. corneus, followed by Planorbis planorbis (Linnaeus) and Segmentina nitida (Müller). The widest spectrum of trematode species was found in P. planorbis and P. corneus. Forty-two cercariae identified to the species level belonging to 15 families, plus an additional 43 taxa recorded under generic or provisional names, were reported from 11 species of planorbids in previous studies carried out in Central Europe. However, the actual number of trematode species occurring in the planorbid snails is probably much lower, because many, if not most, larval stages reported under provisional names or unidentified to the species level may be conspecific with identified adult forms. A key to the cercariae and metacercariae recorded from planorbids in Central Europe, together with illustrations of those species encountered most frequently in the field, is provided to facilitate identification.     

Introduced Schistosomiasis: Evaluation as a Public Health Hazard

Schistosomiasis was found in many agricultural workers from Yemen who are residing in the San Joaquin Valley, and a study was done to evaluate the public health hazard of this imported disease. If the necessary intermediate hosts are present, a local focus of infection could be established.Numerous Biomphalaria obstructa snails collected in the vicinity of the Avenue 82 irrigation canal near Oasis, Riverside County, showed no evidence of natural schistosome infection. Laboratory-reared offspring of these snails were exposed to miracidia of Schistosoma mansoni originating from Yemen immigrants and to miracidia from a standard laboratory strain of Puerto Rican origin obtained from the National Institutes of Health. All exposures of California B. obstructa were made with a laboratory stock of highly susceptible B. glabrata snails also obtained from NIH as controls. Although miracidia penetrated California snails no evidence of infection was detected whereas the B. glabrata controls showed normal, mature infections with numerous cercariae.There appears little likelihood that B. obstructa can serve as intermediate host for schistosomiasis.     

滨鹬马蹄吸虫(复殖目:微茎科)的尾蚴和囊蚴期的季节动态

对科威特湾微茎科滨鹬马蹄吸虫幼虫期的中间宿主双带盾桑椹螺(Clypeomorus bifasciata)及小相手蟹(Nanosesarma minutum)的季节动态进行了研究。调查期超过一年,在检查的1 600只螺和415只蟹中, 11.8 %的螺感染了8种马蹄属线虫中的一种,且以滨鹬马蹄吸虫的感染占优势(9.9 %螺感染) ; 80 %的蟹感染滨鹬马蹄吸虫囊蚴。虽然一年四季两种宿主都会感染,但吸虫的流行和尾蚴(指成熟期感染)在夏季呈现高蜂。从螺体排出的尾蚴具有明显季节性,在此海湾必须要超过最低温度20℃。总的感染率在较大(较老)的螺里有所下降,显示吸虫影响宿主生存并随之影响宿主群体结构。囊蚴的感染丰度与蟹的个体大小有明显相关性;较大的蟹感染较多的囊蚴,显示宿主能耐受更多的吸虫。调查显示,囊蚴的感染率与蟹的大小或性别无相关性。囊蚴体外脱囊以及产卵吸虫的释放证明,成熟虫体终年存在于所有大小和性别不同的蟹里,显示从蟹到鸟的持续感染是可能的。总的来说,滨鹬马蹄吸虫在海湾的传播动态是由这两种无脊椎动物宿主来协调,并似乎是被一系列依赖于温度的活动控制,这些活动影响易感宿主种群及感染性幼虫期尾蚴和囊蚴的存在。     

Tests of ecological equivalence of two species of terrestrial gastropods as second intermediate hosts of Panopistus pricei (Trematoda: Brachylaimidae)

Metacercariae of Panopistus pricei (Brachylaimidae) are common parasites of 2 species of terrestrial gastropods (Neohelix albolabris; Webbhelix multilineata) in southeastern Nebraska. Field data were collected to determine if individuals of N. albolabris and W. multilineata function as ecologically equivalent second intermediate hosts of P. pricei along a transect on the western edge of the Missouri River. Metacercariae were recovered and measured from samples of 30 snails of each species collected at each of 6 sites; whole kidneys from snails of both species were examined to quantify microhabitat use. Microhabitat use of P. pricei did not differ between host species; in both N. albolabris and W. multilineata, metacercariae were concentrated in the primary ureter and occurred throughout the kidney proper. Prevalences and mean abundances did not differ between host species at any site, nor did the relationship between parasite abundance and host size. Prevalences and mean abundances across sites were positively correlated between host species. At 2 sites, demographics of metacercariae differed between host species, suggesting short-term differences in the history of encounters with cercariae in the environment and differences in transmission to shrew definitive hosts. Overall, N. albolabris and W. multilineata appear to be equivalent and required second intermediate hosts of P. pricei in the area studied.     

青海高原牛羊双腔吸虫病病原生物学的初步调查

不文报道在青海高原对牛羊双腔吸虫病病原生物学初步调查的结果。从29个流行地区绵羊收集的标本检查结果,发现有四种双腔吸虫,它们是中华双腔吸虫(Dicrocoelium chinensis)、矛形双腔吸虫(D.lanceatum)、客双腔吸虫(D.hospes)和枝双腔吸虫(D.dendriticum)。从三个流行区检查Helicella candacharica及Cathaica przewalskii二种陆地蜗牛,查到四种双腔吸虫成熟尾蚴,它们是中华双腔吸虫、矛形双腔吸虫及二种未定名的双腔吸虫尾蚴,此二种尾蚴是何种双腔吸虫尾蚴,尚待进一步研究。检查流行区中7种蚂蚁是于9月份从乐都的Formica gagates查到双腔吸虫囊蚴,其形态与中华双腔吸虫囊蚴十分相像。     

Experimental Life History and Biological Characteristics of Fasciola gigantica (Digenea: Fasciolidae)

This study was conducted to investigate the life history, morphology, and maturation of larval stages and adult worms of Fasciola gigantica in experimental mice. Lymnaea auricularia rubiginosa was used as the intermediate host, and Oryza sativa was used for encystment of the metacercariae, while Mus musculus was used as the definitive host for maturation study. Fresh eggs from the gall bladder of water buffaloes fully developed into embryonated ones and hatched out at days 11-12 after incubation at about 29ºC. Free-swimming miracidia rapidly penetrated into the snail host, and gradually developed into the next larval stages; sporocyst, redia, and daughter redia with cercariae. Fully-developed cercariae were separated from the redia and shed from the snails on day 39 post-infection (PI). Free-swimming cercariae were immediately allowed to adhere to rice plants, and capsules were constructed to protect metacercariae on rice plants. Juvenile worms were detected in intestines of mice at days 3 and 6 PI, but they were found in the bile duct from day 9 PI. Juvenile and adult flukes were recovered from 16 mice experimentally infected with metacercariae, with the average recovery rate of 35.8%. Sexually mature adult flukes were recovered from day 42 PI. It could be confirmed that experimentally encysted metacercariae could infect and develop to maturity in the experimental host. The present study reports for the first time the complete life history of F. gigantica by an experimental study in Thailand. The obtained information can be used as a guide for prevention, elimination, and treatment of F. gigantica at environment and in other hosts.     

Life cycle of Stephanoprora uruguayense (Digenea: Echinostomatidae) in Argentina

The life cycle of Stephanoprora uruguayense Holcman et Olagüe, 1989, was experimentally resolved. In an artificial pond in the Zoological Garden in Buenos Aires City, Argentina, Heleobia parchappei (Hydrobiidae) was found to be releasing large-tailed cercariae with a prepharyngeal body, but lacking collar spines and corpuscles in the excretory system. Metacercariae, which encysted on the gills of naturally and experimentally infected Cnesterodon decemmaculatus (Poecilidae), developed collar spines and corpuscles in the excretory system in 7 days. Sexually mature adults were recovered from chicks and immature adults from mice fed metacercariae from C. decemmaculatus. Eggs shed in chick feces developed to miracidia within 10 days; sporocysts were found on the gills of snails. Stephanoprora uruguayense and S. denticulata from Europe are similar in adult morphology, but can be distinguished by morphological and behavioral features of larvae. Likewise, although S. denticulata and S. paradenticulata from Venezuela are similar to S. uruguayense in adult morphology, they differ considerably in larval morphology and intermediate hosts.