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.     

Effect of temperature on emergence, survival and infectivity of cercariae of the marine trematode Renicola roscovita (Digenea: Renicolidae)

Marine bivalves harbour a diversity of trematode parasites affecting population and community dynamics of their hosts. Although ecologically and economically important, factors influencing transmission between first (snail) and second (bivalve) intermediate hosts have rarely been studied in marine systems. In laboratory experiments, the effect of temperature (10, 15, 20, 25 degrees C) was investigated on (1) emergence from snails, (2) survival outside hosts and (3) infectivity in second intermediate hosts of cercariae of the trematode Renicola roscovita (Digenea: Renicolidae), a major parasite in North Sea bivalves. Emergence of cercariae peaked at 20 degrees C (2609 +/- 478 cercariae snail(-1) 120 h(-1)) and was considerably lower at 10 degrees C (80 +/- 79), 15 degrees C (747 +/- 384) and 25 degrees C (1141 +/- 334). Survival time decreased with increasing temperature, resulting in 50% mortality of the cercariae after 32.8 +/- 0.6 h (10 degrees C), 26.8 +/- 0.8 h (15 degrees C), 20.2 +/- 0.5 h (20 degrees C) and 16.6 +/- 0.3 h (25 degrees C ). Infectivity of R. roscovita cercariae in cockles Cerastoderma edule increased with increasing temperature and was highest at 25 degrees C (42.6 +/- 3.9%). However, mesocosm experiments with infected snails and cockle hosts in small aquaria, integrating cercarial emergence, survival and infectivity, showed highest infection of cockles at 20 degrees C (415 +/- 115 metacercariae host(-1)), indicating 20 degrees C to be the optimum temperature for transmission of this species. A field experiment showed metacercariae of R. roscovita to appear in C. edule with rising water temperature in April; highest infection rates were in August, when the water temperature reached 20 degrees C. Since another trematode species (Himasthla elongata; Digenea: Echinostomatidae) occurring at the experimental site showed a similar temporal pattern, trematode transmission to second intermediate bivalve hosts may peak during especially warm (> or = 20 degrees C) summers in the variable climate regime of the North Sea.     

Cercaria patellae Lebour, 1911 developing inPatella vulgata is the cercaria ofEchinostephilla patellae (Lebour, 1911) n. comb. (Digenea,Philophthalmidae)

The echinostome cercaria,Cercaria patellae Lebour, 1911, which develops in the limpetPatella vulgata (Prosobranchia, Diotocardia, Patellidae), was investigated by light and scanning microscopy. The highest prevalence of limpets with rediae/cercariae occurred on bare rocks on the upper part of the shore. The prevalence was higher in larger snails and in those which had an orange foot-sole. The cercariae penetrated into other gastropods,Aplysia punctata (Opisthobranchia, Cephalaspidea, Aplysiidae),Acanthodoris pilosa (Opisthobranchia, Nudibranchiata, Lamellidorididae) andP. vulgata. No differences in size were found between metacercariae which naturally infected onlyP. vulgata and five-day-old metacercariae from experimental infections. Inin vitro experiments excystation of metacercariae from natural infections took place as a passive process after a double treatment with pepsin and trypsin which caused the dissolution of the cyst wall. After metacercariae from natural infections were fed to one-day-old chickens, the young adults developed. Detailed morphometrical data on rediae, cercariae and metacercariae are compared with previously published measurements of living specimens. Chaetotaxy demonstrated a distinct distribution of tegumental papillae of cercariae, resembling that of the Echinostomatidae or Philophthalmidae. Scanning electron microscopy of adults revealed short, blunt spines anteriorly, while flattened serrated spines occurred on the posterior tegument. Morphometry revealed no major differences to previously published data. Considering all ecological data and comparing the morphological data with those of previous publications by different authors, we conclude thatC. patellae Lebour, 1911, the cercaria in the limpet, is a developmental stage of a digenean in the intestine of the oystercatcherHaematopus ostralegus and should be referred toEchinostephilla patellae (Lebour, 1911) n. comb.     

Molecular identification of developmental stages in Opecoelidae (Digenea)

Nuclear ribosomal DNA sequences represent a useful tool for distinction of poorly differentiated developmental stages, such as trematode cercariae or metacercariae. Here, the complete internal transcribed spacer region of the ribosomal DNA (ITS 1 + 5.8S + ITS 2) was sequenced for 29 specimens of the digenean family Opecoelidae, including 16 adult specimens and 13 undescribed larval stages (nine cercariae and four metacercariae) occurring in various marine host organisms. Six cercariae and three metacercariae were found to match their corresponding adult form. This work also revealed that cercariae of the same species are able to infect more than one gastropod host species, suggesting that the specificity for the first intermediate host within the Digenea may be lower than previously thought.     

Temporal variation in prevalence and abundance of metacercariae in the pulmonate snail Lymnaea stagnalis in Chany Lake, West Siberia, Russia: long-term patterns and environmental covariates

Infrapopulations of trematode metacercariae were monitored in the snail Lymnaea stagnalis over 17 yr (1982-1999) at Chany Lake, Novosibirskaya Oblast', Russia. Eighteen trematode species were recorded. Patterns of occurrence varied from 4 species (Echinoparyphium aconiatum, Echinoparyphium recurvatum, Moliniella anceps, and Cotylurus cornutus) that persisted at relatively high prevalence (> 60% of samples) across sites, seasons, and years, to species that were very rare and sporadic in occurrence. The stability of the 4 common species was probably because of their occurrence either in a wide range of definitive hosts or in a host adapted to the extreme abiotic changes that occurred from year to year in these wetlands. The prevalence and mean abundance of C. cornutus were negatively correlated with water level in the wetlands; its prevalence was also correlated with water temperature. The mean abundance of M. anceps was positively correlated with water level. The most probable explanation for the cyclic dynamics of infections of the common species is change in population sizes and densities of definitive and intermediate hosts, which mediated cyclic alterations in water levels.     

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.     

Toxic effects of cadmium and zinc on the transmission of Echinoparyphium recurvatum cercariae

The toxicity of cadmium, zinc and Cd/Zn mixtures to the transmission of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae into the snail second intermediate hosts was investigated at concentrations ranging from 100 microg l-1 to 10 000 microg l-1 in both soft and hard water. A differential response in the infectivity of metal-exposed cercariae into Lymnaea peregra and Physa fontinalis was demonstrated which was dependent on the snail species being infected. Exposure of L. peregra, P. fontinalis, and L. stagnalis to heavy metals caused a differing susceptibility to E. recurvatum cercariae depending on the snail species being exposed. The mechanism and effects of metal toxicity, together with the importance of the parasite/host strain on cercarial transmission are discussed.     

Experimental observations on the specificity of Apatemon (Australapatemon) minor (Yamaguti 1933) (Digenea: Strigeidae) toward leech (Hirudinea) second intermediate hosts

The infectivity of Apatemon (Australapatemon) minor cercariae to 7 species of freshwater leech was examined under laboratory conditions. The leech species exposed to infection were; Erpobdella octoculata, Helobdella stagnalis, Glossiphonia heteroclita, Glossiphonia complanata, Hemiclepsis marginata, Piscicola geometra, and Theromyzon tessulatum. Five species were capable of acting as second intermediate hosts. Despite this broad specificity only Erpobdella octoculata and Helobdella stagnalis could be considered to show a high degree of compatibility with the parasite. In these 2 leech species more than 38% of the cercariae successfully penetrated and established metacercarial cyst infections. The leeches Piscicola geometra and Theromyzon tessulatum did not become infected. Close parallels were found between the species order of host utilization determined experimentally and that revealed by a range of field surveys in Britain, Eastern Europe and Russia. This suggests that the order of second intermediate host utilization in the natural environment has, at least in part, a physiological basis. The degree to which different leeches are utilized as hosts may relate to the opportunity which they afford for transmission of A. minor to wildfowl definitive hosts.     

High Endemicity with Clonorchis sinensis Metacercariae in Fish from Yongjeon-cheon (Stream) in Cheongsong-gun,Gyeongsangbuk-do,Korea

The infection status with Clonorchis sinensis metacercariae (CsMc) was examined in freshwater fishes from Yongjeon-cheon (a branch of Nakdong-gang) located in Cheongsong-gun, Gyeongsangbuk-do, the Republic of Korea (Korea). A total of 750 fishes in 19 species were examined by the artificial digestion method for 2 years (2019 and 2020). CsMc were detected in 378 (51.4%) out of 735 fishes in 14 species (73.7%), and the infection intensity was 666 per fish infected. In 2019, CsMc were found in 172 (68.0%) out of 253 fishes in 10 species, and the infection intensity was 565 per fish infected. In 2020, CsMc were detected in 206 (62.2%) out of 331 fishes in 10 species, and the infection intensity was 751 per fish infected. The other zoonotic trematode, ie. Metagonimus spp., Centrocestus armatus, Echinostoma spp. and Clinostomum complanatum, metacercariae were also detected in fishes from the survey streams, but their endemicities were relatively low. Conclusively, it was first confirmed that CsMc are highly endemic in fishes from Yongjeon-cheon in Cheongsong-gun, Gyeongsangbuk-do, Korea.     

Redescription and life cycle of the monorchiid Postmonorcheides maclovini Szidat, 1950 (Digenea) from the Southwestern Atlantic Ocean: Morphological and molecular data

The adult monorchiid, Postmonorcheides maclovini Szidat, 1950, digenean parasite of the Patagonian blennie Eleginops maclovinus (Cuvier) (Eleginopidae) from Puerto Deseado (47° 45′ S, 65° 55′ W), Argentina, was characterized and its life cycle elucidated. P. maclovinus is the only species of the genus Postmonorcheides, proposed by Szidat (1950) from Tierra del Fuego province (~ 54° S), Argentina. This digenean uses the Patagonian blennie as definitive host, and the intertidal bivalve Lasaea adansoni (Gmelin) (Lasaeidae) as both first and second intermediate hosts (metacercariae encyst inside sporocysts), being the first record of this clam as intermediate host of trematode parasites. The cercaria may, in addition to encysting in the sporocyst, emerge and presumably infect other intermediate hosts. This is the second report of a monorchiid species with metacercariae encysting inside the sporocyst. Adults were found parasitizing the fish stomach, pyloric caeca and intestine with a prevalence of 100%; sporocysts with cercariae and/or metacercariae were found parasitizing the gonad of the bivalve with a prevalence of 2.78%. The cercariae possess a well-developed tail and eye-spots are absent. The ITS1 sequence from the adult digeneans found in the Patagonian blennie, identified as P. maclovini, was found to be identical to the ITS1 sequences obtained both from sporocysts containing cercariae and encysted metacercariae found in L. adansoni.     

Activity of detoxifying and antioxidant enzymes in the pond snail <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> (Gastropoda: Pulmonata) during invasion by trematode cercariae

A comparative analysis of changes of detoxifying and antioxidant enzyme activities was carried out in the body homogenate of the pond snail Lymnaea stagnalis. Molluscs with the shell size of 4–6 mm infested with cercariae of one (Echinopharipjium aconiatum, E. recurvatum or Moliniella anceps) or two (E. aconiatum + M. anceps or E. aconiatum + E. recurvatum or M. anceps + E. recurvatum) trematode species. It has been reveled that activities of detoxifying and antitoxidant enzymes in the mollusc Lymnaea stagnalis body change at invasion with trematodes and that the character of the changes depends on the stage of development of the trematodes in the host body, the variant of infestation (monoinvasion or mixed infestation), and the species of the parasite. The first 2 h after penetration of the trematode cercariae into the mollusc tissues is accompanied by an increase of activities of the detoxifying and antioxidant enzymes. During the long coexistence of metacercariae in the host body (no less than for 13 days), activities of non-specific esterases and glutathione-S-transferase decrease, while the superoxide dismutase activity increases.     

Behavioral responses of Poecilia vivipara (Osteichthyies: Cyprinodontiformes) to experimental infections of Acanthocollaritrema umbilicatum (Digenea: Cryptogonimidae)

The swimming behavior of Poecilia vivipara was evaluated using an image analysis system comparing laboratory-reared uninfected fish before and after experimental infection with different intensities of cercariae of the trematode Acanthocollaritrema umbilicatum. Two experiments were performed, each with 30 fish which were individually exposed to 30 and 50 cercariae, respectively, shed from experimentally infected molluscs, Heleobia australis. Before and after (17–27 days) infection, the behavior of each fish was monitored in terms of Distance travelled, Ambulatory time, Stereotypic time, Resting time and Average speed. At the end of the experiments, the fish were dissected to count the number of metacercariae recovered. In the experiment with 30 cercariae, fish with 2–10 metacercariae did not exhibit any significant differences in their swimming activity, but those with 11–22 metacercariae had a significantly enhanced Stereotypic time and a reduced Time Resting. In the experiment with 50 cercariae, fish with 5–22 metacercariae had an enhanced Distance travelled and a reduced Average speed; highly significant differences occurred with regard to all behavioral parameters when considering the subgroup 23–36 metacercariae: Distance travelled, Stereotypic time, Resting time, Ambulatory time and Average speed. The swimming behavior of P. vivipara changed influenced by an intensity-dependence on metacercariae of A. umbilicatum, supporting the prediction that parasites are able to alter the behavior of their hosts.     

Annotated key to the trematode species infecting Batillaria attramentaria (Prosobranchia: Batillariidae) as first intermediate host

In eastern Asia and western North America at least nine morphologically distinguishable species of digenean trematode infect the mud snail, Batillaria attramentaria (Sowerby 1855) (=B. cumingi (Crosse 1862)), as first intermediate host. Further, molecular and morphological evidence indicates that several of these trematode species comprise complexes of cryptic species. I present an identification key to these nine trematode morphospecies (including four newly reported species). Additionally, I provide an annotated list, which includes further diagnostic information on the larval stages (cercariae, parthenitae, and metacercariae), information on second intermediate host use, links to the previous relevant reports of trematodes infecting B. attramentaria and notes on other aspects of the species' biology.     

The life cycle of Echinochasmus bagulai (Trematoda: Echinostomatidae)

The life history of Echinochasmus bagulai, an echinostomatid trematode in birds, is reported and stages in the life cycle are described. Natural infections with cercariae, which are of the gymnocephalous type, were found in the thiarid snail Thiara tuberculata. Metacercarial cysts were found in the gills of Aplocheilus panchax, Oryzias melastigma, Gambusia affinis and Channa punctata. Adults were obtained in the small intestine of Ardeola grayi. In laboratory experiements development of cercariae into infective metacercariae took 10 days in the gills of A. panchax. Mature flukes were recovered in 10 days in the small intestine of experimentally infected 1-day-old leghorn chicks. The eggs collected from the faeces of these infected chicks were incubated at 39 degrees C. The free-swimming miracidia were found on the fifth day of incubation.     

Infection patterns in white sea blue mussels Mytilus edulis of different age and size with metacercariae of Himasthla elongata (Echinostomatidae) and Cercaria parvicaudata (Renicolidae)

Infection of mussels Mytilus edulis L. by 2 trematode species was studied in a natural intertidal population in the Chupa inlet of the White Sea. The prevalence of metacercariae of Himasthla elongata (Mehlis, 1831) and Cercaria parvicaudata (Stunkard & Shaw, 1931) in mussels reached 100% in 3 to 4 yr old molluscs and remained at this level in older individuals. Infection intensity increased evenly with the age of the molluscan host, showing a tendency to decrease only in the oldest (9 yr old) mussels. These patterns of age dynamics of prevalence and infection intensity were associated with accumulation of trematode larvae in the course of the molluscs' lives. Ability of metacercariae to exist in mussels for long periods (at least 2.5 yr) was verified in the course of an experiment, during which infected molluscs were kept in a subtidal net cage. Decrease of infection intensity in the oldest individuals may reflect selective mortality of the most severely infected molluscs. Among mussels of the same age, higher infection intensity values occurred in larger individuals. This may be due to an enhanced pumping rate in large molluscs, which increases the probability of cercariae, free-living trematode larvae, infecting them via water currents.     

The role of spatial and temporal heterogeneity and competition in structuring trematode communities in the great pond snail, Lymnaea stagnalis (L.)

We assessed how spatial and temporal heterogeneity and competition structure larval trematode communities in the pulmonate snail Lymnaea stagnalis . To postulate a dominance hierarchy, mark-release-recapture was used to monitor replacements of trematode species within snails over time. In addition, we sampled the trematode community in snails in different ponds in 3 consecutive years. A total of 7,623 snails (10,382 capture events) was sampled in 7 fishponds in the Jind?ich?v Hradec and T?eboň areas in South Bohemia (Czech Republic) from August 2006 to October 2008. Overall, 39% of snails were infected by a community of 14 trematode species; 7% of snails were infected with more than 1 trematode species (constituting 16 double- and 4 triple-species combinations). Results of the null-model analyses suggested that spatial heterogeneity in recruitment among ponds isolated trematode species from each other, whereas seasonal pulses in recruitment increased species interactions in some ponds. Competitive exclusion among trematodes led to a rarity of multiple infections compared to null-model expectations. Competitive relationships among trematode species were hypothesized as a dominance hierarchy based on direct evidence of replacement and invasion and on indirect evidence. Seven top dominant species with putatively similar competitive abilities (6 rediae and 1 sporocyst species) reduced the prevalence of the other trematode species developing in sporocysts only.     

Morphological peculiarities of schistosomatid cercariae of Trichobilharzia cf. ocellata group occuring in Moscow and Saint-Petersburg populations

Morphology of schistosomatid cercariae of the genus Trichobilharzia (ocellata group) is described based on original and reference data. The material (Trichobilharzia cercariae) was collected in 1999-2000 from naturally 'infected snails Lymnaea stagnalis from Moscow and Saint-Petersburg megalopolises. The more accurate flame cell formula for these cercariae, as well as some aspects of chaetotaxy and eye morphology are given. The differences between cercariae from these regions were found. Based on a comparison of nef data with the reference data, it was found that the cercariae examined differ from those from the Central Europe.     

Infection by Paragonimus miyazakii cercariae of their crab hosts, Geothelphusa dehaani, by percutaneous penetration

Identification of the transmission routes of the trematode parasite Paragonimus miyazakii into different intermediate hosts would help to explain the natural distribution of the parasite. The behavior of P. miyazakii cercariae released from snails into water and in the presence of a living host or a whole crab leg was observed by stereoscopic or light microscopy at various times after exposure started. On encountering a crab leg or cheliped, the cercariae became entangled with the host via mucoid strands arising from the cercariae. Within 3 hr, most cercariae were attached to the host; cuticular penetration took between 5 and 6 hr, after which cercariae were found in the cavity of the leg. Crabs examined 102-149 days after exposure to the cercariae contained fully developed metacercariae. The metacercariae were fed to 2 rats, and the rats were killed 83 or 111 days later. Some of the metacercariae had reached maturity in the rats. That the cercariae were not ingested by the crabs but penetrated the crabs percutaneously (through hard as well as soft tissue) means that transmission can occur even in areas in which crabs and the host snails do not coexist, as they would if the usual route were oral (when the crabs ate infected snails).     

Temporal variations in the infection of a population of Cerithidea cingulata by larval trematodes in Kuwait Bay

The prosobranch gastropod Cerithidea cingulata (Gastropoda: Potamididae) in Kuwait Bay was examined for larval trematode infections over a 17-month period. A total of 2537 snails were examined and 1265 (49.9%) found to be infected with one or more species of trematodes. The component community in the snail comprised 12 species representing the families Cyathocotylidae (2), Echinostomatidae (2), Haplosplanchnidae (1), Heterophyidae (2), Microphallidae (1), Philophthalmidae (2), Plagiorchiidae (1) and Schistosomatidae (1). Cyathocotylid II (41.6%) was by far the most prevalent species followed by the microphallid (3.9%), the two species comprised 90% of the total trematode fauna. The prevalence of infection increased with shell size and was significantly higher in male (47%) than female (33%) snails. Multiple infections were observed in only 15 (1.2%) of the infected snails; cyathocotylid I and cyathocotylid II combination occurred 14 times and heterophyid I and the microphallid occurred once. Trematode species were more diverse and prevalent in winter, and cercarial shedding peaked in summer. Behaviour of the definitive host and snail population dynamics were probably the major contributors to the detected temporal pattern in the infections.