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.     

Scanning electron microscopy of adult Echinoparyphium recurvatum (Von Linstow, 1873) (Digenea: Echinostomatidae) from Britain

The surface topography of 15-day-old adult Echinoparyphium recurvatum (Von Linstow, 1873) sensu stricto, from an isolate of the parasite utilizing Lymnaea peregra as first intermediate host in southern England, is described and illustrated using scanning electron microscopy, and is compared to those of other Echinoparyphium species from Europe, and with those of E. recurvatum of East Asian origin. The general morphology of tegumental features was found to be very similar to that of worms of the same age observed in a previous study on a Korean isolate of E. recurvatum. Comparison of collar and body spination of E. recurvatum with other 45-collar-spined members of the genus in Europe revealed some significant differences in morphology. Collar spines of E. recurvatum were found to be shorter and more pointed than those of Echinoparyphium mordwilkoi. The body spines of E. recurvatum are rounded and scale-like, extending just beyond the ventral sucker, contrasting with the pointed, thorn-shaped body spines of E. mordwilkoi, extending posteriorly to the level of the second testis. Body spine shape and distribution in E. recurvatum were found to be more similar to those of Echinoparyphium pseudorecurvatum. The value of SEM studies in elucidating the relationship between members of the genus Echinoparyphium in Britain/Europe and those in Africa, Asia and North America is suggested.     

Influence of cadmium exposure on the incidence of first intermediate host encystment by Echinoparyphium recurvatum cercariae in Lymnaea peregra

The effect of cadmium exposure of the snail first intermediate host Lymnaea peregra on the incidence of encystment of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae without emergence from the snail was investigated. Exposure to 100 microg l(-1) Cd for 72 h caused a significant increase in the incidence of first host encystment when compared to controls. In addition, autometallographic staining of E. recurvatum daughter rediae and developing cercariae showed that there was metal accumulation within their body tissues. The significance of these findings to parasite transmission in metal-polluted environments is discussed.     

An ectosymbiotic flatworm, Temnocephala chilensis, as second intermediate host for Echinoparyphium megacirrus (Digenea: Echinostomatidae) in Patagonia (Argentina)

During a survey of parasites of the Patagonian freshwater crab Aegla neuquensis neuquensis, its ectosymbiont Temnocephala chilensis was found parasitized with an echinostomatid metacercaria with 43 collar spines. Of the 414 T. chilensis collected, 106 were parasitized with metacercariae of Echinoparyphium sp. Ovigerous adults of E. megacirrus were obtained from domestic chicks experimentally infected with metacercariae obtained from temnocephalans. This is the first record of temnocephalans being parasitized by digeneans.     

Complete Mitochondrial Genome of Echinostoma hortense (Digenea: Echinostomatidae)

Echinostoma hortense (Digenea: Echinostomatidae) is one of the intestinal flukes with medical importance in humans. However, the mitochondrial (mt) genome of this fluke has not been known yet. The present study has determined the complete mt genome sequences of E. hortense and assessed the phylogenetic relationships with other digenean species for which the complete mt genome sequences are available in GenBank using concatenated amino acid sequences inferred from 12 protein-coding genes. The mt genome of E. hortense contained 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 non-coding region. The length of the mt genome of E. hortense was 14,994 bp, which was somewhat smaller than those of other trematode species. Phylogenetic analyses based on concatenated nucleotide sequence datasets for all 12 protein-coding genes using maximum parsimony (MP) method showed that E. hortense and Hypoderaeum conoideum gathered together, and they were closer to each other than to Fasciolidae and other echinostomatid trematodes. The availability of the complete mt genome sequences of E. hortense provides important genetic markers for diagnostics, population genetics, and evolutionary studies of digeneans.     

Reproduction of the trematode Echinostoma caproni parthenites (Digenea: Echinostomatidae)

Dynamics of the reproduction in the trematode Echinostoma caproni parthenites (Echinostomatidae) was observed. Early laying and maturation of the generative cells are for the first time shown to be characteristic for all parthenogenetic generations. Really the process of reproduction had been finishing to the beginning of the generating of new age by parthenites. Mother sporocysts, as well as redia of different generations, in fact stop producing new generative cells with the beginning of the generating of new age, and assume the function of a brood pouch. This feature was considered previously as peculiar mainly to mother sporocysts. Data on the autotomy of the anterior body end in mother sporocysts are verified. In our opinion, these data are an evidence of an early manifestation of the evolutionary trend to the morpho-functional regress and disintegration of the parasitic stage of mother sporocyst.     

Studies on the attachment response of Isthmiophora melis cercariae (Trematoda: Echinostomatidae)

The cercaria of Isthmiophora melis is relatively specific to suitable intermediate hosts, amphibia and fish, as early as the stage of attachment. Attachment responses are maximally stimulated by intact amphibia, whereas isolated frog tissues have a reduced efficiency. Human urine contains attachment triggering substances. Its effectiveness is reduced, when carbonate systems are removed, and may be restored by addition of carbonate. Aqueous solution systems of dissolved CO2 and H2CO3/HCO-3 stimulate attachment responses, especially at pH 6.0-7.5. The attachment response to frog skin surface seems to be triggered by the buffering system, which uses carbonate solution equilibria as important components.     

Echinoparyphium elegans (Looss, 1899) (Digenea: Echinostomatidae): the life cycle and redescription of the adult with a revision of the 43-spined members of the genusEchinoparyphium

The adult morphology ofEchinoparyphium elegans is redescribed. The life cycle is described. The first intermediate host is the freshwater snail,Bulinus truncatus. This and other freshwater snails, such asBulinus globosus, Physa acuta and albinoBiomphalaria glabrata, serve as second intermediate hosts. Adult worms were obtained experimentally from canaries, chicks, pigeons and albino rats, but not from albino mice. The 43-spinedEchinoparyphium species with a collar spined arrangement 5-5-3-3-27,E. bioccalerouxi, E. ralphaudyi andE. combesi, are placed into synonymy withE. elegans after examination of the paratypes.     

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.     

Surface ultrastructure of juvenile and adult Acanthoparyphium tyosenense (Digenea: Echinostomatidae)

The tegumental ultrastructure of juvenile and adult Acanthoparyphium tyosenense (Digenea: Echinostomatidae) was observed by scanning electron microscopy. One- to 3-day-old juveniles and 10-day-old adults were harvested from chicks experimentally fed metacercariae from a bivalve, Mactra veneriformis. The juvenile worms were minute, curved ventrally, and had 23 collar spines characteristically arranged in a single row. The lips of the oral sucker had 7 single aciliated sensory papillae and 4 grouped uniciliated sensory papillae. The ventral sucker had 25 aciliated round swellings on its lip. The anterolateral surface between the 2 suckers was densely packed with tongue-shaped tegumental spines, and the ventral surface just posterior to the ventral sucker was covered with peg-like spines. Retractile, peg-like spines were seen on the anterolateral surface, whereas scale-like spines with round tips and broad bases were sparsely distributed posterior to the ventral sucker. The cirrus was characteristically protruding and armed with minute spines. The surface ultrastructure of A. tyosenense was unique, especially in the number and arrangement of collar spines, shape, and distribution of tegumental spines and in distribution of sensory papillae.     

Development of the partenitae infrapopulation of Echinostoma caproni (Digenea: Echinostomatidae)

The first generation of Echinostoma caproni partenitae is represented maternal sporocysts developing in the cardium of the Biomphalaria mollusk. During all their life, they produce rediae of maternal generation. Rediae of Echinostoma caproni of all generations are similar. The first generation consists of maternal rediae forming only redoid embryos. Due to this process, the number of partenitae increases very fast. The next generations are represented by daughter rediae. In the beginning of their life they produce redoid embryos but later begin forming cercariae. The number of rediae produced before this shift of production depends on the population density. Further, the partenitae retain their potential ability to form rediae but realize it in exceptional cases. Generative organs of all generations are germinal masses. Proliferation of generative elements and beginning stages of redia and cercaria development take place within these masses. The infrapopulation of E. caproni belongs to the "prolonged type", because it is a complete microhemipopulations; its existence is limited by a lifespan of the mollusk host.     

Diurnal migration of Echinostoma caproni (Digenea: Echinostomatidae) in ICR mice

Twenty-four female ICR mice, 12 acclimated to a 12 ∶ 12 light-dark cycle and 12 to a 12 ∶ 12 dark-light cycle for 7 days, were each infected with 10 metacercariae of Echinostoma caproni. Infected mice were maintained on their respective lighting regimes for 28 days. Six mice (3 from each group) were necropsied at 4-hr intervals beginning at 0700 hr. The small intestine was removed, opened, and the position of individual worms and worm clusters was measured to the nearest 0.1 cm. Each intestine was subsequently divided into 20 equal segments and individual worms and worm clusters were assigned to the appropriate segment based on the original measurements. All worms were found in the posterior 55% of the intestine (ileum). All posterior segments (10-20), with the exception of segment 18, harbored at least 1 worm at some time. A Monte Carlo simulation of worm abundance in segments 10-17 over all time periods indicated a random distribution, while the same analysis of segments 10-20 indicated a non-random distribution due to large numbers of worms in segment 20 and to the absence of worms in segment 18. To analyze temporal changes in worm distribution, mice were grouped by time of necropsy as follows: night (1900 and 2300 hr), morning (0300 and 0700 hr), and day (1100 and 1500 hr). During the night and morning, E. caproni was heavily concentrated in segments 10-17 and, during the day, worms were located more posteriorly, with a heavy concentration in the last segment (20).     

Tegumental ultrastructure of the juvenile and adult Himasthla alincia (Digenea: Echinostomatidae)

The tegumental ultrastructure of juvenile and adult Himasthla alincia (Digenea: Echinostomatidae) was observed by scanning electron microscopy. One-, 5- (juveniles) and 20-day-old worms (adults) were harvested from chicks experimentally fed metacercariae from a bivalve, Mactra veneriformis. The juvenile worms were elongated and curved ventrally. The head crown bore 31 collar spines, arranged in a single row. The lip of the oral sucker had 12 paired, and 3 single type I sensory papillae, and the ventral sucker had about 25 type II sensory papillae. The anterolateral surface between the two suckers was densely packed with tegumental spines with 4-7 pointed tips. The adult worms were more elongated and filamentous, and had severe transverse folds over the whole body surface. On the head crown and two suckers, type I and II sensory papillae were more densely distributed than in the juvenile worms. Retractile brush-like spines, with 8-10 digits, were seen on the anterolateral surface, whereas claw-shaped spines, with 2-5 digits, were sparsely distributed posteriorly to the ventral sucker. The cirrus characteristically protruded out, and was armed with small spines distally. The surface ultrastructure of H. alincia was shown to be unique among echinostomes, especially in the digitation of its tegumental spines, the distribution of sensory papillae and by severe folds of the tegument.     

Echinostoma echinatum (Zeder 1803) sensu Kanev (Digenea: Echinostomatidae): a note of caution

The re-examination of the voucher specimens identified by Kanev et al. (1987) as E. echinatum (Zeder, 1803) showed that two distinct species have been considered as E. echinatum: E. sarcinum Dietz, 1909, possessing 47 collar spines [Lymnaea truncatula strain — set ‘L’ (No. 326 TB)] and Echinostoma sp. (?echinatum) with 37 collar spines [Planorbis planorbis strain — set ‘P’ (No. 327 TB)]. The material of E. sarcinum is described and differentiated from E. revolutum and E. echinatum (sensu Kanev, 1985). The present results suggest that species identity of the material described as E. echinatum in Kanev's (1985) experimental study requires further clarification.     

Production and chronobiology of emergence of the cercariae of Euparyphium albuferensis (Trematoda: Echinostomatidae)

The production and the chronobiology of emergence of the cercariae of Euparyphium albuferensis from Gyraulus chinensis experimentally infected with a single miracidium were established during 28 consecutive days from the first day of cercarial shedding. Moreover, the effect of a sudden change in light-dark cycling was investigated. Although the daily cercarial shedding rates show great variability, a progressive increase in cercarial production was observed in the first weeks of the cercarial shedding periods, probably in relation to the demography of intramolluscan larval stages. Under 12:12 light-dark cycling conditions, E. albuferensis cercariae emerged in the light, and the rhythm was circadian. The sudden change in the light-dark cycle resulted in corresponding alterations in emergence patterns, which shows that cercarial emergence was correlated to light-dark alternation. The ecological consequences of daily emergence by mobile cercariae whose target hosts are organisms that regularly occur in the same habitat with the molluscan emitting host, such as those of E. albuferensis, are discussed with reference to the hypotheses proposed to date, and an alternative hypothesis is proposed.     

Metacercarial utilization of a naturally infected single species (Lymnaea peregra) snail community by Echinoparyphium recurvatum

Utilization of a single-species molluscan community of Lymnaea peregra by metacercariae of Echinoparyphium recurvatum over a summer (July-September) period in south-east England showed an increase in the mean number of cysts per host with host size and time of exposure. Aggregation resulting from host and habitat-related factors increased with host size and time of exposure. Encystment within the host was restricted to the peripheral organs in smaller juvenile snails but as snails increased in size, metacercariae were distributed throughout the tissues.