Attachment and penetration of Centrocestus armatus (Digenea: Heterophyidae) cercariae to gills of secondary intermediate fish hosts

The attachment and penetration of Centrocestus armatus cercariae into the fish host Zacco temmincki are described in this study. Light and scanning electron microscopy (SEM) were used to examine the topographical features and behavior of cercariae. Histochemistry and transmission electron microscopy (TEM) were employed to trace glandular products and secretions released by cercariae during penetration. Cercariae are first carried into the fish gill chambers via the respiratory currents. The frequency of respiratory-current reversals of fish increased when infected with cercariae. The behavior of cercariae, during breaks in current flow that preceded each current reversal, was observed using a specially devised apparatus. Cercariae produce a mucus-like secretion upon attachment, shed their tail, and employ a brief period of leech-like creeping behavior before penetration. In all cases, the site of penetration was via the surface of the primary gill lamellae. SEM revealed a well-developed anterior penetration apparatus, and a highly contractile body region, that created a driving force for penetration. TEM and histochemistry showed that the mucus observed on the surface tegument of cercariae during attachment were glandular secretions from the parasite. The significance of fish respiratory current reversals to the success of cercariae penetration nicely illustrates the exploitation by the parasite of a host response to a stimulus.     

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.     

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.     

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.     

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.     

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.     

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

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

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.     

Distribution of metacercariae of Centrocestus formosanus (Trematoda: Heterophyidae) on the gills of Aplocheilus panchax

The metacercariae of Centrocestus formosanus (Nishigori 1924) occur on the gills of freshwater fishes in association with blood vessels. The microecology of the metacercariae on the gills of the cyprinid fish Aplocheilus panchax (Ham. & Buch.) was investigated. The cysts showed a preference for anterior gills and for the dorsal section of each gill. Variations in the density of infection, season and size of the fish do not affect the spatial distribution of metacercariae. It is presumed that ventilation water currents spread the cercariae onto the anterior gills and then the cercariae actively select the preferred habitat.     

Egg Laying Capacity of Haplorchis taichui (Digenea: Heterophyidae) in Humans

Quantitative fecal egg counts represented as the number of eggs per gram of feces (EPG) are generally a reliable parameter to estimate the worm burden of intestinal and hepatic parasitoses. Although Haplorchis taichui (Digenea: Heterophyidae) is one of the most common minute human intestinal flukes, little is known about the relationship between EPG and the actual worm burden in patients or the severity of the disease. In the present study, fecal samples were collected from 25 villagers in northern Thailand before and after praziquantel treatment. The EPG values of each participant were determined by the modified cellophane thick smear method, and adult worms were collected from the whole stool after the treatment. Eggs per day per worm (EPDPW) of H. taichui were estimated 82 from egg counts and expelled worms. The EPG was not well correlated with the worm burden, and a reverse correlation was observed between the EPDPW and the worm burden.     

Stictodora lari (Digenea: Heterophyidae): the discovery of the first human infections

Stictodora lari Yamaguti, 1939 (Digenea: Heterophyidae) was originally reported as an avian intestinal parasite and is here reported in humans. The patients were adults, aged 46-69 (2 men and 4 women), who resided in coastal villages in Sachon-gun, Kyongsangnam-do, or Shinan-gun, Chollanam-do. in the Republic of Korea. The worms were recovered after praziquantel treatment and purgation with magnesium salts. A total of 15 S. lari specimens (1-10 per individual) was collected, together with 5 other species of intestinal flukes. The flukes were morphologically characterized by a small body size (0.70-0.86 mm long and 0.27-0.36 mm wide), a ventrogenital sac, and a gonotyl armed with 70-80 spines in the form of a comma or reversed comma lying along their lateral margin. The patients had eaten the raw flesh of mullets and gobies that had been caught in an estuary near their villages. To the best of our knowledge, this is the first record of a human S. lari infection.     

Stephanostomum lophii sp. nov. (Digenea: Acanthocolpidae), intestinal parasite of Lophius piscatorius, with reference to seasonal fluctuations of metacercariae in intermediate second hosts (Gadidae)

We describe the adult and metacercaria of a new species of digenetic trematode, Stephanostomum lophii sp. nov., an intestinal parasite of the angler fish, Lophius piscatorius . The species is distinguished from the similar Stephanostomum kovalevi by the numerous peristomial spines in the latter and from S. baccatum by the oral sucker being larger than the ventral sucker and by the fact that the principal intermediate hosts are gadids, not pleuronectids as in the case of Stephanostomum baccatum . We also report the prevalence of S. lophii metacercariae in various potential angler fish prey species caught off the coast of Galicia (NW Spain).     

Population biology of the metacercariae of Centrocestus formosanus (Trematoda: Heterophyidae) on the gills of Aplocheilus panchax

The population biology of metacercariae of Centrocestus formosanus (Nishigori, 1924) on the gills of the larvivorous fish, Aplocheilus panchax (Hamilton) was investigated based on data from examination of monthly samples of fish collected from a stream at Visakhapatnam during the period December 1983 to April 1986. The seasonal dynamics, dispersion pattern and long-term changes were studied. No seasonality in prevalence and density of infection with metacercariae was observed. The recruitment of metacercariae occurred throughout the year. An increase in density of infection with increase in size of the fish was noted. Possible reasons for the observed distribution are discussed. The frequency distribution of the metacercariae followed the negative binomial at different times of the year. The data indicated an increase in degree of overdispersion with increase in size offish except for a slight drop in the largest size class. Long-term changes in the abundance of metacercariae are given and a decline in parasite population abundance is noted.     

From first to second and back to first intermediate host: the unusual transmission route of Curtuteria australis (Digenea: Echinostomatidae)

The trematode Curtuteria australis uses the whelk Cominella glandiformis as first intermediate host and the cockle Austrovenus stutchburyi as second intermediate host before maturing in shorebirds. The whelk also happen to be an important predator of cockles on intertidal mudflats. In this study we show that whelks can act as temporary paratenic hosts for the trematode. A single whelk feeding on 1 cockle can ingest large numbers of metacercariae, which remain within the whelk for 1-3 days before passing out in feces. The viability of these metacercariae assessed as the percentage capable of successfully excysting under conditions simulating those inside a bird's digestive tract, is lower after passage through a whelk (48%) than before (59%). Still, given that shorebird definitive hosts prey on whelks as well as cockles, survival inside the whelk allows C. australis to complete its life cycle: overall, though, whelk predation is likely to be an important sink for the trematode population. To our knowledge, this is the first report of a trematode using a snail as both first intermediate host and paratenic host, offering an alternative transmission route for the parasite as a result of the unusual trophic relationships of its hosts.     

Infection of mollusks and fish by the trematode Apophallus muehlingi (Jagerskiold, 1898) and its interrelations with intermediate hosts

In the Rybinsk Reservoir (Upper Volga basin), a sharp decrease in the occurrence of Apophallus muehlingi (Jagerskiold) parthenites was found in the population of the Ponto-Caspian snail Lithoglyphus naticoides Pfeiffer in 2009–2010. In the muscles of cyprinid underyearlings (ide Leuciscus idus (L.) and dace Leuciscus leuciscus (L.)), interspecific differences in protein content were revealed following their infection by metacercariae in the experimental environment. The helminth infection caused an increase in glycogen content in both fish species. A reduction of motor activity induced by muscle damage might be the main reason for the observed changes in biochemical characteristics.     

Taxonomic study of Ascocotyle (Phagicola) longa Ransom, 1920 (Digenea: Heterophyidae) and related taxa

Species of Ascocotyle Looss, 1899 (subgenus Phagicola Faust, 1920) (Digenea: Heterophyidae) related to A. (P.) longa Ransom, 1920, i.e. those with a bipartite gonotyl and a single crown of 16 circumoral spines, were revised on the basis of examination of type and voucher specimens. The species A. (P.) arnaldoi Travassos, 1929, A. (P.) byrdi (Robinson, 1956) and A. (P.) longicollis (Kuntz & Chandler, 1956), all possessing 16 circumoral spines, are synonymised with A. (P.) longa. A. (P.) ascolonga Witenberg, 1929, the types of which were not available, is provisionally retained as a valid species despite its close similarity to A. (P.) longa (identical number of circumoral spines and morphology of the ventrogenital sac). It differs from A. (P.) longa in the presence of uterine loops posterior to the testes and intestinal caeca ending anterior to the ovary (versus the pretesticular uterus and intestinal caeca reaching to the posterior margin of the ovary in A. (P.) longa). A. (P.) inglei (Hutton & Sogandares-Bernal, 1959), found only once in a single specimen, is also tentatively considered a valid species because it differs in number of circumoral spines (19), slightly larger eggs (23–26×13–16 m) and the distribution of refractile bodies of the gonotyl arranged in four separate groups from A. (P.) longa and A. (P.) ascolonga (16 circumoral spines; eggs 15–20×–12 m; refractile bodies in two groups).     

Phagicola angrense (Digenea: Heterophyidae) as a cause of enteritis in a raccoon (Procyon lotor)

Numerous Phagicola angrense were associated with enteritis in a single male juvenile raccoon (Procyon lotor) live-trapped on Parramore Island, Virginia (USA). The raccoon was weak, ataxic and had melenic soft feces. The carcass was emaciated, pale and had ascites. Mesenteric vessels appeared prominent and the stomach and the intestines contained fetid bloody material. The small intestinal mucosa contained three locally extensive sites of necrosis. Histopathologically, there were numerous small digeneans both attached to the mucosa and free within the lumen. Digeneans were usually found deep within the crypts where the epithelium was markedly attenuated and devoid of epithelial cells at the point of parasite attachment. In the lamina propria there were areas of acute hemorrhage and infiltration with plasma cells and eosinophils. This appears to be the first record of severe enteritis in the raccoon caused by this digenean.