Morphological and molecular differentiation of Clinostomum complanatum and Clinostomum marginatum (Digenea: Clinostomidae) metacercariae and adults

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum , by ～ 1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.     

In vitro Excystation of Metacercariae of Clinostomum complanatum (Rudolphi, 1819) (Digenea: Clinostomatidae)

In vitro excystation of metacercariae of Clinostomum complanatum (Rudolphi, 1819) was studied. Excystation was an active process, initiated by a rise in temperature. The metacercariae excysted in enzymic as well as non enzymic medium, but the rate of excystation was affected by the temperature and/or pH of the medium. The optimal excystation was obtained in pepsin-Tyrode at pH 2.3-3.5 and a temperature of 42°C. Trypsin treatment was not required.     

Parasitism of Cichlasoma paranaense Kullander, 1983 and Gymnotus carapo Linnaeus, 1814 by Clinostomum complanatum (Rudolphi, 1814) metacercariae in the Taquari river.

The infection, known as yellow spot disease, produced by metacercariae of Clinostomum spp. was studied in fishes of the Taquari river, located in Jataizinho, Paraná State, Brazil. A total of 1,582 specimens, belonging to 36 species, were collected between March 1999 and April 2001. Yellow spot disease was observed only in Gymnotus carapo Linnaeus, 1814 (Gymnotiformes, Gymnotidae) and Cichlasoma paranaense Kullander, 1983 (Perciformes, Cichlidae). This parasitism was generated by metacercariae of Clinostomum complanatum (Rudolphi, 1814) (Digenea, Clinostomidae). Among the 88 specimens of G. carapo examined, 7 (prevalence = 8%) had cysts of the parasite. Four of them were captured in July 1999 and three in October 1999. In the other months, no specimen found was infected. Using relative condition factor (Kn) analysis, it was determined that both infected and non-infected specimens had a total weight equal to the theoretically expected value for each total length (Kn = 1.0). Among 56 individuals of Cichlasoma paranaense, 6 (prevalence = 10.7%) had between 1 and 27 metacercariae of C. complanatum (mean intensity of infection = 9.3 +/- 9.6). In March 1999 and April of both 2000 and 2001, the specimens examined were not infected. The infected fish had a total weight higher than the expected value (Kn > 1.0), while the non-infected fish had a weight equal to the expected value (Kn = 1.0).     

Cercarial chaetotaxy of Codonocephalus urniger (Rudolphi, 1819) (Trematoda: Diplostomidae)

The chaetotaxy of Codonocephalus urniger (Rudolphi, 1819) cercariae recovered from Lymnaea palustris in a brackish water lake in Bulgaria is described and figured. Comparisons with Diplostomum spathaceum (Rudolphi, 1819) and D. pseudospathaceum Niewiadomska, 1984 cercariae indicate a similar general pattern in the distribution of sensilla but with marked differences in their number and arrangement on the ventral sucker, tail stem and furcae. Data on the chaetotaxy of other diplostomid species are used to reveal the similarities and variations in some complexes within the members of Diplostomidae with known cercarial chaetotaxy.     

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.     

Life cycle and structure of the fish digenean Brachyphallus crenatus (Hemiuridae).

Cystophorous cercariae from Retusa obtusa (Montagu) (Gastropoda, Opisthobranchia, Retusidae) develop into adults of Brachyphallus crenatus (Rudolphi, 1802) Odhner, 1905 (Hemiuridae). The free-swimming cercariae were ingested by laboratory-reared Acartia tonsa Dana, and the cercarial body was injected into the hemocoel of the copepod. Two-week-old metacercariae held at 15 C were infective to stickleback Gasterosteus aculeatus Linnaeus. The cercariae of B. crenatus are very similar to the cercariae of Hemiurus luehei Odhner, 1905, and Lecithocladium excisum (Rudolphi, 1819) Lühe, 1901 (Hemiuridae), which develop in closely related opisthobranch snails. Scanning electron microscopy of metacercariae and adults of B. crenatus revealed the annular plications of most of the external surface to be scalelike. The area surrounding the genital pore and the presomatic pit was densely plicated.     

Karyological studies on three strigeid digeneans: Ichthyocotylurus erraticus (Rudolphi, 1809), I. variegatus (Creplin, 1825) and Apatemon gracilis (Rudolphi, 1819)

Previous work has shown that karyology may be a valuable technique for discriminating morphologically similar species. In the present study, chromosome number and morphology were described for three strigeid species, Ichthyocotylurus erraticus (Rudolphi, 1809), I. variegatus (Creplin, 1825) and Apatemon gracilis (Rudolphi, 1819). The results show that these species possess the same diploid number of chromosomes (2n = 20) in somatic cells. The relative length and the position of the centromere were determined, and significant inter-specific differences in the karyotypes are demonstrated, enabling the ready discrimination of I. erraticus and I. variegatus cercariae. A review of known karyotypes of the family Strigeidae suggests that possible routes of evolutionary changes involve pericentric inversions within chromosomes rather than Robertsonian translocations between elements.     

Clinostomum complanatum (Trematoda: Clinostomatidae) in five new fish hosts in Japan.

Metacercariae of Clinostomum complanatum were found in six species of natural freshwater fishes collected at Koyama Pond in Tottori City of Tottori Prefecture, Japan. Five of these fish species are reported here as new records for second intermediate hosts of C. complanatum in Japan: the silver crucian carp (Carassius gibelio langsdorfi), the deepbodied crucian carp (Carassius cuvieri), the carp (Cyprinus carpio), the topmouth gudgeon (Pseudorasbora parva) and the rose bitterling (Rhodeus ocelatus).     

Progamotaenia ruficola sp. n. (Cestoda: Anoplocephalidae) from the red kangaroo, Macropus rufus (Marsupialia)

Progamotaenia ruficola sp. n. is described from the red kangaroo (Macropus rufus) from New South Wales, Australia. It is distinguished from other similar species having paired, nondiverticulate uteri in each prolottis: Progamotaenia festiva (Rudolphi 1819), P. diaphana (Zschokke 1907), and P. macropodis Beveridge 1976, by the extremely broad, straight-edged velum, the lack of an external seminal vesicle, the cirrus armature, the number of testes, the lack of vaginal atrophy following insemination, and the morphology of the egg.     

Metacercariae of Clinostomum attenuatum in Ambystoma tigrinum mavortium, Bufo cognatus and Spea multiplicata from west Texas

Tissues from barred tiger salamanders (Ambystoma tigrinum mavortium), Great Plains toads (Bufo cognatus) and New Mexico spadefoots (Spea multiplicata) collected from 16 playa wetlands in Texas during 1999 and 2000 were examined by light microscopy. Digenean cysts were primarily distributed subcutaneously throughout the specimens and occasionally coelomic invasion was noted. The parasites within the cysts were 1.5-2 mm in diameter, with a thin (c. 10 microm wide) eosinophilic-staining tegument, two suckers (oral and ventral), posteriorly located primordial genitalia and paired digestive caeca. These digeneans were identified as the metacercariae of Clinostomum attenuatum. This is the first record of Clinostomum attenuatum in these amphibian species.     

Digenetic trematodes of the genus Clinostomum Leidy, 1856 (Digenea: Clinostomidae) from birds of Queensland, Australia, including C. wilsoni n. sp. from Egretta intermedia

Two species of Clinostomum previously described from Australia, C. hornum from Botaurus poiciloptilus (Australian bittern) and Nycticorax caledonicus (Nankeen night heron) and C. australiense from Pelecanus conspicillatus (Australian pelican), which have previously been synonymised with C. complanatum, are redescribed and recognised as valid species. In addition, C. complanatum is recorded from Egretta alba (large egret), E. garzetta (little egret), E. intermedia (plumed egret), N. caledonicus and Ardea novaehollandiae (white-faced heron). C. wilsoni n. sp. is described from E. intermedia from Queensland. C. wilsoni differs from the other three species in size and shape of the body and in the oral collar, oral sucker, intestinal caeca, caecal diverticula and position of testes. Taxonomic problems within the genus Clinostomum are discussed.     

On Podocotyle scorpaenae,Poracanthium furcatum and Derogenes latus,three poorly known digenean parasites of western Mediterranean teleosts

Three little-known digeneans, two opecoelids and a derogenid, from marine fishes off Corsica are redescribed and their systematics discussed. Distoma scorpaenae Rudolphi, 1819, from Scorpaena porcus, S. scrofa and S. notata, is transferred to the genus Podocotyle as P. scorpaenae (Rudolphi, 1819) n. comb. The authority for the taxon Poracanthium furcatum is amended to Dollfus (1948). This species is described from Mullus surmuletus and distinguished from Opecoeloides furcatus (Bremser in Rudolphi, 1819). Derogenes latus Janiszewska, 1953 is redescribed from Mullus surmuletus and discussed in relation to D. ruber Lühe, 1900.     

Trematodes of the genus Haematoloechus (Digenea: Plagiorchiidae) from frogs in Korea.

Total 242 Rana nigromaculata and 9 R. catesbeiana were collected from the various localities in Korea from February 1989 to July 1991, and their lungs were examined. Five species in genus Haematoloechus, i.e., H. sibiricus japonicus (Yamaguti, 1936), H. nanchangensis Hsiung, 1934, H. variegatus (Rudolphi, 1819), H. lobatus (Seno, 1907), H. lobatus koreanus, were identified in this study. Among them, H. lobatus koreanus was a new subspecies and H. variegatus and H. lobatus were newly recorded from Korea.     

Two species of Synhimantus (dispharynx) railliet, Henry and Sisoff, 1912 (Nematoda: Acuarioidea: Acuariidae), in passerine birds from the Area de Conservacion Guanacaste, Costa Rica

Members of 2 species of Synhimantus (Dispharynx) live under the lining of the gizzard in passerine birds from the Area de Conservacion Guanacaste, Costa Rica. Synhimantus (Dispharynx) nasuta (Rudolphi, 1819) occurs in Thraupis episcopus, Turdus grayi, Caryothraustes poliogaster, Platyrinchus cancrominus, Ramphocaenus melanurus, Vermivora peregrina, and Geothlypis poliocephala. A single male, in Turdus grayi, apparently representing a new species, distinguishable from all other species of Synhimantus (Dispharynx) by having similar shaped left and right spicules, is described but not named.     

Natural infections of Clinostomum complanatum (Trematoda: Clinostomatidae) in wild herons and egrets, Tottori Prefecture, Japan.

Nycticorax nycticorax, Ardea cinerea, Egretta garzetta, and Egretta intermedia were naturally infected with Clinostomum complanatum (Trematoda: Clinostomatidae) among fourteen wild herons, seven wild egrets and one wild bittern evaluated at the Veterinary Hospital of Tottori University, Tottori, Japan. The latter three species of heron and egrets are reported for the first time as definitive hosts of this parasite in Japan.     

Chemical Nature and Mode of Stabilization of Egg-shell in Helicometra pulchella (Rudolphi, 1819) (Digenea: Opecoelidae)

The chemical nature and mode of stabilization of egg-shell protein in digenetic trematode Helicometra pulchella (Rudolphi, 1819) have been investigated using histochemical techniques. It was found that the egg-shell is stabilized by quinone-tanning together with dityrosine. Other structural proteins (elastin, collagen and keratin-like proteins), glycogen and acid mucopolysaccharides were absent in egg-shell. Tyrosine was present in vitelline cells and immature egg-shell indicating that the proteins involved in quinone-tanning were tyrosine rich and tyrosyl residues are modified to form dityrosine in mature egg-shell.     

A new species of Mathevotaenia (Cestoda: Anoplocephalidae) and other tapeworms from marsupials in Argentina

Cestodes are reported from Didelphis albiventris Lund, 1840 and Micoureus cinereus Temminck, 1824 (Marsupialia: Didelphidae) in Argentina. These include a new species of Mathevotaenia Akhumyan, 1946 (Cestoda: Anoplocephalata) as well as M. bivittata (Janicki, 1904) and an unknown hymenolepidid cestode. Mathevotaenia argentinensis n. sp. is characterized by a relatively narrow strobila, 18-37 mm in total length and 1.0-1.5 mm in maximum width, 135-163 craspedote proglottids, 19-27 testes, and a muscular genital atrium. This species differs from M. didelphidis (Rudolphi, 1819) in the disposition of the genital ducts between the excretory canals and in the entrance of the vagina into the genital atrium posterior to the cirrus pouch; from M. paraguayae Schmidt and Martin, 1978 in the disposition of the genital ducts, absence of a seminal receptacle, and presence of an armed cirrus; and from M. boliviana Sawada and Harada, 1986 and M. pennsylvanica Chandler and Melvin, 1951 in the presence of an armed cirrus. Linstowiines appear to be the dominant cestodes in New World marsupials, with M. bivittata representing the most prevalent and widely distributed species. The hymenolepidid is the first record of this family in Neotropical marsupials.     

Bacteria associated with tegument of Clinostomum marginatum (Digenea)

Adults of Clinostomum marginatum freshly collected from a heron, Ardea herodias, were examined using transmission electron microscopy. Specimens from the mouth of the bird were encrusted with bacteria that were not removed by washing unless the saline contained antibiotics. There was no evidence that the attached bacteria were damaging to the trematode tegument. Three species of Gram-negative bacteria were isolated from the worm surfaces and identified; Achromobacter sp. was present in pure culture on 4 of 6 original cultures and in mixed culture with Edwardsiella tarda and Enterobacter agglomerans in 2 cultures. These species and 3 unidentified species of bacteria were isolated from the oral epithelium of the heron. Microorganisms were not seen attached to the surfaces of worms recovered from the esophagus. Because E. tarda and E. agglomerans were the only species isolated from the heron esophagus, the intimate bacterial-worm association in the heron mouth may be due specifically to Achromobacter sp.