Molecular evidence of cryptic diversity in Paracaryophyllaeus (Cestoda: Caryophyllidea), parasites of loaches (Cobitidae) in Eurasia,including description of P. vladkae n. sp.

Molecular phylogenetic analysis of an extensive collection of monozoic tapeworms of the genus Paracaryophyllaeus Kulakovskaya, 1961 (Cestoda: Caryophyllidea), parasites of loaches (Cypriniformes: Cobitidae) in Eurasia, has revealed cryptic species diversity within this long-time monotypic genus, especially in the Paracaryophyllaeus gotoi (Motomura, 1927) species complex [syn. Paracaryophyllaeus dubininorum (Kulakovskaya, 1961); type species]. Three independent, well-supported clades were discovered on the basis of molecular data: (i) specimens from Misgurnus anguillicaudatus and Cobitis lutheri from China, Russian Far East and Japan — called herein P. cf. gotoi 1, which may be conspecific with P. gotoi (Motomura, 1927), although in the absence of sequence data for P. gotoi from its type locality (basin of the River Kumkan in Korea), no certain inferences about their identity can currently be made; (ii) specimens from M. anguillicaudatus from China and Japan — P. cf. gotoi 2, which are morphologically indistinguishable from those of P. cf. gotoi 1; and (iii) morphologically distinct tapeworms from the endemic loach Cobitis bilseli from southwestern Turkey (Beyşehir Lake), which are described herein as a new species. Paracaryophyllaeus vladkae Scholz, Oros and Aydoğdu n. sp. differs from the remaining species of the genus in the following characteristics: the testes begin anterior to the first vitelline follicles (versus posterior), the body is short and robust (versus more elongate and slender), and the scolex is wide, rounded or apically tapered (versus claviform to truncate). Species composition of the genus, host specificity of species and geographical distribution are briefly discussed.     

Spermatological characters of monozoic tapeworms (Cestoda: Caryophyllidea), including first data on a species from the Indomalayan catfish

The ultrastructure of spermiogenesis and mature spermatozoon in Lytocestus indicus (Cestoda: Lytocestidae) is described; this is the first representative of this group of monozoic, presumably most basal, tapeworms (Eucestoda) from the Indomalayan region to be documented in this manner. Similarly, as in other caryophyllideans, its spermiogenesis involves the formation of a conical differentiation zone with 2 centrioles associated with striated roots and an intercentriolar body. In the course of the process, 1 of the centrioles develops a free flagellum, which fuses with a cytoplasmic protrusion, whereas the other remains oriented in a cytoplasmic bud. Spermiogenesis is also characterized by the presence of electron-dense material in the early stages of spermiogenesis and a slight rotation of the flagellar bud. The mature spermatozoon of L. indicus is a filiform cell tapered at both extremities that lacks mitochondria; its nucleus has parallel disposition to the axoneme and does not reach up to the posterior extremity of the spermatozoon, which is typical for spermatozoa of the type III pattern. The new data confirm that caryophyllideans share the same type of spermiogenesis that is considered to be plesiomorphic in the Eucestoda. The existing information on spermatological ultrastructure of 8 members for 3 of 4 caryophyllidean families from different host groups (cyprinids and catostomids, both Cypriniformes, and mochokids and clariids, both Siluriformes) from 4 zoogeographical regions (Palearctic, Neotropic, Ethiopian, and Indomalayan regions) demonstrates great uniformity in spermiogenesis and sperm ultrastructure, which does not reflect different taxonomic position of the species studied.     

A new monozoic tapeworm, Lobulovarium longiovatum n. g., n. sp. (Cestoda: Caryophyllidea), from barbs Puntius spp. (Teleostei: Cyprinidae) in the Indomalayan region

A new caryophyllidean cestode is described from barbs Puntius spp. (Cypriniformes: Cyprinidae), with P. sophore (Hamilton) as its type-host, in the Ganges and Brahmaputra river basins in India and Bangladesh, and a new genus, Lobulovarium n. g., is proposed to accommodate it. The genus belongs to the Lytocestidae because its vitelline follicles are situated in the cortex. It is typified by: (i) a peculiar ovary, which is roughly H-shaped, but with asymmetrical, irregular lobes on its ventral and dorsal sides; (ii) an extensive vitellarium formed by numerous vitelline follicles scattered throughout the cortex; (iii) a long, conical postovarian part of the body with numerous vitelline follicles; (iv) a broadly digitate scolex with a slightly protrusible central cone; (v) a single gonopore (male and female genital ducts open via a single pore and a common genital atrium is absent); and (vi) a small number of testes (< 60). Molecular data (partial sequences of the lsrDNA) indicate that Lobulovarium longiovatum n. sp. belongs among the most basal caryophyllidean cestodes, being unrelated to species from siluriform catfishes in the Indomalayan region. Paracaryophyllaeus osteobramensis (Gupta & Sinha, 1984) Hafeezullah, 1993 (syn. Pliovitellaria osteobramensis Gupta & Sinha, 1984) from another cyprinid fish, Osteobrama cotio (Hamilton), in Uttar Pradesh, India, is tentatively transferred to Lobulovarium as L. osteobramense (Gupta & Sinha, 1984) n. comb. It differs from L. longiovatum by having much smaller eggs (length <50 μm versus >90 μm in L. longiovatum), which are spherical (length/width ratio 1:1 versus 2.5-3:1 in the new species), and the presence of vitelline follicles alongside the ovarian lobes (almost completely absent in L. longiovatum).     

Tour around the globe: The case of invasive tapeworm Atractolytocestus huronensis (Cestoda: Caryophyllidea), a parasite of common carp

The monozoic tapeworm Atractolytocestus huronensis Anthony, 1958 (Cestoda: Caryophyllidea), an intestinal parasite of the common carp, is characterized by its invasive character and potential to colonize new territories. It was initially described from North America and has also been found in several European countries. The most recent findings of A. huronensis originated from China and South Africa; however, no data on genetic relationships of these populations were available. The current study provides the first molecular characterisation of A. huronensis from South Africa and China using a partial sequence of mitochondrial cytochrome c oxidase subunit 1 (cox1) and a complete ribosomal ITS2 spacer. Ribosomal and mitochondrial data were applied for phylogenetic analyses in order to assess the genetic interrelationships among global A. huronensis populations. Divergent intragenomic copies of ribosomal ITS2 were detected in all analysed specimens; the structure and frequency of the ITS2 variants of tapeworms from China and South Africa corresponded with the data on ITS2 paralogues observed previously in A. huronensis from Slovakia, the United States and the United Kingdom. The phylogenetic analysis of cox1 indicated that A. huronensis exist in two slightly differentiated clusters; one cluster was supported by all phylogenetic approaches (NJ, ML, BI) and was represented by samples from China, the USA and the UK. A second cluster was represented by tapeworms from continental Europe (Slovakia, Hungary, Romania, Croatia) and South Africa. Haplotype network analysis revealed that the highest population diversity occurs in China. The results provide useful pilot information about the interrelationships of A. huronensis on four continents and indicate that China, or the eastern Palaearctic, served as the original source population for the global expansion of this invasive tapeworm. Data on the origin and distribution of the common carp, the only specific host of A. huronensis, are also discussed.     

Taxonomic status of cestodes of the genus Paracaryophyllaeus Kulakowskaja, 1961 (Caryophyllidea: Caryophyllaeidae)

A taxonomic revision of cestodes of the genus Paracaryophyllaeus Kulakowskaja, 1961, parasites of cobitid fishes, has been performed. Three species (Paracaryophyllaeus gotoi (Motomura, 1927), P. kulakowskae sp. n., and P. misgurni sp. n.) are considered valid species; two are described as new species. P. dubininorum Kulakowskaja, 1961 is a young synonym of P. gotoi. The taxonomic status for P. gotoi sensu Scholz, 1989, P. gotoi sensu Scholz et al., 2001, and Paracaryophyllaeus sp. 1 and Paracaryophyllaeus sp. 2 (described in present work) needs specification.     

Hydrolytic activity of enzymes produced by symbiotic microflora and its role in digestion processes of bream and its intestinal parasite Caryophyllaeus laticeps (Cestoda, Caryophyllidea)

Bacteria capable to secrete enzymes hydrolyzing proteins and carbohydrates proved to be associated with the digestive-transport surfaces of the bream intestine and tegument of cestode Caryophyllaeus laticeps. Apparently, the contribution of symbiotic digestion to the hydrolysis of polymers increases during active feeding of fishes, while at low feeding rate, bacteria can compete with the host and its parasite for the available monomers. Secretion of bacterial enzymes hydrolyzing both complex carbohydrates and disaccharides reduces the metabolic expenditures of the macroorganisms for the synthesis of their own hydrolases. Glucose resulting from hydrolytic activity of bacterial enzymes can be used by all members of the established community. While bacteria colonizing the digestive-transport surfaces of the host and parasite were generally similar, some specific features were also observed probably due to the structural peculiarities of the intestinal mucosa and cestode tegument.     

Nomimoscolex semenasae n sp (proteocephalidea: Monticelliidae), a cestode parasite of Diplomystes viedmensis (Pisces: Siluriformes) from the Patagonian region of Argentina

Nomimoscolex semenasae n. sp. is described from the primitive fish Diplomystes viedmensis (Siluriformes) from the Patagonian region of Argentina. The new species is placed in Nomimoscolex because of the cortical position of the vitelline follicles, medullary position of the testes, ovary and uterus, and having a scolex with four uniloculate suckers. N. semenasae differs from all other species in the genus by the following combination of characters: (1) apical organ absent; (2) strobila acraspedote; (3) vagina anterior or posterior to cirrus-sac and lacking a sphincter; (4) testes in one irregular layer and in two fields connected anteriorly; (5) uterine stem cortical in immature proglottides, growing from cortical stem into medullary region in mature proglottides; (6) long uteroduct; and (7) presence of spiniform microtriches on all regions of the scolex, proliferation zone and immature proglottis. This is the first record of a proteocephalidean tapeworm in D. viedmensis and in the family Diplomystidae.     

The cestode Atractolytocestus huronensis (Caryophyllidea) continues to spread in Europe: new data on the helminth parasite of the common carp

The caryophyllidean tapeworm Atractolytocestus huronensis Anthony, 1958, originally described from the common carp Cyprinus carpio carpio L. from North America, has recently been introduced into Europe (UK, Hungary). In the present study, the dispersion of the tapeworm in other countries of Central Europe (Slovakia and the Czech Republic) is described, including its first observation in the common carp of the Tisa River, southeastern Slovakia. Because of the transboundary location of the river, there are possible consequences of the rapid distribution of the parasite to other regions throughout the Danube River basin. The parasite has also been found in cultured carp from fishponds in South Bohemia, from where the carp are imported to many European countries. Rapid dissemination of this cestode in Europe indicates its ability to colonise new regions, and represents another example of man-made introduction of potential pathogens of carp and other farmed fish. The morphology of the A. huronensis specimens found is compared with that of specimens from Hungary and North America as well as that of Atractolytocestus sagittatus (Kulakovskaya and Akhmerov, 1965) from the same fish host in eastern Asia.     

Revision of Gangesia (Cestoda: Proteocephalidea) in the Indomalayan Region: Morphology,Molecules and Surface Ultrastructure

Tapeworms of Gangesia Woodland, 1924 (Cestoda: Proteocephalidea) parasitic in freshwater fishes in the Indomalayan Region were critically reviewed. Evaluation of type specimens and newly collected materials from Bangladesh, Cambodia and India, as well as critical examination of extensive literature have shown that only the following four species, instead of 48 nominal species of Gangesia and Silurotaenia Nybelin, 1942 reported from this region (36 new synonymies proposed), are valid: Gangesia bengalensis (Southwell, 1913), type-species of the genus and most common parasite of Wallago attu (Siluridae), G. macrones Woodland, 1924 typical of Sperata seenghala (Bagridae), both species characterized by the possession of two circles of hooks on the rostellum-like organ and several rows of hooklets on the anterior margins of suckers; G. agraensis Verma, 1928 from W. attu (typical host), which has the scolex with only one circle of hooks and 1–3 incomplete rows of tiny hooklets on the suckers; and G. vachai (Gupta and Parmar, 1988) n. comb. from several catfishes, which possesses 4–6 circles of hooks and 5–11 rows of hooklets on the anterior half of suckers. Scolex morphology, including surface ultrastructure (microtriches), of all but one species (G. vachai) is described for the first time using scanning electron microscopy. A phylogenetic analysis based on the partial sequences encoding the large nuclear ribosomal subunit RNA gene has shown that three Indomalayan species, namely G. bengalensis, G. macrones and G. vachai, form a monophyletic group within Gangesia, whereas G. agraensis tends to form a clade with the Palaearctic species of the genus. A table with differential characters of all species from the Indomalayan Region is also provided together with a key to identification of genera of the subfamily Gangesiinae. The present study demonstrates that species of Silurotaenia do not occur in the Indomalayan region.     

Molecular evidence confirms the parasite Congracilaria babae (Gracilariaceae,Rhodophyta) from Malaysia

A phylogenetic study of two parasites found on the common host species Gracilaria salicornia from Japan and Malaysia based on nuclear, plastid, and mitochondrial molecular markers was conducted. The Japanese and Malaysian parasites were placed in the same cluster in the nuclear phylogenies inferred from the LSU rRNA gene and ITS region, supporting the monophyly of these parasites despite the slight anatomical variations observed. In addition to the molecular analyses, shared morphological and reproductive features including the lack of rhizoids, similar coloration to hosts, and the presence of deep spermatangial conceptacles supported the inclusion of both the Japanese and Malaysian parasites in Congracilaria babae. Phylogenetic analyses based on ITS sequences indicated that C. babae most likely evolved directly from its current host species, G. salicornia. C. babae was shown to be a member of Gracilaria sensu stricto along with its host. C. babae growing on G. salicornia is characterized by having plastid rbcL and mitochondrial cox1 gene sequences identical to those of its host, whereas the nuclear LSU rRNA gene and ITS region sequences are different from those of its host.     

Molecular and morphological evidence suggests the reallocation from Parastrigea brasiliana (Szidat, 1928) Dubois, 1964 to Apharyngostrigea Ciurea, 1927 (Digenea: Strigeidae), a parasite of boat-billed heron (Cochlearius cochlearius) from the Neotropical region

Parastrigea brasiliana (Szidat, 1928) Dubois, 1964, was described from (Cochlearius cochlearius) in South America. The taxonomy of this species has been unstable due that it was described as a member of Strigea Abildgaard, 1790. However, the same author one year later transferred it to Apharyngostrigea Ciurea, 1927 and since then, it has been alternatively placed in the genus Apharyngostrigea or Parastrigea Szidat, 1928 from Strigeidae. In the current research, specimens identified as P. brasiliana were collected from type host in southeastern Mexico. We sequenced three molecular markers: the internal transcribed spacers ITS1 and ITS2 including the 5.8S gene (ITS region), the D1-D3 domains of the large subunit (LSU) from nuclear DNA and cytochrome c oxidase subunit I (cox 1) from mitochondrial DNA. These sequences were aligned with other sequences available in the GenBank dataset from Strigeidae. Maximum likelihood and Bayesian analyses inferred with three molecular markers consistently showed that P. brasiliana is not closely related to other members of the genus Parastrigea and are placed in a reciprocal monophyletic clade inside Apharyngostrigea, with very low genetic divergence, varying from 0 to 0.09% for the ITS, from 0 to 0.08% for the LSU and from 0.21 to 0.43% for cox 1. Consequently, we proposed to reallocate it to A. brasiliana. The phylogenetic analyses obtained are key and very useful for re-evaluate the morphology of A. brasiliana because this species share morphological characters with the genera Parastrigea (concentration of vitelline follicles distributed in two lateral expansions on the forebody) and Apharyngostrigea (absence of pharynx). Finally, the current record of A. brasiliana expands its distribution range in four countries, namely, the USA, Mexico, Venezuela and Brazil, in the Neotropical region.     

Ultrastructure of the spermatozoon of Bothriocotyle sp. (Cestoda: Bothriocephalidea), a parasite of Schedophilus velaini (Sauvage, 1879) (Perciformes: Centrolophidae) in Senegal

The mature spermatozoon of Bothriocotyle sp. is filiform and tapered at both extremities. It possesses 2 axonemes of unequal length, showing the 9 + "1" pattern of Trepaxonemata. The anterior extremity exhibits a crest-like body. Thereafter, the crest-like body disappears, and the first axoneme is surrounded by a ring of cortical microtubules (about 27 units) that persist until the appearance of the second axoneme. This ring of cortical microtubules is characteristic only for species of Bothriocephalidea and represents a very useful phylogenetic character. The spermatozoon cytoplasm is slightly electron-dense and contains numerous electron-dense granules of glycogen in several regions. The anterior and posterior extremities of the spermatozoon lack cortical microtubules. The posterior extremity of the spermatozoon of Bothriocotyle sp. possesses a nucleus and a disorganized axoneme, which also characterizes spermatozoa of the Echinophallidae studied to date.     

Morphological polymorphism of the cestode Proteocephalus longicollis Zeder, 1800 (Cestoda: Proteocephalidae) through the ontogeny

The qualitative diversity of Proteocephalus longicollis through the ontogeny was studied: procercoids from copepods, immature and mature cestodes from the vendace Coregonus albula L. The structure of the P. longicollis population was found to be made up of the dominant phenotypes: by the scolex shape--Sc2, by the sucker arrangement type--SI, by the proglottid shape--P3, by the ovary lobe shape--01, by the testes arrangement type -T2. Some rare and scant phenotypes are also present. It is shown that the structure of the parasite phenotypes is relatively stable through the ontogeny. The assumption is made about the high survival rate of the S2 sucker phenotype.     

Spermiogenesis and spermatozoon ultrastructure of the cestode Mosgovoyia ctenoides (Cyclophyllidea: Anoplocephalidae), an intestinal parasite of Oryctolagus cuniculus (Lagomorpha: Leporidae)

Ultrastructural characters in spermiogenesis and spermatozoa are considered important tools to elucidate the phylogenetic relationships within the Platyhelminthes. In the Anoplocephalidae, ultrastructural data refer to the spermatozoon of 14 species, whereas data on spermiogenesis refer to only 7 species. The present study focused on the spermiogenesis and spermatozoon of the anoplocephalid cestode Mosgovoyia ctenoides, as revealed by transmission electron microscopy. Type IV spermiogenesis was detected, beginning with the formation of a differentiation zone containing 2 centrioles, with a centriolar adjunct and vestigial striated rootlets. Different forms of the latter character have been described in other anoplocephalids. This study supports spermiogenesis of type IV as the most frequent in the Anoplocephalidae and confirms the presence of a centriolar adjunct in yet another type IV spermiogenesis species. The spermatozoon of M. ctenoides possesses 1 axoneme of the 9+ '1' trepaxonematan type, 2 crestlike bodies, dense plates, and granules of electron-dense cytoplasmic material, nucleus, and twisted cortical microtubules. It was again confirmed that the presence of granular material and the absence of both a periaxonemal sheath and intracytoplasmic walls are constant characters in the spermatozoa of all the Anoplocephalinae.     

Aploparaksis kornyushini n. sp. (Cestoda: Hymenolepididae), a parasite of the woodcock Scolopax rusticola (L.), and its life-cycle

Aploparaksis kornyushini n. sp. is described from a woodcock Scolopax rusticola L. from Lithuania, Russia (Tver' Region) and the Ukraine. Initially, one specimen of this tapeworm was described and figured by Kornyushin (1975) as A. scolopacis Yamaguti, 1935 together with another specimens belonging to the latter species. A. kornyushini n. sp. and A. scolopacis are morphologically very similar species. They can be distinguished by the slightly different length of the rostellar hooks and by the shape of the cirrus, which lacks basal bulbus in the new species. A. kornyushini can be readily distinguished from the remaining species of Aploparaksis Clerc, 1903 from woodcocks by the structure of its fully-developed embryophore, which has polar thickenings and two large or a few smaller lateral projection; this combination of characters is unknown for embryophores other Aploparaksis spp. (except for A. scolopacis). The life-cycle of A. kornyushini was studied under experimental conditions in Lithuania. The metacestodes were located under the chlorogogenous tissue of the intestine of Dendrobaena octaedra (Lumbricidae). The metacestode exhibits a pattern of postembryonal development typical for the cysticercoid modification termed an 'ovoid diplocyst'.     

Aploparaksis kornyushini n. sp. (Cestoda: Hymenolepididae), a parasite of the woodcock Scolopax rusticola (L.), and its life-cycle

Aploparaksis kornyushini n. sp. is described from a woodcock Scolopax rusticola L. from Lithuania, Russia (Tver' Region) and the Ukraine. Initially, one specimen of this tapeworm was described and figured by Kornyushin (1975) as A. scolopacis Yamaguti, 1935 together with another specimens belonging to the latter species. A. kornyushini n. sp. and A. scolopacis are morphologically very similar species. They can be distinguished by the slightly different length of the rostellar hooks and by the shape of the cirrus, which lacks basal bulbus in the new species. A. kornyushini can be readily distinguished from the remaining species of Aploparaksis Clerc, 1903 from woodcocks by the structure of its fully-developed embryophore, which has polar thickenings and two large or a few smaller lateral projection; this combination of characters is unknown for embryophores other Aploparaksis spp. (except for A. scolopacis). The life-cycle of A. kornyushini was studied under experimental conditions in Lithuania. The metacestodes were located under the chlorogogenous tissue of the intestine of Dendrobaena octaedra (Lumbricidae). The metacestode exhibits a pattern of postembryonal development typical for the cysticercoid modification termed an 'ovoid diplocyst'.     

A redescription of the pinniped parasite Contracaecum ogmorhini (Nematoda, Ascaridoidea), with an assessment of its antiboreal circumpolar distribution

The otariid seal parasite Contracaecum ogmorhini Johnston & Mawson, 1941 is redescribed based upon new material and the type specimens. The geographical records of this species indicate that it has an antiboreal circumpolar distribution with an isolated population on the south-west coast of North America. The species is considered to be morphologically more similar to Contracaecum spp. from birds than to those from phocid seals: its possible evolution is commented upon. The material identified as C. osculatum (Rudolphi) by Flores-Barroeta et al . is considered conspecific with C. ogmorhini , thus suggesting that it is this species, and not C. osculatum , which is responsible for abnormal tissue migration and brain damage in pinnipeds.