Tomáš Scholz

On the Position of Archigetes and Its Bearing on the Early Evolution of the Tapeworms

The tapeworm Archigetes sieboldi Leuckart, 1878 (Platyhelminthes: Cestoda: Caryophyllidea) has been cited as a likely representative of the “protocestode” condition, owing to its lack of segmentation and ability to attain sexual maturity in the invertebrate host (aquatic oligochaetes). The idea has been variously amplified or rejected in the literature, although the actual phylogenetic position of the species has not been investigated until now. New collections of Archigetes sp. from both its vertebrate and invertebrate hosts provided the opportunity to estimate its phylogenetic position with the use of molecular systematics, while prompting new analyses aimed at assessing the early diversification of the Cestoda. Additional collections representing the Amphilinidea, Caryophyllidea, and Gyrocotylidea were combined with published gene sequences to construct data sets of complete 18S (110 taxa) and partial (D1-D3) 28S (107 taxa) rDNA sequences, including 8 neodermatan outgroup taxa. Estimates resulting from Bayesian inference, maximum likelihood, and maximum parsimony analyses of the separate and combined data sets supported a derived position of the genus within the Caryophyllidea, and thus reject the idea that Archigetes sp. may exemplify a “primitive” condition. Topological constraint analyses rejected the hypothesis that Archigetes represents the most basal lineage of the Eucestoda, but did not rule out that it could represent the earliest branching taxon of the Caryophyllidea. In all analyses, the Eucestoda were monophyletic and supported basal positions of the nonsegmented Caryophyllidea and Spathebothriidea relative to other major lineages of the Eucestoda, implying that segmentation is a derived feature of the common ancestor of the di- and tetrafossate eucestodes. However, constraint analyses could not provide unequivocal evidence as to the precise branching patterns of the cestodarian, spathebothriidean, and caryophyllidean lineages. Phylogenetic analyses favor the interpretation that sexual maturity of Archigetes sp. in the invertebrate host, and similar examples in members of the Spathebothriidea, are the result of progenesis and have little if any bearing on understanding the protocestode condition.

Intra-individual internal transcribed spacer 1 (ITS1) and ITS2 ribosomal sequence variation linked with multiple rDNA loci: A case of triploid Atractolytocestus huronensis, the monozoic cestode of common carp

Complete sequences of the ribosomal internal transcribed spacers (ITS1 and ITS2) and karyological characters of the monozoic (unsegmented) tapeworm Atractolytocestus huronensis Anthony, 1958 (Cestoda: Caryophyllidea) from Slovakia were analysed, revealing considerable intra-genomic variability and triploidy in all analysed specimens. Analysis of 20 sequences of each ITS1 and ITS2 spacer yielded eight and 10 different sequence types, respectively. In individual tapeworms, two to four ITS1 and three to four ITS2 sequence types were found. Divergent intra-genomic ITS copies were mostly induced by nucleotide substitutions and different numbers of short repetitive motifs within the sequence. In addition, triploidy was found to be a common feature of A. huronensis. The karyotype of Slovakian A. huronensis possesses three sets of chromosomes (3n=24, n=4m+3st+1minute chromosome), similar to the previously described triploidy in conspecific tapeworms from North America. Fluorescent in situ hybridisation (FISH) with a ssrDNA probe revealed two distinct rDNA clusters for each homologue of the triplet number 2. To date, A. huronensis is the only cestode species in which intra-individual ITS sequence variants were found in parallel with its triploid nature and multiple rDNA loci. Some of these molecular and genetic features were observed in several other species of basal or nearly basal tapeworms of the orders Caryophyllidea and Diphyllobothriidea, which indicates that the phenomena may be characteristic for evolutionarily lower tapeworms and deserve more attention in future studies.

Substitution saturation and nuclear paralogs of commonly employed phylogenetic markers in the Caryophyllidea, an unusual group of non-segmented tapeworms (Platyhelminthes)

Caryophyllidean cestodes (Platyhelminthes) represent an unusual group of tapeworms lacking serially repeated body parts that potentially diverged from the common ancestor of the Eucestoda prior to the evolution of segmentation. Here we evaluate the utility of two nuclear and two mitochondrial molecular markers (ssrDNA and lsrDNA, nad3 and cox1) for use in circumscribing generic boundaries and estimating interrelationships in the group. We show that these commonly employed markers do not contain sufficient signal to infer well-supported phylogenetic estimates due to substitution saturation. Moreover, we detected multiple trnK+nad3+trnS+trnW+cox1 haplotypes within individuals, indicating a history of gene exchange between the mitochondrial and nuclear genomes. The presence of such nuclear paralogs (i.e. numts), to our knowledge described here in cestodes for the first time, together with the results of phylogenetic, saturation and split-decomposition analyses all suggest that finding informative markers for estimating caryophyllidean evolution is unusually problematic in comparison to other major lineages of tapeworms.

Phylogenetic relationships of the monozoic tapeworms (Eucestoda: Caryophyllidea) inferred from morphological characters

Phylogenetic relationships of all genera of the order Caryophyllidea, possibly the earliest branching group of true tapeworms (Platyhelminthes: Eucestoda) and the only one that is monozoic, have been assessed for the first time. Results of this cladistic analysis, inferred from 30 unweighted morphological characters, are only partly congruent with the existing classification, which consists of four families based on the position of the inner longitudinal muscles in relation to the internal genital organs. Whereas all but five genera of the Caryophyllaeidae form a monophyletic clade, members of the Capingentidae are split, occurring within six unrelated groups. The Lytocestidae is also paraphyletic, as some genera appear in four unrelated clades. Archigetes appears in a derived clade, indicating that its direct (monoxenous) life-cycle involving only tubificid oligochaetes is secondarily derived and not plesiomorphic among the Eucestoda, as postulated by some authors.

Population study of Atractolytocestus huronensis (Cestoda: Caryophyllidea), an invasive parasite of common carp introduced to Europe: mitochondrial cox1 haplotypes and intragenomic ribosomal ITS2 variants

The invasive monozoic tapeworm Atractolytocestus huronensis, a specific parasite of common carp, was originally found and described in the North American continent. It has been introduced to Europe and reported in several countries in the last 15xa0years, as well. In the current study, tapeworms from one North American (USA) and five European localities (United Kingdom/UK, Slovakia, Hungary, Croatia, and Romania) were subjected to molecular analyses in order to determine the level of intrapopulation and intraspecific molecular variation and to assess interrelationships among American and European populations of the parasite. Partial sequences (672xa0bp) of the mitochondrial cytochrome c oxidase subunit I (cox1) revealed the presence of only two cox1 haplotypes, in accordance with the nonnative character of the populations. The first haplotype was common for all tapeworms from the Continental Europe (Slovakia, Hungary, Croatia, and Romania); no differences were determined either within or among respective A. huronensis populations. The second cox1 haplotype was characterized in all individuals from the USA and UK, indicating their close genetic relationship. Both haplotypes differed in three nucleotide positions (99.6% identity) which did not change the amino acid sequence. The cox1 data imply that introduction of the parasite to Europe was probably the result of two independent events directed to the UK and Continental Europe. The very close genetic relationship between British and American A. huronensis was reflected also by similar ribosomal internal transcribed spacer 2 (ITS2) sequence structure; considerable intragenomic ITS2 variability was detected in all individuals of both geographic populations. Divergent ITS2 copies were mostly induced by different numbers of short repetitive motifs within the sequences, allowing their assortment into two ITS2 variants.

Tapeworm Khawia sinensis: review of the introduction and subsequent decline of a pathogen of carp, Cyprinus carpio.

The Asian tapeworm Khawia sinensis Hsü, 1935 (Cestoda: Caryophyllidea) is a large-sized (body length up to 11.5 cm) monozoic (unsegmented) parasite of common carp (Cyprinus carpio L.) that may cause mortality of young fish (fry). Since the 1960s, this cestode successfully colonized a large part of Europe, including the British Isles, North America and Japan. However, a review of published records provides evidence that the tapeworm K. sinensis, invasive parasite of carp, has become less common during the last two decades. Decline of K. sinensis may have been related to the recent introduction of another invasive tapeworm, the caryophyllidean Atractolytocestus huronensis Anthony, 1958 to Europe. Other factors that may have caused that K. sinensis is much less common than previously are also briefly discussed. A comparison of K. sinensis from feral and cultured carp, published to date, with those recently found for the first time in wild populations of carp in Slovakia did not reveal any marked differences in their morphology or measurements.

Parasites of Freshwater Fishes In North America: Why So Neglected?

Abstract:u2003 Fish parasitology has a long tradition in North America and numerous parasitologists have contributed considerably to the current knowledge of the diversity and biology of protistan and metazoan parasites of freshwater fishes. The Journal of Parasitology has been essential in disseminating this knowledge and remains a significant contributor to our understanding of fish parasites in North America as well as more broadly at the international level. However, with a few exceptions, the importance of fish parasites has decreased during the last decades, which is reflected in the considerable decline of funding and corresponding decrease of attention paid to these parasites in Canada and the United States of America. After the ‘golden age in the second half of the 20th Century, fish parasitology in Canada and the United States went in a new direction, driven by technology and a shift in priorities. In contrast, fish parasitology in Mexico has undergone rapid development since the early 1990s, partly due to extensive international collaboration and governmental funding. A critical review of the current data on the parasites of freshwater fishes in North America has revealed considerable gaps in the knowledge of their species composition, host specificity, life cycles, evolution, phylogeography, and relationships with their fish hosts. As to the key question, “Why so neglected?” this is probably because: (1) fish parasites are not in the forefront due to their lesser economic importance; (2) there is little funding for this kind of research, especially if a practical application is not immediately apparent; and (3) of shifting interests and a shortage of key personalities to train a new generation (they switched to marine habitats or other fields). Some of the opportunities for future research are outlined, such as climate change and cryptic species diversity. A significant problem challenging future research seems to be the loss of trained and experienced fish parasitologists. This has 2 major ramifications: the loss of expertise in identifying organisms that other biologists (e.g., ecologists, molecular biologists, evolutionists) work with, and an incomplete comprehension of ecosystem structure and function in the face of climate change, emerging diseases, and loss of biodiversity.

Tapeworms (Cestoda: Caryophyllidea), Parasites of Clarias batrachus (Pisces: Siluriformes) in the Indomalayan Region

Abstract Revision of monozoic cestodes (Caryophyllidea) parasitic in commercially important walking catfish Clarias batrachus (L.) in tropical Asia (Indomalayan zoogeographical region) was carried out, based on the evaluation of newly collected material from India, Indonesia, and Thailand, as well as a study of type specimens. Instead of the 59 nominal taxa of 15 genera from 3 caryophyllidean families previously described, only 8 species of the Lytocestidae are considered to be valid: Bovienia indica (Niyogi, Gupta and Agarwal, 1982) n. comb.; Bovienia raipurensis (Satpute and Agarwal, 1980) Mackiewicz, 1994; Bovienia serialis (Bovien, 1926) Fuhrmann, 1931; Djombangia penetrans Bovien, 1926; Lucknowia microcephala (Bovien, 1926) n. comb.; Lytocestus indicus (Moghe, 1925) Woodland 1926; Pseudocaryophyllaeus ritai Gupta and Singh, 1983; and Pseudocaryophyllaeus tenuicollis (Bovien, 1926) n. comb. All valid species are redescribed and SEM photomicrographs of their scolices and photomicrographs of their eggs are provided for the first time. Crescentovitus Murhar, 1963, Heeradevina Srivastav and Khare, 2005, Pseudobatrachus Pathak and Srivastav, 2005, Pseudobilobulata Srivastav and Lohia, 2002, Pseudoclariasis Pathak, 2002, and Pseudoinverta Pathak, 2002 are invalidated and 50 nominal species are newly synonymized, including 4 species described from other fish hosts. Taxonomic status of the remaining caryophyllidean taxa reported from C. batrachus (at least 6 taxa) could not be clarified because of the unavailability of their original descriptions. A key to identification of caryophyllidean tapeworms parasitic in C. batrachus is provided. To avoid current inflation of descriptions of invalid taxa, researchers are strongly encouraged to work only with well-fixed material; damaged, decomposed, or strongly flattened specimens should not be used for taxonomic studies, and type specimens must always be deposited in internationally recognized collections.

Sequence structure and intragenomic variability of ribosomal ITS2 in monozoic tapeworms of the genus Khawia (Cestoda: Caryophyllidea), parasites of cyprinid fish

The sequence structure of the ribosomal internal transcribed spacer 2 (ITS2) was determined for six species of Khawia (Cestoda: Caryophyllidea), parasites of cyprinid fish in the Holarctic Region. Homologous intragenomic ITS2 structure was found in Khawia armeniaca, Khawia baltica, and Khawia rossittensis; whereas divergent intragenomic ITS2 copies were detected in Chinese, Japanese, and Slovak isolates of Khawia sinensis and in Khawia japonensis, both parasitic in common carp, and in Khawia saurogobii, recently described from Chinese lizard gudgeon in China. Despite distinct morphological differences between K. saurogobii and K. sinensis, both species display very high level of molecular homogeneity. Variation in number of short repetitive motifs [(GCCT)n (GCCC)n], [(GTG)n], [(ATAC)n], [ACGTGT (TCGTGT)n], [(GT)n], [(GT)n], and [(ACCT)n (GCCT)n] resulted in assortment of ITS2 sequences in four ITS2 variants in K. saurogobii from China, three in Chinese and Japanese isolates of K. sinensis, and five ITS2 variants in K. sinensis from Slovakia. In K. japonensis, the structure and arrangement of microsatellites was different from those of K. sinensis and K. saurogobii. The heterogeneity in the number of two microsatellite regions [(TG)n; (TTG)n] divided ITS2 clones into two variants—first ITS2 variant (472xa0bp) with (TG)5 and (TTG)6, and second variant with (TG)7 and (TTG)2 (465xa0bp). Sequence identity of K. saurogobii with all but one (K. sinensis) congeneric species ranged between 49.5 and 69.2xa0%, which corresponds to the interspecific differences. In contrast, sequence identity of K. saurogobii and K. sinensis (87.6–95.0xa0%) failed into the range of intraspecific variation determined for K. sinensis samples. This close genetic similarity indicates that recently described K. saurogobii may have undergone morphological divergence as a result of ongoing sympatric speciation by host switching.

Phenotypic plasticity in Caryophyllaeus brachycollis Janiszewska, 1953 (Cestoda: Caryophyllidea): does fish host play a role?

Recent molecular phylogenetic studies on fish tapeworms of the genus Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes in the Palaearctic Region, have revealed unexpected phenotypic plasticity that seems to be related to definitive hosts. In the present paper, Caryophyllaeus brachycollis Janiszewska, 1953 is redescribed and its two morphotypes are circumscribed on the basis of newly-collected specimens. Morphotype 1 from barbels [Barbus spp. including the type-host Barbus barbus (L.); Barbinae] and chubs (Squalius spp.; Leuciscinae) is characterised by a more robust body with spatulate scolex, which is only slightly wider than a very short neck region, and the anterior position of the testes and vitelline follicles, which begin immediately posterior to the scolex. Specimens of Morphotype 2 from breams (Abramis spp., Ballerus spp. and Blicca spp.; Abraminae), which have been previously misidentifed as Caryophyllaeus laticeps (Pallas, 1781), possess a more slender body with a flabellate scolex, which is much wider than a long neck, and the first testes begin at a considerable distance posterior to the first vitelline follicles. Despite conspicuous differences in the scolex morphology and the anterior extent of the testes and vitelline follicles, both morphotypes are identical in the morphology of the posterior end of the body, in particular that of the cirrus-sac, which is large, thick-walled, elongate-pyriform, and contains a long cirrus, and in the distribution of the vitelline follicles, which surround medially vas deferens near the cirrus-sac. A specimen of Morphotype 1 from B. barbus from the Argens River, France, is designated as neotype of C. brachycollis. The presence of phenotypic plasticity in morphological characteristics previously used for differentiation of species of Caryophyllaeus may confound species identification, which is crucial for biodiversity, ecological and evolutionary studies. To avoid these potential problems, combination of morphological and molecular data is strongly recommended.