Search

A study on the life cycle of the trematode Asymphylodora tincae (Modeer 1790) (family Monorchiidae), carried out under both natural and laboratory conditions, revealed as its most remarkable characteristics the absence of a metacercarial stage. The definitive host, the tench [ Tinca tinca (L.)], became directly infected after the ingestion of snails harbouring rediae with mature cercariae. With the exception of the sporocyst, all developmental stages from both naturally and experimentally infected pulmonate snails as well as adults from experimental fish hosts were described and illustrated, including the surface morphology of the cercariae, studied by scanning electron microscopy. Confusion in the identification of larval stages from pulmonate snails and the prosobranchiate genus Bithynia, hitherto identified as A. tincae by other authors, are briefly discussed with regard to the present results. [ABSTRACT FROM AUTHOR]

A new species of Crassicutis Manter, 1936 (Digenea: Megaperidae) is described from the intestine of Satanoperca jurupari (Cichlidae) in the Amazon River basin, Brazil. The genus Crassicutis currently contains eight species. Crassicutis manteri n. sp. is morphologically very similar to Crassicutis cichlasomae Manter, 1936, a parasite of cichlids reported from Mexico, the Antilles, and Central and South America. Molecular data revealed that C. cichlasomae represents a species complex in Middle American cichlids. The new species can be readily distinguished from C. cichlasomae sensu lato, and the other congeners, by a combination of morphological traits such as a narrow, elongate mouth opening (versus spherical in other species), the tandem position of testes (symmetrical or oblique in most congeners), narrow body widening towards its posterior end (versus widely oval, leaf-like in other species), and short intestinal caeca ending close to the posterior end of the posterior testis (versus reaching more posteriorly in other species). Six novel sequences of 28S rDNA, ITS1, and cox1 were generated for two isolates of the new species. Sequences of the 28S rRNA gene were used to corroborate that Crassicutis is sister taxa of Homalometron Stafford, 1904. Mitochondrial DNA corroborated the distinction of the new species with previously sequenced congeners in Middle American cichlids; the interspecific divergence between the new species and the genetic lineages of C. cichlasomae was very high, varying between 23.7 and 27.2%. Biogeographical implications of our findings are briefly discussed including questionable validity of records of C. cichlasomae from South America.

A new genus, Megancestus n. gen., is proposed to accommodate the caryophyllidean tapeworm Biacetabulum carpiodi Mackiewicz, 1969 from carpsuckers and quillback (Carpiodes spp.) in North America. This species is not closely related to other species of Biacetabulum Hunter, 1927 and is transferred to a newly erected genus. This new genus is typified by the possession of a small body (total length of 3.1-7.5 mm) with a scolex that bears a pair of large acetabulum-like loculi, two pairs of shallow lateral loculi, and a slightly convex apical disc, testes arranged in one or two layers, oval, thick-walled cirrus-sac, well-developed external seminal vesicle, separate gonopores, H-shaped ovary, few median vitelline follicles, and the uterus extending by a single loop anterior to the cirrus-sac. Megancestus differs from all Nearctic caryophyllidean genera (family Capingentidae), including Biacetabulum, by vitelline follicles dorsal to the ovary that connect the preovarian and postovarian vitelline fields. The most closely related Hunterella Mackiewicz et McCrae, 1962 differs by shape of the scolex (tholate, i.e., devoid of any loculi), dumbbell-shaped ovary and the uterus not extending anterior to the cirrus-sac. Megancestus carpiodi (Mackiewicz, 1969) n. comb. is the only species of the genus and it is a stenoxenous parasite, which has been found only in the river carpsucker (Carpiodes carpio-type host), quillback (Carpiodes cyprinus) and highfin carpsucker (Carpiodes velifer) (Catostomidae: Ictiobinae) in the lower and middle Mississippi basin.

Tapeworms (Cestoda: Proteocephalidae) are the dominant component of communities of intestinal parasites in pimelodid and other catfishes (Siluriformes) from South America. Even though these parasites have been studied intensively over more than one century, molecular taxonomy and phylogenetics have questioned their morphology-based classification, thus raising doubts about the systematic value of traits commonly used to circumscribe individual taxa. In the present study, members of three morphologically well-characterized genera of proteocephalids from pimelodid (Hemisorubim platyrhynchos and Sorubim lima) and auchenipterid (Ageneiosus inermis) catfishes from the Paraná or Amazon River basins were subjected to DNA sequencing of the large subunit nuclear ribosomal RNA (lsrDNA) and complete mitochondrial cytochrome c oxidase subunit I (COI). Phylogenetic analyses revealed the sister relationship between Manaosia bracodemoca and Mariauxiella piscatorum, and among Mariauxiella pimelodi and Ageneiella brevifilis. As a result, Mar. piscatorum and A. brevifilis are transferred to Manaosia and Mariauxiella, respectively, as Manaosia piscatorum n. comb. and Mariauxiella brevifilis n. comb., and the genus Ageneiella is suppressed. Diagnoses of Manaosia and Mariauxiella are amended. In addition, the present study revealed misidentification of tapeworms whose sequences are deposited in the GenBank database.

A morphological and molecular phylogenetic study of proteocephalid tapeworms of the genus Acanthotaenia von Linstow, 1903, parasites of monitors (Varanidae), was carried out. The type species, A. shipleyi von Linstow, 1903, which was originally described based on an immature specimen from Sri Lanka, is redescribed based on new material from the type host, Varanus salvator, in Sri Lanka, Malaysia, and Vietnam, and its neotype is designated. In addition, Acanthotaenia susanae n. sp. is described from Varanus nebulosus in Vietnam. The new species differs from congeners by the large size of the scolex, width of the rostellum and the number of testes. New molecular data (sequences of lsrDNA and cox1) revealed Acanthotaenia paraphyletic with the inclusion of Australotaenia bunthangi de Chambrier & Scholz, 2012, a parasite of Enhydris enhydris (Ophidia: Homalopsidae) in Cambodia. Molecular data confirm a wide distribution of A. shipleyi (isolates from Malaysia and Vietnam were almost identical) and indicate a strict host specificity (oioxeny) of individual species of the genus. Type specimens of four species made it possible to supplement their morphological descriptions. A survey of all species of Acanthotaenia recognised as valid is presented and the following taxonomic changes are proposed: Acanthotaenia pythonis Wahid, 1968 described from the green python, Morelia viridis, in a zoo, is transferred to Kapsulotaenia as Kapsulotaenia pythonis (Wahid, 1968) n. comb., because it possesses intrauterine eggs grouped in capsules. Acanthotaenia gracilis (Beddard, 1913) from Varanus varius in Australia is considered to be species inquirenda because its original descriptions did not contain sufficient data for adequate circumscription and differentiation from congeners and type material was not available. Generic diagnosis of Acanthotaenia is amended and a key to its seven species is provided.

Metagonimus Katsurada, 1912 is a genus of small intestinal parasites. The genus comprises eight species, primarily from far-eastern Asia, with two exceptions reported from Europe. Metagonimus yokogawai, the most widespread species, is the main agent responsible for the intestinal disease, metagonimiasis, in Japan and some other East Asian countries. On the basis of the ratio of the size of the ventral and oral suckers, Metagonimus has traditionally been morphologically divided into two groups; however, the genus has not been extensively studied using molecular data. To reveal phylogenetic relationships within Metagonimus based on molecular data, we analyzed six of the seven species present in Asia using samples collected in central Japan. Maximum likelihood and Bayesian analyses of a combined 28S ribosomal DNA (rDNA), internal transcribed spacer 2 (ITS2), and mitochondrial cox1 gene sequence dataset separated the six species into two well-supported clades. One clade comprised M. yokogawai, M. takahashii, M. miyatai, and M. hakubaensis, whereas the other consisted of M. otsurui and M. katsuradai. Genetic distances calculated from 28S rDNA and ITS2 nucleotide sequences and a comparison of the predicted amino acid sequences of cox1 gene suggested that M. otsurui and M. katsuradai may have diverged recently. None of the four main morphological characters used to delimit species of Metagonimus (i.e., sucker ratio, positions of the uterus and testes, and distribution of vitelline follicles) was consistent with the distribution of species in the molecular tree.

Biological invasions represent a serious threat for aquaculture because many of introduced parasites may negatively affect the health state of feral and cultured fish. In the present account, the invasive tapeworm Atractolytocestus huronensis Anthony, 1958 (Cestoda: Caryophyllidea), which was originally described from North America and has been introduced to Europe including the British Isles with its specific host, common carp (Cyprinus carpio L.), is reported from Africa for the first time. Its recent introduction to South Africa, where it was found in four localities where common carp is cultured, is another evidence of insufficient prophylactic measures and inadequate veterinary control during transfers of cultured fish, especially common carp, between continents. Together with the Asian fish tapeworm, Bothriocephalus acheilognathi, A. huronensis is another fish tapeworm with ability to spread throughout the globe as a result of man-made introductions of its fish hosts.

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 (472 bp) with (TG)(5) and (TTG)(6), and second variant with (TG)(7) and (TTG)(2) (465 bp). Sequence identity of K. saurogobii with all but one (K. sinensis) congeneric species ranged between 49.5 and 69.2%, which corresponds to the interspecific differences. In contrast, sequence identity of K. saurogobii and K. sinensis (87.6-95.0%) 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.

Sequence structure of complete internal transcribed spacer 1 and 2 (ITS1 and ITS2) of the ribosomal DNA region and partial mitochondrial cytochrome c oxidase subunit I (cox1) gene sequences were studied in the monozoic tapeworm Atractolytocestus sagittatus (Kulakovskaya et Akhmerov, 1965) (Cestoda: Caryophyllidea), a parasite of common carp (Cyprinus carpio carpio L.). Intraindividual sequence diversity was observed in both ribosomal spacers. In ITS1, a total number of 19 recombinant clones yielded eight different sequence types (pairwise sequence identity, 99.7-100%) which, however, did not resemble the structure typical for divergent intragenomic ITS copies (paralogues). Polymorphism was displayed by several single nucleotide mutations present exclusively in single clones, but variation in the number of short repetitive motifs was not observed. In ITS2, a total of 21 recombinant clones yielded ten different sequence types (pairwise sequence identity, 97.5-100%). They were mostly characterized by a varying number of (TCGT)(n) repeats resulting in assortment of ITS2 sequences into two sequence variants, which reflected the structure specific for ITS paralogues. The third DNA region analysed, mitochondrial cox1 gene (669 bp) was detected to be 100% identical in all studied A. sagittatus individuals. Comparison of molecular data on A. sagittatus with those on Atractolytocestus huronensis Anthony, 1958, an invasive parasite of common carp, has shown that interspecific differences significantly exceeded intraspecific variation in both ribosomal spacers (81.4-82.5% in ITS1, 74.4-75.2% in ITS2) as well as in mitochondrial cox1, which confirms validity of both congeneric tapeworms parasitic in the same fish host.

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 15 years, 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 (672 bp) 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.

Spermiogenesis and spermatozoon ultrastructure of the caryophyllidean cestode Breviscolex orientalis Kulakovskaya, 1962, first member of the family Capingentidae studied, a parasite of cyprinid fish Abbottina rivularis, are described using transmission electron microscopy. Spermiogenesis in B. orientalis follows the Type II pattern described by Bâ and Marchand (Mém Mus Natl Hist Nat 166:87-95, 1995) for cestodes. It begins with the formation of a zone of differentiation containing a large nucleus and a pair of centrioles. The centrioles are separated from one another by an intercentriolar body composed of three electron-dense layers. Each centriole is associated with typical striated roots. At the beginning of the spermiogenesis, an electron-dense material is observed in the apical region of the differentiation zone. During the initial stage of spermiogenesis, one of the centrioles gives rise to a free flagellum, which then rotates and undergoes proximodistal fusion with the cytoplasmic protrusion of the differentiation zone. The mature spermatozoon of B. orientalis corresponds to the Type III pattern described by Levron et al. (Biol Rev 85:523-543, 2010). It is characterized by the absence of mitochondrion and crested body. Five regions of the mature spermatozoon are differentiated. The main ultrastructural characteristics are: one axoneme of 9+ "1" trepaxonematan pattern, cortical microtubules and nucleus. The comparison of the spermiogenesis of B. orientalis with those of the other caryophyllidean species demonstrates some variation within the order relative to the presence and morphology of the intercentriolar body, the presence of slight rotation of the flagellar bud and a complete proximodistal fusion of the free flagellum with a cytoplasmic protrusion.

This paper reports results of the first cytogenetic study carried out on a recently described monozoic tapeworm, Khawia saurogobii Xi et al., 2009, from the Chinese lizard gudgeon (Saurogobio dabryi). The karyotype of this species is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16; n = 3m + 5t), metacentric chromosomes representing the first, sixth, and eight pairs. All chromosomes except the largest pair displayed 4',6-diamidino-2-phenylidole (DAPI) positive heterochromatin in centromeric regions. In mitotic preparations stained with Giemsa, one of the homologues of a smaller metacentric chromosome pair (No. 7) showed a distinct secondary constriction, whereas the other did not. Fluorescent in situ hybridization (FISH) with 18S ribosomal DNA (rDNA) probe revealed that the chromosomes No. 7 carry each a cluster of ribosomal genes associated with the centromeric heterochromatin and confirmed that this chromosome pair contains a nucleolar organizer region (NOR). The rDNA-FISH also confirmed heteromorphism in the size of NOR (i.e., secondary constriction) observed after Giemsa staining. The present cytogenetic analysis revealed species-specific characters of K. saurogobii and showed that FISH may represent a new valuable cytogenetic tool suitable for comparative taxonomic or phylogenetic studies within the order Caryophyllidea in the future.

Using transmission electron microscopy, spermiogenesis and the spermatozoon ultrastructure are described in Aspidogaster limacoides (Aspidogastrea, Aspidogastridae), from two cyprinid fishes, Abramis sapa and Blicca bjoerkna, from eastern Slovakia and north-western Russia. Results of the present investigation permit, for the first time in Aspidogastrea, to describe general pattern of spermiogenesis and the organization of the spermatozoon in this presumably the most basal group of parasitic flatworms (Neodermata). Spermiogenesis starts by the formation of a zone of differentiation characterized by long striated rootlets, two centrioles giving rise to flagella, and intercentriolar body composed of different electron-dense plates. Later, a flagellar rotation followed by a proximodistal fusion of the flagella with the cytoplasmic extension takes place. Beside the possession of elements typical of Neodermata, such as two axonemes of the 9 + '1' trepaxonematan pattern, nucleus, mitochondrion and cortical microtubules, the mature spermatozoon of A. limacoides exhibits characteristics unique in Aspidogastrea: (a) a long undulating membrane, (b) an electron-dense zone, and (c) a lamellate body. The terminology and structure of different typical elements of Aspidogastrea are discussed. It is interesting to note that the long undulating membrane and a continuous row of cortical microtubules have already been described in polyopisthocotylean monogeneans, which can be a case of convergence unless future phylogenetic studies confirm relatedness of these groups.

Ultrastructural data are provided on the tegument (or neodermis) of proglottids of the cestode Echinophallus wageneri Monticelli, 1890, the first representative of the family Echinophallidae (Pseudophyllidea), parasites of bathypelagic fish, studied using transmission and scanning electron microscopy. The surface of the ventral concave side of proglottids is covered with filiform microtriches about 1.7 microm long. Their glycocalyx, although very thick, is less extensively developed compared to that on digitiform microtriches localized on the dorsal side of the proglottids, which is also covered with filiform microtriches. The digitiform microtriches of the convex dorsal side are about 0.6 microm long, with a very short, rounded spine and the extraordinarily extensive glycocalyx seen as flocculent material. Short digitiform microtriches of E. wageneri seem to differ from microthrix types previously reported in other cestodes by transmission electron microscopy. The posterodorsal margin of all proglottids is typical in possessing a transverse band of large spiniform, tusk-shaped microtriches with a very long spine (approximately 13 microm in length), merged with filiform microtriches. Marked regional differences found in the morphology and distribution of microtriches on the proglottids of E. wageneri may be related to the different functions of the individual parts of the strobila because of the curling of the tapeworm body within the intestine of its fish host.

This paper provides the first ultrastructural data on the spermatozoon of a pseudophyllidean cestode belonging to the family Echinophallidae, Paraechinophallus japonicus, parasitizing deep-sea fish Psenopsis anomala from Japan. The mature spermatozoon contains two axonemes of the 9+"1" trepaxonematan pattern, nucleus, cortical microtubules, and electron-dense granules. Its anterior extremity exhibits a single electron-dense crested body. One of the most interesting features of the mature spermatozoon is the presence of a ring of tubular structures encircling the axoneme. This character, previously reported in species of the families Bothriocephalidae and Triaenophoridae, may be unique to pseudophyllideans of the group "Bothriocephalidea", thus supporting the assumption that the order Pseudophyllidea is formed in fact by two unrelated clades; "Bothriocephalidea" and "Diphyllobothriidea". The posterior extremity contains the posterior part of the nucleus and the disorganized axoneme--a character previously not described in the Eucestoda.

The fine structure of the mature spermatozoon of the corallobothriine tapeworm Corallobothrium solidum Fritsch, 1886 (Cestoda: Proteocephalidea) from the electric catfish Malapterurus electricus from the Nile River in Egypt was studied by transmission electron microscopy for the first time. The filiform spermatozoon of C. solidum contains two axonemes of unequal length and a typical 9 + "1" trepaxonematan pattern. A single helicoidal crested body (30-200 nm thick) is localized at the anterior extremity of the gamete. The cortical microtubules line the periphery of the cell, largely parallel to the long axis of the spermatozoon and exhibiting signs of twisting at the beginning of region II. The nucleus, in the form of an electron-dense (largely in gametes of testes) and/or fibrous cord (largely in gametes from male reproductive ducts and seminal vesicle), coils in a spiral through the middle part (region III) of the spermatozoon. The cytoplasm contains electron-dense granules in regions II, III and partly in region IV. The cytoplasm of some spermatozoa exhibits an apparently higher electron-density at the end of the nucleated region (III), and continuously toward the middle part of region IV. The anterior and posterior extremities of the spermatozoa have a single axoneme. The ultrastructural features of the mature spermatozoon of C. solidum mostly coincide with those of the spermatozoon of other proteocephalideans, especially the gangesiine Electrotaenia malopteruri parasitizing the same host.

The genetic structure of the most abundant species in the genus Eubothrium, E. crassum and E. salvelini, was studied by allozyme analysis in order to provide a consistent diagnosis for these morphologically similar species, as well as to assess their genetic relatedness. The proportion of fixed allelic differences (56.3%, 9 of 16 loci) between the species falls within the range accepted for valid species. A strict host specificity of E. crassum and E. salvelini for their typical host (lake trout, Arctic charr, respectively) was found in the French localities examined (Lake Annecy, Lake Bourget). For E. salvelini, polymorphism with the established allelic profile was found in the acid phosphatase (ACP) system. No significant deviation from the Hardy-Weinberg equilibrium expectations for ACP were found in either of the E. salvelini populations, which suggests predominantly cross-fertilization of these hermaphrodites. Analysis of the standardized variance in allelic frequencies demonstrated almost no among population differentiation (estimator theta of Fst = -0.03) between E. salvelini populations., (Copyright 2004 Springer-Verlag)

The fine structure of the mature spermatozoon of the tapeworm Electrotaenia malopteruri (Fritsch, 1886), a member of a supposedly primitive group of proteocephalidean tapeworms (Proteocephalidae: Gangesiinae), was studied by transmission electron microscopy for the first time. The mature spermatozoon of E. malopteruri is filiform, tapers at both extremities, and contains two axonemes of 9+"1" trepaxonematan type. A helicoidal crested body (60-150 nm thick) is present at the anterior extremity of the gamete. The twisting cortical microtubules (CM) line the periphery of the spermatozoon continuously. The nucleus, a fine cord of moderately condensed chromatin, occupies the middle part (region III) of the spermatozoon. The slightly electron-dense cytoplasm contains electron-dense granules in regions II and III and becomes more electron-dense at the end of region IV. The anterior and posterior extremities of the spermatozoon contain a single axoneme. Two modes of disorganisation of the axoneme at the posterior end of the mature spermatozoa are described for the first time in cestodes. The present data also indicate that the sperm ultrastructure of E. malapteruri shows some characters typical for onchobothriid tetraphyllideans, but it resembles that of Cyclophyllidea in the arrangement of twisting of the CM.

The reinvestigation of the fine structure of the mature spermatozoon of the proteocephalidean tapeworm Proteocephalus longicollis, a parasite of salmonid fish, has shown new characters not described previously for this species. They include a helicoidal crested body, 60-100 nm thick and localised at the anterior extremity of the spermatozoon. Also new is the finding that cortical microtubules in P. longicollis spermatozoon are of two types. The nucleus situated in the middle part of the gamete is rod-shaped anteriorly, but it is wound around one of the axonemes posteriorly. It is newly observed that both the anterior and posterior extremities of the spermatozoon of P. longicollis contain a single axoneme. The ultrastructural characteristics of the mature spermatozoon of P. longicollis coincide in their main features with those of the spermatozoon of P. torulosus, a parasite of cyprinid fish.

The diploid number 2n = 18 was revealed in Bathybothrium rectangulum, a specific parasite of barbels (Barbus spp.). All chromosome pairs were biarmed (pairs 1, 4, 7, 8, and 9 were metacentric; pairs 2, 3, and 6 were meta-submetacentric; and pair 5 was submetacentric) and small (from 1.49 to 2.95 microm). The C-banding method revealed pericentromeric heterochromatin in pairs 1, 3, 5, and 7; distinct C-bands in pairs 2, 4, 8, and 9; and a slight band in short arms of pairs 1 and 6. The karyotype of B. rectangulum is discussed with respect to the irresolute taxonomic position of B. rectangulum within the Pseudophyllidea.