Speciation and host-parasite relationships in the parasite genus Gyrodactylus (Monogenea, Platyhelminthes) infecting gobies of the genus Pomatoschistus (Gobiidae, Teleostei)

Using species-level phylogenies, the speciation mode of Gyrodactylus species infecting a single host genus was evaluated. Eighteen Gyrodactylus species were collected from gobies of the genus Pomatoschistus and sympatric fish species across the distribution range of the hosts. The V4 region of the ssrRNA and the internal transcribed spacers encompassing the 5.8S rRNA gene were sequenced; by including published sequences a total of 30 species representing all subgenera were used in the data analyses. The molecular phylogeny did not support the morphological groupings into subgenera as based on the excretory system, suggesting that the genus needs systematic revisions. Paraphyly of the total Gyrodactylus fauna of the gobies indicates that at least two independent colonisation events were involved, giving rise to two separate groups, belonging to the subgenus Mesonephrotus and Paranephrotus, respectively. The most recent association probably originated from a host switching event from Gyrodactylus arcuatus, which parasitises three-spined stickleback, onto Pomatoschistus gobies. These species are highly host-specific and form a monophyletic group, two possible ‘signatures’ of co-speciation. Host specificity was lower in the second group. The colonising capacity of these species is illustrated by a host jump from gobiids to another fish order (Anguilliformes), supporting the hypothesis of a European origin of Gyrodactylus anguillae and its intercontinental introduction by the eel trade. Thus, allopatric speciation seems to be the dominant mode of speciation in this host–parasite system, with a possible case of sympatric speciation.     

Initial steps of speciation by geographic isolation and host switch in salmonid pathogen Gyrodactylus salaris (Monogenea: Gyrodactylidae)

To test the hypothesis that host-switching can be an important step in the speciation of gyrodactylid monogenean flatworms, we inferred the phylogeny within a cluster of parasites morphologically close to Gyrodactylus salaris Malmberg 1957, collected from Atlantic, Baltic and White Sea salmon (Salmo salar), farmed rainbow trout (Oncorhynchus mykiss), and grayling (Thymallus thymallus) from Northern Europe. The internal transcribed spacer region of the nuclear ribosomal gene was sequenced for taxonomic identification. Parasites on grayling from the White Sea Basin differed from the others by one nucleotide (0.08%), the remainder were identical to the sequence published earlier from Norway (G. salaris on salmon), England (Gyrodactylus thymalli on grayling), and the Czech Republic (unidentified salaris/thymalli on trout). For increased resolution, 813 nucleotides of the mitochondrial COI gene of 88 parasites were sequenced and compared with 76 published sequences using phylogenetic analysis. For all tree building algorithms (NJ, MP), the parasites formed a star-like phylogeny of six definite sister clades, indicating nearly simultaneous radiation. Average K2P distances between clades were 1.8-2.6%, and internal mean distances 0.2-1.1%. The genetic distance to the nearest known relative, Gyrodactylus lavareti Malmberg, was 24%. A variable salmon-specific mitochondrial Clade I was observed both in the Baltic Basin and in pathogenic populations introduced to the Atlantic and White Sea coasts. An invariable Clade II was common in rainbow trout farms in Sweden, Denmark and Finland; the same haplotype was also infecting salmon in a landlocked population in Russian Karelia, and in Oslo fjord and Sognefjord in Norway. Four geographically vicariant sister clades were observed on graylings: Clade III in the Baltic Sea Basin; Clade IV in Karelian rivers draining to the White Sea; Clade V in Norwegian river draining to Swedish lake V?nern; and Clade VI in rivers draining to Oslo fjord. The pattern fitted perfectly with the postglacial history of grayling distribution. Widely sampled clades from salmon and Baltic grayling had basal haplotypes in populations, which were isolated early during the postglacial recolonisation. The divergence between the six clades was clear and linked with their hosts, but not wide enough to support a species status for them. Parasites from the Slovakian type population of G. thymalli were not available, so this result does not mean that G. salaris and G. thymalli are synonyms. It is suggested that the plesiomorphic host of the parasite cluster was grayling, and the switch to salmon occurred at least once when the continental ice isolated Baltic salmon in an eastern freshwater refugium, 130,000 years ago. At the same time, parasites on grayling were split geographically and isolated into several allopatric refugia. The divergence among the parasite clades allowed a tentative calibration of the evolutionary rate, leading to an estimate of the divergence of 13.7-20.3% per million years for COI coding mtDNA. The results supported the hypothesis that parallel to the allopatric mode, host switch and instant isolation by host specificity can be operated as a speciation mechanism.     

Mitochondrial DNA variation of Gyrodactylus spp (Monogenea, Gyrodactylidae) populations infecting Atlantic salmon, grayling, and rainbow trout in Norway and Sweden

Approximately 800 bp of the mitochondrial cytochrome oxidase I (COI) gene were sequenced from 76 Gyrodactylus specimens of 32 salmonid host populations, i.e. from Salmo salar, Thymallus thymallus, and Oncorhynchus mykiss in Norway, Sweden and Latvia. The COI sequences indicated a substantial intraspecific differentiation of Gyrodactylus salaris and Gyrodactylus thymalli. In total, 12 haplotypes were identified which group into five well supported clades, three clades with parasites from Atlantic salmon and two clades with parasites from grayling. The basal nodes linking the five clades together are only weakly supported. Thus, there is no support for the monophyly of all G. salaris haplotypes and the monophyly of all G. thymalli haplotypes. The lack of monophyly of the mitochondrial haplotypes of G. salaris and G. thymalli may indicate that G. salaris and G. thymalli represent (i). two polytypic species or (ii). one polytypic species, or (iii). refer to a complex of more than two sibling species. The mtDNA data indicate multiple introductions of G. salaris and G. thymalli into Norway. A minimum of three independent introductions of G. salaris and two independent introductions of G. thymalli are supported. This is congruent with earlier hypotheses on the introduction of G. salaris and G. thymalli into Norway.     

Identification of a host-associated species complex using molecular and morphometric analyses,with the description of Gyrodactylus rugiensoides n. sp. (Gyrodactylidae,Monogenea)

Gyrodactylus rugiensis was originally described as a parasite occurring on the marine gobies Pomatoschistus minutus and Pomatoschistus microps. In our preliminary survey this species was also frequently found on Pomatoschistus pictus and Pomatoschistus lozanoi. Subsequent molecular analysis of the internal transcribed spacers rDNA region revealed that this parasite actually represents a complex of two apparently cryptic species, one restricted to P. microps and the other shared by P. minutus, P. lozanoi and P. pictus. Morphometric analyses were conducted on 17 features of the opisthaptoral hard parts of specimens collected from all four host species. Standard discriminant analysis showed a clear separation of both genotypes by significant differences in marginal hook and ventral bar features. Statistical classifiers (linear discriminant analysis and nearest neighbours) resulted in an estimated misclassification rate of 4.7 and 3.1%, respectively. Based on molecular, morphological and statistical analyses a new species, Gyrodactylus rugiensoides is described. This species seems to display a lower host-specificity than generally observed for Gyrodactylus species as it infects three sympatric host species.However, seasonal and host-dependent morphometric variation is shown for G. rugiensoides collected on P. pictus. Host-switching and gene flow might be important factors preventing speciation on closely related and sympatric host species. The presence of host associated species complexes in this Gyrodactylus-Pomatoschistus system is also confirmed by the presence of two host-dependent genotypes within G. micropsi found on P. minutus and P. lozanoi, and P. microps, respectively. By comparing host and parasite phylogeny, phylogenetic and ecological factors influencing host-specificity are discussed.     

Gyrodactylus medaka n. sp. (Monogenea: Gyrodactylidae) parasitic on wild and laboratory-reared medaka Oryzias latipes (Beloniformes: Adrianichthyidae) in Japan

Gyrodactylus medaka n. sp. (Monogenea: Gyrodactylidae) is described from the skin, fins, and gills of medaka Oryzias latipes (Beloniformes: Adrianichthyidae) from Japan. This new species was collected from wild medaka in Hiroshima, Aichi, Saga, and Kumamoto prefectures, and laboratory-reared medaka in Chiba and Aichi prefectures. The small marginal hook sickle (≤4?μm) and the length of the marginal hook of the new species are the diagnostic morphological characters differentiated from other gyrodactylids reported from Asia. The pairwise sequence divergences for the interspecific variation in ITS regions and the phylogenetic analysis suggest that the populations of G. medaka n. sp. may have a similar genetic variation as the medaka populations in Japan. Gyrodactylus medaka n. sp. and Dactylogyrus oryziasi (Monogenea: Dactylogyridae) can maintain their populations in laboratory aquaria using medaka as their hosts, and these monogeneans and medaka have the potential as experimental model animals for clarifying various aspects of their host-parasite relationships. In addition, we report the composition of modified ammonium picrate-glycerin (APG) and show it is advantageous for monogenean taxonomy.     

Evolution and determinants of host specificity in the genus Lamellodiscus (Monogenea)

The evolution and determinants of host specificity in Lamellodiscus species (Monogenea, Diplectanidae) were investigated. The 20 known Mediterranean species were studied, all parasites of fishes from the family Sparidae (Teleostei). An index of specificity, which takes into account the phylogenetic relationships of their fish host species, was defined. The link between specificity and its potential determinants was investigated in a phylogenetic context using the method of independent contrasts. Host specificity in Lamellodiscus species appeared to be highly constrained by phylogeny, but also linked to host size. Mapping specificity onto the parasite phylogenetic tree suggests that specialist species do not represent an evolutionary dead end, and that specialization is not a derived condition. It is hypothesized that the ability to be generalist or specialist in Lamellodiscus is controlled by intrinsic, phylogenetically-related characteristics, and that specialist species tend to use large hosts, which may be more predictable.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77, 431−443.     

Origin and evolution of alternative developmental strategies in amphibious sarcopterygian parasites (Platyhelminthes, Monogenea, Polystomatidae)

Most integrative studies involving phylogenetic, developmental and ecological trends showed that the diversity of developmental modifications among the Platyhelminthes was linked to transmission opportunity pressures. For parasitic flatworms with complex life cycles it was suggested that the evolutionary forces that constrained or enhanced developmental strategies implied heterochronic patterns. Similar patterns were also reported from the Monogenea with direct life cycles, especially for Polystomatidae, which infest amphibious Sarcopterygians. Polystoma, whose members are recovered almost exclusively from anuran hosts of the Neobatrachia, is capable of following two alternative developmental strategies depending on the physiological stage of its host. Processes by which parasites reach maturity are strikingly different, and lead to discrete adult phenotypes within the same parasite species. In the present study, we investigate the origin and evolution of developmental patterns of polystomatids in a phylogenetic framework, using an integrative approach of heterochrony and evolutionary ecology. The results suggest that both phenotypes have coexisted during the early stages of polystome evolution, and that neither of them can be considered as the ancestral one. The two developmental pathways, each associated with one life cycle, may have arisen independently prior to polystome diversification, when strictly aquatic sarcopterygians attempted colonization of temporary freshwater environments. The occurrence of these two patterns within species of the genus Polystoma is suggested to reflect the ancestral condition, and to have allowed both developmental strategies to be successful depending on shifts in transmission opportunities. Thus, host evolutionary ecology may be the main factor in shaping developmental strategies within polystomatids.     

Escape from an evolutionary dead end: a triploid clone of Gyrodactylus salaris is able to revert to sex and switch host (Platyhelminthes, Monogenea, Gyrodactylidae)

Diploid parthenogenesis, with rare sex, is considered as the basic mode of reproduction among the hermaphroditic and viviparous Gyrodactylus. A particular strain of the monogenean parasite Gyrodactylus salaris (RBT clone) was recognized by an invariable, unique mitochondrial DNA haplotype in rainbow trout (Oncorhynchus mykiss) farms. The RBT clone was shown to be triploid and asexual by analyzing a 493 bp sequence of a nuclear DNA marker. Three alleles were present as heterozygous in all 237 individuals sampled in years 2001-2005 from five isolated Finnish farms. The triploid clone probably originated from a diploid oocyte fertilized by a non-self hermaphrodite, most probably in a fish farm. Identical mitochondrial COI gene (1606 bp) was also found in G. salaris parasites on landlocked salmon (Salmo salar) in two rivers draining to the lake Kuitozero, Russian Karelia. In the river Pisto, the clone was triploid, but the diagnostic "short" nuclear allele of the RBT clone was replaced by an allele typical for salmon specific parasites in the Lake Onega. The clone in the river Kurzhma was diploid, having lost the "short" allele, but still heterozygous for the other two alleles of the RBT clone. Evidently, the triploid parthenogenetic RBT clone had produced diploid oocytes, when (as a female) stimulated by a non-self mate in the new environment. The genetic reorganization coincided with a switch to the salmon host. Participation of triploids into the gene pool of the species is rarely reported in animals, and the triploidy is generally considered as an irreversible dead-end of the evolution. Liberalism in ploidy level may significantly add to the evolutionary options available for a parasite in ever-changing environments.     

Seasonal occurrence, topographical distribution and transmission of Gyrodactylus callanatis (Monogenea) infecting juvenile Atlantic cod in the Oslo Fjord, Norway

The seasonal occurrence and topographi0cal distribution of Gyrodactylus callanatis on three year-classes of juvenile Atlantic cod Gadus morhua , has been studied in the Oslo Fjord, Norway over a 2-year period. Both prevalence and abundance increased in spring and peaked in summer on the 1992 year-class and decreased in autumn. The highest intensities recorded in the study were observed on the 1992 year-class in June, with some fish harbouring more than 500 worms each. On the 1993 year-class, prevalence increased throughout the year, until it reached 100% in November; abundance, however, did not differ significantly between monthly samples, but intensities on individual fish were highest in June. The 0 + cod are probably infected when they immigrate to the nursery grounds in autumn and mingle with infected 1 + cod. About 74% of the worms infected the gill arches, oral cavity and pharynx, while 26% infected the body, head and fins. There was a significantly higher percentage of worms on the latter sites at intensities of less than 20 and more than 100 worms per fish. This is possibly related to the mode of transmission in G. callariatis . The initial infection sites are presumably the fins and head, and the most important route of transmission of G. callanatis is believed to be via the substrate. Other routes of transmission are also discussed.     

Water temperature,not fish morph,determines parasite infections of sympatric Icelandic threespine sticklebacks (Gasterosteus aculeatus)

Parasite communities of fishes are known to respond directly to the abiotic environment of the host, for example, to water quality and water temperature. Biotic factors are also important as they affect the exposure profile through heterogeneities in parasite distribution in the environment. Parasites in a particular environment may pose a strong selection on fish. For example, ecological differences in selection by parasites have been hypothesized to facilitate evolutionary differentiation of freshwater fish morphs specializing on different food types. However, as parasites may also respond directly to abiotic environment the parasite risk does not depend only on biotic features of the host environment. It is possible that different morphs experience specific selection gradients by parasites but it is not clear how consistent the selection is when abiotic factors change. We examined parasite pressure in sympatric morphs of threespine stickleback (Gasterosteus aculeatus) across a temperature gradient in two large Icelandic lakes, Myvatn and Thingvallavatn. Habitat‐specific temperature gradients in these lakes are opposite. Myvatn lava rock morph lives in a warm environment, while the mud morph lives in the cold. In Thingvallavatn, the lava rock morph lives in a cold environment and the mud morph in a warm habitat. We found more parasites in fish living in higher temperature in both lakes, independent of the fish morph, and this pattern was similar for the two dominating parasite taxa, trematodes and cestodes. However, at the same time, we also found higher parasite abundance in a third morph living in deep cold–water habitat in Thingvallavatn compared to the cold‐water lava morph, indicating strong effect of habitat‐specific biotic factors. Our results suggest complex interactions between water temperature and biotic factors in determining the parasite community structure, a pattern that may have implications for differentiation of stickleback morphs.     

Variations of opisthaptoral hard parts of Gyrodactylus salaris Malmberg, 1957 (Monogenea: Gyrodactylidae) on rainbow trout Oncorhynchus mykiss (Walbaum, 1792) in a fish farm,with comments on the spreading of the parasite in south-eastern Norway

Studies were carried out on Gyrodactylus salaris Malmberg, 1957 on rainbow trout Oncorhynchus mykiss (Walbaum, 1792) from a fish farm in Lake Tyrifjorden, south-eastern Norway. The anchors were larger, and the shape of the anchors and the ventral bar differed slightly, as compared with the same parts of G. salaris on parr of Atlantic salmon Salmo salar L. in northern and north-western Norwegian rivers. At different water temperatures (0.8°C, 10.0°C, 18.0°C), the opisthaptoral hard parts of G. salaris on rainbow trout showed considerable variation in size, but varied only slightly in shape. It was found that the total length of the anchors of G. salaris on rainbow trout may considerably exceed the previously reported maximum of 80 m for the species. The spread of G. salaris to south-eastern Norway is described and discussed.     

Potential key characters in Opisthobranchia (Gastropoda, Mollusca) enhancing adaptive radiation

Five potential key characters which might have enhanced species radiation in the Opisthobranchia (Gastropoda) are discussed. These are: 3–4 cuticular plates in the gizzard of Cephalaspidea s.str., kleptoplasty in Sacoglossa, kleptocnides in Aeolidoidea, a symbiotic relationship with unicellular algae in Phyllodesmium Ehrenberg, 1831, and mantle dermal formations in Chromodorididae. Interpretation of the characters as key innovations is based on phylogeny and/or comparison of species numbers in subgroups. Possible adaptive zones are discussed, and alternative interpretations indicated.     

The evolution of the Indian Ocean parrots (Psittaciformes): extinction, adaptive radiation and eustacy

Parrots are among the most recognisable and widely distributed of all bird groups occupying major parts of the tropics. The evolution of the genera that are found in and around the Indian Ocean region is particularly interesting as they show a high degree of heterogeneity in distribution and levels of speciation. Here we present a molecular phylogenetic analysis of Indian Ocean parrots, identifying the possible geological and geographical factors that influenced their evolution. We hypothesise that the Indian Ocean islands acted as stepping stones in the radiation of the Old-World parrots, and that sea-level changes may have been an important determinant of current distributions and differences in speciation. A multi-locus phylogeny showing the evolutionary relationships among genera highlights the interesting position of the monotypic Psittrichas, which shares a common ancestor with the geographically distant Coracopsis. An extensive species-level molecular phylogeny indicates a complex pattern of radiation including evidence for colonisation of Africa, Asia and the Indian Ocean islands from Australasia via multiple routes, and of island populations ‘seeding’ continents. Moreover, comparison of estimated divergence dates and sea-level changes points to the latter as a factor in parrot speciation. This is the first study to include the extinct parrot taxa, Mascarinus mascarinus and Psittacula wardi which, respectively, appear closely related to Coracopsis nigra and Psittacula eupatria.     

Molecular phylogeny and evolution of the genus Neoerysiphe (Erysiphaceae, Ascomycota)

The genus Neoerysiphe belongs to the tribe Golovinomyceteae of the Erysiphaceae together with the genera Arthrocladiella and Golovinomyces. This is a relatively small genus, comprising only six species, and having ca 300 species from six plant families as hosts. To investigate the molecular phylogeny and evolution of the genus, we determined the nucleotide sequences of the rDNA ITS regions and the divergent domains D1 and D2 of the 28S rDNA. The 30 ITS sequences from Neoerysiphe are divided into three monophyletic groups that are represented by their host families. Groups 1 and 3 consist of N. galeopsidis from Lamiaceae and N. galii from Rubiaceae, respectively, and the genetic diversity within each group is extremely low. Group 2 is represented by N. cumminsiana from Asteraceae. This group also includes Oidium baccharidis, O. maquii, and Oidium spp. from Galinsoga (Asteraceae) and Aloysia (Verbenaceae), and is further divided into four subgroups. N. galeopsidis is distributed worldwide, but is especially common in western Eurasia from Central Asia to Europe. N. galii is also common in western Eurasia. In contrast, the specimens of group 2 were all collected in the New World, except for one specimen that was collected in Japan; this may indicate a close relationship of group 2 with the New World. Molecular clock calibration demonstrated that Neoerysiphe split from other genera of the Erysiphaceae ca 35–45 M years ago (Mya), and that the three groups of Neoerysiphe diverged between 10 and 15 Mya, in the Miocene. Aloysia citriodora is a new host for the Erysiphaceae and the fungus on this plant is described as O. aloysiae sp. nov.     

Involvement of host recognition by oncomiracidia and post-larval survivability in the host specificity of Heterobothrium okamotoi (Monogenea: Diclidophoridae)

Heterobothrium okamotoi, a monogenean gill parasite, shows high host specificity for tiger puffer, Takifugu rubripes. In the present study, in vivo and in vitro experimental infections were conducted using various fish species, including T. rubripes, to understand the mechanisms of specificity. In in vivo experiments, T. rubripes, grass puffer, Takifugu niphobles, olive flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, were exposed to oncomiracidia of H. okamotoi labelled with a fluorescent dye, 5- (and -6) carboxyfluorescein diacetate succinimidyl ester, and the numbers of parasites on the gills and skin were recorded at intervals. Oncomiracidia were attached to gills and skin of all the experimental fish species immediately after exposure, and the infection intensity on T. rubripes was higher than that on T. niphobles and much higher than those on the other two species. After 2 days, the attached parasites remained on the gills of T. rubripes, but disappeared from the other hosts. During in vitro experiments, gill filaments excised from seven different fish species (four fish species used in the in vivo experiments and panther puffer, Takifugu pardalis, southern flounder, Paralichthys lethostigma and spotted halibut, Verasper variegates) were exposed to oncomiracidia and the attachment to each fish species and subsequent larval behaviour was observed. The percentage of post-larvae that attached to T. rubripes was slightly higher than those which attached to congeneric fish species and much higher than those of non-tetraodontid fish species. In vivo and in vitro experiments demonstrated that oncomiracidia of H. okamotoi have an affinity for their natural host, T. rubripes, and congeneric fish species. The disappearance of attached post-larvae from 'alien' hosts within 2 days during in vivo experiments suggested that host recognition by oncomiracidia and subsequent post-larval survivability are involved in the host specificity of H. okamotoi.     

Phylogeography and host association in a pollinating seed parasite Greya politella (Lepidoptera: Prodoxidae)

Inferring the historical context of ecological diversification is an important step in understanding the way that population-level processes result in a diversity of species and interactions in communities. We performed a phylogeographic analysis of mitochondrial DNA haplotypes from the pollinating seed parasite Greya politella (Lepidoptera: Prodoxidae) in order to determine the degree to which populations were structured according to geographical location and host-plant association. Ninety-eight individuals were sampled from 29 locations ranging from southern California to western Idaho. Restriction-site variation in 87 individuals (27 populations) was screened by digestion with 11 endonucleases, followed by Southern blotting; 38 restriction-site positions were mapped by double digests. Haplotypes were further defined by generating fragments 251 bases in length via PCR, screening them for sequence variation using denaturing gel gradient electrophoresis (DGGE), and sequencing the resulting variants. Parsimony analysis of the resulting 12 restriction-site and 15 sequence haplotypes indicated strong geographical structuring of populations: (i) most populations were monomorphic for haplotype; (ii) haplotypes from California and the Pacific Northwest (Oregon, Washington and Idaho) formed robust monophyletic groups. Population structure was significant both within and between the two regions, as reflected by N _ST. Patterns of host-plant association and haplotype phylogeny suggest that populations have recently undergone host-plant shifts in many different parts of the species range, although the direction and number of host shifts cannot be determined at the present level of sampling resolution.     

Diversity,biogeography and host specificity of kelp endophytes with a focus on the genera Laminarionema and Laminariocolax (Ectocarpales,Phaeophyceae)

Endophytic filamentous brown algae are known to invade stipes and fronds of kelps with potentially negative effects for the hosts. They have simple filamentous thalli and are difficult to identify based on morphology. We investigated the molecular diversity of 56 endophytes isolated from seven different kelp species from Europe, Chile, Korea and New Zealand by sequencing two unlinked molecular markers (5’COI and ITS1). A majority of 49 of the isolated endophytes (88%) belonged to the genera Laminarionema and Laminariocolax. The endophyte Laminarionema elsbetiae was isolated from Saccharina latissima and S. japonica tissues in Europe and Korea, respectively, and showed highly similar sequences in both regions. Three different species of the genus Laminariocolax were identified, the most common of which was L. aecidioides, an endophyte with a worldwide distribution and a broad host range. The other two species, L. tomentosoides and a species described here as Laminariocolax atlanticus sp. nov., were associated with different kelp species in the northern hemisphere and the North Atlantic, respectively. Our results suggest that specific host-endophyte patterns could exist locally, as found in kelps in Brittany where all endophytes isolated from S. latissima were L. elsbetiae, all endophytes isolated from Laminaria digitata were Laminariocolax tomentosoides, and those isolated from Laminaria hyperborea were Laminariocolax atlanticus and L. aecidioides. However, this pattern was not consistent with the results from other places, such as Western Scotland and Helgoland where the same kelp species are present.     

Molecular phylogeny of the Cyathocotylidae (Digenea,Diplostomoidea) necessitates systematic changes and reveals a history of host and environment switches

The Cyathocotylidae is a globally distributed family of digeneans parasitic as adults in fish, reptiles, birds and mammals in both freshwater and marine environments. Molecular phylogenetic analysis of interrelationships among cyathocotylids is lacking with only a few species included in previous studies. We used sequences of the nuclear 28S rRNA gene to examine phylogenetic affinities of 11 newly sequenced taxa of cyathocotylids and the closely related family Brauninidae collected from fish, reptiles, birds and dolphins from Australia, Southeast Asia, Europe, North America and South America. This is the first study to provide sequence data from adult cyathocotylids parasitic in fish and reptiles. Our analyses demonstrated that the members of the genus Braunina (family Brauninidae) belong to the Cyathocotylidae, placing the Brauninidae into synonymy with the Cyathocotylidae. In addition, our DNA sequences supported the presence of a second species in the currently monotypic Braunina. Our phylogeny revealed that Cyathocotyle spp. from crocodilians belong to a separate genus (Suchocyathocotyle, previously proposed as a subgenus) and subfamily (Suchocyathocotylinae subfam. n.). Morphological study of Gogatea serpentum indicum supported its elevation to species as Gogatea mehri. The phylogeny did not support Holostephanoides within the subfamily Cyathocotylinae; instead, Holostephanoides formed a strongly supported clade with members of the subfamily Szidatiinae (Gogatea and Neogogatea). Therefore, we transfer Holostephanoides into the Szidatiinae. DNA sequence data revealed the potential presence of cryptic species reported under the name Mesostephanus microbursa. Our phylogeny indicated at least two major host switching events in the evolutionary history of the subfamily Szidatiinae which likely resulted in the transition of these parasites from birds to fish and snakes. Likewise, the transition to dolphins by Braunina represents another major host switching event among the Cyathocotylidae. In addition, our phylogeny revealed more than a single transition between freshwater and marine environments demonstrated in our dataset by Braunina and some Mesostephanus.