Uncovering the monogenean species diversity of cyprinoid fish in Iraq using an integrative approach.

The freshwaters of Iraq harbour a high diversity of endemic and phylogenetically unique species. One of the most diversified fish groups in this region is cyprinoids, and although their distribution is relatively well known, their monogenean parasites have only rarely been investigated. Herein, we applied an integrative approach, combining morphology with molecular data, to assess the diversity and phylogeny of cyprinoid-associated monogenean parasites. A total of 33 monogenean species were collected and identified from 13 endemic cyprinoid species. The highest species diversity was recorded for Dactylogyrus (Dactylogyridae, 16 species) and Gyrodactylus (Gyrodactylidae, 12 species). Four species of Dactylogyrus and 12 species of Gyrodactylus were identified as new to science and described. Two other genera, Dogielius (Dactylogyridae) and Paradiplozoon (Diplozoidae), were represented only by 4 and 1 species, respectively. Phylogenetic analyses of the Dactylogyrus and Gyrodactylus species revealed that the local congeners do not form a monophyletic group and are phylogenetically closely related to species from other regions (i.e. Europe, North Africa and Eastern Asia). These findings support the assumption that the Middle East served as an important historical crossroads for the interchange of fauna between these 3 geographic regions.

Diversity, phylogeny and intraspecific variability of Paradiplozoon species (Monogenea: Diplozoidae) parasitizing endemic cyprinoids in the Middle East.

Diplozoidae are common monogenean ectoparasites of cyprinoid fish, with the genus Paradiplozoon being the most diversified. Despite recent studies on Diplozoidae from Europe, Africa and Asia, the diversity, distribution and phylogeny of this parasite group appears to be still underestimated in the Middle East. The objective of this study was to investigate the diversity, endemism and host specificity of diplozoids parasitizing cyprinoid fish from the Middle East, considering this region as an important historical interchange of fish fauna, and to elucidate the phylogenetic position of Middle Eastern Paradiplozoon species within Diplozoidae. Four Paradiplozoon species were collected from 48 out of 94 investigated cyprinoid species. Three known species, Paradiplozoon homoion, Paradiplozoon bliccae and Paradiplozoon bingolensis, were recorded on new cyprinoid host species, and a new species, Paradiplozoon koubkovae n. sp., was recorded on Luciobarbus capito and Capoeta capoeta from the Caspian Sea basin in Iran and Turkey. Paradiplozoon bliccae, exhibiting a wide host range in the Middle East, expressed both morphological and genetic intraspecific variabilities. The four Paradiplozoon species collected in the Middle East were placed in divergent clades, showing the rich evolutionary history of diplozoid parasites in the Middle East. Our study also revealed that two lineages of African diplozoids have a Middle Eastern origin. We stress the importance of applying an integrative approach combining morphological, ecological and molecular methods to reveal the real diversity of diplozoids.

Monogeneans and chubs: Ancient host-parasite system under the looking glass.

Host-parasite coevolution is one of the fundamentals of evolutionary biology. Due to the intertwined evolutionary history of two interacting species and reciprocal coadaptation processes of hosts and parasites, we can expect that studying parasites will shed more light onto the evolutionary processes of their hosts. Monogenea (ectoparasitic Platyhelminthes) and their cyprinoid fish hosts represent one of the best models for studying host-parasite evolutionary relationships using a cophylogenetic approach. These parasites have developed remarkably high host specificity, where each host species often serves as a potential host for its own host-specific monogenean species. Here, the cophylogenetic relationships in the Dactylogyrus-Squalius system was investigated, as Squalius is one of several cyprinoid genera with puzzling phylogeography and inhabits all four major peri-Mediterranean peninsulas. Of 29 endemic Squalius species examined for the presence of Dactylogyrus parasites, a total of 13 Dactylogyrus species were collected from the gills of 20 Squalius species across a wide range of distribution. Phylogenetic reconstruction revealed a polyphyletic origin for Dactylogyrus species parasitizing congeneric Squalius, with four major clades being recognized. On the basis of the delimitation of host specificity, strict specialists parasitizing single host species, geographic specialists parasitizing congeners in a limited geographical region, and true generalists parasitizing congeners in various geographical regions were recognized in Dactylogyrus species parasitizing Squalius. The phylogenetic reconstruction of Squalius hosts revealed two major clades, the first encompassing only peri-Mediterranean species and the second including species from other Euro-Asian regions. Distance-based cophylogenetic methods did not reveal a statistically significant global cophylogenetic structure in the studied system; however, several host-parasite links among Iberian endemic species contributed significantly to the overall structure. The widest host range and associated genetic variability were recorded for D. folkmanovae, parasitizing nine Squalius species, and D. vistulae, parasitizing 13 Squalius species. Two different dispersion mechanisms and morphological adaptations to Squalius hosts were clearly reflected in the contrasting cophylogenetic patterns for these two species with different levels of host specificity. While host-parasite cospeciation plays an important role in diversification within D. folkmanovae, diversification within D. vistulae is driven mainly by host switching.

De novo developed microsatellite markers in gill parasites of the genus Dactylogyrus (Monogenea): Revealing the phylogeographic pattern of population structure in the generalist parasite Dactylogyrus vistulae.

Approaches using microsatellite markers are considered the gold standard for modern population genetic studies. However, although they have found application in research into various platyhelminth taxa, they remained substantially underutilized in the study of monogeneans. In the present study, a newly developed set of 24 microsatellite markers was used to investigate the genetic diversity of the generalist monogenean species Dactylogyrus vistulae. The analyzed parasite specimens were collected from 13 cyprinoid species from 11 sites in the Apennine and Balkan peninsulas. A total of 159 specimens were genotyped at each of the loci and the number of alleles per locus ranged from 2 to 16, with a mean number of 6.958 alleles per locus. Exceptionally high genetic diversity was observed among D. vistulae individuals in the southern Balkans (mean N A per locus = 3.917), suggesting that generalist D. vistulae expanded from the south to the north in the Balkans and later into central Europe. The initial clustering analysis divided all investigated specimens into three major clusters; however, the results of the subsequent analyses revealed the existence of various subpopulations, suggesting that the population structure of D. vistulae is associated with the diversification of their cyprinoid hosts. In addition, the partition of the parasite population was observed in regions of the sympatric occurrence of two host species, indicating that these hosts may represent a barrier for gene flow, even for generalist parasite species.

Molecular and morphological phylogeny of host-specific Dactylogyrus parasites (Monogenea) sheds new light on the puzzling Middle Eastern origin of European and African lineages.

BACKGROUND: Freshwater fauna of the Middle East encompass elements shared with three continents-Africa, Asia, and Europe-and the Middle East is, therefore, considered a historical geographic crossroad between these three regions. Even though various dispersion scenarios have been proposed to explain the current distribution of cyprinids in the peri-Mediterranean, all of them congruently suggest an Asian origin for this group. Herein, we investigated the proposed scenarios using monogenean parasites of the genus Dactylogyrus, which is host-specific to cyprinoid fishes. METHODS: A total of 48 Dactylogyrus species parasitizing cyprinids belonging to seven genera were used for molecular phylogenetic reconstruction. Taxonomically important morphological features, i.e., sclerotized elements of the attachment organ, were further evaluated to resolve ambiguous relationships between individual phylogenetic lineages. For 37 species, sequences of partial genes coding 18S and 28S rRNA, and the ITS1 region were retrieved from GenBank. Ten Dactylogyrus species collected from Middle Eastern cyprinoids and D. falciformis were de novo sequenced for the aforementioned molecular markers. RESULTS: The phylogenetic reconstruction divided all investigated Dactylogyrus species into four phylogenetic clades. The first one encompassed species with the "varicorhini" type of haptoral ventral bar with a putative origin linked to the historical dispersion of cyprinids via the North African coastline. The second clade included the majority of the investigated species parasitizing various phylogenetically divergent cyprinid hosts. The morphological and molecular data suggest the ancestral diversification of the species of this clade into two groups: (1) the group possessing the haptoral ventral bar of the "cornu" type, and (2) the group possessing the "wunderi" type. Dactylogyrus diversification apparently occurred in the Middle East, which is indicated by the presence of species with all morphotypes in the region. The last two clades included species parasitizing cyprinids with an East Asian origin, and species possessing the "magnihamatus" type of ventral bar. CONCLUSIONS: The molecular data suggest that some morphological characters of host-specific parasites may undergo convergent evolution in the divergent lineages, and therefore, to fully resolve the phylogenetic relationships among host-specific parasites, an integrative approach combining morphological and molecular data is still needed. In addition, our study indicates that parasite diversity in many regions is still under-explored, and thus we highlight the importance of studies of host-associated parasites, especially in the context of freshwater fish biogeography.

Helminth communities of endemic cyprinoids of the Apennine Peninsula, with remarks on ectoparasitic monogeneans, and a description of four new Dactylogyrus Diesing, 1850 species.

The fauna of the Apennine Peninsula is, in comparison to other southern European peninsulas, relatively species-poor regarding the number of endemic cyprinoid species. Nonetheless, the recent introduction of non-native species has significantly increased the total number of freshwater species in this region. Such invasive species may represent a threat to the native fauna, associated among other things with the introduction of non-native parasites with their original hosts.In the present study, we investigated endemic cyprinoid species for the presence of helminth parasites. A total of 36 ectoparasitic monogenean species and five endoparasitic helminth species were collected from ten cyprinoid species in five localities in northern Italy. Out of 20 Dactylogyrus species (gill monogeneans specific to cyprinoids), four were identified as new to science and herein described: Dactylogyrus opertus n. sp. and Dactylogyrus sagittarius n. sp. from Telestes muticellus, Dactylogyrus conchatus n. sp. from T. muticellus and Protochondrostoma genei, and Dactylogyrus globulatus n. sp. from Chondrostoma soetta. All new Dactylogyrus species appear to be endemic to the Apennine Peninsula; however, they share a common evolutionary history with the endemic Dactylogyrus parasitizing cyprinoids of the Balkans. This common origin of cyprinoid-specific parasites supports a historical connection between these two (currently separated) geographical regions.

Diversity and phylogeny of Paradiplozoon species (Monogenea: Diplozoidae) parasitising endemic cyprinoids in the peri-Mediterranean area, with a description of three new Paradiplozoon species.

The Diplozoidae are monogenean parasites of mainly cyprinoid fishes with a unique life cycle, whereby two larvae undergo anastomosis and fuse into a single cross-shaped specimen. Paradiplozoon is the most species rich and widespread genus of the family, with a distribution range covering Eurasia and Africa; however, some areas remain underexplored and their diplozoid fauna is uncertain. In the present study, the Paradiplozoon diversity was investigated in the peri-Mediterranean region, which has the highest levels of cyprinoid diversity and endemism in Europe. A total of 36 endemic cyprinoid species were sampled from sites in north-west Africa and the southern European peninsulas and investigated for the presence of diplozoid parasites. Of five Paradiplozoon species collected, three were identified as new to science: Paradiplozoon moroccoensis n. sp. from the Moroccan endemic Luciobarbus lepineyi; Paradiplozoon ibericus n. sp. from Iberian endemic cyprinids and leuciscids and Paradiplozoon helleni n. sp. from the Greek endemic Scardinius acarnicus and Tropidophoxinellus hellenicus (descriptions provided herein). In addition, new host records for P. homoion and P. megan are presented, with the former being most prevalent in the investigated region. Phylogenetic analysis supported paraphyly of the genus Paradiplozoon, and suggests the need for a careful taxonomic re-evaluation of this genus. Furthermore, the results showed that endemic Paradiplozoon of the peri-Mediterranean do not form a monophyletic group, suggesting multiple origins of this parasite groups in different peri-Mediterranean regions.

A new species of cyprinoid fish from the Tana River, Kenya (Actinopterygii: Danionidae).

Sampling of streams in the middle reaches of the Tana River Basin in Meru National Park, Kenya, from 2010 to 2012 for an NSF-funded International Research Experiences for Students (IRES) project, resulted in the capture of a number of specimens of what were first thought to be Neobola fluviatilis.  On closer examination the specimens were determined to represent a distinct species, endemic to the Tana River basin, which is herein formally described. The new species is readily diagnosed from N. fluviatilis by higher counts of lateral line, pre-dorsal, and caudal peduncle circumferential scales, higher numbers of pectoral rays, lower numbers of anal fin rays, and a shorter anal-fin base length.

Phylogenetic classification of extant genera of fishes of the order Cypriniformes (Teleostei: Ostariophysi).

The order Cypriniformes is the most diverse order of freshwater fishes. Recent phylogenetic studies have approached a consensus on the phylogenetic relationships of Cypriniformes and proposed a new phylogenetic classification of family-level groupings in Cypriniformes. The lack of a reference for the placement of genera amongst families has hampered the adoption of this phylogenetic classification more widely. We herein provide an updated compilation of the membership of genera to suprageneric taxa based on the latest phylogenetic classifications. We propose a new taxon: subfamily Esominae within Danionidae, for the genus Esomus.

Complete mitochondrial genome of the Pseudogastromyzon jiulongjiangensis Chen (Cypriniformes, Homalopteridae) and phylogenetic analysis of the Cyprinoidei.

The Pseudogastromyzon jiulongjiangensis Chen (Cypriniformes, Homalopteridae) is a promising ornamental and commercial candidate in China. In this study, the complete mitochondrial genome of P. jiulongjiangensis was first determined. It is 16,571 bp length and consists of 22 tRNA genes, 13 protein-coding genes, two rRNA genes, and a control region. Except for eight tRNA and ND6 genes, all other mitochondrial genes are encoded on the heavy strand. Phylogenetic analysis revealed that P. jiulongjiangensis, Formosania lacustris, and other seven fish first clustered into the Homalopteridae clade. Then, the Homalopteridae and Cobitidae formed the sister group. The Catostomoidae and Cyprinidae constituted the sister branch, which is inconsistent with the previous phenotypic report. It is suggested that the taxonomic research might lose some significant evolutionary characters. This study will contribute to phylogenetic analysis of the Homalopteridae and the natural resources conservation of P. jiulongjiangensis.