Polyploidization in the fish family Cyprinidae, order Cypriniformes. I. DNA-content and chromosome sets in various species of Cyprinidae.
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Polyploidization in the fish family Cyprinidae, order Cypriniformes. II. Duplication of the gene loci coding for lactate dehydrogenase (E.C.: 1.1.1.27) and 6-phosphogluconate dehydrogenase (E.C.: 1.1.1.44) in various species of Cyprinidae.
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Animals↗
Molecular evidence for the monophyly of East Asian groups of Cyprinidae (Teleostei: Cypriniformes) derived from the nuclear recombination activating gene 2 sequences.
The family Cyprinidae is one of the largest families of fishes in the world and a well-known component of the East Asian freshwater fish fauna. However, the phylogenetic relationships among cyprinids are still poorly understood despite much effort paid on the cyprinid molecular phylogenetics. Original nucleotide sequence data of the nuclear recombination activating gene 2 were collected from 109 cyprinid species and four non-cyprinid cypriniform outgroup taxa and used to infer the cyprinid phylogenetic relationships and to estimate node divergence times. Phylogenetic reconstructions using maximum parsimony, maximum likelihood, and Bayesian analysis retrieved the same clades, only branching order within these clades varied slightly between trees. Although the morphological diversity is remarkable, the endemic cyprinid taxa in East Asia emerged as a monophyletic clade referred to as Xenocypridini. The monophyly for the subfamilies including Cyprininae and Leuciscinae, as well as the tribes including Labeonini, Gobionini, Acheilognathini, and Leuciscini, was also well resolved with high nodal support. Analysis of the RAG2 gene supported the following cyprinid molecular phylogeny: the Danioninae is the most basal subfamily within the family Cyprinidae and the Cyprininae is the sister group of the Leuciscinae. The divergence times were estimated for the nodes corresponding to the principal clades within the Cyprinidae. The family Cyprinidae appears to have originated in the mid-Eocene in Asia, with the cladogenic event of the key basal group Danioninae occurring in the early Oligocene (about 31-30 MYA), and the origins of the two subfamilies, Cyprininae and Leuciscinae, occurring in the mid-Oligocene (around 26 MYA).
Animals↗
DNA reassociation kinetics in diploid and phylogenetically tetraploid cyprinidae.
Four diploid and three phylogenetically tetraploid Cyprinidae (Ostariophysi) have been characterized as for nuclear DNA content, modal chromosome number and DNA reassociation kinetics (hydroxyapatite chromatography). Among the diploid species nuclear DNA content (10(-12) g DNA/2C) was 1.62 for Tinca tinca, 1.87 for Scardinius erythrophthalmus, 2.53 for Leuciscus cephalus and 2.75 for Alburnus alburnus, while the phylogenetically tetraploid species Carassius auratus, Barbus barbus and Cyprinus carpio attained 3.40, 3.66 and 3.80 respectively. Modal chromosome number was 2n = 48-50 for diploid individuals and 2n = 100-104 for phylogenetically tetraploid ones. In all the species 5--8% of the genome is represented by highly repetitive and foldback DNA. In DNA reassociation kinetics of phylogenetically tetraploid Cyprinidae a distinct plateau separates an intermediate reassociating sequence fraction (about 22% of the genome; with average repetition frequencies between 1,000 and 1,400) from a slow reassociating one (unique DNA; about 72% of the genome). These two genome fractions are not clearly distinguishable from each other in Cot curves of the diploid Cyprinidae, where a similar plateau is not evident. Since simple ploidy changes are not expected to affect DNA reassociation kinetics we suggest a different evolution in the genome organization of the two ploidy groups. Some possible hypotheses are discussed.
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The homologue of mannose-binding lectin in the carp family Cyprinidae is expressed at high level in spleen, and the deduced primary structure predicts affinity for galactose.
Mannose-binding lectin (MBL) participates in the innate immune system as an activator of the complement system and as an opsonin after binding to certain carbohydrate structures on microorganisms. We isolated and characterized cDNA transcripts encoding an MBL homologue from three members of the carp family Cyprinidae, the zebrafish Danio rerio, the goldfish Carassius auratus, and the carp Cyprinus carpio. The carp and zebrafish transcripts contain two polyadenylation sites and RT-PCR on mRNA from carp tissues revealed the carp transcript to be most prominently expressed in the spleen. The deduced mature proteins contain 228 or 233 amino acids with a short N-terminal segment containing a single conserved cysteine expected to form interchain disulfide bridges, a collagen domain interrupted by four amino acids between two glycine residues, a neck region predicted to form an alpha-helical coiled-coil structure, and a C-terminal carbohydrate recognition domain (CRD). Several of the structurally important residues in the CRD are conserved, but the residues known to interact with the calcium ion and hydroxyl groups of the carbohydrate ligand are different. The amino acid motif EPN, important for mannose specificity, was QPD in the Cyprinidae homologue, suggesting specificity for galactose instead. The identity between the deduced amino acid sequences is more than 90% between the carp and the goldfish and 68% and 65% between these two species, respectively, and the zebrafish. The identity with bird and mammalian MBLs ranges from 28 to 33%.
Amino Acid Sequence↗
[Molecular characterization of Diplozoidae populations on 5 species of Cyprinidae: new data on parasite specificity].
The genus Diplozoon (Platyhelminth) exhibits one of the most striking modes of reproduction. Adults reproduce after the permanent fusion of two larval hermaphrodites, which play a symmetrical role. The Diplozoidae are also exceptional among the Monogenea Polyopisthocotylea for two other reasons. They represent the only group really diversified on continental freshwater fishes; however, this diversification is difficult to evaluate since few morphoanatomical criteria are available to distinguish species and their host specificity is atypically variable among the Monogenea. For the first time in the Diplozoidae, the problems of species definition and of host specificity are examined using molecular tools. Two ribosomal markers (ITS2 and 28S rDNA (D1)) have been sequenced in five Diplozoidae, interacting with five Cyprinidae host species: the corresponding parasite-host systems have been well characterised, revealing some contrasting situations in the relations between Diplozoidae and Cyprinidae. Some species are effectively strictly host specific, but Diplozoon scardinii initially considered as a specific species on Scardinius erythrophtalmus and D. homoion on Rutilus rutilus are proposed to be a single species on the basis of their identical ITS2 and 28S rDNA sequences. On the same basis we proposed that D. paradoxum is able to parasitize two fish species, Abramis brama and Blicca bjoerkna, despite the morphological differences observed between the two xenopopulations. Phylogenetic relationships among Diplozoidae species were estimated with ITS2 sequences while cytochrome b sequences were used for their fish hosts. Finally, the comparison between these two molecular phylogenies seems to exhibit the phenomenon of cospeciation.
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Cloning and genetic variability of a HindIII repetitive DNA in Acrossocheilus paradoxus (Cyprinidae).
Thirty clones of a highly repetitive HindIII fragment of DNA from seven populations of Acrossocheilus paradoxus (Cyprinidae) were isolated and sequenced. The fragment represents a tandemly repeated sequence, with a monomeric unit of 270 bp, amounting to 0.08-0.10% of the fish genome. Higher units of this monomer appear as a ladder in Southern blots. The HindIII satellite DNA family is conserved in three genera of the Cyprinidae. Variation in nucleotide sequences of this repetitive fragment, which is A+T-rich, is distributed both within individuals and among populations. High overall nucleotide divergence (dij = 0.056 +/- 0.001) was detected among clones of the HindIII satellite DNAs of Acrossocheilus paradoxus. Based on the molecular clock hypothesis, the maximum evolutionary rate was estimated to be 5.3 x 10(-7) substitutions per site per year. Lineage sorting may have contributed to the genetic heterogeneity within individuals and populations. Cladistic analyses indicated a closer phylogeographic relationship between populations of the central and south regions in Taiwan.
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Susceptibility of several species of Cyprinidae and Salmonidae freshwater fish to larval Gnathostoma nipponicum infection.
Susceptibility of five species of Cyprinidae and Salmonidae freshwater fish to the early third-stage larvae (EaL3) and advanced third-stage larvae (AdL3) of Gnathostoma nipponicum infection were examined. Two fish species inoculated orally with EaL3 were infected, and AdL3 were recovered from them with rate of 21.0% in Tribolodon hakonensis and 0.5% in Cyprinus carpio at 30 days postinoculation (PI). Attempts to infect five fish species with AdL3 were all successful. The recovery rate of AdL3 was 69.0% in T. hakonensis, 47.5% in Carassius auratus subsp., 35.0% in C. carpio, 53.0% in Oncorhynchus masou, and 32.0% in O. mykiss at 10 days PI. These results confirmed that the Cyprinidae and Salmonidae fish species reported here were susceptible to larval G. nipponicum infection and AdL3 had higher infectivity to them than the EaL3.
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Molecular phylogenetic relationships of Eastern Asian Cyprinidae (pisces: cypriniformes) inferred from cytochrome b sequences.
Complete mitochondrial cytochrome b sequences of 54 species, including 18 newly sequenced, were analyzed to infer the phylogenetic relationships within the family Cyprinidae in East Asia. Phylogenetic trees were generated using various tree-building methods, including Neighbor-joining (NJ), Maximum Parsimony (MP) and Maximum Likelihood (ML) methods, with Myxocyprinus asiaticus (family Catostomidae) as the designated outgroup. The results from NJ and ML methods were mostly similar, supporting some existing subfamilies within Cyprinidae as monophyletic, such as Cultrinae, Xenocyprinae and Gobioninae (including Gobiobotinae). However, genera within the subfamily "Danioninae" did not form a monophyletic group. The subfamily Leuciscinae was divided into two unrelated groups: the "Leuciscinae" in East Asia forming as a monophyletic group together with Cultrinae and Xenocyprinae, while the Leucisciriae in Europe, Siberia, and North America as another monophyletic group. The monophyly of subfamily Cyprininae sensu Howes was supported by NJ and ML trees and is basal in the tree. The position of Acheilognathinae, a widely accepted monophyletic group represented by Rhodeus sericeus, was not resolved.
Amino Acid Sequence↗
Diversity of the parasite fauna of cyprinid (Cyprinidae) and percid (Percidae) fishes in the Vistula Lagoon, Poland.
A total of 2398 fish specimens (1091 Percidae and 1307 Cyprinidae) belonging to 16 species (3 Percidae and 13 Cyprinidae), caught in the Polish part of the Vistula Lagoon were examined within December 1994-March 1997. The parasites found were identified as belonging to 63 taxa (61 species as well as Diplostomum spp. flukes and glochidia Unionidae gen. sp. which could not be identified to species) of the Microsporea (1), Protozoa (1), Myxozoa (2), Monogenea (10), Digenea (15), Cestoda (11), Nematoda (11), Acanthocephala (5), Hirudinea (1), Mollusca (1), Copepoda (4), and Branchiura (1). The percids and cyprinids were found to support 37 and 40 parasitic taxa, respectively, the taxon-richest parasite fauna occurring in zander, Sander lucioperca (26 taxa), followed by carp bream, Abramis brama (24), European perch, Perca fluviatilis (24), roach, Rutilus rutilus (19), ruffe, Gymnocephalus cernuus (15), and Prussian carp, Carassius gibelio (11). The remaining fish species hosted less than 10 parasitic species each. Metacercariae of the genus Diplostomumn, found in about 37% of the fish examined, and Tylodelphys clavata, recorded in about 24% of the fish, proved the commonest parasites. The study showed the Vistula Lagoon cyprinid and percid parasite fauna to be dominated by freshwater species, frequencies of their occurrence in the brackishwater lagoon being lower than those in freshwater reservoirs. Frequencies of the 6 marine parasitic species found in the lagoon were, too, lower than those in the sea. It is suggested that some of the parasites (Ancyrocephalus paradoxus, Diplozoon paradoxum of the Monogenea, Diplostomum spp., Tylodelphys clavata of the Digenea, and Achtheres percarum of the Copepoda) prefer brackishwater habitats.
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Structure of the seminal pathway in the European chub, Leuciscus cephalus (Cyprinidae); Teleostei.
The testicular efferent duct system of Leuciscus cephalus (Cyprinidae), is described for three phases of testicular development. Testicular main ducts were analyzed by means of conventional histology and transmission electron microscopy. Additional techniques were applied for lectin histochemistry to determine secretory activity, as well as immunohistochemistry for cell proliferation activity and for muscle actin to demonstrate the distribution and amount of contractile cells. The contribution of the main ducts' epithelia and of degenerating spermatocytes to seminal fluid composition was confirmed, with the former being a source of carbohydrates and the latter that of phospholipids. The apical glycocalyx of epithelial cells, which is important in cell recognition and potentially involved in sperm storage, was marked by RCA I, LCA, and WGA lectin. Higher numbers of proliferating epithelial cells were ascertained during spawning phase compared to pre- and postspawning phases. In the ducts' stroma, a large number of cells expressed muscle actin and tropomyosin, indicating the ducts' contractile potential for the transport of seminal fluid towards release. Adjacent to these contractile cells, numerous nerves were found, indicating neuronal control of sperm fluid flow.
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Molecular phylogeny of Cyprinidae inferred from cytochrome b DNA sequences.
Phylogenetic relationships between European cyprinids (Teleostei:Cypriniformes:Cyprinidae) were investigated by comparing cytochrome b gene sequences from 29 species, among which 20 were newly sequenced. Results were in general agreement with previous morphologically-based studies, but new interesting relationships were found. The classical barbelled/lacking barbels split is dubious. Genus Leuciscus appears paraphyletic. The phylogenetic location of some American cyprinid species was recovered; at least two distinct invasions of the New World are likely. Finally, the problem of intergeneric cyprinid hybrids is addressed. The genus rank for these interbreeding entities is supported and hybrids are seen as the consequence of a high genetic flexibility. This is the first molecularly based study of cyprinid diversity. It sheds light on the evolution and taxonomy of this major freshwater fish family.
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Use of Central stonerollers (Cyprinidae: Campostoma anomalum) from Tennessee as a bioindicator of metal contamination.
We compared the levels of arsenic, cadmium, chromium, lead, manganese, mercury and selenium in a small species of fish (Central stonerollers, Campostoma anomalum) collected from East Fork Poplar Creek (EFPC) and a reference site in eastern Tennessee. Stonerollers are minnows in the Cyprinidae family that serve as prey for many carnivores in aquatic systems. Fish were collected from East Fork Poplar Creek within the U.S. Department of Energy's Y-12 National Security Complex, part of the Oak Ridge Reservation, and from a reference stretch of the Little River in East Tennessee. Whole fish were homogenized for analysis. Concentrations of all metals (except arsenic) were significantly higher in stonerollers from EFPC compared to the reference site. Mercury levels in minnows from EFPC averaged 0.4 ppm (microg/g), four times higher than the average for fish in the U.S. in general. This was higher than levels in fish from the nearby Clinch River and higher than fillets of white bass (Morone chrysops) from the same creek. Most metal levels were inversely related to size and weight of the stonerollers, perhaps due to growth dilution.
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Separation and partial characterization by high-performance liquid chromatography and radioimmunoassay of different forms of melanocyte-stimulating hormone from fish (Cyprinidae) neurointermediate lobes.
From neurointermediate lobe (NIL) extracts of two species of Cyprinidae, Carassius auratus and Cyprinus carpio, several peptides were separated by high-performance liquid chromatography (HPLC) on a C18 muBondapak column eluted with a methanol/acetic acid/triethylamine mixture. Monitoring all fractions by radioimmunoassay (RIA) with an antibody against melanocyte-stimulating hormone (MSH) C terminal gave positive reactions for fractions 7, 11-12, 15-16, 23-24, and 25-27. For further characterization, the elution positions of these peaks were compared to those of known synthetic reference substances. Peak 7 elutes in the same position as oxidized alpha MSH, whereas peak 15-16 matches the elution position of des-acetyl alpha MSH and 23-24 that of alpha MSH. The product from peak 26-27 has several characteristics of the diacetylated form of alpha MSH: its immunoreactivity in RIA, its sensitivity to weak bases and to HCl and its mass spectrum which is identical with that of mammalian diacetyl alpha MSH. In both species, the diacetylated form is predominant in the intracellular pool. This study establishes the coexistence of three different forms of alpha MSH, a des-acetylated, monoacetylated, and diacetylated in the cyprinid NIL extracts.
Adrenocorticotropic Hormone↗
Pollution correlated modifications of liver antioxidant systems and histopathology of fish (Cyprinidae) living in Seyhan Dam Lake, Turkey.
Specimen of freshwater fish (Cyprinidae) was collected from polluted and clean areas in dam Lake of Seyhan. Ten (n=10) fish were taken from each site. Liver preparations of fish from the two sources were comparatively assayed for glutathione-S-transferase (GST), lactic dehydrogenase (LDH), catalase (CAT), glucose-6-phosphate dehydrogenase (G6PDH), superoxide dismutase (SOD) and malondialdehyde (MDA). All systems were significantly (p<0.1) affected at the polluted site. The microscopic examination of the liver tissues of the fish living in the polluted area showed mononuclear inflammatory cell infiltration, cloudy degenerations, congestions and also micro- and macrovesicular fatty degenerations. These results provide evidence that both biochemical parameters and histopathological observations can be assumed as sensitive indicators of fish liver metabolism for polluted areas.
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The influence of translational selection on codon usage in fishes from the family Cyprinidae.
In this paper, the main factors shaping codon usage in three species of fishes that belong to the family Cyprinidae (namely Brachidanio rerio, Cyprinus carpio, and Carassius auratus) are reported. Correspondence analysis (COA), a commonly used multivariate statistical approach, was used to analyze codon usage bias. Our results show that the main trend is strongly correlated with the GC(3) content at silent sites of each sequence. On the other hand, the second axis discriminates between presumed highly and lowly expressed genes, a result that is confirmed by the distribution of matching expressed sequence tags (ESTs) along that axis. Translational selection appears, therefore, to influence synonymous codon usage in these fishes. The comparison of codon usages of the sequences displaying the extreme values on the second axis indicates that several codons are significantly incremented among the heavily expressed sequences. Interestingly, several of these triplets are not only shared by the three fishes but also by Xenopus laevis, another cold-blooded vertebrate in which translational selection influences codon choices. We postulate that natural selection was operative for codon usage in the last common ancestor of these fishes and Xenopus, and will probably be detected in cold-blooded vertebrates in general. Finally, we raise the possibility that the same phenomena will be found among warm-blooded vertebrates.
Amino Acids↗
Identifying main evolutionary mechanisms shaping genetic variation of Leuciscus cephalus L. 1758 (Cyprinidae) in western Greece: discordance between methods.
Numerous methods can be used in intraspecific phylogeographic studies to infer the evolutionary mechanisms that shaped observed genetic variation in populations. However, these methods are scarcely used jointly, and the evolutionary outcomes they could propose are not fully compared. In this study, using a chub (Leuciscus cephalus; Cyprinidae) mitochondrial DNA data set (13 populations in Western Greece, 14 haplotypes), we compare three distinct 'historical' methods that could possibly infer relative importance of basic evolutionary mechanisms (isolation vs migration) shaping genetic variation: the nested clade analysis, the psi-test and the 'mismatch distributions'. Taking together, interpretations of these analyses allow to draw a picture of the evolutionary history of chub in Western Greece based on isolation and genetic drift for higher clades. However, results issued each method can differ for low differentiated clades. We discuss such differences and suggest that methods should be used jointly in phylogeographic studies for a better evaluation of the evolutionary mechanisms that shaped genetic variation.
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Multiple origins of polyploidy in the phylogeny of southern African barbs (Cyprinidae) as inferred from mtDNA markers.
The cyprinid genus Barbus, with more than 800 nominal species, is an apparently polyphyletic assemblage to which a number of unrelated species, groups and/or assemblages have been assigned. It includes species that exhibit three different ploidy levels: diploid, tetraploid and hexaploid. Several lineages of the family Cyprinidae constitute a major component of the African freshwater ichthyofauna, having about 500 species, and fishes assigned to the genus 'Barbus' have the most species on the continent. We used complete sequences of the mitochondrial cytochrome b gene in order to infer phylogenetic relationships between diploid, tetraploid and hexaploid species of 'Barbus' occurring in southern Africa, the only region where representatives of all of the three ploidy levels occur. The results indicate that most of the lineages are incorrectly classified in the genus 'Barbus'. The southern African tetraploids probably originated from southern African diploids. They constitute a monophyletic group distinct from tetraploids occurring in the Euro-Mediterranean region (Barbus sensu stricto). The 'small' African diploid species seem to be paraphyletic, while the 'large' African hexaploid barbs species are of a single, recent origin and form a monophyletic group. The evidence of multiple, independent origins of polyploidy occurring in the African cyprinine cyprinids thus provides a significant contribution to the knowledge on the systematic diversity of these fishes, and warrants a thorough taxonomic reorganization of the genus.
Africa, Southern↗