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PubMed2001

Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH).

When surveying the karyotype diversity of European loaches of the genus Cobitis to identify species involved in hybrid polyploid complexes, an extensive polymorphism in number and location of NORs was discovered in C. vardarensis using Ag-staining, C-banding, CMA3-fluorescence and fluorescence in situ hybridization (FISH). This species had 2n = 50, the karyotype contained 13 pairs of metacentric, 10 pairs of submetacentric and two pairs of subtelocentric chromosomes. The NOR-bearing chromosomes included one medium-sized metacentric pair with a large CMA3-positive heterochromatic pericentromeric block, one small metacentric as well as one large submetacentric pairs. Ribosomal sites were always located in telomeres of these chromosomes. Each of the pair of NOR-bearing chromosomes occurred in three variants - (1) presence and/or (2) absence of NORs on both homologues and (3) heterozygous combination where only one of the homologues bears NORs. Altogether, 10 different NOR cytotypes from 27 theoretically possible ones were discovered among 20 indviduals examined. The number of NORs ranged from two to five per specimen. The results regarding the number and locations of NORs as revealed by banding techniques were confirmed using FISH with rDNA probe. NOR sites were of CMA3-positive, suggesting that ribosomal sites are associated with GC-rich DNA. Very similar structural polymorphism with multiple NORs is expressed in the Danubian loach C. elongatoides indicating a close relationship between both species.

Animals↗
PubMed2001

Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis.

Transgenic mud loaches (Misgurnus mizolepis), in which the entire transgene originated from the same species, have been generated by microinjecting the mud loach growth hormone (mlGH) gene fused to the mud loach beta-actin promoter. Out of 4,100 eggs injected, 7.5% fish derived from the injected eggs showed dramatically accelerated growth, with a maximum of 35-fold faster growth than their non-transgenic siblings. Many fast-growing transgenic individuals showed extraordinary gigantism: their body weight and total length (largest fish attained to 413 g and 41.5 cm) were larger and longer than even those of 12-year-old normal broodstock (maximum size reached to 89 g and 28 cm). Of 46 transgenic founders tested, 30 individuals transmitted the transgene to next generation with a wide range of germ-line transmission frequencies ranging from 2% to 33%. The growth performance of the subsequent generation (F1) was also dramatically accelerated up to 35-fold, although the levels of enhanced growth were variable among transgenic lines. Three transgenic germ-lines up to F4 were established, showing the expected Mendelian inheritance of the transgene. Expression of GH mRNA in many tissues was detected by RT-PCR analyses. The time required to attain marketable size (10 g) in these transgenic lines was only 30-50 days after fertilization, while at least 6 months in non-transgenic fish. Besides growth enhancement, significantly improved feed-conversion efficiency up to 1.9-fold was also observed.

Actins↗
PubMed2003

Biomagnification factors (fish to Osprey eggs from Willamette River, Oregon, U.S.A.) for PCDDs, PCDFs, PCBs and OC pesticides.

A migratory population of 78 pairs of Osprey (Pandion haliaetus) nesting along the Willamette River in western Oregon was studied in 1993. The study was designed to determine contaminant concentrations in eggs, contaminant concentrations in fish species predominant in the Ospreys diet, and Biomagnification Factors (BMFs) of contaminants from fish species eaten to Osprey eggs. Ten Osprey eggs and 25 composite samples of fish (3 species) were used to evaluate organochlorine (OC) pesticides, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). Mercury was also analyzed in fish. Geometric mean residues in Osprey eggs were judged low, e.g., DDE 2.3 microg g(-1) wet weight (ww), sigma PCBs 0.69 microg g(-1), 2,3,7,8-TCDD 2.3 ng kg(-1), and generally well below known threshold values for adverse effects on productivity, and the population was increasing. Osprey egg residue data presented by River Mile (RM) are discussed, e.g., higher PCDDs were generally found immediately downstream of paper mills and eggs from the Willamette River had significantly elevated PCBs and PCDDs compared to reference eggs collected nearby in the Cascade Mountains. Prey remains at nest sites indicated that the Largescale Sucker (Catostomus macrocheilus) and Northern Pikeminnow (Ptychocheilus oregonensis) accounted for an estimated 90.1% of the biomass in the Osprey diet, and composite samples of these two species were collected from different sampling sites throughout the study area for contaminant analyses. With the large percentage of the fish biomass in the Osprey diet sampled for contaminants (and fish eaten by Ospreys similar in size to those chemically analyzed), and fish contaminant concentrations weighted by biomass intake, a mean BMF was estimated from fish to Osprey eggs for the large series of contaminants. BMFs ranged from no biomagnification (0.42) for 2,3,7,8-TCDF to 174 for OCDD. Our findings for the migratory Osprey were compared to BMFs for the resident Herring Gull (Larus argentatus), and differences are discussed. We believe a BMF approach provides some basic understanding of relationships between contaminant burdens in prey species of fish-eating birds and contaminants incorporated into their eggs, and may prove useful in understanding sources of contaminants in migratory species although additional studies are needed.

Animals↗
PubMed2003

Replication banding patterns in the spined loach, Cobitis taenia L. (Pisces, Cobitidae).

The chromosomal complement of Cobitis taenia was analysed by replication banding techniques to determine whether there were specific patterns that could allow distinction of the different chromosomes. The diploid chromosome number of 2n = 48 is diagnostic of this species. In vivo 5-bromodeoxyuridine (5-BrdU) incorporation induced highly reproducible replication bands. Most of the chromosome pairs were distinguishable on the base of their banding patterns. The karyotype, consisting of five pairs of metacentrics, nine pairs of submetacentrics and 10 pairs of subtelocentrics and acrocentrics, was confirmed. C-banding and replication banding patterns were compared, and heterochromatin was both early and later replicating. C-positive heterochromatin in centromeric regions was mainly early replicating, but that located in pericentromeric regions was late replicating. Most of the late-replicating regions found interstitially were C-band negative. The results obtained so far for combined chromosomal staining methods of C. taenia and other Cobitis fish species are discussed.

Animals↗
PubMed2000

Isogenic transgenic homozygous fish induced by artificial parthenogenesis.

As a model system for vertebrate transgenesis, fish have many attractive advantages, especially with respect to the characteristics of eggs, allowing us to produce isogenic, transgenic, homozygous vertebrates by combining with chromosome-set manipulation. Here, we describe the large-scale production of isogenic transgenic homozygous animals using our experimental organism, the mud loach Misgurnus mizolepis, by the simple process of artificial parthenogenesis in a single generation. These isogenic fish have retained transgenic homozygous status in a stable manner during the subsequent 5 years, and exhibited increased levels of transgene expression. Furthermore, their isogenic nature was confirmed by cloned transgenic homozygous offspring produced via another step of parthenogenic reproduction of the isogenic homozygous transgenic fish. These results demonstrate that a combination of transgenesis and artificial parthenogenesis will make the rapid utilization of genetically pure homozygous transgenic system in vertebrate transgenesis possible.

Animals↗
PubMed2003

Resurrection of Proteocephalus sagittus (Grimm, 1872) (Cestoda: Proteocephalidea) based on morphological and molecular data.

This paper presents a redescription of the proteocephalidean cestode Proteocephalus sagittus (Grimm, 1872) based on freshly collected specimens from the type-host, the stone loach Barbatula barbatula (L.) (family Balitoridae), from the Czech Republic. This species has recently been synonymised with P. torulosus (Batsch, 1786), a parasite of cyprinid fishes in the Holarctic Region, but it differs in the following characters: (i) scolex morphology (scolex rounded, with more apically situated suckers and lacking longitudinal wrinkles in P. sagittus versus large and club-shaped, with more laterally situated suckers and longitudinal wrinkles in P. torulosus); (ii) position of the vagina (opens anterior to the cirrus-sac in P. sagittus versus overlapping its distal region dorsally in P. torulosus); (iii) length of the cirrus-sac (shorter in P. sagittus); (iv) osmoregulatory canals (more distinct and situated more laterally, but almost indistinguishable and more medial in P. torulosus); and (v) sequences of the partial 18S rRNA (V4 region - 96.9% similarity) and 5.8S-ITS2 genes (65.9% similarity). P. pamirensis Dzhalilov & Ashurova, 1971, a poorly described species from the Tibetan stone loach Noemacheilus stoliczkai[= Tryplophysa stoliczkae (Steindachner)] in Tadjikistan, is synonymised with P. sagittus.

Animals↗
PubMed2003

Clestobothrium neglectum (Lönnberg, 1893) n. comb. (Cestoda: Bothriocephalidae) from the tadpole fish Raniceps raninus (L.) (Gadidae) from Sweden.

Clestobothrium neglectum (Lönnberg, 1893) n. comb. is redescribed from specimens previously deposited in The Natural History Museum, London, UK by Dr David I. Gibson obtained from the intestine of the tadpole fish Raniceps raninus (Gadidae) collected from off the western coast of Sweden, near Kristineberg. C. neglectum resembles C. crassiceps (Rudolphi, 1819), the type- and only species currently recognised in the genus, but differs in having fine spiniform-like structures that are likely microtriches covering the posterior fourth of the scolex and all proglottids, a highly folded tegument forming numerous longitudinal ridges on both the dorsal and ventral surfaces of the proglottids giving them a scalloped appearance, a somewhat smaller egg (68 x 35 vs 75 x 40 microm), a U-shaped rather than H-shaped ovary and more testes (70-85 vs 40-50 per proglottid). C. neglectum is also compared to two species with similar scoleces that were previously assigned to Clestobothrium, Bothriocephalus acheilognathi Yamaguti, 1934 and B. kivuensis Baer & Fain, 1958 (considered to be synonymous with B. acheilognathi by Pool, 1987). This is the first report of a species of Clestobothrium Lühe, 1899 (Bothriocephalidae) from a gadid fish.

Animals↗
PubMed2026

High-Resolution Chromosome-Level Genome Assembly and Annotation of Triplophysa stewarti, an Endemic Plateau Loach from the Qinghai-Tibet Plateau.

The bottom-dwelling fish Triplophysa stewarti, endemic to the Qinghai-Tibet Plateau, is a valuable model for studying high-altitude adaptation in aquatic ecosystems. However, the lack of a high-quality reference genome has hindered comparative genomic and evolutionary studies within this genus. Here, we present a chromosome-level genome assembly for T. stewarti, generated using PacBio HiFi long-read sequencing and Hi-C scaffolding. The 697.9 Mb assembly is highly continuous (scaffold N50 of 253.58 Mb) and encompasses 25 chromosomes, representing 92.65% of the genome. BUSCO analysis indicated a 98.4% completeness, supporting the high quality of the assembly. We annotated 28,009 protein-coding genes, with 97.04% being functionally assigned across multiple databases (NR, UniProt, KEGG, GO, Pfam and InterPro). Additionally, repetitive elements constituted 42.47% of the genome, and we identified 52,709 non-coding RNAs. This high-quality reference genome provides a fundamental resource for exploring the adaptive evolution, population structure, and conservation genetics of T. stewarti and related species on the Qinghai-Tibet Plateau.

Animals↗
PubMed2001

River basin-related genetic structuring in an endangered fish species, Chondrostoma lusitanicum, based on mtDNA sequencing and RFLP analysis.

Chondrostoma lusitanicum is a Portuguese endemic cyprinid with a restricted distribution and reduced numbers in some basins, justifying its status as a threatened species. We examined genetic population structure using samples from throughout its geographical range in Portugal, using sequencing of b cytochrome and restriction fragment length polymorphism analysis of the NADH subunits 5 and 6. There was reduced within-population genetic variability but considerable among-population differentiation, particularly marked between both the Mira and Arade basins in the extreme south and other populations. These results confirm phylogeographic relationships suggested by previous fragmentary allozyme studies for C. lusitanicum, and are in accordance with allozyme and mitochondrial DNA data on phylogeography of coexisting cyprinid species of the genus Leuciscus. The levels of genetic divergence revealed by sequence and RFLP data showed strongly concordant patterns: geographical genetic structuring, with the definition of three distinct groups, was observed. The high values of nucleotide divergence and pairwise sequence divergence of the Mira and Arade groups, when compared with all other samples, support a distinct taxonomic status probably at the species level. Results are also discussed in relation to conservation of this highly fragmented species, in terms of Evolutionary Significant Units and Management Units.

Analysis of Variance↗
PubMed2004

Raphidascaris acus (Bloch, 1779) larvae infections of the stone loach, Barbatula barbatula (L.), from the River Haná, Czech Republic.

The stone loach, Barbatula barbatula (L.), is a typical and dominant intermediate host of Raphidascaris acus in lowland streams of Central Europe. The prevalence of infection of R. acus in B. barbatula from the River Haná ranged throughout the year from 73.3 to 100%. The abundance and the mean intensity of infection also varied throughout the year with a peak in September. Larvae were located mainly in the liver parenchyma. High numbers of larvae and their migration through the tissue caused cyst- or abscess-like formations in the host parenchyma. The severity of the disease condition ranged from mild to severe. We speculate that the infection of stone loach by R. acus larvae regulates the population density and abundance of the intermediate host in lowland streams where natural predators are absent.

Animals↗
PubMed2003

Genetic variation among interconnected populations of Catostomus occidentalis: implications for distinguishing impacts of contaminants from biogeographical structuring.

Exposure to contaminants can affect survivorship, recruitment, reproductive success, mutation rates and migration, and may play a significant role in the partitioning of genetic variation among exposed and nonexposed populations. However, the application of molecular population genetic data to evaluate such influences has been uncommon and often flawed. We tested whether patterns of genetic variation among native fish populations (Sacramento sucker, Catostomus occidentalis) in the Central Valley of California were consistent with long-term pesticide exposure history, or primarily with expectations based on biogeography. Field sampling was designed to rigorously test for both geographical and contamination influences. Fine-scale structure of these interconnected populations was detected with both amplified fragment length polymorphisms (AFLP) and microsatellite markers, and patterns of variation elucidated by the two marker systems were highly concordant. Analyses indicated that biogeographical hypotheses described the data set better than hypotheses relating to common historical pesticide exposure. Downstream populations had higher genetic diversity than upstream populations, regardless of exposure history, and genetic distances showed that populations from the same river system tended to cluster together. Relatedness among populations reflected primarily directions of gene flow, rather than convergence among contaminant-exposed populations. Watershed geography accounted for significant partitioning of genetic variation among populations, whereas contaminant exposure history did not. Genetic patterns indicating contaminant-induced selection, increased mutation rates or recent bottlenecks were weak or absent. We stress the importance of testing contaminant-induced genetic change hypotheses within a biogeographical context. Strategic application of molecular markers for analysis of fine-scale structure, and for evaluating contaminant impacts on gene pools, is discussed.

Analysis of Variance↗
PubMed2003

Evolutionary history of asexual hybrid loaches (Cobitis: Teleostei) inferred from phylogenetic analysis of mitochondrial DNA variation.

Reconstruction of the evolutionary history of asexual lineages undermines their suitability as models for the studies of evolutionary consequences of sexual reproduction. Using molecular tools we addressed the origin, age and maternal ancestry of diploid and triploid asexual lineages arisen through the hybridization between spiny loaches Cobitis elongatoides, C. taenia and C. tanaitica. Reconstructions of the phylogenetic relationships among mitochondrial DNA (mtDNA) haplotypes, revealed by sequence analyses, suggest that both hybrid complexes (C. elongatoides-taenia and C. elongatoides-tanaitica) contained several asexual lineages of independent origin. Cobitis elongatoides was the exclusive maternal ancestor of all the C. elongatoides-tanaitica hybrids, whereas within the C. elongatoides-taenia complex, hybridization was reciprocal. In both complexes the low haplotype divergences were consistent with a recent origin of asexual lineages. Combined mtDNA and allozyme data suggest that the triploids arose through the incorporation of a haploid sperm genome into unreduced ova produced by diploid hybrids.

Animals↗
PubMed1994

Structure, function, and phylogeny of [Arg8]vasotocin receptors from teleost fish and toad.

[Arg8]Vasotocin (AVT) is considered to be the most primitive known vertebrate neurohypophyseal peptide of the vasopressin/oxytocin hormone family and may thus be ancestral to all the other vertebrate peptide hormones. The molecular evolution of the corresponding receptor family has now been studied by cloning an AVT receptor, consisting of 435 amino acid residues, from the teleost fish, the white sucker Catostomus commersoni. Frog oocytes injected with the AVT receptor-encoding cRNA respond to the application of AVT, but not to its structural and functional counterpart isotocin, by an induction of membrane chloride currents indicating the coupling of the AVT receptor to the inositol phosphate/calcium pathway. The pharmacological properties of the expressed AVT receptor show that it represents, or is closely related to, an ancestral nonapeptide receptor: oxytocin, aspargtocin, mesotocin, and vasopressin activated the receptor, but other members of the vasopressin/oxytocin family tested showed little or no potency; antagonists of the mammalian vasopressin V1 and oxytocin receptors blocked the AVT response. Comparison of AVT receptor sequences spanning transmembrane domains two to five, deduced by cloning cDNAs from the Pacific salmon Oncorhynchus kisutch, the cave-dwelling fish Astyanax fasciatus, and the anuran Xenopus laevis, with those of their mammalian counterparts emphasizes amino acid residues that are involved in hormone binding. The presence of a 5.0-kb transcript in various teleost tissues (pituitary, liver, gills, swim bladder, and lateral line) points to a physiological role for the fish AVT receptor in metabolic, osmoregulatory, and sensory processes.

Amino Acid Sequence↗
PubMed1988

Experimental infection in a monkey with Gnathostoma hispidum larvae obtained from loaches.

Since the 1908s gnathostomiasis has occurred sporadically in Japanese who have eaten loaches, fresh-water fish imported from southeast Asia. Symptoms include creeping skin eruptions and eosinophilia. The symptoms disappeared in most patients within three months after the onset and the patients then remained symptom-free. To elucidate the causative factors, we fed one monkey with 80 gnathostome larvae collected from loaches and autopsied the animal 466 days later. Fifty-one viable larvae (64%) were recovered from muscle; these were in the advanced third stage and were encapsulated.

Animals↗
PubMed1986

Organophosphate dichlorvos induced dose-related differential alterations in lipid levels and lipid peroxidation in various regions of the fish brain and spinal cord.

The effect of dichlorvos (DDVP) (0-0, dimethyl 2:2-dichlorovinyl phosphate), on various lipid fractions and lipid peroxidation in the discrete areas of the brain and spinal cord were studied in the fresh water teleost (Heteropneustes fossilis). Fishes were exposed to three different doses (3.0, 6.0 and 9.0 ppm) of DDVP daily for 7 days. Dose-related increase in the levels of total lipids, cholesterol and esterified fatty acids was detected in the fore brain, optic lobes, cerebellum, medulla oblongata and spinal cord. However, phospholipids were significantly decreased in the aforementioned regions of the central nervous system. The rate of lipid peroxidation was significantly increased in all the regions of the CNS.

Animals↗
PubMed1999

Radionuclides and trace elements in fish collected upstream and downstream of Los Alamos National Laboratory and the doses to humans from the consumption of muscle and bone.

The purpose of this study was to determine radionuclide and trace element concentrations in bottom-feeding fish (catfish, carp, and suckers) collected from the confluences of some of the major canyons that cross Los Alamos National Laboratory (LANL) lands with the Rio Grande (RG) and the potential radiological doses from the ingestion of these fish. Samples of muscle and bone (and viscera in some cases) were analyzed for 3H, 90Sr, 137Cs, totU, 238Pu, 239,240Pu, and 241Am and Ag, As, Ba, Be, Cr, Cd, Cu, Hg, Ni, Pb, Sb, Se, and Tl. Most radionuclides, with the exception of 90Sr, in the muscle plus bone portions of fish collected from LANL canyons/RG were not significantly (p < 0.05) higher from fish collected upstream (San Ildefonso/background) of LANL. Strontium-90 in fish muscle plus bone tissue significantly (p < 0.05) increases in concentration starting from Los Alamos Canyon, the most upstream confluence (fish contained 3.4E-02 pCi g-1 [126E-02 Bq kg-1]), to Frijoles Canyon, the most downstream confluence (fish contained 14E-02 pCi g-1 [518E-02 Bq kg-1]). The differences in 90Sr concentrations in fish collected downstream and upstream (background) of LANL, however, were very small. Based on the average concentrations (+/- 2SD) of radionuclides in fish tissue from the four LANL confluences, the committed effective dose equivalent from the ingestion of 46 lb (21 kg) (maximum ingestion rate per person per year) of fish muscle plus bone, after the subtraction of background, was 0.1 +/- 0.1 mrem y-1 (1.0 +/- 1.0 microSv y-1), and was far below the International Commission on Radiological Protection (all pathway) permissible dose limit of 100 mrem y-1 (1000 microSv y-1). Of the trace elements that were found above the limits of detection (Ba, Cu, and Hg) in fish muscle collected from the confluences of canyons that cross LANL and the RG, none were in significantly higher (p < 0.05) concentrations than in muscle of fish collected from background locations.

Animals↗
PubMed2006

CYP1A induction and blue sac disease in early life stages of white suckers (Catostomus commersoni) exposed to oil sands.

The objectives of this study were to evaluate the influence of natural oil sands on the early developmental stages of white sucker (Catostomus commersoni) and to determine whether biochemical responses in this species were similar to native fish caught in the Athabasca Oil Sands area. Early life stage (ELS) sediment toxicity tests were conducted using controls, reference sediments, natural oil sands, and industrially contaminated (wastewater pond) sediments collected from sites along the Athabasca River, Alberta (Canada). Eggs and larvae were observed for mortality, hatching, deformities, growth, and cytochrome P-4501A (CYP1A) activity using immunohistochemistry. E-Nat-, S-Nat-, and wastewater pond sediment-exposed groups showed significant premature hatching, reduced growth, and exposure-dependent increases in ELS mortality and larval malformations relative to controls. The most common larval deformities included edemas (pericardial, yolk sac, and subepidermal), hemorrhages, and spinal defects. Juveniles exposed to oil sands and wastewater pond sediments (96 h) demonstrated significantly increased 7-ethoxyresorufin-O-deethylase (EROD) activity (30- to 50-fold) as compared to controls. Reference sediment-exposed groups and water controls demonstrated reliable embryo and larval survival, minimal malformations, and negligible CYP1A staining. These observed signs of blue sac disease (ELS mortality, malformations, growth reductions, CYP1A activity induction) may produce deleterious reproductive effects in natural fish populations exposed to oil sands mixtures.

Alberta↗