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PubMed2002

Toxicity of selenium and other elements in food organisms to razorback sucker larvae.

Elevated selenium concentrations documented in water, sediment, and biota in irrigation drain water studies by U.S. Department of the Interior agencies and academia have raised concerns that selenium may be adversely affecting endangered fish in the upper Colorado River basin. The objective of the study was to determine the effects on endangered razorback sucker (Xyrauchen texanus) larvae from exposure to selenium and other trace elements in water and zooplankton collected from sites adjacent to the Colorado River near Grand Junction, CO. A 30-day study was initiated with 5-day-old larvae exposed in a 4 x 4 factor experiment with four food and four water treatments, and the biological endpoints measured were survival, growth, development, and whole-body residues of selenium. Mean selenium concentration in reference water (24-Road) was <0.7 microg/l, in reference food (brine shrimp) was 3.2 microg/g, at Horsethief was 1.6 microg/l in water and 6.0 microg/g in zooplankton, at Adobe Creek was 3.4 microg/l in water and 32 microg/g in zooplankton, and at Walter Walker was 13 microg/l in water and 52 microg/g in zooplankton. Although there were differences in concentrations of inorganic elements in water and biota among the three sites, selenium was apparently the only element elevated to concentrations of concern. Effects on survival were more prominent from dietary exposure compared to waterborne exposure. Selenium concentrations of >or=4.6 microg/g in food organisms adversely affected the survival of razorback sucker larvae. The onset of mortality in larvae exposed to food and water from Walter Walker seemed delayed compared to mortality in larvae exposed to food and water from Horsethief, which has been observed in two other studies. Elevated arsenic in one food source seemed to interact with selenium to reduce the toxic effects of selenium.

Animals↗
PubMed2003

Utility of the TBARS assay in detecting oxidative stress in white sucker (Catostomus commersoni) populations exposed to pulp mill effluent.

Recent evidence indicates that contaminant-stimulated free radical production and resulting oxidative damage may be an important mechanism of toxicity in organisms exposed to water-borne contaminants. This study tested the hypothesis that increases in oxidative stress and associated biochemical alterations would be present in white sucker (Catostomus commersoni) living in an environment receiving pulp mill effluent. Consistent increases in liver TBARS and frequent increases in gonadal TBARS were observed immediately downstream of the pulp mill effluent discharge; observable effects were attenuated with distance until they were not significantly different from reference values. Increases detected with the TBARS assay were commensurate with increases detected using the lipid hydroperoxides assay, a mechanistically independent technique. Fish exposed to pulp mill effluent also exhibited significant increases in hepatic free iron and ascorbic acid and a reduced free radical scavenging capacity in the livers of fish downstream of the effluent discharge relative to reference fish. Increases in oxidative stress are not necessarily dependent on increases in lipid substrate or related to reductions in ascorbic acid. TBARS values similar to those observed in fish resident below pulp mill effluent discharges were observed in white sucker 2 h after intraperitoneal injection using 15 mg/kg body weight ferric nitrilotriacetate (Fe(3+) NTA) as a positive control. This study indicates oxidative stress could be a mechanism of toxicity in fish exposed to pulp mill effluent and demonstrates the utility of TBARS in delineating zones of exposure to pulp mill effluent.

Animals↗
PubMed2006

Cloning of black carp beta-actin gene and primarily detecting the function of its promoter region.

A 3 338 bp DNA fragment including the open reading frame and 5'-flanking region of beta-actin gene for black carp genome was obtained through PCR amplification. Analysis of the sequencing results indicated the ORF of black carp beta-actin gene encoding a 375 amino acid protein that shares a high degree of conservation to other known actins. The black carp beta-actin sequence showed 100% identity to common carp, grass carp, and zebrafish, 99.2% identity to human and Norway rat beta-actin gene, 98.9% and 98.1% identity to chicken and Kenyan clawed frog beta-actin gene, respectively. The promoter region of black carp beta-actin gene was inserted into the promoterless pEGFP1 vector. The recombinant plasmid was microinjected into the fertilized eggs of mud loach before two-cell stage as well as transfected into HeLa cell line. GFP expression was found in 50% of mud loach embryos and 2/3 HeLa cells. The GFP expression could be observed in every part of the mud loach embryos, and in some embryos, the GFP was expressed in the whole body. Thus, the usefulness of black carp beta-actin promoter as a ubiquitous expression promoter was confirmed using the EGFP as a reporter gene.

5' Flanking Region↗
PubMed1998

Functional coordination of microtubule-based and actin-based motility in melanophores.

The fish melanophore has been considered the exemplar of microtubule-based organelle transport. In this system, a radial array of uniformly polarized microtubules [1] provides a framework on which dynein-related and kinesin-related motors drive pigment granules toward the minus or plus ends, respectively [2-4]. Stimulation of minus-end motors accounts satisfactorily for aggregation of granules at the cell center. Rapid dispersion is clearly microtubule-dependent; however, the uniform distribution of granules throughout the cytoplasm is paradoxical because stimulation of plus-end motors is predicted to drive the granules to the cell margin. This paradox suggested that the transport system was incompletely understood. Here, we report the discovery of a microtubule-independent motility system in fish melanophores. The system is based on actin filaments and is required for achieving uniform distribution of pigment granules. When it is abrogated, granules accumulate at the cell's margin as predicted for microtubule plus-end motors acting alone. The results presented here demonstrate the functional coordination of microtubule and actin filament systems, a finding that may be of general significance for organelle motility in cytoplasm.

Actin Cytoskeleton↗
PubMed2002

The preparation and characterization of an antimicrobial polypeptide from the loach, Misgurnus anguillicaudatus.

Using Sephadex G-50 gel filtration, DEAE-52 cellulose ion-exchange chromatography, and an improved polyacrylamide gel electrophoresis together with electroelution, a novel polypeptide with antimicrobial activity in vitro was isolated and characterized from loach, Misgurnus anguillicaudatus. The polypeptide, named MAPP, contains about 94 residues containing l0 different amino acids, of which cysteine was the most abundant. No alkaline residue was found in MAPP. MAPP is a single-chain polypeptide with Mw of about 9800Da and pI of about 4.78; the N-terminus of MAPP was CFGWN. MAPP showed good inhibition of various bacteria including Bacillus subtilis, Escherichia coli, and Staphylococcus aureus. MAPP is thermally stable with more than 70% inhibitory bioactivity remaining after treatment at 60 degrees C for 30min. In addition, MAPP could inhibit the autoxidation of pyrogallol with a high efficiency. Similarity searches by comparing amino acid composition, MS-fingerprint, and the N-terminus of MAPP demonstrated that no protein exactly matched MAPP in databases around the world.

Animals↗
PubMed2003

Lateral line pore diameters correlate with the development of gas bubble trauma signs in several Columbia River fishes.

Gas bubble trauma (GBT) caused by gas supersaturation of river water continues to be a problem in the Columbia River Basin. A common indicator of GBT is the percent of the lateral line occluded with gas bubbles; however, this effect has never been examined in relation to lateral line morphology. The effects of 115, 125 and 130% total dissolved gas levels were evaluated on five fish species common to the upper Columbia River. Trunk lateral line pore diameters differed significantly (P<0.0001) among species (longnose sucker>largescale sucker>northern pikeminnow>/=chinook salmon>/=redside shiner). At all supersaturation levels evaluated, percent of lateral line occlusion exhibited an inverse correlation to pore size but was not generally related to total dissolved gas level or time of exposure. This study suggests that the differences in lateral line pore diameters between species should be considered when using lateral line occlusion as an indicator of gas bubble trauma.

Animals↗
PubMed2001

Massaliatrema misgurni n. sp. (Trematoda: Heterophyidae) whose metacercariae encyst in loaches (Misgurnus anguillicaudatus).

Hitherto unknown metacercariae were found encysted in loaches (Misgurnus anguillicaudatus) from China. They were experimentally fed to golden hamsters, and gravid adults were recovered 1 week post-infection from their small intestines. A new species, Massaliatrema misgurni n. sp. (Heterophyidae), is described from the adults. This new species is different from M. gyrinicola Dollfus and Timon-David, 1960, in having a smaller acetabulum/oral sucker ratio, less branched vitellaria widely entering the intercecal anteroacetabular area and an almost median seminal receptacle; and from M. yamashitai Kamiya and Ohbayashi, 1975, in having a larger acetabulum/oral sucker ratio, with the seminal vesicle situated in the uterine loop and the vitellaria entering the intercecal anteroacetabular area. This report is the first record of M. anguillicaudatus as a second intermediate host of the genus Massaliatrema Dollfus and Timon-David, 1960.

Animals↗
PubMed2003

The effects of pulp mill effluent on the sex steroid binding protein in white sucker (Catostomus commersoni) and longnose sucker (C catostomus).

The objective of this study was to characterize the effects of pulp mill effluent on the properties of the sex steroid binding protein (SBP) in the plasma of white sucker (Catostomus commersoni) and longnose sucker (C. catostomus). SBPs which specifically bind estradiol and testosterone with high affinity (k(D) approximately 3 nM) and low capacity (B(max) approximately 73-81 nM) were identified in both species. Subsequent studies determined if the properties of the SBP in white sucker exposed to bleached kraft mill effluent (BKME) at Terrace Bay, ON, and in longnose sucker exposed to BKME at Grande Prairie, AB. differed from appropriate reference fish. There were no effects of BKME exposure on the binding affinity (k(D)) of the SBP in either species, but there was a significant increase in the binding capacity (B(max)) of longnose sucker SBP exposed to BKME. The livers of nai;ve white sucker exposed to effluent at Terrace Bay or a bleached sulfite/groundwood mill in Edmundston, NB, rapidly accumulated compounds of differing hydrophobicity that bound to both the white sucker and goldfish (Carassius auratus) SBP. Conversely, there was reduced accumulation of SBP ligands in the bile of effluent-exposed fish. We have demonstrated that constituents present within pulp mill effluent bind to both the white sucker and goldfish SBP, and that native species residing downstream of pulp mill effluents may experience modifications in the properties of their SBP.

Animals↗
PubMed2003

Structural changes in gills of Lost River suckers exposed to elevated pH and ammonia concentrations.

The Lost River sucker (Deltistes luxatus) is a federally listed, endangered fish that occurs primarily in Upper Klamath Lake-a hypereutrophic lake in southern Oregon, USA. A decline of the sucker population in the lake over the past few decades has been partly attributed to adverse water quality conditions, including elevated pH and ammonia concentrations that occur during summer cyanobacterial blooms. We quantitatively analyzed structural changes in gills of larval Lost River suckers after they were exposed to elevated pH and ammonia concentrations for 30 d. Exposure to pH as high as 10 caused no observed structural changes. However, lamellar thickness and O(2) diffusion distance increased significantly (P<0.05) at ammonia concentrations that did not significantly decrease survival, growth, whole-body ion concentrations, or swimming performance. Additionally, we qualitatively observed increases in the frequency of hyperplasic and hypertrophic mucous cells, tissue damage, epithelial lifting, and infiltration of white blood cells into paracellular lymphatic spaces at the highest sublethal ammonia concentration. These observed gill changes typically indicate compromised respiratory and ionoregulatory capacity, although such effects were not manifested in the assays we performed. Regardless, these structural gill changes appear to be a more sensitive indicator of exposure to elevated ammonia concentrations than are more traditional sublethal indices. Therefore, gill histopathology might be a relevant early-warning monitoring tool of the health of Lost River suckers in Upper Klamath Lake, and other species in similar eutrophic systems.

Ammonia↗
PubMed2003

Membrane chromatographic method for the rapid purification of vitellogenin from fish plasma.

A membrane chromatographic method was developed for the rapid purification of vitellogenin (Vtg) from the plasma of 17beta-estradiol induced loach (Misgurnus angaillicaud atus) and carp (Cyprinus carpio). The time required for the proposed procedure is less then 10 min at a flow-rate of 5 ml/min of the mobile phase, and 0.5 ml of fish plasma could be separated in one cycle. Multistep gradient elution was more suitable for the separation than linear gradient elution. Under optimized conditions, a single Vtg peak can be obtained and its identity was confirmed by SDS-PAGE and gel-permeation chromatography assessment. This method is rapid and easy to operate compared to conventional HPLC and FPLC columns for Vtg separation.

Animals↗
PubMed1992

Life cycle of Gnathostoma nipponicum Yamaguti, 1941.

The life cycle of Gnathostoma nipponicum was examined by field survey and by experimental infection of animals with the larvae. Naturally infected larval G. nipponicum were found in loaches, catfish, and snakes. Experimentally, loaches, killifishes, frogs, salamanders, mice, and rats were successfully infected with the early third-stage larvae of G. nipponicum obtained from copepods (the first intermediate host), whereas snakes, quails, and weasels were not. Frogs, snakes, quails, and rats were experimentally infected with the advanced third-stage larvae (AdL3) from loaches. These results reveal that some species of fishes, amphibians and mammals can act as the second intermediate host and that some species of reptiles, birds and mammals can act as a paratenic host. The life cycle was completed in weasels, the definitive host, which were infected with AdL3 from loaches and started to evacuate eggs of G. nipponicun in faeces on days 65-90 postinfection.

Animals↗
PubMed1991

Culture characteristics of Trypanosoma catostomi and Trypanosoma phaleri from North American freshwater fishes.

Culture characteristics of two species of piscine trypanosomes were compared. Trypanosoma catostomi from Catostomus commersoni was isolated in hypo-osmotic blood agar media but not in hyper-osmotic media. However, T. phaleri from Amia calva was isolated in both media. Only T. catostomi was stimulated to divide by diluting infected blood with water. The maximum number of T. phaleri and percentage of trypomastigotes were not affected by NaCl concentration. The maximum number of T. catostomi was inversely related to osmotic pressure and the percentage of trypomastigotes was directly related to osmotic pressure. This was confirmed by using equi-osmotic concentrations of NaCl, NaBr, Na acetate, KCl, choline-Cl or sucrose. Supplementation with urea had no effect on the number or morphology of trypanosomes. The survival of T. catostomi following cryopreservation was enhanced by prior incubation in hyper-osmotic media. Differential culture characteristics of these species may reflect adaptation to different host-vector systems.

Animals↗
PubMed2001

Morphometric correlates of host specificity in Dactylogyrus species (Monogenea) parasites of European Cyprinid fish.

We test the hypothesis that living on larger fish may impose constraints, i.e. the need to develop large attachment organs, related to the necessity to remain attached on large gills. For this, we compiled data on body size and morphometric measurements of attachment organs of 44 Dactylogyrus species (ectoparasites with direct life-cycle) from 19 cyprinid species. Nineteen dactylogyrid species were considered as specialists (infecting only 1 host species) and 25 as generalists (infecting more than 1 species). The lack of phylogenetic information lead us to perform comparative analyses using raw values and independent contrasts obtained by random phylogenies. Our results show that rich parasite communities are formed by specialists and generalists whereas poor communities are composed mainly of generalist parasites. Moreover, specialists are found on larger hosts, which may reflect a specialization on a predictable resource, as larger fish live longer and offer large gills for parasite colonization. Parasite specialization is shown to be linked with adaptation of attachment organs to their fish hosts. Two morphometric variables of the attachment organ, the total length of anchor and length of base of anchor, were positively correlated with host length for specialists.

Animals↗
PubMed2002

Lead concentrations in fish and river sediments in the old lead belt of Missouri.

Although active mining activities in the Old Lead Belt of Missouri ceased in 1968, old tailings piles remain and continue to impact the lead (Pb) concentrations in sediments and fish in the Big River and Flat River Creek in Missouri. A 3-year study was conducted that examined the Pb concentrations in organic-rich sediments, fish fillets, and fish whole bodies at 13 sites in these two rivers. The results showed that the sediments had significantly elevated Pb concentrations near and beyond the tailings piles compared with control sites upstream from the mining activity. The results also showed the Pb concentration in sediments correlated well with Pb concentrations in suckers (r = 0.86, p < 0.0001) and small sunfish (r = 0.82, p < 0.0001). Annual average Pb concentrations in sucker fillets ranged up to 0.67 microg/g (wet weight basis) and exceeded the World Health Organization guidelines of 0.3 microg/g (wetweight basis) at many of the sample sites. Fillets of bass, which feed at a higher trophic level, had much lower Pb concentrations than either suckers or small sunfish, and no bass fillet samples exceeded the WHO guideline. Whole body Pb concentrations in small sunfish were also determined and correlated strongly (r = 0.92, p = 0.0004) with sediment concentrations before leveling at a maximum of approximately 20 microg/g (wet weight basis).

Animals↗
PubMed2003

Rapidly increasing polybrominated diphenyl ether concentrations in the Columbia River system from 1992 to 2000.

Concentrations and congener patterns of 32 individual PBDE congeners from mono- through hexa-brominated were investigated in two fish species occupying similar habitats--but having different diets and trophic levels--and surficial sediments from several locations on the major river system of western North America, the Columbia River, in southeastern British Columbia, Canada. Total PBDE concentrations have increased by up to 12-fold over the period from 1992 to 2000 in mountain whitefish from the Columbia River, with a doubling period of 1.6 years. The rate at which PBDE concentrations are increasing in whitefish is greater than has been previously reported worldwide. At the current rate of increase, sigmaPBDE will surpass those of sigmaPCB by 2003 to become the most prevalent organo-halogen contaminant in this region. SigmaPBDE in whitefish from the mainstem of the Columbia River range up to 72 ng/g wet weight, concentrations that are 20-50-fold higher than in a nearby pristine watershed affected only by atmospheric contaminant transport. Conversely, sigmaPBDE in largescale suckers were approximately an order of magnitude lower than in whitefish, demonstrating the influence of biomagnification and feeding habits. Congener patterns in whitefish from the Columbia River directly correlated with the two major commercial penta-BDE mixtures in use and represent the first time free-swimming aquatic biota such as fish have been found to contain PBDE congener patterns so similar to commercial mixtures. PBDE concentrations in sediments were not linked to a variety of investigated point sources but were instead inversely correlated with the ratio of organic carbon:organic nitrogen in surficial sediments with a pattern suggesting the dominant influence of septic field inputs from the primarily rural population.

Animals↗
PubMed2001

Identification of DNA polymerase delta in eggs of a teleost fish (loach).

DNA polymerase found in an extract from eggs of the teleost fish Misgurnus fossilis (loach) has been identified as an enzyme of the delta type. The enzyme was purified 4000- to 5000-fold from the extract by liquid chromatography. The DNA polymerase activity was sensitive to the inhibiting action of aphidicolin but resistant to N2-(p-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate (BuPdGTP). The enzyme activity correlates with the presence of a polypeptide with molecular mass of 120-130 kD that interacts specifically with polyclonal antibodies against calf thymus DNA polymerase delta as revealed by Western blotting and is presumably the catalytic subunit of the enzyme. The loach DNA polymerase possesses the 3'-->5'-exonuclease activity specific to single-stranded DNA and catalyzes distributive elongation of primers in primer-template complexes.

Animals↗
PubMed2001

Interaction of loach DNA polymerase delta with DNA duplexes with single-strand gaps.

The interaction of DNA polymerase delta purified from eggs of the teleost fish Misgurnus fossilis (loach) with DNA duplexes with single-strand gaps of 1-13 nucleotides was studied. In the absence of template-restricting DNA, the enzyme elongated primers on single-stranded DNA templates in a distributive manner. However, in the presence of the proximal 5;-terminus restricting the template, the enzyme activity significantly increased. In this case, the enzyme was capable of processive synthesis by filling gaps of 5-9 nucleotides in DNA duplexes. These data indicate that DNA polymerase delta can interact with both the 3;- and 5;-termini located upstream and downstream from the gap. Analysis of the complexes formed by DNA polymerase delta and different DNA substrates by electrophoretic mobility shift assay confirmed the assumption that this enzyme can interact with the proximal 5;-terminus restricting the gap. DNA polymerase delta displayed much higher affinity in duplexes with gaps of approximately 10 nucleotides compared to the standard template-primer complexes. Maximal affinity was observed in experiments with DNA substrates containing unpaired 3;-tails in primers. The results of this study suggest that DNA polymerase delta exerts high activity in the cell nuclei during repair of DNA intermediates with single-strand gaps and unpaired 3;-termini.

Animals↗
PubMed2001

Microsatellite-centromere mapping in the loach, Misgurnus anguillicaudatus.

Primer sets for 15 polymorphic microsatellite loci were developed in the loach, Misgurnus anguillicaudatus (Cobitidae) by molecular cloning and sequencing techniques. Mendelian inheritance was confirmed for the 15 loci by examining the genotypic segregation produced with the primer sets in two full-sib families. The loci were mapped in relation to their centromere in four gynogenetic diploid lines, which were induced by inhibition of the second meiotic division after fertilization with genetically inert sperm. Microsatellite-centromere recombination rates ranged between 0.06 and 0.95 under the assumption of complete interference. Thus, these loci are distributed from the centromeres to the telomeres of their respective chromosomes. The success of mitotic gynogenesis, produced by suppression of the first cleavage, was verified by homozygosity at three diagnostic microsatellite loci that exhibited high gene-centromere meiotic recombination rates in the same family. The differences in heterozygosity levels observed with these markers were attributed to differences in the temporal application of heat shock following inert sperm activation.

Animals↗