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PubMed1996

Capture and confinement stress in white sucker exposed to bleached kraft pulp mill effluent.

This study evaluates the effects of handling and confinement stress and a 3-day recovery period on a number of biochemical parameters used to monitor exposure of fish to bleached kraft mill effluent (BKME). Plasma was collected at four times of the day from male and female white sucker subjected to four levels of handling stress during their spawning migration at a BKME and a reference site. Indicators of a general response to stress (plasma cortisol, glucose, lactate, and total protein) and of reproductive fitness (plasma testosterone, 11-ketotestosterone, and 17 beta-estradiol) were measured. With the exception of 17 beta-estradiol in females, all of the parameters measured varied with time of day and stress level. The general stress indicators ranged from being highly to marginally responsive to handling and confinement stress and were inconsistent in terms of indicating a site difference. The reproductive steroids were moderately responsive to stress and gave the most consistent site difference with testosterone and 11-ketotestosterone levels reduced in BKME-exposed fish under low levels of stress. There was limited evidence for recovery from capture, handling, and transport in the spring for plasma cortisol, lactate, and 17 beta-estradiol at the BKME site, plasma glucose at the reference site, and plasma testosterone at both sites. Fish were also more responsive to an additional acute stress on Day 1 and Day 3 of recovery compared to that on Day 0. This study emphasizes the need for standardized methods in field collections and sampling with the least amount of stress possible, and suggests that holding white sucker for 1 or 3 days does not allow them to recover from the stress of capture.

Analysis of Variance↗
PubMed1995

A peptide from the caudal neurosecretory system of the dogfish Scyliorhinus canicula that is structurally related to urotensin I.

Using reversed-phase HPLC in combination with a radioimmunoassay for ovine corticotropin-releasing hormone (CRH), a peptide with CRH-like immunoreactivity was isolated in pure form from an extract of the caudal spinal cord region of the spotted dogfish, Scyliorhinus canicula. The primary structure of the peptide was established as Pro-Ala-Glu-Thr-Pro-Asn-Ser-Leu10-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg- Glu-Met-Ile- Glu20-Ile-Ala-Lys-His-Glu-Asn-Gln-Gln-Met-Gln30-Ala-Asp-Ser- Asn-Arg-Arg-Ile-Met - Asp-Thr40-Ile.NH2. This amino acid sequence shows moderate structural similarity to Catostomus urotensin I (51%) and to human CRH (56%). The data provide, therefore, chemical evidence to support the conclusions of earlier immunohistochemical studies that the diffuse caudal neurosecretory system of elasmobranchs produces a peptide that is immunochemically related to teleost urotensin I peptides. However, the primary structure of urotensin I has been poorly conserved during evolution.

Amino Acid Sequence↗
PubMed2001

Cloning of two thyrotropin-releasing hormone receptor subtypes from a lower vertebrate (Catostomus commersoni): functional expression, gene structure, and evolution.

A PCR approach was used to clone thyrotropin-releasing hormone receptors (TRH-R) from the brain and anterior pituitary of the teleost Catostomus commersoni (cc), the white sucker. Two distinct TRH-R, designated ccTRH-R1 and ccTRH-R2, were identified. ccTRH-R1 was similar to mammalian TRH-R of the subtype 1, whereas ccTRH-R2 exhibited the highest identity (61% at the amino acid level) with the recently discovered rat TRH-R2. It is postulated that ccTRH-R2 and rat TRH-R2 are members of the same TRH-R subfamily 2. Functional expression of ccTRH receptors in human embryonic kidney cells and in Xenopus laevis oocytes demonstrated that both ccTRH receptors were fully functional in both systems. Oocytes expressing either receptor responded to the application of TRH by an induction of membrane chloride currents, indicating that ccTRH-R of both subtypes are coupled to the inositol phosphate/calcium pathway. The analysis of genomic clones revealed, for the first time, both similarities and differences in the structure of TRH-R subtype genes. Both ccTRH-R genes contained an intron within the coding region at the beginning of transmembrane domain (TM) 6. The position of this intron is highly conserved, as it was found at an identical position in the human TRH-R1 gene. The ccTRH-R2 gene contained an additional intron at the end of TM 3 that was not found in any of the TRH-R1 genes identified so far. The analysis of the gene structure of ccTRH-R and the amino acid sequence comparisons of mammalian and teleost TRH-R of both subtypes suggest that TRH receptors have been highly conserved during the course of vertebrate evolution. A common ancestral TRH receptor gene that could be found much earlier in evolution, possibly in invertebrates, might be the origin of ccTRH-R genes.

Amino Acid Sequence↗
PubMed2001

KDN-containing glycoprotein from loach skin mucus.

It has been widely recognized that the mucus coat of fish plays a variety of important physical, chemical, and physiological functions. One of the major constituents of the mucus coat is mucus glycoprotein. We found that sialic acids in the skin mucus of the loach, Misgurnus anguillicaudatus, consisted predominantly of KDN. Subsequently, we isolated KDN-containing glycoprotein from loach skin mucus and characterized its chemical nature and structure. Loach mucus glycoprotein was purified from the Tris-HCl buffer extract of loach skin mucus by DEAE-cellulose chromatography, Nuclease P1 treatment, and Sepharose CL-6B gel filtration. The purified mucus glycoprotein was found to contain 38.5 KDN, 0.5% NeuAc, 25.0% GalNAc, 3.5% Gal, 0.5% GlcNAc and 28% amino acids. Exhaustive Actinase digestion of the glycoprotein yielded a glycopeptide with a higher sugar content and higher Thr and Ser contents. The molecular size of this glycopeptide was approximately 1/12 of the intact glycoprotein. These results suggest that approximately 11 highly glycosylated polypeptide units are linked in tandem through nonglycosylated peptides to form the glycoporotein molecule. The oligosaccharide alditols liberated from the loach mucus glycoprotein by alkaline borohydride treatment were separated by Sephadex G-25 gel filtration and HPLC. The purified sugar chains were analyzed b --> 6GalNAc-ol, KDNalpha2 --> 3(GalNAcbeta1 --> 14)GalNAc-ol, KDNalpha2 --> 6(GalNAcalpha1 --> 3)GalNAc-ol, KDNalpha2 --> 6(Gal3alpha1--> 3)GalNAc-ol, and NeuAcalpha2 --> 6Gal NAc-ol. It is estimated that one loach mucus glycoprotein molecule contains more than 500 KDN-containing sugar chains that are linked to Thr and Ser residues of the protein core through GalNAc.

Amino Acids↗
PubMed1994

Selenium in eggs and milt of razorback Sucker (Xyrauchen texanus) in the middle Green River, Utah.

Eggs from three female and milt from five male endangered razorback suckers (Xyrauchen texanus) were collected from the Razorback Bar (about 20 km upstream of Ashley Creek) in the Green River of northeastern Utah. Eggs, but not milt, had concentrations of selenium that were above the range of selenium concentrations in control fish from laboratory studies or reference fish from field studies. The concentrations, however, were below those reported in selenium-exposed fish that had reproductive problems in laboratory studies or field investigations. Tests with three streamside spawned pairs of razorback suckers, which were sampled for eggs and milt in this study, resulted in no hatching of fertilized eggs. Concentrations of selenium in eggs and milt were significantly correlated with selenium concentrations in muscle plugs taken from the same fish, but egg and milt concentrations were not significantly different from muscle plugs. Selenium concentrations in eggs of razorback suckers in the Green River may be sufficiently elevated to cause reproductive problems that are contributing to the decline of this species in the upper Colorado River Basin.

Animals↗
PubMed1989

Innervation and gap junctions of intestinal striated and smooth muscle cells in the loach. Thin section and freeze-fracture study.

The tunica muscularis of the proximal intestine of the loach consisted of intermingling striated and smooth muscle cells without forming any distinct sublayers. Close contacts devoid of intervention by a basal lamina sometimes occurred between these different types of muscle cells. Gap junctions were occasionally found between heterologous as well as homologous muscle cells. In freeze-fracture replicas, striated muscle cells were distinguished from smooth muscle cells by numerous, evenly distributed subsurface caveolae. These were relatively rare and linearly arranged in smooth muscle cells. Variously-sized and -formed aggregations of connexon particles were found in the protoplasmic fracture-face of both muscle cells. Striated muscle cells had aggregates of connexon particles taking the form of either a small solid polygon or an annulus with a particle-free central region. In smooth muscle cells, the particles were arranged either in variously-sized patches or in straight lines. Topologically, heterologous gap junctions observed in ultrathin section were thought to correspond to the small patchy aggregations. Striated muscle cells in the gut had neuromuscular junctions, which differed morphologically from "cholinergic" nerve terminals at neuromuscular junctions of typical skeletal muscle cells. The smooth muscle cells had close apposition with axonal terminals containing many granular vesicles and a variable number of small, clear vesicles. Occasionally, a "cholinergic"-type axonal terminal with a presynaptic active site was found close to a smooth muscle cell.

Animals↗
PubMed1995

Comparative study of DNA adducts levels in white sucker fish (Catostomus commersoni) from the basin of the St. Lawrence River (Canada).

The levels of DNA adducts in the hepatic tissue of the white sucker fish species Catostomus commersoni were determined by 32P-postlabelling. The fish were caught at four sites: two sites near the city of Windsor (Québec, Canada) on the St. François River, a downstream tributary of the St. Lawrence River, and two sites in the St. Lawrence River itself, near the city of Montréal (Québec, Canada). The latter sites are known to be contaminated by many pollutants including polycyclic aromatic hydrocarbons. Total adduct levels in all fish ranged from 25.1-178.0 adducts per 10(9) nucleotides. White sucker from the selected sites of the St. Lawrence River had a significantly higher mean level of DNA adducts than those of the St. François River (129.4 vs 56.8, respectively). These results suggest that the effluents of many heavy industries (e.g. from a Soderberg aluminium plant) flowing in the St. Lawrence River are more likely to produce genotoxic damage to fish than those released in one of its tributary, and mainly associated to the activities of a small town and a nearby pulp and paper mill.

Animals↗
PubMed1992

Treatment of fish parasites. 9. Effects of a medicated food containing malachite green on Ichthyophthirius multifiliis Fouquet, 1876 (Hymenostomatida, Ciliophora) in ornamental fish.

For systemic therapy against trophozoites of the skin-inhabiting stage of Ichthyophthirius multifiliis in ornamental fish, the latter were fed medicated food flakes containing malachite green once daily for 1-11 days ad libitum. Naturally or artificially infected cardinal tetras (Paracheirodon axelrodi), blue gouramis (Trichogaster trichopterus), or clown loach (Botia macracantha) were used in the trials. The fish were maintained in aerated 12.5- or 60-1 aquaria at 23 degrees C. Ultrastructural investigations (scanning and transmission electron microscopy) revealed clear deleterious effects of malachite green on the parasitic stages. Following the initial application, the inner membrane of the mitochondria was destroyed. In fish fed for 2 days, aggregation of the mucocysts and polymerization the microtubules within the macronucleus occurred. Finally, the trophozoite's membrane was completely destroyed. In fish fed for 4 days, the medicated food killed all trophozoites of I. multifiliis. Sensitive ornamental fish (e.g., P. axelrodi) showed no adverse effects after they had been fed with only the medicated food flakes for 2 months. Therefore, the oral administration of malachite green using this newly developed medicated food considerably reduces the risk of toxic effects on the fish hosts, which are sometimes caused by malachite green following its application by immersion therapy. The feeding of flakes medicated with malachite green provides and easy-to-handle and highly effective treatment of I. multifiliis in ornamental fish.

Animal Feed↗
PubMed2005

Survival of Lost River suckers (Deltistes luxatus) challenged with Flavobacterium columnare during exposure to sublethal ammonia concentrations at ph 9.5.

The Lost River sucker (Deltistes luxatus) is a federally listed, endangered species inhabiting the hypereutrophic waters of Upper Klamath Lake in southern Oregon, USA. High pH (> or =10) and elevated ammonia concentrations (> or =1 mg NH(3)-N/L) often occur during blooms of cyanobacteria (Aphanizomenon flos-aquae) in the lake, with major fish kills sometimes following a mid- or late-summer "crash" of the cyanobacterial population. Previous histopathology analyses and bacterial sampling indicated that infections of the pathogenic bacterium Flavobacterium columnare might contribute to the fish kills. We hypothesized that prior exposure to adverse water quality conditions increases the susceptibility of Lost River suckers to F. columnare infections. To test this, we exposed juvenile Lost River suckers to four sublethal ammonia concentrations at pH 9.4 for 62 d. On day 31, fish in half of the aquaria were exposed to F. columnare. As expected, survival of the Lost River suckers decreased in aquaria inoculated with F. columnare. Ninety-four percent of the fish that died were infected by F. columnare in the gills, kidney, or skin, whereas none of the survivors or unexposed control fish was infected. However, contrary to our hypothesis, survival of the fish exposed to F. columnare increased significantly (p < 0.05) as unionized ammonia concentrations increased. Our results suggest that complex interactions can complicate prediction of the responses of fish to concurrent chemical stressors and bacterial pathogens.

Ammonia↗
PubMed2005

Chronic toxicity of un-ionized ammonia to early life-stages of endangered Colorado pikeminnow (Ptychocheilus lucius) and razorback sucker (Xyrauchen texanus) compared to the surrogate fathead minnow (Pimephales promelas).

Ammonia-contaminated groundwater enters the Upper Colorado River from beneath the abandoned Moab Uranium Mill Tailings Pile near Moab, Utah. This reach of the Upper Colorado River was designated as critical habitat for four endangered fish species because it is one of the few existing areas with known spawning and rearing habitats. Un-ionized ammonia (NH3) concentrations frequently exceed 1.00 mg/L in backwaters adjacent to the tailings pile, which exceeds the Utah 30-d average chronic water quality criterion for un-ionized ammonia (0.07 mg/L NH3; temperature 20 degrees C; pH 8.2) by a factor of more than 10. However, there is little published information regarding the sensitivity of endangered fishes to ammonia. We conducted 28-d static renewal studies with post-swim-up larvae to determine the relative sensitivity of Colorado pikeminnow (Ptychocheilus lucius), razorback sucker (Xyrauchen texanus), and the standard surrogate fathead minnow (Pimephales promelas) to NH3. Chronic values (ChVs) for mortality and growth were determined as the geometric mean of the no observed effect concentration and the lowest observed effect concentration based on analysis of variance. The ChVs for growth of fathead minnow, Colorado pikeminnow, and razorback sucker were 0.43, 0.40, and 0.67 mg/L NH3, respectively. The ChVs for mortality of fathead minnow, Colorado pikeminnow, and razorback sucker were 0.43, 0.70, and 0.67 mg/L NH3, respectively. Therefore, the ChVs for mortality and growth were similar for fathead minnow and razorback sucker; however, the ChV for growth was lower than the ChV for mortality for Colorado pikeminnow. Maximum likelihood regression was used to calculate 28-d lethal concentrations (LCx) for each species. The 28-d LC50, LC20, and LC1 values for fathead minnow were 0.69, 0.42, and 0.13 mg/L NH3, respectively. The 28-d LC50, LC20, and LC1 values for Colorado pikeminnow were 0.76, 0.61, and 0.38 mg/L NH3, respectively. The 28-d LC50, LC20, and LC1 values for razorback sucker were 0.54, 0.38, and 0.25 mg/L NH3, respectively. The fathead minnow, Colorado pikeminnow, and razorback sucker are relatively similar in sensitivity and rank at the 35th, 49th, and 31st percentiles, respectively, of the theoretical chronic fish sensitivity distributions for NH3. The existing water quality criteria for NH3, if met by remediation activities at the Moab site, would be protective of these endangered fishes even if fish sensitivity is based on the conservative LC1 value.

Ammonia↗
PubMed2002

Assessment of exposure of larval razorback sucker to selenium in natural waters.

This investigation evaluated effects of exposure of larval razorback sucker to waterborne and dietary selenium and other contaminants that occur in nursery habitats. Site waters were collected from three localities on the Colorado River near Grand Junction, CO; a total of five test waters (including control) were studied. Razorback sucker larvae were exposed to site-water contaminants via waterborne and dietary exposure using a laboratory food chain (algae, rotifer, razorback sucker). Fish were exposed for 28 days to site waters and food organisms cultured in site waters. Survival data were analyzed by inspection. Growth data were analyzed using analysis of variance to describe the response of fish in each site water and to describe the relative contribution of waterborne versus dietary exposure to constituents in site waters. Selenium concentrations in test-water treatments ranged from < 1 to 20.3 microg/L in water, < 0.702 to 21.8 microg/g in diet, and 2.34 to 42.0 microg/g in fish. Negative effects from dietary exposure to site-water constituents were detected, but the data suggest that they were caused by cocontaminants in the diet, not selenium exposure. Lack of detection of adverse effects from exposure does not imply that razorback sucker populations are not affected by increased environmental selenium concentrations. There are a variety of factors not included in this investigation that may influence sensitivity of razorback sucker populations to selenium.

Animals↗
PubMed2003

Lethal effect of ammonia on metacercariae of Clonorchis sinensis.

The effect of endogenous ammonia produced in dead fish or ammonia in ammonium bicarbonate solutions was evaluated on the viability of Clonorchis sinensis metacercariae. Viability was determined by worm recovery in rats infected with metacercariae exposed to ammonia in decaying fish or in vitro solutions. The recovery rates were 58%, 48%, 44% and 2% in fish that had been allowed to decay for 5, 10, 20 and 30 days at 4 degrees C, respectively, and this rate ( Y) was inversely correlated with the ammonia concentration ( X) in fish muscle extract ( Y=80.62-0.0667 X, r(2)=0.905, P=0.049). C. sinensis metacercariae barely survived in ammonia solutions with concentrations exceeding 1 g N/l. Our results indicate that the ammonia produced in dead fish is lethal to the viability of C. sinensis metacercariae.

Ammonia↗
PubMed2004

Associations between fish reproductive cycle and the dynamics of metazoan parasite infection.

The parasite fauna of the stone loach (Barbatula barbatula) was investigated monthly from May to December 2001. Four parasite species were recorded: Gyrodactylus spp. (Monogenea), Tylodelphys clavata (Digenea), Proteocephalus sagittus (Cestoda), and Raphidascaris acus (Nematoda). The changes in the seasonal dynamics of parasite infection were investigated throughout the 8 months. The potential effect of the reproductive investment of the host, measured by gonad mass and gonado-somatic index (GSI), on the parasite infection was tested against the prediction that, during periods of high reproductive investment (beginning of the breeding period or forming gonads after breeding), the fish are more susceptible to parasite infection. Differences between parasite loads between genders were also hypothesized. Seasonal differences in infection were observed for all parasite species studied. The values of GSI showed a pattern of energy accumulation in the pre-reproductive period and at the beginning of breeding, a decrease during breeding, and an increase in the post-breeding period. A similar pattern was observed for parasite abundance, a strong or weak increase in spring and/or autumn and a decrease during summer (July and August). Positive correlations between the abundance of Gyrodactylus spp. and R. acus and both gonad mass and GSI were found in females after eliminating the effect of fish weight. Our results suggest that stone loach females are more susceptible to parasite infection in periods of higher reproductive investment. The main factor determining the infection of T. clavata was fish size. The abundance of P. sagittus was positively correlated with GSI in the total fish sample with no detectable effect of sex or fish weight. The increase in cestode infection in spring supports the hypothesis that the parasite life cycle could by synchronized with the beginning of host reproduction, probably induced by increasing fish hormone levels in the spring.

Animals↗