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PubMed1988

Evidence for a maternal yolk factor associated with increased tolerance and resistance of feral white sucker (Catostomus commersoni) to waterborne copper.

White suckers, collected from lakes containing elevated levels of copper (12 micrograms liter-1) and zinc (250 micrograms liter-1), were evaluated for reproductive performance, growth and survival of the larvae, and tolerance of the larvae to waterborne copper. Fertilization success was not impaired in white suckers from contaminated sites; metal-exposed males performed better than control males in fertilization trials with control eggs. Larvae hatched from eggs collected at contaminated sites and fertilized in clean water were smaller, developed at a slightly increased rate, and exhibited poorer growth and survival than larvae from control sites. Larvae showed significant changes in resistance and tolerance to copper with age. During the period of endogenous nutrition, larvae from eggs taken at contaminated sites showed increased resistance and tolerance to waterborne copper relative to controls. The effect was not seen in larvae at first feeding, at ages older than 4 days after the onset of feeding (25 days posthatch) or in larvae hatched from control eggs fertilized with sperm from males taken at contaminated sites. This study provides evidence for a maternal yolk factor associated with increased tolerance and resistance of larvae to copper.

Animals↗
PubMed1988

Branchial pathogenesis in a freshwater fish, Puntius conchonius Ham., chronically exposed to sublethal concentrations of cadmium.

Effects of sublethal concentrations, 630 and 840 micrograms/liter (0.05 and 0.066 fractions of the 96-hr LC50), of cadmium chloride on the gills of a freshwater fish, Puntius conchonius, were examined light microscopically during a 12-week exposure. The secondary gill lamellae showed disrupted epithelium, necrosis, accumulation of cellular debris, capillary congestion, and wilting of the pillar cell system. Hypertrophy and hyperplasia of chloride cells as well as partial or complete fusion of secondary lamellae also occurred in the Cd-exposed fish. Branchial lesions together with coagulation film anoxia are likely to result in serious respiratory distress and related tissue hypoxia.

Animals↗
PubMed1989

Effects of natural exposure to copper and zinc on egg size and larval copper tolerance in white sucker (Catostomus commersoni).

Sucker eggs were collected from a metal-contaminated site where previous collections had documented a maternal yolk factor associated with an increased larval copper tolerance during yolk nutrition. In this study, white sucker eggs were divided into groups which were water hardened and/or incubated at the contaminated site or a reference (control) site. Incubation at the contaminated site was associated with a decreased egg size and an increased deformity rate, but had no effect on fertilization rate or larval size. Incubation in contaminated streams increased the larval whole-body metal content, and decreased the tolerance and resistance of larvae exposed to copper in laboratory bioassays. An increase in water temperature accelerated the developmental rate of the larvae, but had no detectable effect on copper tolerance. Although metals did enter the eggs during water hardening, there was no difference in egg metallothionein content, and the lack of obvious hepatic activity until several days after hatching may prevent the formation of protective proteins. The authors hypothesize that the transfer of metals to white sucker eggs in association with yolk precursors, and the subsequent mobilization of these metals during yolk utilization, can account for the observed differences in larval tolerance and growth.

Abnormalities, Drug-Induced↗
PubMed1989

Lead and cadmium in stone loach (Noemacheilus barbatulus L.) from three rivers in Derbyshire.

Fish Noemacheilus barbatulus L. (stone loach) were caught at about 4-week intervals from single sites in three Derbyshire rivers, with different concentrations of cadmium and lead in sediments and water, during a 1-year sampling program. Fish were classified by age, site, and sampling occasion. Growth was allometric and affected by temperature. A steady state of cadmium burden was reached by fish of 2 years old or more but not by younger fish. For lead, fish rarely showed an increase in body burden. Differences in body size accounted for most of the variation in cadmium levels between loach of different age groups but were less important for lead levels. The exponent for body weight was not affected by age of fish and was about 0.79 +/- 0.06 for cadmium and 0.13 +/- 0.21 for lead. There was some correlation between cadmium levels in fish of different age groups taken at the same time from any site; levels of significance were higher when differences due to body size were discounted. Then, sampling time did not explain a significant part of the residual variation. Fluctuations in the cadmium and lead burdens for fish in the same age group from each of the sites were correlated for some comparisons. Loach from sites with higher metal concentrations had higher levels of both cadmium and lead. It is suggested that cadmium uptake from food contributed considerably to the body burden of loach.

Aging↗
PubMed1990

A mathematical model for cadmium in the stone loach (Noemacheilus barbatulus L.) from the River Ecclesbourne, Derbyshire.

A mathematical model which linked metabolism of stone loach with cadmium dynamics was developed using information from laboratory experiments and field studies. Three possible sources of cadmium were distinguished: water, food, and sediment. Predicted results over 1 year were compared with field observations from three sites in Derbyshire. The model adequately predicted growth of fish for all three sites. Predicted cadmium levels in loach were in good agreement with measured levels in fish from all three sites despite the fact that concentrations of cadmium in the environment were kept constant during each simulation. The weight of fish affected the relative importance of different pathways of cadmium intake under similar conditions. Uptake from water contributed substantially to body burden even though the concentration in water was lower than that in food or in sediment. However, uptake from both food and sediment could not be ignored given the measured levels of cadmium in the field. The relative importance of uptake from the three sources also differed with site. The model showed that metabolism, affected by temperature, is important to the dynamics of cadmium in the stone loach.

Animals↗
PubMed1991

Altered performance of white sucker populations in the Manitouwadge chain of lakes is associated with changes in benthic macroinvertebrate communities as a result of copper and zinc contamination.

White sucker (Catostomus commersoni) collected from the Manitouwadge chain of lakes show a lower growth rate and fecundity in lakes contaminated with copper and zinc from a mixed metal mine. This study evaluated whether the changes in performance of the fish were related to direct impacts of the metals or indirect impacts associated with changes in food availability. Concentrations of metals in the water and sediment of lakes in the Manitouwadge chain were elevated, relative to reference sites. The concentrations of Cu and Zn in the digesta of white sucker were significantly higher, as were the levels of both Cu and Zn in liver, kidney, and gill tissue. Muscle and spleen levels of Cu and Zn were significantly lower or not different from controls. Tissue levels were within the homeostatic range for Cu and Zn. However, the total density of invertebrates varied from greater than 25,000 m-2 at control sites to less than 13,000 m-2 at contaminated sites, and the number of genera recorded was more than 50% lower in shallow water samples. There was almost a complete absence of several invertebrate taxa at contaminated sites, including Plecoptera, Ephemeroptera, Odonata, Trichoptera, Amphipoda, and Unionidae. Diptera accounted for 78 to 96% of the total numbers of individuals at metal-contaminated sites as compared with 40 to 75% at the control sites. An analysis of white sucker stomach contents showed that the contents closely reflected the benthic composition observed in the natural substrate. Changes in food availability and feeding activity were correlated with previous changes documented in the growth, fecundity, and lipid levels of white sucker.

Animals↗
PubMed1992

Milt characteristics, reproductive performance, and larval survival and development of white sucker exposed to bleached kraft mill effluent.

White sucker from a Lake Superior bay which receives bleached kraft mill effluent (BKME) show increased hepatic mixed-function oxygenase (MFO) activity, reduced plasma sex steroid levels, decreased egg and gonad size, a decrease in the occurrence of secondary sexual characteristics, and an increased age to maturation. This study evaluated the reproductive performance of that white sucker population relative to a similar reference population. Spawning male white sucker from the BKME site had reduced spermatozoan motility but no significant differences in milt volume, spermatocrit levels, or seminal plasma constituents. BKME male and female fish had equal or greater fertilization potential compared to both male and female fish at the reference site. There was no difference either in the hatchability of the eggs or in larval size at hatch. BKME larvae did show reduced growth rates by 24 days posthatch but showed equal rates of yolk utilization. No difference in larval MFO activity was detected between sites at 21 days posthatch, indicating no parental transfer of induction to the progeny.

Animals↗
PubMed1993

Epizootic haematopoietic necrosis virus (EHNV): improved ELISA for detection in fish tissues and cell cultures and an efficient method for release of antigen from tissues.

An improved antigen capture ELISA for the detection of epizootic haematopoietic necrosis virus (EHNV) is described. Affinity purified rabbit anti-EHNV immunoglobulin was used as capture antibody, enabling the detection of EHNV at a concentration of 10(3.5) TCID50/ml cell culture supernatant. The sensitivity and specificity of the ELISA, determined by examination of clarified tissue homogenates from 55 infected fish and 348 uninfected fish in relation to virus isolation, were 81.2% and 98.9%, respectively. Specificity for redfin perch (Perca fluviatilis) tissue samples was 100%, however, high background optical density (OD) in samples from uninfected rainbow trout (Oncorhynchus [corrected] mykiss) resulted in lower specificity (98.5%). Specificity for rainbow trout tissues was increased to 100% by testing clarified tissue homogenates that had been diluted 1:10. Sensitivity and specificity estimates for cell culture supernatants were 96.2% and 99.9% based on the results obtained for 158 infected and 355 uninfected samples. Incubation steps in the ELISA can be shortened to enable its completion in less than 4 h if a rapid qualitative diagnosis is required. Of eight methods of releasing EHNV from infected tissue that were evaluated, manual grinding of tissue with a fitted pestle in a 1.5-ml plastic disposable tube, followed by vortexing in the same tube with 3 mm diameter glass beads and clarification in a microcentrifuge was the most efficient. Samples from 150-200 individual fish could be prepared for ELISA and cell culture by a single operator in a day. Antigen capture ELISA can replace virus isolation as the method of choice for diagnosis of EHNV.

Animals↗
PubMed1991

Elucidation of cDNA sequences for metallothioneins from rainbow trout, stone loach and pike liver using the polymerase chain reaction.

Metallothionein cDNAs were generated from the livers of three fish species and amplified by PCR. Two distinct coding sequences (A and B) were elucidated for rainbow trout metallothioneins but single isoforms were encoded by genes isolated from the stone loach and pike. Different sized transcripts were observed both with stone loach and pike but these were accounted for by altered lengths of 3' untranslated regions.

Amino Acid Sequence↗
PubMedDate not supplied

Urotensin I- and CRF-like peptides in Catostomus commersoni brain and pituitary--HPLC and RIA characterization.

Two distinct neuronal systems, containing urotensin I-like immunoreactivity (UI-LI) and CRF-LI respectively, have previously been demonstrated immunocytochemically in the brain and pituitary of the teleost Catostomus commersoni. In the present studies, we used HPLC followed by UI- and CRF-RIA to further characterize these UI- and CRF-LI substances. HPLC of Catostomus brain extracts (including extracts modified by dilute H2O2 oxidation or dilute acid cleavage) suggests that the major form of brain UI-LI is very likely identical to the previously characterized urophysial UI; the small amount of pituitary UI-LI is not yet fully characterized. The behavior of brain CRF-LI on reversed phase (RP) extraction suggests that it may exist largely in precursor form. Pituitary CRF-LI, however, behaved as expected for a peptide on RP extraction, and eluted from HPLC as essentially a single (though somewhat broad) peak. The HPLC behavior of pituitary CRF-LI, and its crossreactivity with various CRF antisera, suggest that it represents authentic fish CRF, apparently similar to (but not identical with) human/rat CRF.

Animals↗
PubMed1993

Immunoreactive atrial natriuretic peptides in the oocyte.

1. The presence and partial characterization of immunoreactive ANP (irANP) in eggs were investigated using high performance liquid chromatography (HPLC), immunohistochemistry and Northern-blot hybridization in mammalian and nonmammalian vertebrates. 2. Serial dilution curves of egg extracts from frogs and freshwater teleostean fishes (silver crusian carp and loach) were parallel to the standard curve of atriopeptin III. Rat oocytes also contained irANP (25-30 pg/egg). 3. The HPLC profile of irANP showed two main peaks corresponding to low and high molecular weight of irANP. 4. ANP mRNA was detected in the fish eggs. With immunohistochemical analysis of the rat ovarian follicles, this ANP-like material was localized mainly at the oocytes. 5. We, therefore, suggest that the vertebrate oocytes synthesize ANP and that the presence of ANP in the oocytes may be linked with the cell differentiation.

Animals↗
PubMed1986

The distribution of 'extraurophyseal' urotensin I-immunoreactivity in the central nervous system of Catostomus commersoni after urophysectomy.

The occurrence and distribution of the fish neuropeptide urotensin I-immunoreactive (IR-UI) perikarya and fibres was investigated by peroxidase-anti-peroxidase immunohistochemistry in paraffin and vibratome sections of the central nervous system of urophysectomized and control fishes. IR-UI neurones not described previously were found in the area ventralis telencephali pars dorsalis, in the area pretectalis and in two unidentified brainstem nuclei of control and urophysectomized fishes. In urophysectomized fishes a 4-fold increase in the number of parvocellular IR-UI neurons was observed in the nucleus lateralis tuberis, whereas no significant differences were seen in other IR-UI neuronal groups. A functional relationship between IR-UI 'extraurophyseal' (i.e. brain) and caudal neurosecretory systems is suggested.

Animals↗
PubMed1990

The 5'-flanking sequence of the loach oocyte 5S rRNA gene contains a signal for effective transcription.

Deletion mutants of loach oocyte 5S rRNA genes were injected and transcribed in vivo in the nuclei of loach (Misgurnus fossilis) and Xenopus laevis. A control region was found in the 5'-flanking sequence, the elimination of which greatly decreases in vivo transcription of 5S rRNA genes. This cis-acting element is located in the region between nt-18 and the transcription start point. We propose that the oocyte nucleus contains (a) specific transcriptional factor(s), NTFO, which interacts with the cis-acting element we described. We also propose that NTFO is inactivated in maturing oocytes when nucleoplasm interacts with oocyte cytoplasm after germinal vesicle breakdown. The residual activity of this factor(s) may be responsible for low-level synthesis of oocyte 5S rRNA at the beginning of embryogenesis. We consider the disappearance of NTFO during gastrulation to be responsible for the total inactivation of oocyte 5S rRNA genes in embryonic and somatic tissue.

Animals↗
PubMed1987

Sequestration of environmental cadmium by metallothionein in the roach (Rutilus rutilus) and the stone loach (Noemacheilus barbatulus).

Two species of coarse fish that are relatively resistant to cadmium poisoning were exposed to sub-lethal concentrations of the metal in their aquarium water. Thus, roach were exposed to cadmium concentrations between 30 and 500 micrograms/l for periods of 14-70 days whereas stone loach were exposed to 1250 micrograms Cd/l for 21-77 days. Under all conditions of exposure, it was found upon analysis of the major organs of accumulation of cadmium in the two species that the toxic metal was sequestered by a single isoform of metallothionein. The amino acid compositions of roach and stone loach metallothionein were determined and found to be similar to those reported for other piscine metallothioneins. The two proteins were found to contain Cd:Zn:Cu in approximate ratios of 4:1:2 per mole of protein. The sequestration of Cd by metallothionein in the two resistant species of fish is contrasted with the situation observed previously in a cadmium-sensitive species, the rainbow trout.

Amino Acids↗
PubMed1990

The role of glutathione S-transferases in the hepatic metabolism of benzo[a]pyrene in white suckers (Catostomus commersoni) from polluted and reference sites in the Great Lakes.

1. Orally administered 3H-benzo[a]pyrene (3H-BaP) was excreted in the bile of White Suckers predominantly as water soluble metabolites some of which were hydrolyzed by arylsulfatase or beta-glucuronidase. 2. Non-hydrolysible polar metabolites comprised a substantial proportion of biliary metabolites. 3. HPLC analysis revealed fluorescent and 3H-labelled peaks which co-eluted with standards of the glucuronide and sulfate conjugates of BaP. 4. The most polar peak co-chromatographed with a double-radiolabelled metabolite produced in vitro with 3H-BaP and 35S-glutathione. 5. Inhibition of epoxide hydrolase in vitro reduced all water soluble metabolites except the glutathione conjugate of BaP. 6. Glutathione conjugation represents a major hepatic detoxication pathway of BaP in White Suckers.

Animals↗
PubMed1990

Phalloidin inhibits cortical granule exocytosis and ooplasmic segregation in loach eggs.

Injections of phalloidin under the surface of loach eggs, followed by activation of the eggs in tap water, result in local inhibition of cortical granule (CG) exocytosis. Light and electron microscopy revealed that in the region where exocytosis is inhibited the thickness of the microfilamentous cortex (MC) separating CGs from the plasma membrane (PM) is increased significantly, and many CGs are detached and have moved away from the MC. Injections of phalloidin also inhibit ooplasmic segregation in fertilized eggs. The experiments suggest that in intact eggs the MC represents a physical barrier to CG exocytosis, and that interactions of the MC with the PM and CGs are crucial for the retention of CGs near the sites of fusion.

Actins↗
PubMed1993

Transduction of Ca2+ signals upon fertilization of eggs; identification of an S-100 protein as a major Ca(2+)-binding protein.

A transient increase in the level of free cytosolic Ca2+ is observed upon fertilization of the eggs of many species and is thought to represent a key event in the initiation of development. To identify components in the egg which could be involved in mediating such Ca2+ signals we searched for Ca(2+)-binding proteins in eggs of the fresh-water fish Misgurnus fossilis (loach). We show that loach eggs contain two major Ca(2+)-binding proteins which can be purified through their Ca(2+)-dependent interaction with a hydrophobic matrix. Protein sequencing revealed that the larger 18 kDa protein is calmodulin, while the smaller polypeptide of 10 kDa is a member of the S-100 protein family. This is the first report of the presence of an S-100 protein in vertebrate eggs and shows that this protein is found in two fold higher concentration than calmodulin. Since the 10 kDa protein shares 68% sequence identity with S-100 alpha from bovine brain, it can be considered as the loach homologue of mammalian S-100 alpha. During early embryonic development, de novo protein synthesis of calmodulin is observed at the earliest stages analyzed (mid-blastula), while de novo protein synthesis of the S-100 alpha homologue begins with the mid-gastrula stage.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed2004

Oxidative stress responses in longnose sucker (Catostomus catostomus) exposed to pulp and paper mill and municipal sewage effluents.

While recent evidence indicates that the generation of reactive oxygen species (ROS) and associated oxidative damage are frequently observed in fish populations with exposure to pulp and paper mill effluents, the potential for ROS generation from municipal sewage effluents has not been addressed. This study investigates the utility of measures of oxidative stress in delineating the effects of both pulp and paper mill and municipal sewage discharges. Longnose sucker (Catostomus catostomus) were collected below three pulp and paper mill and two municipal sewage effluent discharges over a 3-year period within the Wapiti and Athabasca River systems in northern Alberta. Biochemical responses in longnose sucker varied between the two rivers systems, with more pronounced changes occurring within the Wapiti River. Of the suite of biochemical parameters examined, fatty acyl-CoA oxidase (FAO) activity was the most sensitive indicator of pulp and paper mill exposure, but was only infrequently induced with exposure to municipal sewage effluent. Hepatic and gonadal 2-thiobarbituric acid reactive substances, lipid hydroperoxides, and hepatic free iron were less consistently elevated with exposure to pulp and paper mill effluent than FAO activity, and were also only infrequently altered with sewage effluent exposure. Hepatic ascorbic acid, liver somatic index, and condition factor were consistently altered with exposure to both sewage and pulp and paper mill effluents. While specific biochemical and organismal responses varied with effluents and time, the collective suite of oxidative stress endpoints proved to be useful tools in identifying relative influences of municipal sewage and pulp and paper mill effluent on fish populations in adjacent receiving waters.

Acyl-CoA Oxidase↗