Zinc content of the gills of rainbow trout (S. gairdneri) after treatment with zinc solutions under normoxic and hypoxic conditions

Forty specimens of rainbow trout (54–127 g) were divided into 4 groups which were treated as follows: (a) normoxic clean water; (b) hypoxic clean water; (c) normoxic water with 10 ppm zinc for 10 h; (d) hypoxic water with 10 ppm zinc for 10 h. The zinc content was determined separately for each of the 4 gill arches on each side of the fish. Values for the zinc concentration were greater following the zinc treatments, but no significant difference between hypoxia and normoxia was observed. Differences in concentrations of zinc were found in different arches whether expressed per gram dry weight or per unit surface area of the secondary lamellae.     

Sub-lethal plasma ammonia accumulation and the exercise performance of salmonids

The proposal that plasma ammonia accumulation might impair the swimming performance of fish was first made over a decade ago, and has now proven to be the case for a number of salmonid species. The first experimental evidence was indirect, when a negative linear relationship between plasma ammonia concentrations and maximum sustainable swimming speed (U(crit)) was found following the exposure of brown trout (Salmo trutta) to sub-lethal concentrations of copper in soft acidic water. Since then, negative linear relationships between plasma ammonia concentration and U(crit) have been demonstrated following exposure of brown trout, rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch) to elevated water ammonia. For brown trout, the relationships between plasma ammonia and U(crit) were remarkably similar following either exposure to elevated water ammonia or to sub-lethal copper. This indicates that the impairment of swimming performance resulting from exposure to sub-lethal concentrations of heavy metals may be attributable in large part to an accumulation of endogenous ammonia. The negative relationship between plasma ammonia concentration and U(crit) was similar in size-matched rainbow and brown trout but, under similar regimes of ammonia exposure, rainbow trout were able to maintain a significantly lower plasma ammonia concentration, revealing inter-specific differences in ammonia permeability and/or transport. One primary mechanism by which ammonia accumulation may impair exercise performance is a partial depolarisation of membrane potential in tissues such as the brain and white muscle. This may prejudice the co-ordination of swimming movements and reduce or abolish the development of muscle tension, thus, compromising swimming efficiency and performance at the top end of the range.     

The toxicity to fish of mixtures of poisons: II. Copper-ammonia and zinc-phenol mixtures

Tests with rainbow trout in mixtures of ammonium chloride with copper sulphate and of phenol with zinc sulphate have shown that the threshold of toxic concentration for a 50% mortality occurs in solutions for which a value of I is obtained by summing the concentrations of the individual poisons expressed as fractions of their individual threshold concentrations. With ammonia-copper mixtures this method of predicting the threshold concentration becomes progressively less adequate as lower percentage mortalities are considered.     

Copper exposure and the degeneration of olfactory receptors in rainbow trout (Oncorhynchus mykiss)

Abstract

Sublethal concentrations of copper in water cause the degeneration of olfactory receptors in rainbow trout (Oncorhynchus mykiss). Receptor cell loss has been correlated to the loss of olfaction in fish and may cause difficulties in olfactory mediated behaviors such as migration. This study investigated the effects of three levels of copper (100, 75 and 50 mg L^?1) on the olfactory epithelium of rainbow trout. Twenty fish randomly allocated between three exposure groups and one control were exposed for 24 hours under static renewal conditions. Light and scanning electron microscopic observations of olfactory tissue were taken to determine the extent of degeneration of receptors. In addition, levels of copper and zinc in the brain tissues were analyzed to determine if the olfactory route was a significant route of copper exposure and transfer to fish brain tissue. Results indicate that degeneration of receptors is related to the concentration of copper. Levels of copper in brain were found to be below detection of the instrument. Levels of zinc were extremely variable ranging from 52 to 132 ng zinc g^?1 brain tissue.     

The acute lethal toxicity to rainbow trout of mixtures of copper,phenol, zinc and nickel†

The acute lethal toxicities to rainbow trout, Salmo gairdneri Richardson, of mixtures of copper and phenol, of copper, zinc and phenol, and of copper, zinc and nickel, over an exposure period of 48 h were determined. The concentrations selected for testing were such that each poison theoretically contributed equally to the toxicity of the mixture. It was found that the toxicities of these mixtures could be adequately predicted by summation of the fractional toxicities of the particular poisons which were present.     

Changes in circulating blood cell levels of rainbow trout, Salmo gairdneri Richardson, following acute and chronic exposure to copper

Significant leucopenia, predominantly the result of a large reduction in lymphocyte concentration, was observed in rainbow trout exposed to 301 μg1⁻¹ copper for 24h. A strong trend ( P = 0–085) toward reduced erythrocyte concentration was also apparent. The pattern of haematological change after chronic exposure (118 μg 1⁻¹ copper for 16 weeks) was substantially different. With the exception of a significant increase in neutrophil concentration (neutrophilia), all blood cell concentrations had returned to control levels.     

The influence of biological and environmental factors on metallothionein concentration in the blood

The concentration of metallothionein (MT), a low-molecular-weight protein, is regulated by many factors, primarily metals (zinc, cadmium, copper), cytokines, glucocorticoides and free radicals. These factors are determined by such aspects of human biology as gender, pregnancy and age, as well as by environmental factors including the use of oral contraceptives and cigarette smoking, all which may affect MT levels in the body.The aim of this study was to investigate the influence of these biological and environmental factors on MT concentrations in erythrocyte lysate and in plasma.MT concentrations were determined by a two-step direct enzyme-linked immunosorbent assay. Evaluation of exposure to cigarette smoking was performed by checking cotinine levels in the plasma of subjects.The studies showed higher MT concentrations in both the erythrocyte lysate and plasma of women when compared to men. Furthermore, pregnancy causes an increase of MT concentration in plasma, while oral contraceptives cause an elevated concentration of MT in erythrocyte lysate. Age impacts plasma MT concentrations in men, whereas it does not affect concentrations of MT in erythrocyte lysate.     

Zinc status in human immunodeficiency virus infection

Plasma zinc and copper concentrations, erythrocyte zinc concentration, copper-zinc superoxide dismutase activity and urinary zinc concentrations were determined for control subjects and individuals with AIDS, ARC, or asymptomatic HIV infection. Significant differences among the population groups were not noted for the above parameters with the exception of plasma copper which was higher in the AIDS group than in other patient groups. These results do not support the idea that zinc deficiency is a common contributory factor of HIV infectivity or clinical expression, nor that HIV infection induces a zinc deficiency.     

Effect of end-stage renal disease and diabetes on zinc and copper status

The aim of this study was to compare the nutritional status of zinc and copper in patients with and without diabetes submitted to chronic hemodialysis. Thirty-three patients with type 2 diabetes (DM group), 30 nondiabetic patients (NDM group), and 20 healthy individuals (control group) were studied. Plasma, erythrocyte, and urinary zinc and plasma copper were obtained from atomic absorption spectrophotometry and ceruloplasmin by immunonephelometry. The anthropometric parameters were similar among the groups. Plasma zinc was lower and erythrocyte zinc was higher in the DM and NDM groups in relation to the control group. No difference in urinary zinc was observed comparing the groups. Plasma copper was higher in the DM group when compared to the NDM and control groups. Ceruloplasmin was similar in the three groups. Serum urea was a positive independent determinant of plasma zinc concentrations. The determinants of erythrocyte zinc were MAMC midarm nuscle circumference and Kt/V dialysis adequacy. The determinants of plasma copper concentration were serum creatinine and serum glucose. The results of this study demonstrate an alteration in the distribution of zinc of patients with chronic kidney disease (CKD) independently of the presence of DM. Also, the status of copper seems not to be influenced by CKD, but only by the metabolic derangements associated with diabetes.     

Trace element concentrations and superoxide dismutase and catalase activities in benign and malignant larynx tumors

The plasma and erythrocyte levels of zinc, copper, and magnesium and the activities of red-cell copper-zinc superoxide dismutase (Cu/Zn-SOD) and catalase (CAT) were determined in patients with benign and malignant tumors of the larynx. Blood samples from patients and healthy controls were drawn using heparinized tubes. The erythrocyte Cu/Zn-SOD and CAT activities were determined spectrophotometrically and the zinc, copper, and magnesium concentrations were determined in erythrocyte and plasma by atomic absorption spectrometry. Variance analysis was employed in the statistical evaluation of the findings. There was a significant increase in red-cell Cu/Zn-SOD activity in the subjects with malignant and benign tumors compared to controls (p<0.001). The CAT activity increased only in the benign tumor group (p<0.01). The plasma zinc concentrations were significantly lower in the malignant tumor group (p<0.05) and significantly higher in the benign tumor group (p<0.01). The erythrocyte copper concentrations were significantly lower in both benign and malignant tumor groups (p<0.001). The plasma copper and magnesium and the erythrocyte magnesium concentrations did not show significant differences relative to controls (p>0.05). The increases in the activities of SOD and CAT activities and the changes in trace elements concentrations can indicate the presence of increased reactive oxygen species that might play a part in the pathogenesis larynx tumors. Presented at the IX Asian-Pacific Congress of Clinical Biochemistry, March 9–14, 2002, New Delhi, India.     

A field investigation of water quality, fish and invertebrates in the Mawddach river system, Wales

A 3-year study was made of the soft, acid waters of the Rivers Mawddach, Wen and Gain, which contained copper, zinc and iron from natural outcrops and abandoned mines. Invertebrates, typical of acid to neutral streams, were present but there was no pattern in overall numbers or in biomass between stations. However, the abundance and distribution of Ephemeroptera were related to pH and water hardness although not to copper and zinc. Brown trout, Salmo trutta , were present and there was little difference in rate of growth between stations, whereas biomass and population density varied considerably between stations, being related to water quality such that, where annual median and 95-percentile of the summed proportions ( p ) of the threshold LC50 concentrations of copper and zinc to rainbow trout, S. gairdneri were 0·3 and 0·7 respectively, biomass was half that in the absence of the metals and, where they were 0·6 and 1·3 respectively, no fish would be expected to occur. Low pH was an additional adverse factor at some locations. Salmon, S. salar , failed to populate a tributary where the sum of the p t LC50 values of copper and zinc was 0·45. High aqueous concentrations of copper resulted in elevated concentrations of copper in the liver of trout, especially in older fish, but not in the muscle. Concentrations of zinc in fish liver and muscle were low despite high ambient concentrations.     

Selenium-induced autometallographic demonstration of endogenous zinc in organs of the rainbow trout, Salmo gairdneri

Autometallographic (AMG) silver enhancement of endogenous zinc was studied in seven organs of the rainbow trout Salmo gairdneri. Groups of trout were injected intraperitoneally with sodium selenite in doses ranging from 0.08 to 25 ppm, administered 1 h before being killed. The concentration of selenium obtained by each organ was determined by gamma-spectrometry, and compared with the autometallographic deposition of silver grains. The relative accumulation of selenium in the organs was: liver greater than spleen greater than kidney greater than intestine greater than gills greater than brain greater than muscle. In the fish labelled with 10 and 25 ppm Se, AMG-deposits were found (1) within lysosomes of liver cells, (2) within the granules and on the nuclear membrane of melanophores in the spleen, (3) on the microvilli and in the apical cytoplasm of renal proximal tubular cells, (4) within the granules and along the plasma membrane of intestinal eosinophilic granule cells, and in the apical portion of the intestinal epithelium, and (5) in the gills, within granule cells and on the surface of the ionocytes. In the trouts injected with 5 ppm Se, silver grains were still observed in the liver, the intestine, and the gills, whereas, no such grains were found in preparations from fish having received 1 ppm Se. The use of selenium for the histochemical demonstration of endogenous zinc versus exogenous metals is discussed. Also, consideration is given to the question of which part of the total tissue zinc that is histochemically reactive.     

The expression of a synthetic rainbow trout metallothionein gene in E. coli

A synthetic gene for rainbow trout metallothionein was constructed and inserted into a dual origin plasmid where expression was induced by a temperature shift in a proteinase-deficient strain of Escherichia coli. The recombinant protein was purified to homogeneity, and a partial amino acid sequence was determined to confirm its identity. Its immunochemical characteristics were similar to those of native metallothionein from rainbow trout. The amounts of recombinant metallothionein produced were quantified in soluble cell extracts by ELISA. Low concentrations were detected when growth was performed either in L-broth or defined (GMM-II) medium. Supplementation of the medium with zinc or copper had no effect on the amount of metallothionein produced. By contrast, when cadmium was included in either L-broth or GMM-II medium, much higher concentrations of the protein within the cells (approx. 13 micrograms/mg soluble cell protein) were detected. This stabilisation of the protein by metal reconstitution in vivo is considered in relation to the selective uptake/exclusion of metals by the cells and its significance for the scavenging of certain precious or toxic heavy metals is discussed.     

Isolation and preliminary characteristics of β-N-acetylglucosaminidase in the sperm of Siberian sturgeon (Acipenser baerii) and rainbow trout (Oncorhynchus mykiss)

The aim of this study was to characterize the enzyme β- N -acetyglucosaminidase (β-NAGase) in the milt and spermatozoa extracts from Siberian sturgeon and rainbow trout. After ion exchange chromatography one protein peak showed β-NAGase activity in sturgeon milt plasma and sperm extracts of both species. Surprisingly, two protein peaks showing β-NAGase activity were found in rainbow trout milt plasma. The molecular mass of β-NAGase was estimated by gel filtration as 127 kDa for rainbow trout spermatozoa, 271 kDa for sturgeon spermatozoa, and 74 kDa for milt plasma from both species. The kinetic parameters were determined for milt plasma and sperm extracts. The optimum pH of the β-NAGases was 3.8 for sturgeon milt plasma, 4.4 for sturgeon sperm extract, and 4.4–4.8 for milt plasma and sperm extract from rainbow trout. K _m value of the β-NAGases was 0.212, 0.563, 0.779 m m for sturgeon milt plasma, sturgeon sperm extract or rainbow trout extract, respectively. The β-NAGase from sperm extracts in both species showed 100% activity even after incubation at 56°C by 20 min, whereas its activity was decreased to 23% in sturgeon milt plasma and to 2% in trout milt plasma.     

Selenium-induced autometallographic demonstration of endogenous zinc in organs of the rainbow trout,Salmo gairdneri

Summary Autometallographic (AMG) silver enhancement of endogenous zinc was studied in seven organs of the rainbow trout Salmo gairdneri. Groups of trout were injected intraperitoneally with sodium selenite in doses ranging from 0.08 to 25 ppm, administered 1 h before being killed. The concentration of selenium obtained by each organ was determined by gamma-spectrometry, and compared with the autometallographic deposition of silver grains. The relative accumulation of selenium in the organs was: liver > spleen > kidney > intestine > gills > brain > muscle. In the fish labelled with 10 and 25 ppm Se, AMG-deposits were found (1) within lysosomes of liver cells, (2) within the granules and on the nuclear membrane of melanophores in the spleen, (3) on the microvilli and in the apical cytoplasm of renal proximal tubular cells, (4) within the granules and along the plasma membrane of intestinal eosinophilic granule cells, and in the apical portion of the intestinal epithelium, and (5) in the gills, within granule cells and on the surface of the ionocytes. In the trouts injected with 5 ppm Se, silver grains were still observed in the liver, the intestine, and the gills, whereas, no such grains were found in preparations from fish having received 1 ppm Se. The use of selenium for the histochemical demonstration of endogenous zinc versus exogenous metals is discussed. Also, consideration is given to the question of which part of the total tissue zinc that is histochemically reactive.     

Comparative study of copper and zinc metabolism in cattle and camel

At an experimental farm, five camels and five cows were fed a similar basal diet for 6 mo. They received oral trace element supplementation for 3 mo (day 22–112). This supplementation included zinc, copper, selenium, managanese, iodine, and cobalt, and corresponded to twice the requirements generally recommended for cows. Plasma copper and zinc concentrations were significantly lower in the camels (61 μg/100 mL for copper and 38 μg/100 mL for zinc) than in the cows (111 and 83, respectively). The supplementation had no effect on plasma zinc concentration in the camels in spite of the low observed values in this species. Liver copper concentration at the beginning of the trial was lower in the camels (9 ppm) than in the cows (35 ppm), and stayed at lower levels during the entire supplementation period. There was no clear difference in fecal excretion of copper and zinc between the camels and the cows. The results suggested that trace element requirements are lower in camels than in cows and that camels regulate their plasma zinc concentration at a very low level (<40 μg/100 mL).     

Selenium status,plasma zinc,copper, and magnesium in vegetarians

Plasma zinc (Zn), copper (Cu), and magnesium (Mg) concentrations, copper/zinc ratio, and selenium (Se) status were studied in 44 vegetarians (22 males and 22 females) and their age- and sex-matched nonvegetarians in the Bratislava region (Slovakia). Vegetarians had statistically significant lower levels of plasma Zn and Cu than nonvegetarians, which may be the result of lower bioavailability of Zn and Cu from this type of diet. No differences in plasma Mg levels were found between vegetarians and nonvegetarians. Se status, as expressed by plasma and erythrocyte concentrations and plasma and erythrocyte glutathione peroxidase activities (GPx), was significantly lower in vegetarians when compared to nonvegetarians. In the series as a whole, there were significantly higher correlations between plasma and erythrocyte Se concentrations and between plasma and erythrocyte GPx activities. Significant positive correlations were also found between plasma Se concentrations and erythrocyte GPx activities, and between erythrocyte Se concentrations and erythrocyte GPx activities. A vegetarian diet does not provide a sufficient supply of essential antioxidant trace elements, like Zn, Cu, and especially Se. Se supplementation should be recommended to this risk group of the population.     

Zinc concentration in erythrocyte membranes in normal volunteers and in patients with taste and smell dysfunction

Erythrocyte membrane zinc concentration was measured in 14 normal volunteers and in 36 patients with taste and smell dysfunction, and compared with zinc concentration in erythrocyte cytosol and blood plasma in the normal volunteers and with zinc concentration in erythrocyte cytosol, blood serum, urine, and parotid saliva in the patients. A A significant negative correlation was found between age and membrane zinc concentration in the normals. A significant positive correlation was found between parotid saliva zinc and erythrocyte membrane zinc concentrations in the patients. Concentration of zinc in the erythrocyte membrane is 50% greater than its concentration in cytosol, and three times greater than the zinc concentration in plasma or serum; membrane zinc concentration in humans is approximately one-half that reported in the dog or in the mouse.     

Plasma binding of 1-butanol, phenol, nitrobenzene and pentachlorophenol in the rainbow trout and rat: a comparative study

1. The in vitro binding of 1-butanol, phenol, nitrobenzene, and pentachlorophenol in trout plasma and rat plasma was determined. 2. Binding to rainbow trout plasma proteins agreed within 9% of that observed in rat plasma. 3. Percentage bound to rainbow trout (2-99%) or rat (10-99%) plasma proteins increased as the log octanol/water partition coefficient of the chemicals increased within the Log P 1-3 range, and was suggestive of hydrophobic interactions in binding.     

Plasma cortisol concentrations before and after social stress in rainbow trout and brown trout

Two related experiments examined the relationship between plasma cortisol concentrations and the development of social hierarchies in fish. In the first, rainbow trout, Oncorhynchus mykiss, and brown trout, Salmo trutta, were observed for dominance interactions when confined within single-species pairs for 4, 48, or 168 h. Subordinate members of a pair exhibited significantly higher cortisol concentrations than dominant and single fish, but the pattern of cortisol elevation differed between the two species, being quicker to rise and increasing to a higher level in rainbow trout. Cortisol concentrations were correlated with behavioural measurements; the more subordinate the behaviour exhibited by a fish, the higher its cortisol concentration. Social stress was a chronic stressor, and no acclimation to social status occurred during the week. In the second experiment, measurements of plasma cortisol were made before pairing of rainbow trout and then after 48 h of confinement in pairs. Subordinate fish demonstrated significantly higher concentrations of plasma cortisol both before and after social stress. It therefore appears that in addition to cortisol being elevated during periods of social stress, an association may exist between initial cortisol levels and the likelihood of a fish becoming subordinate.