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.     

Studies on the toxicity of zinc sulphate and of cadmium sulphate to stone loach Noemacheilus barbatulus (L.) in hard water

The toxicity of zinc sulphate and cadmium sulphate to stone loach Noemacheilus barbatulus was examined in an attempt to explain the distribution of the species in two polluted streams. Stone loach in hard water were more sensitive than rainbow trout to zinc, but much more resistant to cadmium. Their disappearance from the Willow Brook, Northants, in 1971–72 could be accounted for by zinc toxicity. A behavioural change, the loss of the instinct to hide during daylight, occurred in subsequently lethal concentrations of cadmium. The accumulation of cadmium in several tissues was also determined.     

THE SURVIVAL OF RAINBOW TROUT (SALMO GAIRDNERII RICHARDSON) AND PERCH (PERCA FLUVIATILIS L.) AT VARIOUS CONCENTRATIONS OF DISSOLVED OXYGEN AND CARBON DIOXIDE

The relative importance of dissolved oxygen and dissolved carbon dioxide in determining the lethal effect of an environment for rainbow trout and perch has been investigated with an apparatus which controls the concentrations of these gases in a body of water. It is shown that concentrations of carbon dioxide which sometimes occur in polluted streams can more than double the minimum concentration of dissolved oxygen necessary for the survival of half a population of rainbow trout fingerlings for 24 hr. Increase in temperature between 12·5 and 19·5°C. shortens period of survival in solutions containing up to 67 p. p. m. CO₂. Within the range of dissolved oxygen concentration which is lethal in the presence of 59 p. p. m. CO₂ or more, perch are more resistant than rainbow trout in the lower, but less resistant in the higher, oxygen concentrations. The relation between carbon dioxide concentration and the oxygen tension at which rainbow trout blood is half saturated with oxygen is similar to the relation between carbon dioxide concentration and the oxygen tension at which the median period of survival of this species is I hr.     

The interactions of water hardness and pH with the acute toxicity of zinc to the brown trout, Salmo trutta L.

Exposure of brown trout, Salmo trutta , to zinc under continuous flow conditions over 96 h showed that both water hardness and pH exert major influences on the toxicity of the metal. 96-h LC50 values for total zinc ranged from <0.14mg 1⁻¹ in alkaline soft water (pH 8; lOmg 1⁻¹ as CaCO₃) to 3.20 mg 1⁻¹ in acidic hard water (pH 5; 204 mg 1⁻¹ as CaCO₃). A variable reduction in zinc toxicity in hard water compared with soft water over the pH range 4–9 was attributed to high external calcium. Zinc toxicity was positively correlated with decreasing acidity over the pH range 5–7, the metal being most toxic at pH 8–9 where metal complexes predominate. Below pH 5 metal toxicity also increased, irrespective of hardness. Water hardness and pH interacted with zinc toxicity in a complex manner, apparently dependent on physical and chemical transformations of the metal, and as changes in uptake. detoxification and excretion by the fish.     

Toxicity to rainbow trout and minnows of some substances known to be present in waste water discharged to rivers

Toxicity to fish (rainbow trout or minnows) of solutions of several pure substances has been measured under controlled conditions. The substances (sodium arsenite, sodium arsenate, sodium picrate, sodium dinitrophenate, zinc sulphate, potassium chromate, potassium dichromate, ammonium chloride, and ammonium sulphate) were dissolved in distilled water, in Watford tap water, or in mixtures of distilled water and tap water.     

Radiotracer Studies on Waterborne Copper Uptake,Distribution, and Toxicity in Rainbow Trout and Yellow Perch: A Comparative Analysis

Rainbow trout (Oncorhynchus mykiss) are often used to estimate important biotic ligand model (BLM) parameters, such as metal-binding affinity (log K) and capacity (B_max). However, rainbow trout do not typically occupy metal-contaminated environments, whereas yellow perch (Perca flavescens) are ubiquitous throughout most of North America. This study demonstrates that dynamic processes that regulate Cu uptake at the gill differ between rainbow trout and yellow perch. Rainbow trout were more sensitive to acute aqueous Cu than yellow perch, and toxicity was exacerbated in soft water relative to similar exposures in hard water. Whole body Na loss rate could account for acute Cu toxicity in both species, as opposed to new Cu uptake rate that was not as predictive. Time course experiments using radiolabelled Cu (⁶⁴Cu) revealed that branchial Cu uptake was rather variable within the first 12 h of exposure, and appeared to be a function of Cu concentration, water hardness, and fish species. After 12 h, new branchial Cu concentrations stabilized in both species, suggesting that metal exposures used to estimate BLM parameters should be increased in duration from 3 h to 12+ h. In rainbow trout, 71% of the new Cu bound to the gill was exchangeable (i.e., able to either enter the fish or be released back to the water), as opposed to only 48% in yellow perch. This suggests that at equal exposure concentrations, proportionally more branchial Cu can be taken up by rainbow trout than yellow perch, which can then go on to confer toxicity. These qualitative differences in branchial Cu handling between the two species emphasize the need to develop BLM parameters for each species of interest, rather than the current practice of extrapolating BLM results derived from rainbow trout (or other laboratory-reared species) to other species. Data reported here indicate that a one-size-fits-all approach to predictive modeling, mostly based on rainbow trout studies, may not suffice for making predictions about metal toxicity to yellow perch—that is, a species that inhabits metal-contaminated lakes around northern Canadian industrial operations.     

THE EFFECT OF TENSION OF DISSOLVED OXYGEN ON THE TOXICITY OF UN-IONIZED AMMONIA TO SEVERAL SPECIES OF FISH

The resistance to rapidly lethal concentrations of un-ionized ammonia ranging from about 2.0 to 8.8 p. p. m. N was determined for rainbow trout ( Salmo gairdnerii Richardson), perch ( Perca fluviatilis L.), roach ( Rutilus rutilus (L.)), and gudgeon ( Gobio gobio (L.)) in tensions of dissolved oxygen 53.4 and 96.7% of air-saturation value at 15.2°C.
The relation between concentration of un-ionized ammonia and period of survival from a few hours to 13 days was investigated for rainbow trout in two tensions of dissolved oxygen, 45.7 and 100.3% of air-saturation value, at 20.1°C.
Period of survival of all the species tested decreased with rise in concentration of un-ionized ammonia.
Decrease in tension of dissolved oxygen increased the toxicity of un-ionized ammonia to all species except gudgeon, for which there was no significant difference in period of survival at the two levels of oxygen.
The effect of oxygen tension on period of survival was greatest in the lowest concentrations of un-ionized ammonia.
Resistance of perch and roach to lack of oxygen was not significantly affected, but that of rainbow trout was reduced significantly, by the presence of a small concentration of ammonia (not toxic to trout within at least 13 days).     

Uptake and accumulation of zinc in juvenile rainbow trout,Salmo gairdneri

Synopsis The toxicity of zinc to rainbow trout was determined and the 72 h median lethal concentration was found to be 2.00 mg l^–1 in freshwater, hardness 7.50 mg l^–1 as calcium. An insignificant increase in zinc concentration of internal tissues occurred in fish exposed to 1.52 mg l^–1 in freshwater for 72 h. However, there was a significant uptake of zinc by gills and the body surface. Fish exposed to 10 mg l^–1 zinc for 72 h in two-thirds sea water showed significant zinc uptake by liver, rectum and muscle, when compared to control fish. Drinking rate decreased from 1.43 to 0.26 ml kg^–1 h^–1 when zinc sulphate was added to freshwater. Trout adapted to two-thirds sea water showed no decrease in drinking, about 7 ml kg^–1 h^–1 when zinc was added to the water.     

A FURTHER STUDY OF THE TOXICITY OF POTASSIUM CYANIDE TO RAINBOW TROUT (SALMO GAIRDNERII RICHARDSON)

The distributions of the periods of survival of individual rainbow trout in solutions of potassium cyanide are asymmetrical and approximately log-normal within the range of concentration where the relation between log time and log concentration is linear, whereas they are symmetrical and approximately normal at higher concentrations.
Some inherent, persistent features possessed by the fish influence survival time in both the higher and lower ranges of concentration, but other such features exert a significant influence only in the lower concentrations.
Some of the features influencing survival time in the lower concentration range only, are associated with body size.     

The distribution of zinc and copper in plasma, erythrocytes and erythrocyte membranes of rainbow trout (Salmo gairdneri)

1. The zinc and copper concentration of plasma was determined in rainbow trout, lake trout, walleye and whitefish. 2. These fish had mean plasma zinc concentrations ranging from 9.3 to 15.1 ppm and copper concentrations from 0.6 to 1.3 ppm. 3. In rainbow trout, the concentration of zinc and copper is greater in the erythrocyte membrane than in the total erythrocyte. 4. Ultrafilterable plasma zinc and copper concentration in rainbow trout was determined to be 0.03 and 0.019 ppm, respectively. 5. Dialysis of rainbow trout plasma against 20 mM EDTA results in removal of 99% of the zinc and 88% of the copper from plasma proteins.     

A comparison of cadmium-induced metallothionein gene expression and Me2+ distribution in the tissues of cadmiumsensitive (rainbow trout; Salmo gairdneri) and tolerant (stone loach; Noemacheilus barbatulus) species of freshwater fish

1. The accumulation of cadmium in the liver, kidney and gills of rainbow trout and stone loach was measured during exposure of the fish to the metal at 3 smg/l in their aquarium water. The pattern of accumulation of the toxic metal in the individual organs was different between the two species.2. The tissue concentrations of metallothionein-specific mRNA and metallothionein protein were also determined in these organs from the same fish. In rainbow trout, the induction of metallothionein gene expression resulted in a gradual increase in metallothionein concentration in gill over the course of the experiment whereas increases in metallothionein in the liver and kidney were detected only at the later time points of analysis (beyond 19 weeks). By contrast, in the same tissues from stone loach, relatively minor changes were quantified in specific mRNA and metallothionein concentrations.3. Throughout the experimental period, tissue concentrations of zinc and copper were determined in the liver, kidney and gills of the rainbow trout and stone loach. Subtle decreases were observed in the zinc concentration of gills in rainbow trout and substantial increases were observed in the hepatic copper concentrations in both species at the later time points of analysis.4. The ability of cadmium to induce metallothionein gene expression and its subsequent ability to compete for the sequestration sites on the newly-synthesized protein is discussed with regard to the relative levels of cadmium, zinc and copper in the organs studied and differing regimes of cadmium administration.     

Intestinal zinc uptake in freshwater rainbow trout: evidence for apical pathways associated with potassium efflux and modified by calcium

Understanding the mechanisms of intestinal zinc uptake in fish is of considerable interest from both nutritional and toxicological perspectives. In this study, properties of zinc transport across the apical membrane of freshwater rainbow trout intestinal epithelia were examined using right-side-out brush border membrane vesicles (BBMV's). Extravesicular calcium was found to have complex actions on zinc uptake. At a low zinc concentration of 1 microM, calcium (0.1-2 mM) significantly stimulated zinc uptake. In contrast, calcium inhibited zinc uptake at higher zinc levels (100 microM). Lanthanum and cadmium in the external medium did not block zinc uptake, suggesting that interactions between zinc and calcium were not exerted at a calcium channel. Copper also failed to exercise any inhibitory action. Zinc association with the BBMV's was enhanced by an outward potassium gradient. This stimulatory effect was only present at a zinc concentration of 100 microM. The potassium channel blocker, tetraethylammonium chloride inhibited zinc uptake at this relatively high zinc concentration, suggesting the presence of a low affinity zinc uptake pathway linked to potassium efflux. The present study provides evidence that the mechanism of intestinal zinc uptake in rainbow trout is pharmacologically very different from that of the piscine gill and the mammalian intestine.     

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.     

Toxicity of cadmium and zinc to Diplostomum spathaceum (Trematoda: Diplostomidae) cercarial survival

The toxicity of cadmium and zinc at concentrations ranging from 0.1 to 10000 microg/l was investigated with cercariae of Diplostomum spathaceum at three temperatures (12, 20, 25 degrees C) and three levels of water hardness (distilled water, soft water, hard water). Under most environmental conditions survival of cercariae was reduced by increasing metal concentration. Increasing water hardness and decreasing water temperature caused an increase in the survival of both control and metal exposed cercariae. However, with decreasing temperature increased survival of metal-exposed cercariae above the control occurred at a number of low metal concentrations. Differences in the relative toxicity of cadmium and zinc to cercariae was dependent on the environmental conditions of exposure. The two heavy metals demonstrated little toxicity to cercariae during the period of maximum cercarial infectivity (0-5 h).The mechanisms of metal toxicity and their effects on cercarial survival are discussed.     

Mortality patterns in trout (Salmo trutta and S. gairdneri) exposed to an anionic detergent in relation to concentration and mechanisms of toxic action

SUMMARY. Histological and electron microscopical study of the gills of fish poisoned by the anionic detergent sodium lauryl sulphate indicates that the nature of the toxic action changes at a concentration of about 120 mg I⁻¹. Toxicity tests were conducted to determine whether this change could be detected in the test data, using brown trout ( Salmo trutta ) and rainbow trout ( S. gairdneri ).
Toxicity curves showed no indication of the change in toxic action, but at concentrations of 120 and 130 mg I⁻¹ of sodium lauryl sulphate 'split' probit lines occurred, and the lines for 150 mg I⁻¹ consistently differed in slope from other lines.     

Application of XANES spectroscopy in understanding the metabolism of selenium in isolated rainbow trout hepatocytes: insights into selenium toxicity

Selenium (Se) is an essential element, but causes toxic effects in fish at a slightly elevated level beyond the threshold. However, the degree of Se toxicity differs depending on the chemical forms of Se (e.g., organic vs. inorganic) to which fish are exposed to. The mechanisms of Se metabolism and toxicity in fish, particularly at cellular level, are poorly understood. The present study was designed to examine the metabolic fate of different seleno-compounds, both inorganic and organic, in isolated hepatocytes of rainbow trout (Oncorhynchus mykiss) in primary culture using XANES spectroscopy. In cells exposed to 100 μM of selenate and selenite for 6-24 h, elemental Se was found to be the primary metabolite. Whereas, selenocystine appeared to be the major metabolite in cells exposed to 100 μM seleno-L-methionine for 6-24 h. Interestingly, we recorded L-methionine-γ-lyase activity in S9 fraction of cell lysate-an enzyme that directly catalyzes selenomethionine into methylselenol. We also found concurrent reduction of glutathione (GSH) concentration following reaction of seleno-L-methionine with cellular S9 fraction. Moreover, we observed a rapid increase in cellular reactive oxygen species (ROS) generation with increasing seleno-L-methionine exposure dose (100-1000 μM). These findings indicated the rapid cellular metabolism of seleno-L-methionine into methylselenol at higher exposure dose (≥100 μM), and the occurrence of GSH mediated redox cycling of methylselenol--a process that is known to produce reactive oxygen species (ROS). Overall, our results suggest that inorganic and organic selenium are metabolized through different metabolic pathways in rainbow trout hepatocytes. The findings of our study have important implications for understanding the chemical species-specific differences in Se toxicity to fish.     

Induction of metallothionein gene expression by cadmium and the retention of the toxic metal in the tissues of rainbow trout (Salmo gairdneri)

1. When rainbow trout were exposed to cadmium by intraperitoneal injection, there was a rapid (within 3hr) and significant (approx. 63%) loss of the metal from the whole bodies of the fish.2. Of the metal retained in the bodies of the fish (approx. 37% of the injected dose), more than 98% was accounted for collectively among the liver, kidney and gills.3. Subsequent maintenance of the rainbow trout in fresh water for up to 98 days post-metal administration, indicated that there was no further loss of the cadmium accumulated in the organs studied and that the distribution of the metal among the liver, kidney and gills remained unchanged over that period.4. During this 98-day period of maintenance of the fish, tissue concentrations of metallothionein-specific mRNA and metallothionein protein were quantified using riboprobe and ELISA systems respectively. Metallothionein-specific mRNA concentrations increased rapidly (within 24 hr) before falling back to levels similar to, or slightly greater than, those found in control animals. The concentration of metallothionein protein also increased significantly (within 3 days) then remained elevated thereafter.5. Throughout the experimental period, the concentrations of zinc and copper were also monitored in the liver, kidney and gills of the rainbow trout. The concentrations of each ion differed between each of the organs but did not change during the experiment.6. The induction of metallothionein gene expression by cadmium in the liver, kidney and gill of rainbow trout and the subsequent sequestration of the toxic metal is discussed with regard to the relative levels of these other essential metal ions.     

Effects of pH and/or calcium-enriched freshwater on plasma levels of vitellogenin and Ca2+ and on bone calcium content during exogenous vitellogenesis in rainbow trout (Salmo gairdneri)

1. Three year old rainbow trout were exposed to low pH (5.1) and/or calcium-enriched (1.52 mM) freshwater for 10 weeks. 2. Plasma was collected periodically from individually-marked fish for analysis of total calcium and alkaline-labile phosphate (vitellogenin). 3. After the last sample gonadosomatic and hepatosomatic indices were measured and the caudal vertebrae centra were analysed for total calcium content. 4. Female trout exposed to calcium-enriched freshwater had increased plasma vitellogenin levels compared to females in soft water, whereas there was a tendency for low pH to decrease plasma vitellogenin in these fish. 5. The gonadosomatic and hepatosomatic indices were reduced in female trout exposed to acidified water. 6. There was no evidence of bone demineralization in trout exposed to low pH.     

Zn^2＋对罗非鱼呼吸运动的影响和解毒的初探

将罗非鱼分别置于含有1、4、8、12ppm ZnSO_4的软、硬质水中,研究15d 中不同亚致死浓度 Zn~(2+)对鱼呼吸和咳嗽反应频率的影响;观测比较加入 EDTA 络合剂的变化,探索解除鱼Zn~(2+)中毒的方法。结果如下:1)呼吸频率的变化和咳嗽反应明显受 Zn~(2+)浓度影响,浓度越大,呼吸频率曲线越偏离对照曲线,咳嗽反应的频率亦越高,它们可作为鱼中毒和解毒的生理学监测指标。2)硬水组鱼受 Zn~(2+)毒害明显小于软水组。3)EDTA 可解除或减轻 Zn~(2+)对鱼的毒害,但其在软、硬水中的解毒效应无明显差异。     

The effects of various cations on the survival of brown trout, Salmo trutta at low pHs

The survival times of hatchery reared year-old brown trout ( Salmo trutta ) were tested in solutions containing various ions at pHs between 3.5 and 4.0. The effect of calcium in prolonging the survival time compared with that in deionized water at all pHs tested, was more marked than that of sodium which was only effective at pH 4.0. Potassium and magnesium had no effect individually, but aluminium prolonged the survival time in sodium solutions. Fish from a naturally acid river (Tovdal-Norway) survived significantly longer than hatchery reared fish, indicating the importance of the previous history of the fish to their sensitivity to low pH.