Effects of cadmium chloride on the development of rainbow trout Oncorhynchus mykiss early life stages

The sub-chronic (28–56 days) effects of exposure to low concentrations of cadmium (Cd; 0·05, 0·25, 0·50 and 2·50 μg l⁻¹) shortly following fertilization on embryos, larvae and juvenile rainbow trout Oncorhynchus mykiss were examined. Premature hatching occurred at lower concentrations (0·05 and 0·25 μg l⁻¹ Cd), however, delayed hatching was seen in the 2·50 μg l⁻¹ Cd group, with >90% of hatching occurring on the last day of the hatching period. Larval growth was negatively affected by Cd exposure in a concentration-dependent manner. Larvae exposed to 2·50 μg l⁻¹ Cd were 13·9 ± 0·8% shorter in total length ( L _T) and weighed 22·4 ± 3·5% (mean ± s . e .) less than controls at the end of the exposure period. Plasma sex steroid concentrations (oestradiol in juvenile females and 11-ketotestosterone in juvenile males) were elevated (four- to 10-fold over controls) in exposed fish in both males and females, following 28 days of exposure to 0·05, 0·25 and 0·50 μg l⁻¹ Cd, respectively. These results suggest that environmentally realistic concentrations (in the μg l⁻¹ range) of Cd can affect the development of O. mykiss impacting embryos, larvae and juvenile fish.     

Morphological changes in the gills of Lophiosilurus alexandri exposed to un-ionized ammonia

The average lethal concentration of un-ionized ammonia (48-h LC₅₀NH₃) has been determined by the static assay for larvae (0.48 mg l⁻¹) and alevins (0.92 mg l⁻¹) of 'pacamã' Lophiosilurus alexandri. Studies by light and scanning electron microscopes at the greatest concentration of NH₃ (0.99 mg l⁻¹ for larvae and 1.5 mg l⁻¹ for alevins) have shown that the changes in the cells and branchial tissue were more intense in the alevins.     

The effect of sodium and calcium concentrations on the hatching of eggs and the survival of the yolk sac fry of brown trout, Salmo trutta L. at low pH

The effect of various external concentrations of sodium and calcium on the survival and hatching of brown trout, Salmo trutta , eggs at pH 4.5 was tested. A calcium concentration of approximately 10 ppm (500 μE l⁻¹) enables freshly fertilized eggs to survive whereas eyed ova are tolerant of deionized water acidified by sulphuric acid with no other ions added. Concentrations of sodium and calcium of 1 ppm (44 and 50 μE l⁻¹, respectively) are sufficient to ensure the successful hatching of eyed ova and subsequent survival of the alevins. At pH 4.5 hatching is prolonged by the alevins passing through a temporary encapsulated stage.     

Dissolved oxygen concentration and emergence of sea trout fry from natural redds in tributaries of the River Rhine

A total of 4289 sea trout Salmo trutta alevins swam up from six of eleven covered redds. They emerged only from those redds where the mean dissolved oxygen level was >6·9 mg l⁻¹ and the Sorting coe.cient, an indicator of intragravel void space, was <5. These results are discussed in relation to field studies using artificially buried salmonid eggs but also in relation to the ongoing programme for the reintroduction of Atlantic salmon in tributaries of the River Rhine.     

The effects of acid and acid/aluminum exposure on circulating plasma cortisol levels and other blood parameters in the rainbow trout, Salmo gairdneri

Softwater (Ca²⁺=50, Na⁺= 50(μequiv. l⁻¹) acclimated rainbow trout were fitted with chronic arterial catheters to allow for repetitive blood sampling. After 48 h recovery they were then exposed to either control (pH 6.5, Al = 0μg l⁻¹), acid (pH 4.8, Al = 0μg l⁻¹) or acid plus aluminum (pH 4.8, A1 = 112 μg l⁻¹) conditions for 72 h. Parameters measured included blood glucose, lactate, haemoglobin, haematocrit and plasma Na⁺, Cl⁻, protein and cortisol.
Exposure to pH 4'8 alone caused no mortality, a moderate ionoregulatory disturbance and a transient elevation in plasma cortisol. All other parameters were not significantly different from controls. Addition of aluminum to this exposure caused 100% mortality with a mean survival time of only 27.0 h. There was a marked decrease in plasma ions, hyperglycemia, lactate accumulation, haemoconcentration, red cell swelling, and a sharp rise in plasma cortisol becoming greatly increased as the fish neared death. The mechanism of toxicity of acute acid/aluminum exposure, the role for cortisol under such conditions, and the validity of cortisol and glucose as indicators of stress in fish are discussed.     

The effect of humic substances on the toxicity of aluminium to adult rainbow trout, Oncorhynchus mykiss (Walbaum)

Some physiological parameters were measured in adult rainbow trout during a 10-day exposure to 180 μg Al_total l⁻¹ in acid water (pH 4.7) with or without humic substances (10 mg l⁻). The fish were acclimatized to pH 5.0 for 7 days prior to the experimental treatments.
Chemical analyses revealed that, in the presence of human substances, 74–80% of the A1 was organic bound, while in the absence of humic substances most of the Al(987percnt;) occurred in the inorganic form.
Al bound to humic substances (13–150 μg l⁻¹) did not alter the plasma NaCl-concentration, nor the haematocrit value, of rainbow trout during an exposure period of 10 days. This contrasts with the high death rate obtained within 2–3 days when most of the A1 (175 μg l⁻¹) was in the inorganic form. The lethality was accompanied by a 25% decrease in the plasmaconcentration of NaCl and a doubling of the haematocrit value. Bulk analysis revealed that when the metal was present in inorganic forms the total Al content of the gills (75 μg A1 g⁻¹ wet weight) was 15 times higher than when it was present as bound to the humic substances. These experiments showed that the accumulation of A1 at the gills was accompanied by physiological disturbances, both being a function of the chemical speciation of Al.     

Growth, mineral uptake and skeletal calcium deposition in brown trout, Salmo trutta L., yolk-sac fry exposed to aluminium and manganese in soft acid water

In 30-day exposures in artificial soft water medium, survival of brown trout alevins was not affected by low pH (4.5,4.8, 5.4), by low calcium concentration (10.25 μmol l⁻¹) or by manganese (≤20 μmol l⁻¹), but was impaired by aluminium (6–8 μmol l⁻¹) at low calcium concentration (10 μmol l⁻¹) irrespective of pH (4.5 or 5.4). Manganese (6.6, 20 μmol l⁻¹) impaired net calcium uptake and calcium deposition in the skeleton at low calcium concentration (25 μmol l⁻¹) irrespective of pH. Aluminium (2–8 μmol l⁻¹) impaired gross development, net uptake of calcium, potassium and sodium, and calcium deposition in the skeleton, and slightly increased the net loss of magnesium, some of these effects being more severe at calcium concentration 10 μmol l⁻¹ than 50 μmol l⁻¹, and some more severe at pH 5.4 than pH 4.5. Net uptake of calcium and sodium were impaired at low pH (4.5, 4.8), and skeletal calcium deposition was impaired at low calcium concentration (10 μmol l⁻¹), but these effects of low pH and low calcium concentration were slight compared with those of the trace metals. The possible role of trace metals in reports of the deleterious effects on fish of low pH levels is discussed.     

The effect of oxygen on the growth of Oncorhynchus mykiss embryos with and without a chorion

Growth was measured in dechorionated and chorionated embryos of rainbow trout Oncorhynchus mykiss exposed to three oxygen treatments: hypoxia, normoxia and hyperoxia (nominally 5, 10 and 15 mg l⁻¹). Dechorionated embryos grew faster than chorionated ones in all three oxygen treatments. Similar patterns, however, were found in both dechorionated and chorionated groups; embryos exposed to hypoxia grew less than normoxic and hyperoxic exposed embryos. Moreover, there was no significant interaction between the effect of oxygen levels and presence or absence of the chorion on growth after hatching. These results suggest that the effect of oxygen on growth was not affected by the presence of the chorion.     

Cadmium uptake by rainbow trout, Salmo gairdneri eggs and alevins

The uptake of cadmium by eggs and alevins of rainbow trout from water concentrations of between 0.01 and 50.0 mg Cd 1⁻¹ was investigated. The cadmium content of eggs and alevins increased with time and with exposure concentration. Lower cadmium levels were detected in alevins than in eggs. Most of the cadmium (98%) in the eggs was found to be associated with the egg membrane or chorion. This explains the considerable reduction in cadmium concentration observed in alevins after hatching. Alevins hatching from eggs which had been exposed to cadmium survived longer in cadmium than alevins not exposed as eggs. This suggests that the pretreatment of eggs with cadmium serves some protective function. Behavioural and pathological signs of cadmium poisoning such as erratic swimming and blood clotting in alevins were observed.     

Cadmium toxicity to rainbow trout, Salmo gairdneri Richardson: a study of eggs and alevins

Cadmium toxicity to rainbow trout was evaluated at two cadmium concentrations (0.01, 0.10 mg1⁻¹) and in water without added cadmium using eggs reared through the larval (alevin) phase to swim-up fry. Exposure to 0.1 mg Cd 1⁻¹ promoted premature hatching, retarded growth, increased rates of mortality and the occurrence of developmental abnormalities such as spinal curvature and blood clots. Three methods of holding the eggs and alevins during the toxicity test were compared. Hatching time and success, alevin mortality, growth and development were not influenced by the method of containment but of the three methods, a subdivided perforated box system proved particularly useful for monitoring the responses of specific animals throughout the toxicity test.     

Effect of rearing density on thyroid and interrenal gland activity and plasma and hepatic metabolite levels in rainbow trout, Salmo gairdneri Richardson

There were significant inverse correlations between rearing density of rainbow trout, Salmo gairdneri Richardson, and final body weight, plasma L-thyroxine (T₄), trüodo-L-tryronine (T₃), cortisol and protein concentrations, plasma T₄/T₃ ratios and thyroid epithelial cell height. In addition, hepatosomatic indices and plasma free fatty acid concentrations were higher in fish reared at low (134 g 1⁻¹) density compared with groups reared at medium (210g1⁻¹) or high density (299g 1⁻¹), and the post-feeding (3.5-4h) elevation in plasma glucose and triglyceride levels evident in trout maintained at low rearing density was not found in those fish reared at higher densities. There were no significant effects of rearing density on hematocrit, carcass composition, hepatic glycogen and lipid levels and interregnal nucleus size.     

Resistance to cadmium by pretreated rainbow trout alevins

A toxicity test with cadmium concentrations ranging from 0·1 to 100·0 mg Cd 1⁻¹ was used to assess the effect of cadmium pretreatment on rainbow trout ( Salmo gairdneri Richardson) alevins. The median period of survival for fish pretreated at 0·01 mg Cd 1⁻¹ was found to be increased at test concentrations up to 10mg Cd 1⁻¹ compared with alevins pretreated with dilution water. However, at concentrations above 10mg Cd 1⁻¹ pretreatment at 0·01 mg Cd 1⁻¹ reduced the median period of survival.     

Intracellular ion levels in erythrocytes and hepatocytes isolated from three teleost species

Intracellular concentrations of Na⁺ and K⁺ were similar (∼75 mmol l⁻¹) in rainbow trout Oncorhynchus mykiss hepatocytes directly following isolation by collagenase digestion, but partial recovery occurred over 6 h with K⁺ levels increasing to 110 mmol l⁻¹ and Na⁺ levels decreasing to 42 mmol l⁻¹. Black bullhead Ameiurus melas hepatocytes exhibited higher intracellular concentrations of K⁺ (90 mmol l⁻¹) than Na⁺ (55 mmol l⁻¹) with no recovery occurring over 6 h following cell isolation. Concentrations of Na⁺, K⁺ and Cl⁻ in eel Anguilla rostrata hepatocytes were similar (∼ 55 mmol l⁻¹) following isolation, with no recovery occurring over time. Erythrocytes from all species apparently did not experience an intracellular ion imbalance following isolation as indicated by high K⁺ levels (<140 mmol l⁻¹) and low Na⁺ levels (<40 mmol l⁻¹) during the entire 24-h monitoring period. Although hepatocytes from all species exhibited an ion imbalance post-isolation, comparison of their in vitro intracellular Na⁺ and K⁺ concentrations with those in plasma demonstrated that directionally correct ion gradients still exist across the cell membrane, albeit differing from those that would be found in the tissue in vivo .     

Effect of Cytochalasin B on Eggs of Atlantic Salmon and Rainbow Trout

Cytochalasin B action has been investigated in polyploidisation and gynogenesis in Atlantic salmon and rainbow trout. Solutions of 10, 20, 30 and 50 mg/1 were used to treat eggs from 35 h° to the 2-, 4- and 8-cell stages for rainbow trout, from 35 h° to the 2- and 4-cell stages, and 50 h° to the 8-cell stage for salmon. Survival rate was low in all treated groups. Low frequencies of polyploid and gynogenetic individuals were observed in embryos and alevins by means of two methods: DNA analysis by flow cytometry and chromosome counting. All polyploids observed were triploids without evidence of mosaicism. A few spontaneous diploid gynogenetic rainbow trout were registrated, as well as spontaneous triploid salmon, in controls.     

Effect of excess vitamin D3 on calcium metabolism in rainbow trout Salmo gairdneri Richardson

The effect of excess vitamin D₃ on calcium metabolism in rainbow trout was investigated. Trout were reared for 24 weeks on diets containing 4000, 104 000 or 1 004 000 iu of supplemental vitamin D₃ kg^-1 dry diet. No effect of excess vitamin D₃ was detected in the weight gain, feed efficiency or total mortalities of trout in the different groups. None of the fish showed any signs of hypercalcaemia and no significant difference was detected in the calcium, magnesium, phosphorus and sodium content of the trout vertebrae or total carcass. However, there was a significant increase in the calcium, magnesium and phosphorus content of the skin in trout reared on the highest supplemental level of vitamin D₃ (1 004 000 iu kg^-1 diet). No overt signs of renal calcinosis were detected in any of the trout. This study indicates that trout are resistant to excess vitamin D₃, and that excess vitamin D₃ does not appear to be related to the increased incidence of renal calcinosis in rainbow trout.     

The effects of environmental factors on rainbow smelt Osmerus mordax embryos and larvae

Experiments were conducted to identify environmental factors that influence the survival of rainbow smelt Osmerus mordax during their early life stages. Developing rainbow smelt embryos and yolk-sac larvae were cultured under controlled conditions with different dissolved oxygen (DO; 1·09, 2·18, 4·37 and 6·55 mg l⁻¹, pH (4·0, 4·5, 5·0, 5·5, 6·0 and 7·0), nitrate ( 0·7, 3·6, 7·3, 14·6 and 29·2 mg l⁻¹), phosphate (0·04, 0·21, 0·42, 2·08 and 4·17 mg l⁻¹) and salinity (0, 5, 10, 15, 20 and 30) levels. Rainbow smelt embryos were also incubated with simulated tidal salinity fluctuations (2–28), ultraviolet radiation (irradiances of 2·8, 6·2 and 5·1 W m⁻²) and under natural conditions in two rainbow smelt spawning rivers. In the laboratory, hatch was only impaired under the lowest DO and pH conditions (0 and 13% hatch, respectively) and at highest constant salinity levels (0% hatch). Larval survival was only affected by pH levels ≤5·0. The experiment that compared hatch under natural conditions was terminated when embryos became covered with silt and fungus. These results suggest that water acidification, sediment and fungal growth may affect rainbow smelt survival during their early life stages.     

Antifungal effect of gaseous nitric oxide on mycelium growth, sporulation and spore germination of the postharvest horticulture pathogens, Aspergillus niger, Monilinia fructicola and Penicillium italicum

Aim:  To evaluate the antifungal activity of nitric oxide (NO) against the growth of the postharvest horticulture pathogens Aspergillus niger , Monilinia fructicola and Penicillium italicum under in vitro conditions.
Methods and Results:  Different volumes of NO gas were injected into the Petri dish headspace to obtain the desired concentrations of 50–500  μ l l⁻¹ . The growth of the fungi was measured for 8 days of incubation in air at 25°C . All concentrations of NO were found to produce an antifungal effect on spore germination, sporulation and mycelial growth of the three fungi, with the most effective concentration for A. niger and P. italicum being 100 and 500  μ l l⁻¹ for M. fructicola .
Conclusions:  Short-term exposure to a low concentration of NO gas was able to inhibit the subsequent growth of A. niger , M. fructicola and P. italicum .
Significance and Impact of the Study:  NO gas has potential use as a natural fungicide to inhibit microbial growth on postharvest fruit and vegetables.     

Effect of acute and chronic ammonia and nitrite exposure on oxygen consumption and growth of juvenile big bellied seahorse

Juvenile big bellied seahorse Hippocampus abdominalis were exposed acutely and chronically to elevated ammonia and nitrite {24 h exposure: 0·01, 5·0, 10·1, 14·8 and 19·9 mg l⁻¹ total ammonia-nitrogen [TA-N] and <0·001, 74·4, 99·2 and 123·6 mg l⁻¹ [NO₂-N] nitrite-nitrogen and 35 days exposure: 0·11, 0·55, 1·67 and 3·07 mg l⁻¹ TAN and <0·001, 0·92, 4·67 and 9·10 mg NO₂-N l⁻¹}. Significant ( P <0·001) increases in oxygen consumption rate and ventilation frequency occurred at 14·8, 19·9 mg l⁻¹ TA-N and 99·2, 123·6 mg l⁻¹ NO₂-N for acutely exposed fish. Oxygen consumption rate was significantly ( P <0·05) elevated at 1·67 and 3·07 mg l⁻¹ TA-N in chronically treated fish and ventilation frequency increased significantly ( P <0·05) at 0·55, 1·67, 3·07 mg l⁻¹ TA-N and 4·59, 9·10 mg l⁻¹ NO₂-N. There were no significant differences in growth between controls and ammonia exposed fish. Mortalities occurred at 14·8, 19·9 mg l⁻¹ TA-N.     

Studies on the relationship between osmotic or ionic regulation and thyroid gland activity in two salmonid fishes, Salmo gairdneri Richardson and Oncorhynchus kisutch Walbaum

In freshwater-acclimated rainbow trout a single intraperitoneal injection of ovine TSH significantly elevated plasma thyroxine (T₄) levels within 1 h after the injection. In seawater adapted trout the increase in T₄ after TSH-treatment was not evident until 6 h after the injection. TSH caused a transient fall in plasma Na⁺ and Cl^- between 3 h and 9 h after the injection in seawater-adapted fish and plasma Na⁺ was lowered in freshwater-adapted trout 24 h after the injection. Although there were clear histological changes in the thyroid gland of freshwater-adapted trout after TSH-injection, no such changes were evident in seawater-adapted fish.
Plasma thyroid hormone levels and thyroid histology in freshwater-adapted rainbow trout and coho salmon transferred to sea water, and seawater-adapted trout transferred to fresh water showed no consistent changes compared with controls.
The data are interpreted to indicate that although ambient salinity may have indirect effects on thyroid activity there is no direct involvement in ionic or osmotic regulation in the two species.     

Kinetic characteristics of the sodium uptake mechanism during the development of embryos and fry of Atlantic salmon, Salmo salar L., in an improved water quality

The kinetics of active sodium uptake in dechorionated embryos, yolk-sac fry and start-feed fry of Atlantic salmon were compared in two groups reared either in low conductivity, untreated, river water (conductivity ∼ 46 μS cm⁻¹, pH 5.75), or in 'improved' river water buffered with sea water (conductivity ∼2200 μS cm ¹, pH 6.56), the latter treatment often being used in commercial hatcheries to avoid problems associated with periodic acidification.
Maximal transport rate ( V _max) increased during development in both groups but was always significantly higher in embryos and fry maintained in untreated river water. Values for K _m were not seen to vary during development up to 12 weeks after hatching and were not significantly different between groups, or from values reported for adult Atlantic salmon in fresh water.
The results are discussed with respect to the influence of Na⁺ concentrations in the perivitelline fluid of developing eggs and in the external medium surrounding fry on V _max and K _m. The ability of fry reared entirely in buffered river water to maintain sodium balance following transfer to untreated river water is also considered.