Influence of dissolved humic acid on hydrophobic chemical uptake in juvenile rainbow trout

This study measured the chemical uptake of three hydrophobic chemicals (1,2,4-trichlorobenzene (1,2,4-TCB), 1,2,3,4,5-pentachlorobenzene (PeCB) and 2,2',4,4',6,6'-hexachlorobiphenyl (HCBP) with differing octanol-water partition coefficients (log K(ow) values of 3.95, 5.05 and 7.55, respectively) in juvenile rainbow trout (Oncorhynchus mykiss) after 2-day and 4-day aqueous exposures. Because of the affinity of hydrophobic compounds for dissolved organic carbon (DOC) and previous work demonstrating that fish gills take up these three hydrophobic chemicals, we predicted that chemical uptake into the fish would be lowered by the addition of humic acid to the water (1.54, 4.81 and 14.3 mg/l) compared with control fish (no humic acid added to the water). As predicted, humic acid concentrations of >or=4.81 mg/l significantly reduced the whole body concentrations of all three chemicals when compared with 1.54 mg/l humic acid. This effect of humic acid was greatest for HCBP, the chemical with the highest log K(ow), such that chemical uptake was reduced by 3.4-fold for 14.3 mg/l humic acid compared with the control exposure. However, an unexpected finding was that, compared with the control exposure, the lowest concentration of humic acid tested (1.54 mg/l humic acid) significantly increased chemical uptake by up to 112% for the two chemicals with the lower log K(ow), PeCB and 1,2,4-TCB, and did not affect uptake of the high log K(ow) chemical HCBP. We conclude that the ability of DOC to inhibit aqueous uptake of hydrophobic chemicals was dependent on both the concentration of DOC and the log K(ow) of the chemical, but that low humic acid concentrations of approximately 1.5 mg/l can significantly increase uptake of certain chemicals with a log K(ow) between 4 and 5.     

Intersexual diVerences in aggression in juvenile rainbow trout

Agonistic interactions were observed in dyadic contests between size-matched rainbow trout. The results suggest that male juveniles are more aggressive than females.     

The distribution kinetics of waterborne silver-110m in juvenile rainbow trout

Juvenile rainbow trout (Oncorhynchus mykiss) were subjected to a 2-day radioactive pulse of 110mAg at 11.9 microg/l (as AgNO3), followed by a 19-day post-tracer exposure to non-radioactive Ag(I) (3.8 microg/l). The distribution of 110mAg in the gills, liver, intestine, kidney, brain and remaining carcass was investigated over a 19-day post-tracer period. Initially, the intestine contained the highest proportion of the 110mAg burden (34%), however, by day 8, less than 5% of the total radioactivity remained in this tissue. The majority of the 110mAg eliminated from the intestine appeared to distribute to the liver. Eventually, the 110mAg content in the liver accounted for as much as 65% of the total radioactivity in the fish. Apart from the liver and intestine, only the gills and carcass contained any appreciable amount (>5%) of the total body 110mAg content. Liver and gill samples were fractionated using differential centrifugation techniques to discern the subcellular distribution of 110mAg in these tissues. In the liver, the 110mAg levels in the cytosolic fraction increased from 35% to 72% of the total cellular burden between days 8 and 19, respectively. The radioactive pulse in the gills was predominantly found in a membrane compartment termed the nuclear fraction ( approximately 60% of the total). Little change was observed over time (day 8 to day 19) to the subcellular distribution of Ag in the gills. Using size-exclusion chromatography, most ( approximately 70%) of the 110mAg content in the liver cytosol eluted at a molecular weight characteristic of metallothionein. The cytosolic distribution of 110mAg in gills was quite diffuse, occurring primarily in the heavy molecular weight fractions.     

Creatine supplementation affects sprint endurance in juvenile rainbow trout

Fingerling rainbow trout were supplemented with equal amounts of creatine (Cr) by two routes: dietary (12.5 mg Cr per g food); or intraperitoneal injection (0.5 mg Cr per g fish). Endurance in a fixed velocity sprint test (at a speed of 7 BL s(-1)), and resting levels of white muscle metabolites (total creatine [a measure of free creatine plus phosphocreatine (PCr), ATP, lactate and glycogen] were assessed following 7 days of supplementation and compared to controls. None of the treatments had a significant effect on growth, muscle total creatine, percent phosphorylation of creatine, ATP or lactate. However, resting muscle glycogen was elevated in creatine-supplemented fish. Higher muscle glycogen corresponded to significantly greater endurance in creatine-supplemented fish. Although fish do not actively transport additional creatine into the muscle, a mechanism whereby circulating creatine acts to enhance muscle glycogen is present. These results suggest that the improved endurance may be due to an insulin-dependent mechanism (similar to that elucidated in mammalian studies) that allows fish to supercompensate muscle glycogen stores, thus extending endurance through enhanced glycolytic flux.     

Influence of surfagon on rheoreaction of juvenile rainbow trout

The rheoreaction of juvenile rainbow trout (aged 2 months) was studied 30–40 days after a single injection of surfagon (synthetic analogue of gonadotropin-releasing hormone). Individuals subjected to surfagon preferred to move against the water current both in normal and migratory state, while control (not injected) individuals preferred to move downstream. Surfagon strongly modified the rheoreaction of individuals in normal and migratory state.     

Stress does not inhibit induced vitellogenesis in juvenile rainbow trout

Vitellogenin (Vtg) is a widely used biomarker for xenoestrogen exposure in male fishes. In female fishes Vtg can be negatively affected by stress independent of declines in estrogen. However, few data are available on the effect of stress in male fish abnormally producing Vtg, such as when exposed to xenoestrogens. The objective for these studies was to determine the effects of stress on fish forced to produce Vtg. Three weeks prior to the experiment immature juvenile rainbow trout, Oncorhynchus mykiss, were acclimated to the experimental tanks and fed a maintenance ration. We induced Vtg synthesis by injecting 17β-estradiol (E₂) 7 days prior to experimentation. Treatments in duplicate tanks were: (1) no stressor; (2) stressor; (3) E₂; (4) E₂ and stressor. Plasma was collected at time = 0 for baseline measurements from eight fish per tank and Vtg was significantly elevated in treated fish compared to uninjected controls. Water was drained from the stressor tanks then refilled to a level that just covered the backs of the fish. Eight fish were sampled again at 4 and 9 h, and 1, 7, and 14 days of continuous stress. Stressor tanks were refilled with water to pre-stress levels and the fish were sampled after another 2 weeks. Cortisol was significantly elevated from the unstressed fish at 4 h; however, plasma Vtg in the E₂-stimulated fish was not affected by the stressor at any timepoint. These results indicate that fish capture procedures employed in the field or caging experiments likely do not lead to false negative results when plasma Vtg is used as a biomarker for xenoestrogen exposure. It also suggests that the energetic load induced by stress is insufficient to cause a reduction in Vtg, during a continuous E₂ administration, at least within the timepoints examined in this study.     

Relating intramuscular fuel use to endurance in juvenile rainbow trout

This study examined fuel depletion in white muscle of juvenile rainbow trout sprinted to fatigue to determine whether the onset of fatigue is associated with a measurable metabolic change within the muscle and whether muscle glycogen levels influence endurance. In this study, "fuels" refer to any energy-supplying compounds and include glycogen, phosphocreatine (PCr), and ATP. Fuel depletion in white muscle was estimated by the calculation of the anaerobic energy expenditure (AEE; in micromol ATP equivalents g(-1)) from the reduction of PCr and ATP and the accumulation of lactate. Progression of fuel use during sprinting was examined by sampling fish before they showed signs of fatigue and following fatigue. Most of the AEE before fatigue was due to PCr depletion. However, at the first signs of fatigue, there was a 32% drop in ATP. Similarly, when fish were slowly accelerated to a fatiguing velocity, the only significant change at fatigue was a 30% drop in ATP levels. Muscle glycogen levels were manipulated by altering ration (1% vs. 4% body weight ration per day) combined with either daily or no exercise. Higher ration alone led to significantly greater muscle glycogen but had no effect on sprint performance, whereas sprint training led to higher glycogen and an average threefold improvement in sprint performance. In contrast, periodic chasing produced a similar increase in glycogen but had no effect on sprint performance. Taken together, these observations suggest that (i) a reduction in ATP in white muscle could act as a proximate signal for fatigue during prolonged exercise in fish and (ii) availability of muscle glycogen does not limit endurance.     

Effect of AC electroshock on blood features in juvenile rainbow trout

Brief AC electroshock caused significant elevations in plasma cortisol, glucose, osmolality, and chloride levels in rainbow trout within 15min, but plasma protein was reduced at 3h. Circulating granulocytes had declined by 15min, but other haematological features remained unchanged. Thus, AC electrofishing would not be suitable for monitoring physiological conditions in trout if short delays are encountered before blood is sampled.     

Competitive ability is related to metabolic asymmetry in juvenile rainbow trout

Of 55 size–matched pairs of juvenile rainbow trout Oncorhynchus mykiss , each consisting of one high and one low relative metabolic rate fish (relative size difference <5%), the high relative metabolic rate fish was dominant in 36 pairs (65·5%), significantly more often than expected by chance. The probability of a fish winning was related to its relative metabolic rate (measured prior to introduction into the test arena); the higher the relative metabolic rate of a fish compared to its opponent, the greater its probability of being dominant. In the 36 pairs of fish where the low relative metabolic rate fish was the subordinate, its competitive ability was significantly correlated with how closely the two fish were matched in terms of their relative metabolic rates. The smaller the difference between the relative metabolic rates of the dominant high relative metabolic rate fish and subordinate low metabolic rate fish, the greater the competitive ability and feeding success of the low relative metabolic rate subordinate. However, no such relation was found in the 19 pairs of fish where the high relative metabolic rate fish was the subordinate. In these pairs, the competitive ability of the subordinate high relative metabolic rate fish was correlated with its size relative to its dyad partner. The larger the dominant low relative metabolic rate fish, the lower the competitive ability of the subordinate high relative metabolic rate fish.     

Respiratory gas exchange,nitrogenous waste excretion,and fuel usage during aerobic swimming in juvenile rainbow trout

 The types of fuel burned by juvenile rainbow trout (17 g) during a 58-h period of aerobic sustained exercise were studied by respirometry. Attempts to measure fuel usage by depletion (the compositional approach) in these same fish were unsuccessful due to lack of detectable changes in proximate body composition. O₂ consumption, CO₂ excretion, and nitrogenous waste excretion (ammonia-N plus urea-N) were measured in individual fish swum continuously at 55% and 80% of maximum sustainable swimming speed and in non-swimming controls. O₂ consumption and CO₂ excretion increased with swimming speed, and decreased over time. Absolute rates of N excretion were independent of swimming speed and time. Instantaneous aerobic fuel use, as calculated from the respiratory quotients and nitrogen quotients, was approximately 47% lipid, 30% protein, and 23% carbohydrate in non-swimmers at the start of the experiment. With increased swimming speed there was no change in absolute rates of protein oxidation, while lipid and carbohydrate oxidation both increased. Therefore, the relative protein contribution decreased with increasing speed but increased with swimming duration as the oxidation of other fuels declined over time. However, lipid oxidation predominated at all speeds and at all times. The relative contribution of carbohydrate increased with swimming speed and decreased over time. These results suggest that swimming becomes more efficient over time and help resolve uncertainties in the literature. We conclude that lipid is the main fuel of aerobic exercise, that protein catabolism is kept at minimum levels necessary for maintenance, and that carbohydrate oxidation becomes more important with increased white muscle recruitment at higher speed. Accepted: 3 July 1996     

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.     

Skin-penetrating parasites and the release of alarm substances in juvenile rainbow trout

Juvenile trout Oncorhynchus mykiss did not react to the odours of Diplostomum sp. Cercariae alone, indicating that they were incapable of detecting the parasites directly. However, they increased the number of random darts as well as the amount of time spent motionless when exposed to the odours of a conspecific that was being infected by Diplostomum cercariae. These results suggest that even the minor damage inflicted by the cercariae to the fish's skin was enough to cause the release of alarm substances. The effectiveness of the fish's response with respect to the avoidance of parasites remains to be demonstrated.     

The recovery of locomotory activity following exhaustive exercise in juvenile rainbow trout (Oncorhynchus mykiss)

This study investigated the recovery of locomotory activity in exhausted juvenile rainbow trout (Oncorhynchus mykiss, approximately 6-10-cm fork length) in response to two conditions: (1) direct transfer to a range of velocities (0-15 cm s(-1)) in a swim flume (forced swimming) and (2) direct transfer to a pool downstream of a swim channel where a choice of velocities was presented: 2-3 cm s(-1) in the lower half of the pool, a range of velocities from 7 to 40 cm s(-1) in the upper half the pool near the channel entrance, and a velocity of 57 cm s(-1) in a swim channel emptying into the pool (volitional swimming). Exhausted trout showed a pronounced delay in the recovery of normal locomotory activity. With forced swimming, the time required to resume swimming was inversely proportional to water velocity. At 15 cm s(-1), almost all exhausted fish recovered immediately, whereas it took about 1 h for recovery at a current of 5 cm s(-1). In contrast, nonexhausted fish responded to imposed velocity with immediate rheotactic responses (orientation and station holding) at all test velocities. In voluntary swim trials, exhausted trout showed a marked preference for holding station in current in the downstream pool (approximately 11 cm s(-1)) but took, on average, 2 h longer than nonexhausted fish to make transits in the swim channel. Moreover, their ground speed in the swim channel was significantly slower. We conclude that swimming performance is impaired for at least 6 h by exhaustive exercise. Maladaptive behaviors during this time include a preference for current near the surface over cover and a reduced capacity for burst activity, both of which would translate into greater predation risk and reduced ability to forage.     

Respiratory gas exchange,nitrogenous waste excretion,and fuel usage during starvation in juvenile rainbow trout,Oncorhynchus mykiss

Oxygen consumption, CO₂ excretion, and nitrogenous waste excretion (75% ammonia-N and 25% urea-N) were measured daily in 4-g rainbow trout over a 15-day starvation period. Oxygen consumption and CO₂ excretion declined while N excretion increased transiently in the mid-part of the starvation period but was unchanged from control levels at the end. Component losses (as percentage of total fuel used) of protein, lipid, and carbohydrate were 66.5, 31.1, and 2.4% respectively, as measured from changes in body weight and body composition, the latter relative to a control group at day 0. Instantaneous fuel use, as calculated from the respiratory quotients and nitrogen quotients, indicated that relative protein use rose during starvation, but contributed at most 24% of the aerobic fuel (as carbon). Lipid metabolism fell from about 68 to 37%, and was largely replaced by carbohydrate metabolism which rose from 20 to 37%. We conclude that the two approaches measure different processes, and that the instantaneous method is preferred for physiological studies. The compositional method is influence by greater error, and measures the fuels depleted, not necessarily burned, because of possible interconversion and excretion of fuels.Department of Biology, St. Francis Xavier University, P.O. Box 5000, Antigonish, Nova Scotia, Canada B2G 2W5     

Dietary Na does not reduce dietary Cu uptake by juvenile rainbow trout

Rainbow trout Oncorhynchus mykiss fry in moderately hard water were exposed to control or high levels of dietary Cu (c. 6 and 580 ug Cu g food^?1) at one of three levels of Na (1·5, 3·0 or 4·5%) in the diet, i.e. six experimental groups. Fish were fed a 4% body mass ration daily for 28 days and 10 individuals from each group were sampled every 7 days. Concentrations of Cu and Na were measured in the gills, liver, gut and remaining carcass of sampled fish. Growth was not affected and no consistent differences were found in mass, total lengths (L_T) or indices of body condition among any of the groups on any sampling day. Copper concentration was significantly higher in tissues of Cu‐exposed groups, although within treatment types (control Cu v. high Cu diet), it did not differ consistently among groups that received different levels of dietary Na. Tissue Na concentration did not differ among any of the groups and did not show any marked changes over time. In Cu‐exposed groups, the proportion of total body Cu burden contained in the liver approximately doubled over time, from c. 30% on day 7 to c. 60% on day 28. In unexposed fish, the liver maintained c. 25% of the total Cu burden throughout the experiment. In contrast, the proportion of the total body Cu burden contained in the gut decreased somewhat over time in Cu‐exposed fish, from c. 40% on day 7 to c. 30% on day 28, and remained fairly stable at c. 25–30% in control groups, i.e. approximately equal to liver values. In all groups, the carcass contained by far the largest portion of the total Na content (>80%). Measurements made 36 h post‐feeding indicated that all six groups had much higher Na efflux relative to influx, suggesting that the fish were eliminating excess Na taken up from the diet, and differences in Na influx rates were small. Na efflux rate was significantly higher in the high Cu and high Na group than in the high Cu and low Na group. The results indicate that at the concentrations used in this experiment, dietary Na has little effect on dietary Cu uptake by juvenile rainbow trout, and dietary Cu has little effect on Na homeostasis.     

The role of hepatic, renal and intestinal gluconeogenic enzymes in glucose homeostasis of juvenile rainbow trout

Rainbow trout is unable to utilize high levels of dietary carbohydrates and experiences hyperglycemia after consumption of carbohydrate-rich meals. Carbohydrates stimulate hepatic glycolytic activity, but gene expression of the rate-limiting gluconeogenic enzymes glucose-6-phosphatase (G6Pase), fructose-1,6-bisphosphatase (FBPase) and phosphoenolpyruvate carboxykinase (PEPCK) remains high. Although there is significant mRNA expression and activity of gluconeogenic enzymes in trout intestine and kidney, the regulation of these enzymes by diet is not known. We tested the hypothesis that dietary carbohydrate modulates intestinal and renal G6Pase, FBPase and PEPCK. Fish were either fasted or fed isocaloric carbohydrate-free (CF) or high carbohydrate (HC) diets for 14 days. As expected, fish fed HC exhibited postprandial hyperglycemia and enhanced levels of hepatic glucokinase mRNA and activity. Dietary carbohydrates had no significant effect on the expression and activity of PEPCK, FBPase and G6Pase in all three organs. In contrast, fasting enhanced the activity, but not the mRNA expression of both hepatic and intestinal PEPCK, as well as intestinal FBPase. Therefore, the activity of rate-limiting gluconeogenic enzymes in trout can be modified by fasting, but not by the carbohydrate content of the diet, potentially causing hyperglycemia when fed high levels of dietary carbohydrates. In this species consuming low carbohydrate diets at infrequent intervals in the wild, fasting-induced increases in hepatic and intestinal gluconeogenic enzyme activities may be a key adaptation to prevent perturbations in blood glucose during food deprivation. Presented in part at Experimental Biology, April 2006, San Francisco, CA [Kirchner S., Panserat S., Kaushik S. and Ferraris R. FASEB-IUPS-2006 A667.6].     

Effect of total dissolved gas supersaturation on the survival of common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix)

Flood discharge results in total dissolved gas (TDG) supersaturation downstream of a dam during the flood period. Fish suffer death from gas bubble disease (GBD) caused by TDG supersaturation. Nonetheless, current studies mainly attach importance to the survival of benthic fish affected by TDG supersaturation in the Yangtze River in China. Few studies have attempted to investigate the survival of pelagic fish influenced by TDG supersaturated water and compare the tolerance characteristics to TDG supersaturation between benthic and pelagic fish. To identify the survival of fish species that inhabit the various water layers affected by TDG supersaturation, silver carp (Hypophthalmichthys molitrix) (pelagic fish) and common carp (Cyprinus carpio) (benthic fish) were chosen to conduct an acute exposure experiment of four different TDG supersaturation levels (125%, 130%, 135% and 140%). The findings illustrated that the two fish species both exhibited evident aberrant behaviours of maladjustment in TDG supersaturated water. Obvious GBD symptoms were also found in the test fish. The survival probability of silver carp and common carp decreased with increasing levels of TDG supersaturation. The median survival time (ST₅₀) values of the silver carp exposed to four levels of TDG supersaturated water (125%, 130%, 135% and 140%) were 26.84, 7.96, 5.56 and 3.62 h, respectively, whereas the ST₅₀ values of common carp were 53.50, 26.00, 16.50 and 11.70 h, respectively. When compared with common carp, silver carp had a weaker tolerance to TDG-supersaturated water and were vulnerable to GBD. It shows that levels above 125% are not safe for common carp survival. In terms of the tolerance threshold value, silver carp merits further investigation because it showed lower tolerance to TDG than did common carp.     

Nocturnal emergence of juvenile rainbow trout from winter concealment relative to light intensity

This study examined the relationship between light intensity and the number of juvenile rainbow trout (Oncorhynchus mykiss) visible to a snorkeler during February in the Henrys Fork of the Snake River, Idaho, USA. Fish were concealed in the substratum during daylight. Emergence from concealment was observed from 30 to 80 min after real sunset time and began when stars were first visible (pyranometric irradiance, 4.5 × 10^–3 W^–2). Densities of visible fish were negatively correlated with light intensity (r ²=0.81,P<0.001). Later at night, densities decreased in the presence of moonlight and artificial light. Fish were observed to feed at night.     

Differences in metabolic response to Loma salmonae infection in juvenile rainbow trout Oncorhynchus mykiss and brook trout Salvelinus fontinalis

Routine and post-exercise metabolic rates were measured for juvenile rainbow trout Oncorhynchus mykiss and brook trout Salvelinus fontinalis infected with the microsporidium gill parasite Loma salmonae under laboratory conditions. Rainbow trout increased routine and post-exercise metabolic rate in response to infection compared with controls. Brook trout, on the other hand, lowered routine metabolic rate without effecting post-exercise metabolic rate compared to controls. The result of these 2 different strategies may either reflect defense of metabolic scope or a difference in the rate of recovery of the excess post-exercise oxygen consumption between the 2 species in response to the same infection.