How does elevated water temperature affect fish brain? (A neurophysiological and experimental study: Assessment of brain derived neurotrophic factor, cFOS, apoptotic genes, heat shock genes, ER-stress genes and oxidative stress genes).

Water temperature is one of the most important environmental factors affecting the growth and survival of fish. Increased water temperature became a global problem and it is estimated that there will be an increase in water temperature due to global climate change. The physiological mechanism for the effects of high water temperature on the fish brain is not fully known. In the present study, fish were exposed to different temperatures (10 °C/15 °C/20 °C/25°) and brain tissues were sampled 2 h-4h-6h-8h per hour respectively and then we investigated transcriptional changes of BDNF, cFOS, apoptotic genes (caspase 3, Bax, Bcl2), heat shock genes (Hsp70 and Hsp 90) ER-Stress genes (grp78, atf6, and ire1) and oxidative stress genes (CAT, SOD, and GPx) and also immunoflourescence changes of BDNF and cFOSin rainbow trout brain. The results indicated that high temperature stress lead to physiological changes in the fish brain by causing a decrease in mRNA expression levels of CAT, SOD, GPx and Bcl2 and by causing an increase in mRNA expression of BDNF, cFOS, apoptotic genes (caspase 3, Bax), heat shock genes (Hsp70 and Hsp 90) ER-Stress genes (grp78, atf6, and ire1). This study will provide important information to elucidate the physiological mechanisms related to the effects of high water temperature on the fish brain.

Efficacy of a Modified Health Assessment Utilized on Two Genetically Distinct Stocks of Rainbow Trout.

The Rainbow Trout Oncorhynchus mykiss is a salmonid fish that is maintained in aquaculture facilities and used as a source of protein around the world. Veterinarians need accurate, efficient tools and straightforward assessments to evaluate the health status of fish, like Rainbow Trout, that are cultivated on farms. In this study, two genetically distinct broodstock lines of Rainbow Trout originally obtained from two different U.S. states but propagated at Rushing Waters Trout Farm (Palmyra, Wisconsin) were evaluated. Groups originating from both states (State A: n = 20; State B: n = 20) were surveyed with a modified health assessment in a prospective, randomized, single-blind study. The adapted health assessment included external and internal gross evaluations, skin scrapes, gill clips, intestinal scrapes, and calculated indices, including condition factor and the hepatosomatic index. Generally, there were few significant differences in health assessment values between State A and State B fish. When this information was presented to the producer, it was discovered that perceived differences between groups were due to management errors in calculated growth rates. The results of this study add validity to the use of standardized health assessments for evaluating fish within aquaculture.

Climate vulnerability assessment for Pacific salmon and steelhead in the California Current Large Marine Ecosystem.

Major ecological realignments are already occurring in response to climate change. To be successful, conservation strategies now need to account for geographical patterns in traits sensitive to climate change, as well as climate threats to species-level diversity. As part of an effort to provide such information, we conducted a climate vulnerability assessment that included all anadromous Pacific salmon and steelhead (Oncorhynchus spp.) population units listed under the U.S. Endangered Species Act. Using an expert-based scoring system, we ranked 20 attributes for the 28 listed units and 5 additional units. Attributes captured biological sensitivity, or the strength of linkages between each listing unit and the present climate; climate exposure, or the magnitude of projected change in local environmental conditions; and adaptive capacity, or the ability to modify phenotypes to cope with new climatic conditions. Each listing unit was then assigned one of four vulnerability categories. Units ranked most vulnerable overall were Chinook (O. tshawytscha) in the California Central Valley, coho (O. kisutch) in California and southern Oregon, sockeye (O. nerka) in the Snake River Basin, and spring-run Chinook in the interior Columbia and Willamette River Basins. We identified units with similar vulnerability profiles using a hierarchical cluster analysis. Life history characteristics, especially freshwater and estuary residence times, interplayed with gradations in exposure from south to north and from coastal to interior regions to generate landscape-level patterns within each species. Nearly all listing units faced high exposures to projected increases in stream temperature, sea surface temperature, and ocean acidification, but other aspects of exposure peaked in particular regions. Anthropogenic factors, especially migration barriers, habitat degradation, and hatchery influence, have reduced the adaptive capacity of most steelhead and salmon populations. Enhancing adaptive capacity is essential to mitigate for the increasing threat of climate change. Collectively, these results provide a framework to support recovery planning that considers climate impacts on the majority of West Coast anadromous salmonids.

Proactive coping style in early emerging rainbow trout carries a metabolic cost with no apparent return.

The timing with which salmonid larvae emerge from their gravel nests is thought to be correlated with a particular suite of behavioural and physiological traits that correspond to the stress coping style of the individual. Among these traits, aggressiveness, dominance and resilience to stress, are potentially interesting to exploit in aquaculture production. In the present study a series of experiments were performed, with the purpose of characterising behavioural, metabolic and production related traits in rainbow trout juveniles from different emergence fractions. Newly hatched rainbow trout were sorted according to their emergence time from an artificial redd. The early, middle, and late fractions were retained and assessed for their physiological response to stress, growth performance, metabolism, fasting tolerance, and potential for compensatory growth. The early emerging fraction showed proactive behavioural traits; they were faster to reappear following startling, showed a reduced cortisol response following stress, and a reduced metabolic cost of recovery. Emergence time was not correlated with any differences in standard or maximum metabolic rates, but was however, correlated with higher routine metabolic rates, as demonstrated by significantly bigger weight losses during fasting in the early emerging group. Growth rates and feed conversion efficiencies were not significantly different when fish were co-habitated under a restrictive feeding regime, suggesting that early emerging fish are not able to monopolise food resources. The intermediate emerging group, which makes up the bulk of a population and is often ignored, appears to possess the best growth performance traits, possibly because they do not expend excessive energy on dominance behaviour such as the early emerging group, while they are also not overly timid or stress prone such as the late emerging group.

Acute toxicity of Corexit EC9500A and assessment of dioctyl sulfosuccinate as an indicator for monitoring four oil dispersants applied to diluted bitumen.

The present study investigated oil dispersant toxicity to fish species typical of the cooler regions of Canada, together with less well-documented issues pertaining to oil dispersant monitoring. The oil dispersant toxicity of Corexit EC9500A was assessed for the freshwater fish species rainbow trout and the seawater species coho, chinook, and chum, with a final median lethal concentration (LC50) acute lethality range between 35.3 and 59.8 mg/L. The LC50 range was calculated using confirmed 0-h dispersant concentrations that were justified by fish mortality within the first 24 h of exposure and by variability of the dispersant indicator dioctyl sulfosuccinate (DOSS) used to monitor concentrations at later time points. To investigate DOSS as an oil dispersant indicator in the environment, microcosm systems were prepared containing Corexit EC9500A, Finasol OSR52, Slickgone NS, and Slickgone EW dispersants together with diluted bitumen. The DOSS indicator recovery was found to be stable for up to 13 d at 5 °C, 8 d at 10 °C, but significantly less than 8 d at ≥15 °C. After 3 d at temperatures ≥15 °C, the DOSS indicator recovery became less accurate and was dependent on multiple environmental factors including temperature, microbial activity, and aeration, with potential for loss of solvents and stabilizers. A final assessment determined DOSS to be a discrepant indicator for long-term monitoring of oil dispersant in seawater. Environ Toxicol Chem 2018;37:1309-1319. © 2018 SETAC.

Risk assessment of a formamidine pesticide, Amitraz, focusing on thyroid hormone receptors (TRs) in rainbow trout, Oncorhynchus mykiss.

Amitraz, a formamidine pesticide, and their metabolites have the potential to disrupt endocrine homeostasis in a variety of organisms, nevertheless there is a lack of information concerning such effects and underlying mechanisms in any fish species.To evaluate the potential impacts of Trasil (EC; active constituent 200 g amitraz/L), a commercial product of amitraz, on thyroid hormone (TH) homeostasis of rainbow trout (Oncorhynchus mykiss); mRNA levels of thyroid hormone receptors (TRs), TRα and TRß, were determined by RT-PCR soon after sub-lethal administration in a static bio-assay system. The sub-lethal exposure of 0.84 mg/L amitraz resulted in upregulation of both TRαand TRßgenes for muscle and liver, respectively in a tissue-manner, though the differences were found statistically insignificant (P>0.05). The present results emerged an endocrine interaction between amitraz based formulation and TH homeostasis, but still needs further detail studies to a better understanding of TH mechanism in teleosts in response to environmental compounds.

Lactate kinetics of rainbow trout during graded exercise: do catheters affect the cost of transport?

Changes in lactate kinetics as a function of exercise intensity have never been measured in an ectotherm. Continuous infusion of a tracer is necessary to quantify rates of lactate appearance (Ra) and disposal (Rd), but it requires double catheterization, which could interfere with swimming. Using rainbow trout, our goals were to: (1) determine the potential effects of catheters and blood sampling on metabolic rate (O2), total cost of transport (TCOT), net cost of transport (NCOT) and critical swimming speed (Ucrit), and (2) monitor changes in lactate fluxes during prolonged, steady-state swimming or graded swimming from rest to Ucrit. This athletic species maintains high baseline lactate fluxes of 24 µmol kg(-1) min(-1) that are only increased at intensities >2.4 body lengths (BL) s(-1) or 85% Ucrit. As the fish reaches Ucrit, Ra is more strongly stimulated (+67% to 40.4 µmol kg(-1) min(-1)) than Rd (+41% to 34.7 µmol kg(-1) min(-1)), causing a fourfold increase in blood lactate concentration. Without this stimulation of Rd during intense swimming, lactate accumulation would double. By contrast, steady-state exercise at 1.7 BL s(-1) increases lactate fluxes to ~30 µmol kg(-1) min(-1), with a trivial mismatch between Ra and Rd that only affects blood concentration minimally. Results also show that the catheterizations and blood sampling needed to measure metabolite kinetics in exercising fish have no significant impact on O2 or TCOT. However, these experimental procedures affect locomotion energetics by increasing NCOT at high speeds and by decreasing Ucrit.

Effects of ibuprofen on the viability and proliferation of rainbow trout liver cell lines and potential problems and interactions in effects assessment.

Under some conditions ibuprofen was either cytotoxic or cytostatic to rainbow trout cell lines: RTL-W1 (liver) and RTH-149 (hepatoma). Ibuprofen at up to 15 microg/mL was not cytotoxic, regardless of dosing protocols, exposure conditions, viability endpoints, or cell lines. Responses to higher ibuprofen concentrations depended on the test methodology. No cytotoxicity was seen when stock ibuprofen solutions had been prepared in ethanol. For stock solutions in dimethylsulfoxide (DMSO), ibuprofen from 50 to 1500 microg/mL elicited little cytotoxicity in cultures in which the final DMSO concentration was 0.05% (v/v), but was consistently cytotoxic after 24 h for cultures with 0.5% DMSO (v/v). Cytotoxicity was evaluated with alamar Blue (AB) and carboxyfluoroscein diacetate acetoxymethyl ester (CFDA-AM) as measures respectively of metabolic activity and membrane integrity. Effective concentrations (EC50s) for ibuprofen with AB and CFDA-AM depended on whether the stock solution was dosed directly into a culture well or mixed in medium prior to being added to a well. For indirect dosing, ibuprofen was more cytotoxic in medium without fetal bovine serum (FBS), whereas for direct dosing ibuprofen was equally cytotoxic in medium with or without FBS. As judged by AB and CFDA-AM EC50s, dosing ibuprofen was directly 10 to 30 times more cytotoxic. In FBS-containing cultures, which was dosed with increasing ibuprofen and DMSO at 0.05% (v/v), cell proliferation was impaired at 50 and 150 microg/mL ibuprofen. Lipopolysaccharide (LPS) at 50 microg/mL had little influence on these cytotoxic and cytostatic effects of ibuprofen in medium with FBS.

Assessment of thermal stratification within stream pools as a mechanism to provide refugia for native trout in hot, arid rangelands.

Native trout species, such as the redband trout (Oncorhynchus mykiss), occupy thermally harsh stream habitats in hot, arid rangeland basins of the western United States. Declines in the distribution and abundance of these species has generated interest in understanding how these cold water species survive in these systems, as well as in identifying opportunities to restore these species to their former ranges. The purpose of this study was to assess the potential for thermal stratification to provide thermal refuge for redband trout in stream pools characterized by warm intermittent flow conditions on arid rangelands. We studied vertical thermal stratification in two pools during three summers on Boles Creek located on the Modoc Plateau in extreme northeastern California. Water and air temperature data were collected on a 0.5 h time step from 15-Jun through 15-Sep during 1996, 1997, and 2000 using commercial temperature data-loggers. Water temperature was measured at the top (0.3 m below pool surface) and bottom (0.3 m above pool substrates) of each pool. Vertical thermal stratification occurred within these pools creating conditions as much as 7.6 C cooler and consistently more constant at the bottom of pools compared to pool surface waters. Thermal stratification was dependent upon air temperature with the magnitude of stratification increasing as air temperature increased. The magnitude of thermal stratification varied significantly from year to year, likely reflecting variation in annual weather conditions. The thermal regime in the study pools was often near the upper lethal limit reported for redband trout, but temperatures at the bottom of these pools did offer refuge from lethal temperatures realized near the pool surface. Temperatures at pool bottom were consistently above optimal levels published for redbands.

Neuromuscular control of trout swimming in a vortex street: implications for energy economy during the Karman gait.

Approximating the complexity of natural locomotor conditions provides insight into the diversity of mechanisms that enable animals to successfully navigate through their environment. When exposed to vortices shed from a cylinder, fishes hold station by adopting a mode of locomotion called the Kármán gait, whereby the body of the fish displays large, lateral oscillations and the tail-beat frequency matches the vortex shedding frequency of the cylinder. Although field studies indicate that fishes often prefer turbulent flows over uniform currents, the effect of hydrodynamic perturbations on the mechanics, control and energetics of locomotion is still poorly understood. In this study, electromyography is used to measure red and white axial muscle activity for rainbow trout (Oncorhynchus mykiss) holding station in a vortex street. When trout Kármán gait, they show a significantly reduced but still rhythmic pattern of muscle activity compared with that seen when they swim steadily in uniform flow. Specifically, trout selectively activated only their anterior red axial muscles and abandoned the antero-posterior wave of red muscle activity that drives undulatory locomotion in uniform flow. This supports a previously proposed hypothesis that trout are not just swimming in the reduced flow behind a cylinder (drafting). Anterior axial muscle activity was correlated to head amplitude during steady swimming but not during the Kármán gait, indicating that while activation of muscles during the Kármán gait may aid in stability and control, vortices determined overall head motion. Furthermore, anterior red axial muscle activity, the only region of muscle activity shared between both the Kármán gait and steady swimming, had a lower intensity and longer duration during the Kármán gait. At times when paired fins were active during the Kármán gait, there was no axial muscle activity measured, lending support to a passive mechanism of thrust generation in oscillating flows. Comparisons with dead trout towed behind a cylinder confirm this intriguing observation that live trout may temporarily adopt the Kármán gait with no axial muscle activity, revealing paradoxically that at times fish can passively move against turbulent flow. To Kármán gait for prolonged periods, however, trout must adapt to the demands of turbulence by eliciting a shift in neural control strategy. By decoupling motor output both down and across the body, the pattern of rhythmic Kármán gait muscle activity may reflect the entrainment of a central pattern generator to environmental vortices.

The mechanisms and costs of physiological and toxicological acclimation to waterborne silver in juvenile rainbow trout (Oncorhynchus mykiss).

Juvenile rainbow trout were exposed to 0, 0.1, 1, 3, and 5 micro g/l silver (Ag, as AgNO3) for 23 days. Specific growth rate, cumulative food consumption, food-conversion efficiency, and critical swimming speed (U(crit)) were significantly reduced during 5 micro g/l Ag exposure, demonstrating a physiological cost of silver acclimation. Only the 5 microg/l Ag treatment had significant cumulative mortality (5.2%). Fish were most susceptible to silver on days 5 and 15. Exposure to 5 microg/l Ag significantly lowered plasma Na+ and Cl- on days 5 and 10, but plasma ion concentration recovered thereafter. Unidirectional Na+ uptake and gill Na/K-ATPase activity were significantly inhibited by 3 and 5 microg/l Ag exposure. Na+ uptake was inhibited by 3 micro g/l Ag at day 5 alone, whereas the effects at the highest Ag exposure persisted until day 15. Gill Na/K-ATPase was inhibited on day 5 in both the 3 and 5 microg/l Ag treatments but increased to approx. 1.5 times of control levels by day 23. Only the 3 and 5 microg/l Ag treatments produced toxicological acclimation (at least twofold elevations in 168-h LC50 values in fish subsampled on days 15 and 23). We conclude that physiological acclimation results from compensatory changes in Na+ transport at the gills, and that these changes may eventually lead to toxicological acclimation.

Assessment of thyroid function in adult medaka (Oryzias latipes) and juvenile rainbow trout (Oncorhynchus mykiss) using immunostaining methods.

The effect of injected bovine TSH on the pattern of anti-T(4) and anti-T(3) immunostaining of the thyroid tissue was examined in rainbow trout (Oncorhynchus mykiss) and medaka (Oryzias latipes) to determine if the previously reported immunostaining of the cytoplasm of thyrocytes is due to the presence of colloid pinocytotic vesicles and is thus indicative of thyroid hormone release. We hypothesized that the number of immunostained thyrocytes should increase following a TSH challenge, and this should parallel other indicators of increased thyroid activity. In medaka, immunostained thyrocytes were only found following the TSH challenge, and were most marked after 24 to 72 hours; the immunostaining was associated with large colloid-filled cytoplasmic vesicles. In trout, the number and staining intensity of immunostained thyrocytes were increased after the TSH challenge; the immunostaining was present throughout the cytoplasm of the thyrocytes. These findings support the working hypothesis that the immunostaining of the thyrocytes is associated with the pinocytosis of thyroglobulin by the thyrocytes in parallel with an increase in release of thyroid hormone, and that this investigational approach provides a reliable indicator of thyroid hormone release activity.

Toxicity of boron to rainbow trout: a weight-of-the-evidence assessment.

From the large data set available on the toxicity of boron to aquatic organisms, the toxicity of boron to the early life stages of rainbow trout (Oncorhyncus mykiss) is the seminal issue relative to setting water quality criteria and effluent standards. Issues associated with the early life stage studies are the flat concentration-response curve, the low threshold of toxicity, and teratogenic effects observed. Recent laboratory and field studies offer new experimental data that make a weight-of-the-evidence assessment timely. In a re-examination of the effect of boron on the embryo-larval stage in rainbow trout and zebrafish, adverse effects due to boron deficiency are observed which decrease with increasing dose. It was found that low concentrations of boron stimulate embryonic growth in rainbow trout and increase the viability and survival of embryonic zebrafish. As boron concentration is further increased, the dose-response curve becomes flat as homeostatic processes are active; this is followed at higher doses by a new adverse response that increases with increasing dose. As a result, the dose-response relationship is U shaped, consistent with the characteristic shape of an essential micronutrient. Thus, effects originally reported to be toxicity at low exposures rather may be due to boron deficiency. Water analyses in trout hatcheries and field studies in wild trout streams add additional information on the toxicity of boron to trout. Of particular note is a controlled field study carried out in the Firehole River in Yellowstone Park (WY, USA), where trout populations survive and reproduction successfully occurs in natural water containing boron concentrations up to and in some cases greater than 1.0 mg B/L. Teratogenic effects due to boron exposure were not observed in any of these more recent studies.

Assessment of the bioactivity of creosote-contaminated sediment by liver biotransformation system of rainbow trout.

A sediment site in the Lake Jämsänvesi (municipality of Petäjävesi, Finland) contaminated by creosote was investigated to assess the possible ecotoxicological risks it may cause to benthic animals, including ones which may arise due to physical measures in remediating the site. It is suggested that polycyclic aromatic hydrocarbons (PAHs) are bioavailable to fish and other aquatic animals during exposure to contaminated water, sediment, and food. In order to assess toxicological risks of sediment contents to fish, juvenile rainbow trout (Onchorhynchus mykiss) were intraperitoneally dosed with extracts of the creosote-contaminated sediments and their elutriates. This was compared to pristine lake sediment spiked with creosote. Activity of CYP1A1 in trout liver was measured as ethoxyresorufin O-deethylase (EROD). When compared to vehicle controls and the pristine reference sediment (0.9-1.3 pmol/min/mg PMS protein), the extract of creosote-contaminated sediment of Lake Jämsänvesi induced EROD activity up 20-30 times with a dose of 100 mg/kg [total PAHs (mg)/(kg) in fish]. The rise of EROD was associated with increasing levels of PAH metabolites in bile, analyzed as 1-OH pyrene equivalents.

Evaluation of thyroid hormone economy in elasmobranch fishes, with measurements of hepatic 5'-monodeiodinase activity in wild dogfish.

This paper reviews the current understanding of thyroid hormone economy and homeostasis in elasmobranch fishes and considers those measures of the activity of the hypothalamic-pituitary gland-thyroid gland-peripheral tissue axis that are necessary for adequate assessment of thyroid hormone physiology. In particular, we focus on the value of measuring hepatic 5'-monodeiodinase (5'-MDA) activity as an indicator of the animal's cellular production rate of the active thyroid hormone, triiodo-L-thyronine (T(3)). We also examine the characteristics of hepatic 5'-MDA activity, in vitro, in adult female dogfish (Squalus acanthias) collected from Passamaquoddy Bay, New Brunswick, Canada, and in the embryos that they were carrying. T(3) production from T(4) by hepatic homogenates in vitro was time- and temperature-dependent, and was enhanced by the presence of a thiol donor. Michaelis constant (K(m)) and maximum reaction velocity (V(max)) values were 3.8 x 10(-7) M and 0.29 nM T(3)/mg protein/hr, respectively. The inclusion of trimethylamine-N-oxide (TMAO) or a mixture of urea, TMAO, betaine and sarcosine significantly enhanced T(3) production. Hepatic 5'-MDA activity was depressed in fish fasted for 7 days. J. Exp. Zool. 284:492-499, 1999.

Assessment of PCBs and hydroxylated PCBs as potential xenoestrogens: In vitro studies based on MCF-7 cell proliferation and induction of vitellogenin in primary culture of rainbow trout hepatocytes.

In the present study, four structurally diverse polychlorinated biphenyls (PCBs) were chosen from a set of 20 PCBs selected to represent the 154 tetra- through hepta-chlorinated biphenyls. The purpose was to determine estrogenic activities of the chosen PCBs and five of their hydroxylated derivatives (OH-PCBs). A human breast cancer cell line (MCF-7) and primary cultures of rainbow trout (Oncorhyncus mykiss) hepatocytes were used to determine estrogenic effects. The PCBs 2,2',4,6,6'-pentachlorobiphenyl (104) and 2,2',3, 4', 5,6,6'-heptachlorobiphenyl (188), and the hydroxylated PCBs 2,2', 4',6'-tetrachloro-4-biphenylol (4'-50), 2',4', 6'-trichloro-4-biphenylol (4'-30), 2',3,5, 5'-tetrachloro-4-biphenylol (4'-72), 2',3,3',5', 6'-pentachloro-4-biphenylol (4'-112), and 2',3,4',5, 6'-pentachloro-4-biphenylol (4'-121) significantly increased MCF-7 cell proliferation. The coaddition of hydroxytamoxifen, an estrogen antagonist, inhibited increased cell proliferation. The activity of the hydroxylated PCBs 4'-50 and 4'-30 was significantly higher at all nominal concentrations tested as compared to the corresponding PCB, viz., PCB 104. The hydroxylated PCBs 4'-50, 4'-30, 4'-72 and 4'-112 induced vitellogenin synthesis in rainbow trout hepatocytes. Significant differences were found in the MCF-7 system between the parent PCB and its hydroxylated derivative, viz., for 4'-50/4'-30 and 104, and in the rainbow trout hepatocyte assay between 4'-112 and 112, respectively. No activity was observed for PCB 58 in any of the two assays in the present study. Even though cells from two different species (human and fish) are used in the present study, the results obtained by the two methods agree fairly well. In both studies the hydroxylated PCBs were more active than the PCBs, and 4'-30 was the most active compound second only to 17beta-estradiol. http://link.springer-ny. com/link/service/journals/00244/bibs/37n2p145.html