Towards science-based sediment quality standards-Effects of field-collected sediments in rainbow trout (Oncorhynchus mykiss).

Sediments can act as long-term sinks for environmental pollutants. Within the past decades, dioxin-like compounds (DLCs) such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs) have attracted significant attention in the scientific community. To investigate the time- and concentration-dependent uptake of DLCs and PAHs in rainbow trout (Oncorhynchus mykiss) and their associated toxicological effects, we conducted exposure experiments using suspensions of three field-collected sediments from the rivers Rhine and Elbe, which were chosen to represent different contamination levels. Five serial dilutions of contaminated sediments were tested; these originated from the Prossen and Zollelbe sampling sites (both in the river Elbe, Germany) and from Ehrenbreitstein (Rhine, Germany), with lower levels of contamination. Fish were exposed to suspensions of these dilutions under semi-static conditions for 90 days. Analysis of muscle tissue by high resolution gas chromatography and mass spectrometry and of bile liquid by high-performance liquid chromatography showed that particle-bound PCDD/Fs, PCBs and PAHs were readily bioavailable from re-suspended sediments. Uptake of these contaminants and the associated toxicological effects in fish were largely proportional to their sediment concentrations. The changes in the investigated biomarkers closely reflected the different sediment contamination levels: cytochrome P450 1A mRNA expression and 7-ethoxyresorufin-O-deethylase activity in fish livers responded immediately and with high sensitivity, while increased frequencies of micronuclei and other nuclear aberrations, as well as histopathological and gross pathological lesions, were strong indicators of the potential long-term effects of re-suspension events. Our study clearly demonstrates that sediment re-suspension can lead to accumulation of PCDD/Fs and PCBs in fish, resulting in potentially adverse toxicological effects. For a sound risk assessment within the implementation of the European Water Framework Directive and related legislation, we propose a strong emphasis on sediment-bound contaminants in the context of integrated river basin management plans.

Determination of Metabolic Stability Using Cryopreserved Hepatocytes from Rainbow Trout (Oncorhynchus mykiss).

Trout provide a relatively easy source of hepatocytes that can be cryopreserved and used for a range of applications including toxicity testing and determination of intrinsic clearance. Standard protocols for isolating, cryopreserving, and thawing rainbow trout hepatocytes are described, along with procedures for using fresh or cryopreserved hepatocytes to assess metabolic stability of xenobiotics in fish by means of a substrate depletion approach. Variations on these methods, troubleshooting tips, and directions for use of extrapolation factors to express results in terms of in vivo intrinsic clearance are included. These protocols have been developed for rainbow trout, but can be adapted to other fish species with appropriate considerations.

Effect of temperature and diet on wound healing in Atlantic salmon (Salmo salar L.).

Compromised skin integrity of farmed Atlantic salmon, commonly occurring under low temperature and stressful conditions, has major impacts on animal welfare and economic productivity. Even fish with minimal scale loss and minor wounds can suffer from secondary infections, causing downgrading and mortalities. Wound healing is a complex process, where water temperature and nutrition play key roles. In this study, Atlantic salmon (260 g) were held at different water temperatures (4 or 12 °C) and fed three different diets for 10 weeks, before artificial wounds were inflicted and the wound healing process monitored for 2 weeks. The fish were fed either a control diet, a diet supplemented with zinc (Zn) or a diet containing a combination of functional ingredients in addition to Zn. The effect of diet was assessed through subjective and quantitative skin histology and the transcription of skin-associated chemokines. Histology confirmed that wound healing was faster at 12 °C. The epidermis was more organised, and image analyses of digitised skin slides showed that fish fed diets with added Zn had a significantly larger area of the epidermis covered by mucous cells in the deeper layers after 2 weeks, representing more advanced healing progression. Constitutive levels of the newly described chemokines, herein named CK 11A, B and C, confirmed their preferential expression in skin compared to other tissues. Contrasting modulation profiles at 4 and 12 °C were seen for all three chemokines during the wound healing time course, while the Zn-supplemented diets significantly increased the expression of CK 11A and B during the first 24 h of the healing phase.

Prediction of Benthic Impact for Salmon Net-Pens Based on the Balance of Benthic Oxygen Supply and Demand

The ratio between oxygen supply and oxygen demand was examined as a predictor of benthic response to organic enrichment caused by salmon net-pen aquaculture. Oxygen supply to the benthos was calculated based on Fickian diffusion and near-bottom flow velocities. A strong linear correlation was found between measured carbon sedimentation rates and rates of benthic metabolism. This relationship allowed an estimation of oxygen demand based on sedimentation rates. Comparison of several production sites in Maine (USA) coastal waters showed that for sites where oxygen demand exceeded supply benthic impacts were high and for sites where oxygen supply exceeded demand benthic impacts were low. These findings were summarized in the form of a predictive model that should be useful in siting salmon production facilities.

qPCR counts of mRNA for hepatic reference gene selection in juvenile female Atlantic salmon from perturbation experiments under artificial temperature conditions

The dataset contains raw data (quantification cycle) for a study which determined the most suitable hepatic reference genes for normalisation of qPCR data orginating from juvenile Atlantic salmon (14 days) exposed to 14 and 22 degrees C. These results will be useful for anyone wanting to study the effects of climate change/elevated temperature on reproductive physiology of fish (and perhaphs other vertebrates).

qPCR counts of mRNA for hepatic reference gene selection in adult female Atlantic salmon from perturbation experiments under artificial temperature conditions

The dataset contains raw data (quantification cycle) for a study which determined the most suitable hepatic reference genes for normalisation of qPCR data orginating from adult (entire reproductive season) Atlantic salmon (14 days) exposed to 14 and 22 degrees C. These results will be useful for anyone wanting to study the effects of climate change/elevated temperature on reproductive physiology of fish (and perhaphs other vertebrates). In addition, a target gene (vitellogenin) has normalised using an inappropriate and an 'ideal' reference gene to demonstrate the consequences of using an unstable reference gene for normalisation. For the adult experiment, maiden and repeat adult females were held at the Salmon Enterprises of Tasmania (SALTAS) Wayatinah Hatchery (Tasmania, Australia) at ambient temperature and photoperiod in either 200 (maidens) or 50 (repeats) m3 circular tanks at stocking densities of 12-18, and 24-36 kg m-3 for maidens and repeats, respectively, until transfered to the experimental tanks.

Effect of short-term fasting prior to slaughter on welfare and meat quality or rainbow trout (Oncorhynchus mykiss)

El manejo pre-sacrificio es de vital importancia en acuicultura, ya que afecta tanto a las reacciones fisiológicas como a los procesos bioquímicos post mortem, y por tanto al bienestar y a la calidad del producto. El ayuno pre-sacrificio se lleva a cabo de forma habitual en acuicultura, ya que permite el vaciado del aparato digestivo de restos de alimento y heces, reduciendo de esta manera la carga bacteriana en el intestino y la dispersión de enzimas digestivos y potenciales patógenos a la carne. Sin embargo, la duración óptima de este ayuno sin que el pez sufra un estrés innecesario no está clara. Además, se sabe muy poco sobre la mejor hora del día para realizar el sacrificio, lo que a su vez está regido por los ritmos diarios de los parámetros fisiológicos de estrés. Finalmente, se sabe que la temperatura del agua juega un papel muy importante en la fisiología del estrés pero no se ha determinado su efecto en combinación con el ayuno. Además, las actuales recomendaciones en relación a la duración óptima del ayuno previo al sacrificio en peces no suelen considerar la temperatura del agua y se basan únicamente en días y no en grados día (ºC d). Se determinó el efecto del ayuno previo al sacrificio (1, 2 y 3 días, equivalente a 11,1-68,0 grados día) y la hora de sacrificio (08h00, 14h00 y 20h00) en trucha arco iris (Oncorhynchus mykiss) de tamaño comercial en cuatro pruebas usando diferentes temperaturas de agua (Prueba 1: 11,8 ºC; Prueba 2: 19,2 ºC; Prueba 3: 11,1 ºC; y Prueba 4: 22,7 ºC). Se midieron indicadores biométricos, hematológicos, metabólicos y de calidad de la carne. En cada prueba, los valores de los animales ayunados (n=90) se compararon con 90 animales control mantenidos bajo condiciones similares pero nos ayunados. Los resultados sugieren que el ayuno tuvo un efecto significativo sobre los indicadores biométricos. El coeficiente de condición en los animales ayunados fue menor que en los controles después de 2 días de ayuno. El vaciado del aparato digestivo se produjo durante las primeras 24 h de ayuno, encontrándose pequeñas cantidades de alimento después de 48 h. Por otra parte, este vaciado fue más rápido cuando las temperaturas fueron más altas. El peso del hígado de los animales ayunados fue menor y las diferencias entre truchas ayunadas y controles fueron más evidentes a medida que el vaciado del aparato digestivo fue más rápido. El efecto del ayuno hasta 3 días en los indicadores hematológicos no fue significativo. Los niveles de cortisol en plasma resultaron ser altos tanto en truchas ayunadas como en las alimentadas en todas las pruebas realizadas. La concentración media de glucosa varió entre pruebas pero mostró una tendencia a disminuir en animales ayunados a medida que el ayuno progresaba. En cualquier caso, parece que la temperatura del agua jugó un papel muy importante, ya que se encontraron concentraciones más altas durante los días 2 y 3 de ayuno en animales mantenidos a temperaturas más bajas previamente al sacrificio. Los altos niveles de lactato obtenidos en sangre parecen sugerir episodios de intensa actividad muscular pero no se pudo encontrar relación con el ayuno. De la misma manera, el nivel de hematocrito no mostró efecto alguno del ayuno y los leucocitos tendieron a ser más altos cuando los animales estaban menos estresados y cuando su condición corporal fue mayor. Finalmente, la disminución del peso del hígado (índice hepatosomático) en la Prueba 3 no se vio acompañada de una reducción del glucógeno hepático, lo que sugiere que las truchas emplearon una estrategia diferente para mantener constantes los niveles de glucosa durante el periodo de ayuno en esa prueba. En relación a la hora de sacrificio, se obtuvieron niveles más bajos de cortisol a las 20h00, lo que indica que las truchas estaban menos estresadas y que el manejo pre-sacrificio podría resultar menos estresante por la noche. Los niveles de hematocrito fueron también más bajos a las 20h00 pero solo con temperaturas más bajas, sugiriendo que las altas temperaturas incrementan el metabolismo. Ni el ayuno ni la hora de sacrificio tuvieron un efecto significativo sobre la evolución de la calidad de la carne durante los 3 días de almacenamiento. Por el contrario, el tiempo de almacenamiento sí que parece tener un efecto claro sobre los parámetros de calidad del producto final. Los niveles más bajos de pH se alcanzaron a las 24-48 h post mortem, con una lata variabilidad entre duraciones del ayuno (1, 2 y 3 días) en animales sacrificados a las 20h00, aunque no se pudo distinguir ningún patrón común. Por otra parte, la mayor rigidez asociada al rigor mortis se produjo a las 24 h del sacrificio. La capacidad de retención de agua se mostró muy estable durante el período de almacenamiento y parece ser independiente de los cambios en el pH. El parámetro L* de color se incrementó a medida que avanzaba el período de almacenamiento de la carne, mientras que los valores a* y b* no variaron en gran medida. En conclusión, basándose en los resultados hematológicos, el sacrificio a última hora del día parece tener un efecto menos negativo en el bienestar. De manera general, nuestros resultados sugieren que la trucha arco iris puede soportar un período de ayuno previo al sacrificio de hasta 3 días o 68 ºC d sin que su bienestar se vea seriamente comprometido. Es probable que con temperaturas más bajas las truchas pudieran ser ayunadas durante más tiempo sin ningún efecto negativo sobre su bienestar. En cualquier caso, se necesitan más estudios para determinar la relación entre la temperatura del agua y la duración óptima del ayuno en términos de pérdida de peso vivo y la disminución de los niveles de glucosa en sangre y otros indicadores metabólicos. SUMMARY Pre-slaughter handling in fish is important because it affects both physiological reactions and post mortem biochemical processes, and thus welfare and product quality. Pre-slaughter fasting is regularly carried out in aquaculture, as it empties the viscera of food and faeces, thus reducing the intestinal bacteria load and the spread of gut enzymes and potential pathogens to the flesh. However, it is unclear how long rainbow trout can be fasted before suffering unnecessary stress. In addition, very little is known about the best time of the day to slaughter fish, which may in turn be dictated by diurnal rhythms in physiological stress parameters. Water temperature is also known to play a very important role in stress physiology in fish but the combined effect with fasting is unclear. Current recommendations regarding the optimal duration of pre-slaughter fasting do not normally consider water temperature and are only based on days, not degree days (ºC d). The effects of short-term fasting prior to slaughter (1, 2 and 3 days, between 11.1 and 68.0 ºC days) and hour of slaughter (08h00, 14h00 and 20h00) were determined in commercial-sized rainbow trout (Oncorhynchus mykiss) over four trials at different water temperatures (TRIAL 1, 11.8 ºC; TRIAL 2, 19.2 ºC; TRIAL 3, 11.1 ºC; and TRIAL 4, 22.7 ºC). We measured biometric, haematological, metabolic and product quality indicators. In each trial, the values of fasted fish (n=90) were compared with 90 control fish kept under similar conditions but not fasted. Results show that fasting affected biometric indicators. The coefficient of condition in fasted trout was lower than controls 2 days after food deprivation. Gut emptying occurred within the first 24 h after the cessation of feeding, with small traces of digesta after 48 h. Gut emptying was faster at higher water temperatures. Liver weight decreased in food deprived fish and differences between fasted and fed trout were more evident when gut clearance was faster. The overall effect of fasting for up to three days on haematological indicators was small. Plasma cortisol levels were high in both fasted and fed fish in all trials. Plasma glucose response to fasting varied among trials, but it tended to be lower in fasted fish as the days of fasting increased. In any case, it seems that water temperature played a more important role, with higher concentrations at lower temperatures on days 2 and 3 after the cessation of feeding. Plasma lactate levels indicate moments of high muscular activity and were also high, but no variation related to fasting could be found. Haematocrit did not show any significant effect of fasting, but leucocytes tended to be higher when trout were less stressed and when their body condition was higher. Finally, the loss of liver weight was not accompanied by a decrease in liver glycogen (only measured in TRIAL 3), suggesting that a different strategy to maintain plasma glucose levels was used. Regarding the hour of slaughter, lower cortisol levels were found at 20h00, suggesting that trout were less stressed later in the day and that pre-slaughter handling may be less stressful at night. Haematocrit levels were also lower at 20h00 but only at lower temperatures, indicating that higher temperatures increase metabolism. Neither fasting nor the hour of slaughter had a significant effect on the evolution of meat quality during 3 days of storage. In contrast, storage time seemed to have a more important effect on meat quality parameters. The lowest pH was reached 24-48 h post mortem, with a higher variability among fasting durations at 20h00, although no clear pattern could be discerned. Maximum stiffening from rigor mortis occurred after 24 h. The water holding capacity was very stable throughout storage and seemed to be independent of pH changes. Meat lightness (L*) slightly increased during storage and a* and b*-values were relatively stable. In conclusion, based on the haematological results, slaughtering at night may have less of a negative effect on welfare than at other times of the day. Overall, our results suggest that rainbow trout can cope well with fasting up to three days or 68 ºC d prior to slaughter and that their welfare is therefore not seriously compromised. At low water temperatures, trout could probably be fasted for longer periods without negative effects on welfare but more research is needed to determine the relationship between water temperature and days of fasting in terms of loss of live weight and the decrease in plasma glucose and other metabolic indicators.

Atlantic Salmon (Salmo salar L.) as a Marine Functional Source of Gamma-Tocopherol

Gamma tocopherol (gT) exhibits beneficial cardiovascular effects partly due to its anti-inflammatory activity. Important sources of gT are vegetable oils. However, little is known to what extent gT can be transferred into marine animal species such as Atlantic salmon by feeding. Therefore, in this study we have investigated the transfer of dietary gT into salmon. To this end, fish were fed a diet supplemented with 170 ppm gT for 16 weeks whereby alpha tocopherol levels were adjusted to 190 ppm in this and the control diet. Feeding gT-rich diets resulted in a three-fold increase in gT concentrations in the liver and fillet compared to non-gT-supplemented controls. Tissue alpha tocopherol levels were not decreased indicating no antagonistic interaction between gamma- and alpha tocopherol in salmon. The concentration of total omega 3 fatty acids slightly increased in response to dietary gT. Furthermore, dietary gT significantly decreased malondialdehyde in the fillet, determined as a biomarker of lipid peroxidation. In the liver of gT fed salmon we observed an overall down-regulation of genes involved in lipid homeostasis. Additionally, gT improved the antioxidant capacity by up-regulating Gpx4a gene expression in the pyloric caeca. We suggest that Atlantic salmon may provide a marine functional source capable of enriching gT for human consumption.

Seal and salmon fisheries and general resources of Alaska.

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Atlantic salmon (Salmo salar) parr as a model to predict the optimum inclusion of air classified faba bean protein concentrate in feeds for seawater salmon

Peer reviewed

Expression of multiple insulin and insulin-like growth factor receptor genes in salmon gill cartilage

In mammals, one of the major actions of insulin-like growth factor I (IGF-I) is to increase skeletal growth by stimulating new cartilage formation. IGF-I stimulates chondrocytes in vitro to synthesize new cartilage matrix, measured by enhanced uptake of 35S-sulfate, but the addition of insulin does not produce a similar effect except when added at high concentrations. However, recent studies have shown that, in teleosts, both insulin and IGF-I are potent activators of 35S-sulfate uptake in gill cartilage. To further characterize the growth-promoting activities of these hormones in fish, we have used reverse transcriptase-linked PCR to analyze the expression of insulin receptor family genes in salmon gill cartilage. Partial cDNA sequences encoding the tyrosine kinase domains from six distinct members of the IR gene family were obtained, and sequence comparisons revealed that four of the cDNAs encoded amino acid sequences that were highly homologous to human IR whereas the encoded sequences from two of the cDNAs were more similar to the human type I IGF receptor (IGF-R). Furthermore, a comparative reverse transcriptase-linked PCR assay revealed that the four putative IR mRNAs expressed in toto in gill cartilage were 56% of that found in liver whereas the expressed amount of the two IGF-R mRNAs was 9-fold higher compared with liver. These results suggest that the chondrogenic actions of insulin and IGF-I in fish are mediated by the ligands binding to their cognate receptors. However, further studies will be required to characterize the binding properties and relative contribution of the individual IR and IGF-R genes.

The Decline of Wild Pacific Salmon

As threatened and endangered species, wild Pacific salmon are in peril. This paper discusses the differences of the five species of wild Pacific salmon. As salmon go through several stages of their lifecycles, they face a myriad of threats to their existence. Threats from humans in the form of hydropower dams, habitat destruction, harvesting issues, and hatcheries are explained. A draft recovery plan for salmon in the Puget Sound area of Washington State is used as a case study. Strengths and weaknesses of this plan are discussed. The paper then discusses the need for growth management laws supporting salmon habitat and a change in individual behaviors if wild Pacific salmon sustainability is to become a reality.