Long-term persistence of piscine orthoreovirus-1 (PRV-1) infection during the pre-smolt stages of Atlantic salmon in freshwater.

Piscine orthoreovirus (PRV) causes heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon. During salmon production cycles, HSMI has predominantly been observed after seawater transfer. More recently, better surveillance and longitudinal studies have detected occurrences of PRV-1 in freshwater broodstock farms and hatcheries. However, very little is known about the viral kinetics of PRV-1 or disease development of HSMI during these pre-smolt stages. In this study, we conducted a long-term PRV-1 challenge experiment to examine the profile of viral load, infectiousness and/or clearance in Atlantic salmon during their development from fry to parr stage. Atlantic salmon fry (mean weight: 1.1 ± 0.19 g) were infected with PRV-1 (high virulent variant) via intraperitoneal (IP) injection. The viral load reached a peak at 2-4 weeks post-challenge (wpc) in heart and muscle tissues. The virus was detected at relatively high levels in whole blood, spleen, and head kidney tissues until 65 wpc. Heart and muscle lesions typical of HSMI were clearly observed at 6 and 8 wpc but then subsided afterwards resolving inflammation. Innate and adaptive immune responses were elicited during the early/acute phase but returned to basal levels during the persistent phase of infection. Despite achieving high viremia, PRV-1 infection failed to cause any mortality during the 65-week virus challenge period. Cohabitation of PRV-1 infected fish (10 and 31 wpc) with naïve Atlantic salmon fry resulted in very low or no infection. Moreover, repeated chasing stress exposures did not affect the viral load or shedding of PRV-1 at 26 and 44 wpc. The present findings provide knowledge about PRV-1 infection in juvenile salmon and highlight the importance of continued monitoring and management to prevent and mitigate the PRV-1 infection in freshwater facilities.

Experimental studies on combined production of CH4 and safe long-term storage of CO2 in the form of solid hydrate in sediment.

The production of the confirmed enormous resources of CH4 trapped in permafrost and deep ocean sediments in the form of hydrates has been hampered by the lack of an extraction procedure that is both effective and environmentally sensitive. This research explores experimentally the dynamic rate limiting steps in the dissociation of methane hydrates and the formation of CO2 hydrates in a sediment matrix. The use of CO2 injection and substitution for hydrate extraction takes advantage of novel thermodynamics and also provides a safe storage option for greenhouse gas. This experimental work incorporates a high-pressure facility dedicated for CH4 hydrates exchange with CO2 that replicates creation of natural gas hydrate from incoming gas below water in the pore space. The hydrate formation/exchange chamber follows the state-of-art hydrate science and is equipped with sensors distributed in several sections: the top section for gas release, a CH4 hydrate section, and a subsequent injection of CO2 from the bottom section, which also mimics hydrate dissociation towards incoming seawater through fracture systems connected from the seafloor. Four experimental conditions were examined. They comprise pure CO2 injection, and 10, 20, and 30 mole% N2 added to the CO2. We observed an increase in CH4 release from pure CO2 injection to 10 mole% N2 addition. A significant extra release of CH4 occurred by stepping up to 20 mole% N2 addition but no significant change was observed from 20 to 30 mole% N2 addition. Maximum conversion in this study is 34 mole% of CO2, and 2 mole% N2 taking the place of methane hydrate in large and small cavities. The results also show that effective substitution for hydrate production cannot rely on pure carbon dioxide injection.

Methanol as a hydrate inhibitor and hydrate activator.

During their transport to processing plants, produced hydrocarbon streams are always accompanied by water appearing as a separate phase in contact with the hydrocarbons. Dramatic temperature and pressure changes during processing can lead to condensation of water and possibly hazardous formation of ice and hydrates. Historically, methanol has been the dominating chemical agent added to prevent the formation of solid water phases. In this work, the technique of molecular dynamics simulation has been utilized to investigate and illustrate the impact of methanol as a surfactant in a water-methane system. We have found that adding 5% of methanol boosted the diffusion of methane through the interface by more than 40% compared to the reference system. The amount of methane accumulated in the aqueous phase was also significantly higher. This effect will likely also result in a significant increase in homogeneous and heterogeneous hydrate formation in these regions in the case of the methanol-stimulated system, and thus necessitate the application of classical nucleation theory. In particular, our analysis emphasised the fact that several different hydrates may form in this scenario. In case of the homogeneous hydrate formation, there will theoretically exist an infinite number of hydrate phases corresponding to concentrations spanning the range between methane's solubility and its hydrate stability limit.

Aggressive melanoma in an infant with congenital melanocytic nevus syndrome and multiple, NRAS and BRAF mutation-negative nodules.

We report the case of a newborn boy with multinodular NRAS and BRAF mutation-negative congenital melanocytic nevi and cerebral lesions compatible with congenital intraparenchymal melanosis. Histopathology from skin lesions showed atypical nodular melanocytic proliferation with marked melanocytic atypia and a large number of mitoses and apoptosis, indicating aggressive proliferation. The child developed several new subcutaneous tumors and multiple internal lesions, which were confirmed to be metastases, and died at 5 months of age. This case may represent an infantile melanoma developing from a giant congenital melanocytic nevus or a congenital melanoma.

High nervous necrosis virus (NNV) diversity in wild wrasse (Labridae) in Norway and Sweden.

Wild goldsinny wrasse Ctenolabrus rupestris, corkwing wrasse Symphodus melops and ballan wrasse Labrus bergylta were collected at 8 sampling sites in Sweden and Norway during summer 2014. Brain tissue from 466 wrasses were analyzed for nervous necrosis virus (NNV) infections by real-time RT-PCR, and positive samples were subjected to sequencing and phylogenetic analysis of partial segments of the RNA2 and RNA1 genes. This study shows that NNV is present in wild ballan, corkwing and goldsinny wrasse along the coastline of Sweden and Norway. The overall prevalence in the sampled labrids was 6.7%. Prevalence was 6.4% in goldsinny, 6.3% in corkwing and 18% in ballan wrasse. The wrasse RNA2 NNV sequences revealed high genetic variability and were divided into 3 clusters within the cold water barfin flounder NNV (BFNNV) and warm water cluster red-spotted grouper NNV (RGNNV) genogroups. Within the BFNNV genogroup, wrasse NNVs clustered in 2 sub-genogroups, with grey mullet NNV (GMNNV) and with Atlantic halibut NNV (AHNNV). These groups were previously dominated by virus originating from Atlantic cod Gadus morhua and Atlantic halibut Hippoglossus hippoglossus from the northeast Atlantic. The presence of NNV in wild wrasse and the surprising high genetic variability observed in this study should be considered before moving wild-caught wrasse between geographically distant sites. The results show that use of wild-caught wrasse as brood fish in wrasse farming represents a risk of introducing NNV into aquaculture.

Adsorption Properties of Triethylene Glycol on a Hydrated {101Ì 4} Calcite Surface and Its Effect on Adsorbed Water.

Molecular dynamics (MD) and Born-Oppenheimer MD (BOMD) simulations were employed to investigate adsorption of aqueous triethylene glycol (TEG) on a hydrated {101̅4} calcite surface at 298 K. We analyzed the orientation of TEG adsorbed on calcite, as well as the impact of TEG on the water density and adsorption free energy. The adsorption energies of TEG, free energy profiles for TEG, details of hydrogen bonding between water and adsorbed TEG, and dihedral angle distribution of adsorbed TEG were estimated. We found that while the first layer of water was mostly unaffected by the presence of adsorbed TEG, the density of the second water layer was decreased by 71% at 75% surface coverage of TEG. TEG primarily attached to the calcite surface via two adjacent adsorption sites. Hydrogen bonds between water and adsorbed TEG in the second layer almost exclusively involved the hydroxyl oxygen of TEG. The adsorption energy of TEG on calcite in a vacuum environment calculated by classical MD amounted to 217 kJ/mol, which agreed very well with estimates found by using BOMD. Adsorption on hydrated calcite yielded a drastically lower value of 33 kJ/mol, with the corresponding adsorption free energy of 55.3 kJ/mol, giving an entropy increase of 22.3 kJ/mol due to adsorption. We found that the presence of TEG resulted in a decreased magnitude of the adsorption free energy of water, thus decreasing the calcite wettability. This effect can have a profound effect on oil and gas reservoir properties and must be carefully considered when evaluating the risk of hydrate nucleation.

Phase field modelling of spinodal decomposition in the oil/water/asphaltene system.

In this paper the quantitative applicability of van der Sman/van der Graaf type Ginzburg-Landau theories of surfactant assisted phase separation [van der Sman et al., Rheol. Acta, 2006, 46, 3] is studied for real systems displaying high surfactant concentrations at the liquid-liquid interface. The model is applied for the water/heptane/asphaltene system (a model of heavy crude oil), for which recent molecular dynamics (MD) simulations provide microscopic data needed to calibrate the theory. A list of general requirements is set up first, which is then followed by analytical calculations of the equilibrium properties of the system, such as the equilibrium liquid densities, the adsorption isotherm and the interfacial tension. Based on the results of these calculations, the model parameters are then determined numerically, yielding a reasonable reproduction of the MD density profiles. The results of time-dependent simulations addressing the dynamical behaviour of the system will also be presented. It will be shown that the competition between the diffusion and hydrodynamic time scales can lead to the formation of an emulsion. We also address the main difficulties and limitations of the theory regarding quantitative modelling of surfactant assisted liquid phase separation.

Water-wetting surfaces as hydrate promoters during transport of carbon dioxide with impurities.

Water condensing as liquid drops within the fluid bulk has traditionally been the only scenario accepted in the industrial analysis of hydrate risks. We have applied a combination of absolute thermodynamics and molecular dynamics modeling to analyze the five primary routes of hydrate formation in a rusty pipeline carrying dense carbon dioxide with methane, hydrogen sulfide, argon, and nitrogen as additional impurities. We have revised the risk analysis of all possible routes in accordance with the combination of the first and the second laws of thermodynamics to determine the highest permissible content of water. It was found that at concentrations lower than five percent, hydrogen sulfide will only support the formation of carbon dioxide-dominated hydrate from adsorbed water and hydrate formers from carbon dioxide phase rather than formation in the aqueous phase. Our results indicate that hydrogen sulfide leaving carbon dioxide for the aqueous phase will be able to create an additional hydrate phase in the aqueous region adjacent to the first adsorbed water layer. The growth of hydrate from different phases will decrease the induction time by substantially reducing the kinetically limiting mass transport across the hydrate films. Hydrate formation via adsorption of water on rusty walls will play the decisive role in hydrate formation risk, with the initial concentration of hydrogen sulfide being the critical factor. We concluded that the safest way to eliminate hydrate risks is to ensure that the water content of carbon dioxide is low enough to prevent water dropout via the adsorption mechanism.

Consequences of CO2 solubility for hydrate formation from carbon dioxide containing water and other impurities.

Deciding on the upper bound of water content permissible in a stream of dense carbon dioxide under pipeline transport conditions without facing the risks of hydrate formation is a complex issue. In this work, we outline and analyze ten primary routes of hydrate formation inside a rusty pipeline, with hydrogen sulfide, methane, argon, and nitrogen as additional impurities. A comprehensive treatment of equilibrium absolute thermodynamics as applied to multiple hydrate phase transitions is provided. We also discuss in detail the implications of the Gibbs phase rule that make it necessary to consider non-equilibrium thermodynamics. The analysis of hydrate formation risk has been revised for the dominant routes, including the one traditionally considered in industrial practice and hydrate calculators. The application of absolute thermodynamics with parameters derived from atomistic simulations leads to several important conclusions regarding the impact of hydrogen sulfide. When present at studied concentrations below 5 mol%, the presence of hydrogen sulfide will only support the carbon-dioxide-dominated hydrate formation on the phase interface between liquid water and hydrate formers entering from the carbon dioxide phase. This is in contrast to a homogeneous hydrate nucleation and growth inside the aqueous solution bulk. Our case studies indicate that hydrogen sulfide at higher than 0.1 mol% concentration in carbon dioxide can lead to growth of multiple hydrate phases immediately adjacent to the adsorbed water layers. We conclude that hydrate formation via water adsorption on rusty pipeline walls will be the dominant contributor to the hydrate formation risk, with initial concentration of hydrogen sulfide being the critical factor.

Modulation of innate immune responses in Atlantic salmon by chronic hypoxia-induced stress.

Atlantic salmon post-smolts were exposed to either chronic hypoxic (Hy) or normal oxygen (No) conditions in seawater tanks for 58 days, mimicking conditions typical of sea cages for farmed salmon at some periods of the year. By day 29 head kidney macrophages were isolated and subjected to in vitro poly I:C stimulation to simulate viral infection, and samples were collected over 48 h. By day 58 fish were subjected to in vivo stimulation using poly I:C or a Vibrio water-based vaccine to simulate viral or bacterial infection, respectively. The fish were monitored for stress responses and expression of several pro-inflammatory genes in head kidney and intestinal tissue up to five days post-injection. Stress load was monitored by plasma cortisol estimation at days 29 and 58, and on days 1, 2, 3 and 5 post-injection in the in vivo trial. Hy exposure resulted in elevated plasma cortisol levels on day 29 compared to No, while on day 58 cortisol levels were higher in the control group. Additionally, both poly I:C and the Vibrio vaccine gave significantly increased cortisol levels one day post-injection compared to PBS treated controls, irrespective of previous oxygen exposure. In vitro stimulation of macrophages with poly I:C revealed higher IFNα mRNA levels at 6, 12 and 24 h and for Mx at 12 and 24 h post-stimulation, for both No and Hy individuals. Moreover, IFNα levels were higher in No than in Hy individuals at all time points, and a similar difference was seen in Mx at 48 h. In vivo stimulation with poly I:C elicited strong elevation of the IL-1ß, IFNγ, Mx and IP10 mRNA transcripts in head kidney, while TNFα1 and IFNα were found unaffected. The Vibrio vaccine elicited a strong up regulation of IL-1ß, IFNγ and IP10 mRNA, whereas Mx, TNFα1 and IFNα appeared unchanged. Significant differences in expression between different oxygen exposure groups were found for all genes and both stimuli. The overall trend suggests that long-term hypoxia either reduces or delays the expression of these genes in head kidney. Expression of IFNγ and Mx in intestinal tissues also showed a strong up regulation of the genes following poly I:C stimulation, and also here the overall trend suggests that chronic hypoxia results in a lower or delayed expression of the measured genes. In summary, our results indicate that chronic hypoxia modulates the expression of important immune related genes putatively altering the immune response. As the effect is present in isolated macrophages as well as head kidney and intestinal tissue the modulation appears to be affecting local as well as systemic responses.

Can hydrate form in carbon dioxide from dissolved water?

Transport of carbon dioxide in offshore pipelines involves high pressures and low temperatures, which may lead to formation of hydrate from the residual dissolved water and carbon dioxide. While thermodynamics is able to tell us whether the hydrate phase will be stable, the question of whether its formation will actually occur under given pipeline conditions does not have a straightforward answer. In this work, we have made use of water properties obtained from molecular simulations to examine the thermodynamics of hydrate formation from water dissolved in carbon dioxide. This paper proposes a method that allows estimation of absolute thermodynamic properties and thus makes it possible to compare free energy changes due to several possible phase transitions and determine the most probable transition. This information can be used directly to choose the optimum hydrate prevention strategy. We have found that hydrate formation from a carbon dioxide solution will be thermodynamically viable at water concentration exceeding a certain level; a conclusion also supported by several previous studies. We have also extended the quantitative analysis of the thermodynamics and the kinetics of formation through a modified version of phase field theory (PFT). The work presents the way to obtain parameters required for the practical implementation of the PFT in the case of hydrate formation, as well as outlines the estimation of thermodynamic properties for systems unable to reach true equilibrium.

Optimization of an oil leaching process to reduce the level of dioxins and dioxin-like PCBs in fishmeal.

BACKGROUND: Fishmeal produced from fish caught in polluted fishing areas might contain dioxins and dioxin-like polychlorinated biphenyls (PCBs) above maximum permitted levels (MPL) for use in feed. Decontamination of the fishmeal can be achieved by hexane extraction. The principal objective of this study was to optimize a more environmentally friendly alternative based on oil leaching of the moist presscake intermediate product during fishmeal manufacturing. RESULTS: A central composite design and response surface methodology was used to study the influence of the process variables temperature (T), presscake moisture content (MC) and leaching time (LT) on the decontamination process. A significant squared MC effect was observed, resulting in an optimum leaching rate at 27% MC. This corresponds to 5% improved dibenzo-p-dioxin/dibenzo furan (PCDD/F)-PCB toxic equivalent (TEQ) reduction compared to normal presscake (55% MC). The initial leaching rate was fast, with a TEQ reduction of 69% after only 2 min at 87 °C and 55% MC. Under the best experimental conditions (87 °C, 38% MC, 12 min LT) a TEQ reduction of 82% was achieved. Excess oil in the presscake after the leaching operation could be removed by use of a water washing step. No reduction in protein quality measured by mink digestibility could be observed. CONCLUSION: The results confirm that the oil leaching process is robust and offers easily achievable TEQ levels well below present MPLs based on process conditions normally used by the industry. Comparative effects on non-dioxin-like PCBs are expected.

Slaughter of Atlantic salmon (Salmo salar L.) in the presence of carbon monoxide.

The different stunning methods for Atlantic salmon can still be improved with regard to animal welfare. Salmon exposed to carbon monoxide expressed no aversive reactions towards CO as such. CO exposed fish showed an earlier onset of rigour mortis and a faster decrease in muscle pH due to depletion of oxygen during the treatment. Exposure to CO did increase the level of cortisol compared to undisturbed control fish, but the increase was less than in the water only control group. Neuroglobin, a CO binding globin, was found in salmon brain and Saccus vasculosus, a richly vascularized sac connected to the fish brain. Binding of CO to neuroglobin during sedation might possibly improve animal welfare.

Black Sea hydrate production value and options for clean energy production.

Natural gas hydrates of Bulgaria and Romania in the Black Sea have been subject to studies by several European research projects. The current understanding of the hydrate distribution, and the total amounts of hydrate in the region, makes it interesting to evaluate in terms of commercial potential. In this study, we have evaluated some well-known hydrate production methods. Thermal stimulation and adding chemicals are considered as not economically feasible. Pressure reduction may not be efficient due to the endothermic dissociation of hydrates and long-term cooling of the sediments. Chemical work due to pressure reduction is an additional mechanism but is too slow to be commercially feasible. Adding CO2/N2, however, has a dual value. In the future, CO2 can be stored at a price proportional to a CO2 tax. This is deducted from the value of the released natural gas. The maximum addition of N2 is around 30 mol% of the CO2/N2 mixture. A minor addition (in the order of 1 mol%) of CH4 increases the stability of the hydrate created from the injection gas. The maximum N2 amount is dictated by the demand for the creation of a new hydrate from injection gas but also the need for sufficient heat release from this hydrate formation to dissociate the in situ CH4 hydrates. An additional additive is needed to accelerate the formation of hydrate from injection gas while at the same time reducing the creation of blocking hydrate films. Based on reasonable assumptions and approximations as used in a verified kinetic model it is found that CH4/CO2 swapping is a feasible method for Black Sea hydrates. It is also argued that the technology is essentially conventional petroleum technology combined with learning from projects on aquifer storage of CO2, and a thermodynamic approach for design of appropriate injection gas. It is also argued that the CH4/CO2 swap can be combined with well-known technology for steam cracking of produced hydrocarbons to H2 and CO2 (for re-injection).

Cloning, expression and purification of Atlantic salmon (Salmo salar, L.) neuroglobin.

Neuroglobin (Ngb) exists only in small amounts in salmon brain. In order to study the protein in more detail salmon neuroglobin (sNgb) was cloned, heterologously expressed in Escherichia coli and purified. The protein had red color and showed the characteristic peaks at 411nm (metNgb), 415nm (carboxyNgb) and 424nm (deoxyNgb). Western analysis showed that sNgb reacted weakly against a rabbit anti human neuroglobin (hNgb) and strongly to a sNgb specific antibody. Our 3D-homology model of the sNgb indicated modifications adjacent to and in the O(2)/CO binding site. This may correlate to differences in substrate affinities for the sNgb compared to the hNgb. Also sNgb contained shorter helixes and longer interhelical loops typical for psychrophilic proteins.

Adsorption of water and carbon dioxide on hematite and consequences for possible hydrate formation.

The interest in carbon dioxide for enhanced oil recovery is increasing proportional to the decline in naturally driven oil production and also due to the increasing demand for reduced emission of carbon dioxide into the atmosphere. Transport of carbon dioxide in offshore pipelines involves high pressure and low temperatures, conditions which may lead to formation of hydrates from residual water dissolved in carbon dioxide and carbon dioxide. The critical question is whether the water at certain temperatures and pressures will drop out as liquid droplets first, and then form hydrates, or alternatively, adsorb on the pipeline surfaces, and subsequently form hydrates heterogeneously. In this work, we used several different basis sets of density functional theory in ab initio calculations to estimate the charge distribution of hematite (the dominating component of rust) crystals. These rust particles were embedded in water and chemical potential for adsorbed water molecules was estimated through thermodynamic integration and compared to similar estimates for water clusters of the same size. While the generated charges were not unique, the use of high order approximations and different basis sets provides a range of likely charge distributions. Values obtained for the chemical potential of water in different surroundings indicated that it would be thermodynamically favorable for water to adsorb on hematite, and that evaluation of potential carbon dioxide hydrate formation conditions and kinetics should be based on this formation mechanism. Depending on the basis set and approximations, the estimated gain for water to adsorb on the hematite surface rather than condense as droplets varied between -1.7 kJ mole(-1) and -3.4 kJ mole(-1). The partial charge distribution on the hematite surface is incompatible with the hydrate structure, and thus hydrates will be unable to attach to the surface. The behavior of water outside the immediate vicinity of hematite (beyond 3-4 diameters) closely resembled that of bulk liquid. This indicates that it will be able to form hydrates with readily available carbon dioxide. Thus hematite may still act as adsorption medium in heterogeneous hydrate formation just a few water diameters away from its surface. Theoretical estimates were compared to experimental measurements of water content in carbon dioxide co-existing with liquid water, adsorbed water and hydrate.

Double Life of Methanol: Experimental Studies and Nonequilibrium Molecular-Dynamics Simulation of Methanol Effects on Methane-Hydrate Nucleation.

We have investigated systematically and statistically methanol-concentration effects on methane-hydrate nucleation using both experiment and restrained molecular-dynamics simulation, employing simple observables to achieve an initially homogeneous methane-supersaturated solution particularly favorable for nucleation realization in reasonable simulation times. We observe the pronounced "bifurcated" character of the nucleation rate upon methanol concentration in both experiments and simulation, with promotion at low concentrations and switching to industrially familiar inhibition at higher concentrations. Higher methanol concentrations suppress hydrate growth by in-lattice methanol incorporation, resulting in the formation of "defects", increasing the energy of the nucleus. At low concentrations, on the contrary, the detrimental effect of defects is more than compensated for by the beneficial contribution of CH3 in easing methane incorporation in the cages or replacing it altogether.

Molecular dynamics simulation studies of absorption in piperazine activated MDEA solution.

Development of more efficient solvent solutions for removal of CO(2) from natural gas and flue gases is a major task, which contributes to improved design of process plants and leads to decreased costs for its removal. Understanding the mechanisms of CO(2) absorption as well as analysis of undesired simultaneous processes is crucially important in this regard. In this work, we have applied Molecular Dynamics (MD) to investigate the absorption of CO(2) from a binary mixture of CO(2) and CH(4) into aqueous piperazine activated MDEA solution. The MD simulations were performed at a constant temperature of 298 K for five different systems with a loading factor of 0.07 to provide insight into molecular distribution in the amine solution and to enhance understanding of absorption mechanisms on the molecular scale. Force field parameters that were missing from the OPLS-AA force field, as well as charge distribution of piperazine (PZ), protonated piperazine (PZH(+)), piperazine carbamate (PZCOO(-)) and MDEA were obtained by QM calculations. The results of our simulations emphasize the importance of piperazine and piperazine carbamate in accelerating the absorption process. For the first time, we have shown the undesirable trapping of CH(4) by the amine solution and revealed that amine groups are mainly responsible for both absorption of CO(2) and the undesired trapping of CH(4).

Intestinal barrier function of Atlantic salmon (Salmo salar L.) post smolts is reduced by common sea cage environments and suggested as a possible physiological welfare indicator.

BACKGROUND: Fish farmed under high intensity aquaculture conditions are subjected to unnatural environments that may cause stress. Therefore awareness of how to maintain good health and welfare of farmed fish is important. For Atlantic salmon held in sea cages, water flow, dissolved oxygen (DO) levels and temperature will fluctuate over time and the fish can at times be exposed to detrimentally low DO levels and high temperatures. This experimental study investigates primary and secondary stress responses of Atlantic salmon post smolts to long-term exposure to reduced and fluctuating DO levels and high water temperatures, mimicking situations in the sea cages. Plasma cortisol levels and cortisol release to the water were assessed as indicators of the primary stress response and intestinal barrier integrity and physiological functions as indicators of secondary responses to changes in environmental conditions. RESULTS: Plasma cortisol levels were elevated in fish exposed to low (50% and 60% saturation) DO levels and low temperature (9°C), at days 9, 29 and 48. The intestinal barrier function, measured as electrical resistance (TER) and permeability of mannitol at the end of the experiment, were reduced at 50% DO, in both proximal and distal intestine. When low DO levels were combined with high temperature (16°C), plasma cortisol levels were elevated in the cyclic 1:5 h at 85%:50% DO group and fixed 50% DO group compared to the control (85% DO) group at day 10 but not at later time points. The intestinal barrier function was clearly disturbed in the 50% DO group; TER was reduced in both intestinal regions concomitant with increased paracellular permeability in the distal region. CONCLUSIONS: This study reveals that adverse environmental conditions (low water flow, low DO levels at low and high temperature), that can occur in sea cages, elicits primary and secondary stress responses in Atlantic salmon post smolts. The intestinal barrier function was significantly affected by prolonged hypoxic stress even when no primary stress response was observed. This suggests that intestinal barrier function is a good experimental marker for evaluation of chronic stress and that it can be a valuable tool to study the impact of various husbandry conditions on health and welfare of farmed Atlantic salmon.

Properties of aging FeCl(2) clusters grown in supercritical water investigated by molecular dynamics simulations.

We have investigated the growth and properties of FeCl(2) nanoparticles from supercritical water by molecular dynamics simulations. After particle growth had finished after less than 250 ps, selected properties of the aging clusters such as structure, shape, and amount of crystal water content were analyzed for up to 1.5 ns simulation time. Very quick growth and shape relaxation of the clusters are observed. The developments of cluster size, radius of gyration, and moments of inertia with time have been analyzed. The number of water molecules that are part of the clusters is found to increase almost linearly with cluster size. The shape relaxation of the clusters happens faster after cluster-cluster collisions and faster than the time needed for the clusters to reorganize their structure and attain a highly ordered ground state structure. The development of the cluster structure with time as analyzed by radial distribution functions shows hardly any changes over time, but shows significant variations with system temperature and system density.