Modulation of cellular innate immunity by Lepeophtheirus salmonis secretory products.

Lepeophtheirus salmonis produces pharmacologically active substances that have been shown to modify genetic expression of inflammatory mediators in SHK-1 cells and head kidney macrophages of salmon. Differences in genetic expression among genera of Oncorhynchus and Salmo reflect different susceptibilities to L. salmonis. This study was conducted to determine if the presence of L. salmonis secretory products (SEPs)(1) alters the cellular innate immune response (specifically macrophage function) among several salmonids. Phagocytic assays were performed using SHK-1 cells and macrophages isolated from pink (Oncorhynchus gorbuscha), chum (Oncorhynchus keta) and Atlantic (Salmo salar) salmon following incubation with SEPs and Aeromonas salmonicida. Respiratory burst assays were analyzed using pink, chum and Atlantic salmon macrophages after exposure to SEPs. For SHK-1 cells, incubation with SEPS led to dose-dependent increases in phagocytosis. Following incubation with SEPs, chum salmon macrophages had the highest phagocytic index (55.1%) followed by Atlantic (26.4%) and pink (15.8%) salmon. In contrast, respiratory burst response was greatest in pink salmon and minimal in the other two species. Our results suggest that the cellular innate immune response of salmon is modified in the presence of L. salmonis secretions and differences observed among species provide insight into species-specific consequences of sea lice infection.

Does river of origin influence the early marine migratory performance of Salmo salar?

The early marine migratory behaviour of two populations of hatchery-reared Atlantic salmon Salmo salar was compared in a common-garden experiment. Post-smolts from a river in a long fjord (Laerdal River, 144 km from the open coastline, n = 79) and a short fjord (Flekke River, 20 km from the open coastline, n = 80) in western Norway were tagged with acoustic transmitters and released during the spring of 2005 and 2006 in the inner part of the Hardangerfjord system (Opo River mouth, 179 km from the open coastline). The migratory behaviour of the tagged fish was monitored by acoustic listening stations in the fjord system up to 167 km from the release site. The Laerdal fish began migrating before the Flekke fish and had higher progression rates in the middle part of the fjord system. A greater number of Laerdal fish was detected along the most direct migratory route and in the outermost part of the Hardangerfjord system, which is indicative of a higher survival. The results from this study demonstrate differences in early marine migratory behaviour between S. salar from two different stocks and suggest that the distance a S. salar population travels to reach the open coastline may influence its early marine migratory behaviour and performance. The selective pressures of marine predation and arrival time at feeding areas in the ocean may be stronger for stocks with a longer inshore migration, creating more efficient migrants over time.

Effects of the sea louse Lepeophtheirus salmonis on temporal changes in cortisol, sex steroids, growth and reproductive investment in Arctic charr Salvelinus alpinus.

Groups of mature (5+ year old) Arctic charr Salvelinus alpinus held in sea water were exposed for 34 days to either a high (mean +/-s.e. 0.15 +/- 0.01 sea lice Lepeophtheirus salmonis g(-1) fish mass) (HI), medium (0.07 +/- 0.00 sea lice g(-1) fish mass) (MI) or no [control (C)] sea-lice infection during early stages of gonad development (June to July). Infection with sea lice resulted in increased plasma cortisol concentrations and this was related to intensity of infection; females tended to have higher cortisol concentrations than males at high infection intensities (HI group: female c. 130 ng ml(-1); male c. 80 ng ml(-1)). Plasma osmolality (C c. 330, MI c. 350 and HI c. 415 mOsm) and chloride concentrations (C c. 135, MI c. 155 and HI c. 190 mM) increased significantly with infection intensity, indicating osmoregulatory problems in infected fish. A strong positive relationship between plasma osmolality and cortisol concentration was recorded. Plasma sex-steroid concentrations were influenced negatively by sea-lice infection, particularly in the HI group, and were inversely related to plasma cortisol concentrations. The most heavily infected fish postponed the initiation of reproductive development until exposed to fresh water and timing of ovulation tended to be delayed in these fish. Growth rate and condition were negatively influenced by sea-lice infection and growth rate was inversely related to plasma cortisol concentrations. Sea-lice infection resulted in mortality among females in the HI group, and the proportion of maturing females was lower in the MI group (46%) than in the controls (85%). Egg production in the MI and HI groups was c. 50 and 30% of the C group. Egg size, embryonic survival and fry mass did not differ across groups. Sea lice influence reproductive development and egg production in S. alpinus, and consequently these parasites may influence populations via sublethal effects on broodfish, affecting growth and condition, and their reproductive output.

Concurrent injection of a rhabdovirus-specific DNA vaccine with a polyvalent, oil-adjuvanted vaccine delays the specific anti-viral immune response in Atlantic salmon, Salmo salar L.

Vaccines are commonly used in salmonid aquaculture as a method of disease prevention. Although there is a substantial amount of published research regarding the immunological and physiological effects following the injection of different polyvalent vaccines and DNA vaccines, there are no published reports examining the physiological and immunological effects of concurrent vaccine injection, which is the situation encountered in aquaculture. Using key immunological parameters such as lysozyme activity and specific antibody titres we examined the short-term activation of the immune response of cultured Atlantic salmon (Salmo salar L.) following concurrent injection with a traditional, polyvalent, oil-adjuvanted vaccine (AV) and an IHNV-specific DNA vaccine (DV). Our results indicate that different aspects of the innate and adaptive immune responses are influenced in either a positive or negative manner. While concurrent vaccine injection elicited an increase in lysozyme activity, changes in antibody titre (Ab) were antigen specific. The production of anti-Aeromonas salmonicida Abs was significantly greater in the combined vaccine group at 296 degree days post-vaccine injection (dd pvi), while the production of anti-Listonella anguillarum Abs was significantly greater at 106 dd pvi in the combined vaccine group. Of even greater interest was the apparent delay in production of IHNV-specific neutralizing antibodies (NAb) when the DV was injected concurrently with the polyvalent AV. The results indicated that concurrent injection of a polyvalent oil-AV and a DV can be beneficial to the production of antibodies; however, the specific anti-viral response may be delayed.

The association between metabolic rate, immune parameters, and growth performance of rainbow trout, Oncorhynchus mykiss (Walbaum), following the injection of a DNA vaccine alone and concurrently with a polyvalent, oil-adjuvanted vaccine.

This research demonstrates a significant increase in routine metabolic rate (RMR) following injection of a DNA vaccine concurrently with a polyvalent, oil-adjuvanted vaccine. The increase in RMR was transient and associated with increased activity of both the non-specific and specific immune responses. Rainbow trout (Oncorhynchus mykiss) were injected with a DNA vaccine (DV), a commercially available polyvalent, oil-adjuvanted vaccine (AV), or the two vaccines in combination and sampled at 203, 305, and 406 days (dd) post-vaccine injection (pvi) for RMR and key immune parameters (serum lysozyme activity, serum neutralization antibody titres). The RMR of fish that received both the DV and the AV was significantly higher at 203 dd pvi, compared to fish from all other treatment groups which included the control, the AV, and the DV groups. The increased RMR corresponded to elevated levels of serum lysozyme activity and an earlier seroconversion of virus-specific neutralizing antibodies. To determine if growth performance was affected by the transient increase in RMR, specific growth rate (SGR), percent daily weight gain (WG), and feed conversion ratio (FCR) were determined at 798, 1204, and 1610 dd pvi. Although fish in all three vaccine groups showed significant increases in SGR and WG at 798 and 1610 dd pvi compared to the control group, the overall weight of the fish was not different at the end of the experiment. In summary, this study shows that concurrent injection of a DV and an AV transiently increases the RMR of rainbow trout and changes the manner in which the immune response occurs, but does not affect the overall growth performance of the fish.

Metabolic adaptations of oxidative muscle during spawning migration in the Atlantic salmon Salmo salar L.

The adaptability/plasticity of the highly oxidative red muscle in Atlantic salmon was demonstrated during spawning migration. Substrate concentrations and the enzymatic pathways of ATP production were examined in red muscle obtained from Atlantic salmon at different sites along their migratory route in the Exploits River, Newfoundland, Canada. Individuals were chronologically sampled from a seawater site, two sites upstream, and at spawning. The 20% decrease in salmon body weight during the later stages of migration was accompanied by large decreases (mg dry weight(-1)) in both glycogen (P < 0.01) and total muscle lipid (P < 0.01). In contrast, water content and protein concentration (mg dry weight(-1)) of the red muscle increased by 25 and 34%, respectively, at spawning. Enzymes of the glycolytic pathways demonstrated a significant (P < 0.001) decrease in maximal activity as migration proceeded whereas enzymes of the oxidative phosphorylation pathways, specifically the citric acid cycle enzymes, exhibited an increase (P < 0.001) in maximal activity at spawning. The antioxidant enzyme superoxide dismutase also demonstrated an increase (P < 0.001) in maximal activity during the latter stages of migration. These adaptations imply that the red epaxial muscle of Atlantic salmon has a more efficient means of oxidizing lipids, while minimizing free radical damage, during the later stages of migration and spawning, thereby potentially increasing post spawning survival.

Effects of soft-water acclimation on the physiology, swimming performance, and cardiac parameters of the rainbow trout, Oncorhynchus mykiss.

Rainbow trout acclimated to soft water were submitted to an incremental velocity trial, and exhibited a 14% decrease in critical swimming speed (U(crit) * 1.37 +/- 0.055 vs. 1.54 +/- 0.044 m s(-1)) compared to fish kept in hard water. After a standardized swimming protocol, soft-water-acclimated fish had higher blood lactate concentrations (6.5 +/- 0.66 and 6.0 +/- 0.64 mmol L(-1) (soft water) vs. 5.0 +/- 0.46 and 3.9 +/- 0.32 mmol L(-1) (hard water)), revealing a greater use of anaerobic metabolism for the same exercise. Cardiovascular parameters were investigated while fish were swimming at increasing water velocities, revealing that soft-water-acclimated fish had lower increases in heart rate (105% vs. 118% of pre-exercise values), due to higher heart rates observed during acclimation and during the first 10 min of the swimming trial. This was also reflected by the plateau in heart rate and stroke volume observed during the swimming protocol, which can be attributed to increased cardiovascular function in response to soft-water acclimation. These results are in accord with previously reported increases in blood-to-water diffusion distance, due to proliferation of chloride cells at the gills in response to soft-water conditions, and underscore the costs and limitations of soft-water acclimation.

Growth and performance of Atlantic salmon, Salmo salar L., following administration of a rhabdovirus DNA vaccine alone or concurrently with an oil-adjuvanted, polyvalent vaccine.

This research demonstrates for the first time an absence of growth-related side effects in Atlantic salmon, Salmo salar L., following the injection of a DNA vaccine alone or concurrently with a commercially available, polyvalent, oil-adjuvanted vaccine. Using weight and specific growth rate measurements, individually tagged Atlantic salmon were monitored for 2028 degree days (dd) post-vaccination. During this time, DNA-vaccinated fish did not differ in weight, length, condition factor or specific growth rate compared to unvaccinated control fish. While differences in weight were observed between unvaccinated control and concurrently vaccinated fish, there were no significant differences in weight, length, condition factor or specific growth rate between concurrently vaccinated fish and adjuvant-vaccinated fish, suggesting that only adjuvant vaccination affected growth. To further determine if concurrent injection of a DNA vaccine and a polyvalent, oil-adjuvanted vaccine had a physiological impact on the Atlantic salmon, swimming performance tests were performed at 106 dd post-vaccination with U(crit,1), U(crit,2), the U(crit) recovery ratio (RR) and the normalized RR being similar to values obtained from unvaccinated control fish. In summary, this study shows that concurrent injection of a DNA vaccine and a polyvalent, oil-adjuvanted vaccine does not negatively influence the growth or swimming performance of Atlantic salmon compared to adjuvant vaccination alone.

Structure and regulation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in killifish: A comparative genomics approach.

The structure, regulation and evolution of the cystic fibrosis transmembrane conductance regulator (CFTR) gene were characterized in common killifish (Fundulus heteroclitus). Killifish CFTR (kfCFTR) structure was conserved with other CFTR homologues, but was more compact than those of mammals. A motif in intron 1 was conserved across all teleost CFTR homologues except zebrafish, and was similar to a functionally important site in human CFTR. The sequence of the CFTR promoter was highly conserved across nine species within the genus Fundulus, but contained additional glucocorticoid responsive elements in seawater species. The promoters of the seawater species also contained a putative osmotic responsive element that differed by a single base in the freshwater species. The kfCFTR promoter was only active in cell lines that express the endogenous CFTR gene. Transcription from the kfCFTR promoter was unaffected by application of dexamethasone or cortisol in cell culture, but increased by 1.5-fold in response to high osmolarity. Cortisol injection in vivo increased CFTR mRNA but there was no increase in luciferase expression driven by the kfCFTR promoter in transiently transgenic fish. Administration of the cortisol blocker RU486 resulted in a significant reduction in luciferase activity driven by the kfCFTR promoter in vivo.

Physiological effects of thermomechanical newsprint mill effluent on Atlantic salmon (Salmo salar L.).

Anadramous Atlantic salmon (Salmo salar L.) returning to Exploits River, Newfoundland, Canada, to spawn encounter low concentrations of thermomechanical pulp (TMP) effluent as they migrate upstream past an integrated newsprint mill. Various physiological responses of adult Atlantic salmon from the Exploits River were studied under laboratory conditions. The effects of a 6-h exposure to 0%, 6%, 12%, or 25% (v/v) TMP effluent or an increasing concentration gradient of effluent (0-25%) on cardiac output (Q ), critical swimming performance (U(crit)), hematocrit, and blood glucose, cortisol, lactate, and osmolality were examined. Relative to other treatment groups, Q during routine and low-level activity was 7-10% higher in fish exposed to at least 12% effluent. The 25% exposure group had a distinctly lower U(crit) and scope for increase in Q than the other treatment groups. These findings suggest that effluent exposure elevates physiological maintenance and repair costs, resulting in a reallocation of energetic resources.

Effects of subchronic exposure to aluminium in acidic water on bioenergetics of Atlantic salmon (Salmo salar).

Atlantic salmon (Salmo salar) were exposed for 36 days to water from the acidified (pH 5.2) Fossbekk River (Norway) with 50 micro x L(-1) of aluminium (Al) added as AlCl(3), or to circumneutral water (pH 6.6) from Ims River (Norway). Food consumption, cardiac output, swimming activity, and growth rate were measured to determine the effects of sublethal levels of Al in acidic water on the bioenergetics of the fish. Food consumption was reduced in acidic water and Al during the first 12 days, but was not significantly different from the control group during the rest of the exposure. A significant decrease in body weight was concomitant with the reduction in food consumption. The fish also kept losing weight during the rest of the exposure despite the return of their appetite to normal. Cardiac output of the fish exposed to acidic water and Al was not significantly different from the control group after 36 days of exposure. Swimming activity remained significantly elevated throughout the exposure to acidic water and Al. These results indicate that sublethal levels of Al can alter the energy budget of Atlantic salmon living in acidified surface waters.

Inhibition of oxygen consumption by pentachlorophenol and tetrachloroguaiacol in rainbow trout (Oncorhynchus mykiss).

Rainbow trout (Oncorhynchus mykiss) were exposed for 24 h to concentrations representing 100, 50 and 25% of the 96 h-LC50 of pentachlorophenol (PCP) or tetrachloroguaiacol (TCG), and their oxygen consumption, cardiac output, heart rate and stroke volume were measured at regular intervals. Oxygen consumption either remained stable at basal levels (PCP), or increased to 130% of basal levels (TCG) when fish were exposed to the 96 h-LC50 of each chemical. However, oxygen consumption decreased to about 50-60% of basal levels when fish were exposed to concentrations of PCP or TCG representing 50 and 25% of the 96 h-LC50. This decrease in oxygen consumption did not appear to affect cardiac function since cardiac output, heart rate and stroke volume remained stable. PCP is best known for its capacity to uncouple oxidative phosphorylation and increase oxygen consumption. However, this study showed that it can also decrease oxygen consumption, and that the effects of PCP and TCG on fish metabolism are similar.