Pathology and immunohistochemistry in three species of salmonids after experimental infection with Flavobacterium psychrophilum

Rainbow trout, Oncorhynchus mykiss (Walbaum), sea trout, Salmo trutta L., and Atlantic salmon, Salmo salar L., were experimentally infected with Flavobacterium psychrophilum in order to evaluate any species differences in susceptibility to the bacterium. Furthermore, differences in pathological changes and distribution of the bacteria in internal organs were studied. The bacteria were injected intraperitoneally in two doses, high dose (Hd) 1 x 10(7) colony forming units (CFU) fish(-1) and low dose (Ld) 1 x 10(6) CFU fish(-1). The mortalities in the Ld groups varied between 0 and 7.5% and in the Hd groups between 55-70%. No significant differences in mortality between the species were recorded. Clinical signs and pathological findings were similar in the three species and in accordance with those of rainbow trout fry syndrome. Rainbow trout showed more pronounced lesions in the spleen compared with the other species. Necrosis of renal tubular epithelium and haematopoietic tissue was most prominent in rainbow trout and Atlantic salmon. Intracellular eosinophilic droplets in the kidney tubular epithelium were a prominent finding in rainbow trout and sea trout surviving the infection. The distribution of the bacteria in internal organs was similar in the three species, as studied with immunohistochemistry.     

Inclusion of camelina meal as a protein source in diets for farmed salmonids

Camelina meal (Camelina sativa) (CM) is a potential protein source for aquaculture feeds, on account of its crude protein level (380 g kg^?1) and inclusion of most indispensable amino acids. Two experiments were conducted with rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). Rainbow trout (44.9 g fish^?1) were fed diets with CM at 0 g kg^?1 (0% CM), 70 g kg^?1 (7% CM), 140 g kg^?1 (14% CM) or 210 g kg^?1 (21% CM) for 12 weeks at 14 °C in freshwater, and salmon (241.8 g fish^?1) were fed diets with CM at 0 g kg^?1 (0% CM), 80 g kg^?1 (8% CM), 160 g kg^?1 (16% CM) or 240 g kg^?1 (24% CM) for 16 weeks at 14 °C in sea water. Growth, lipid and amino acid tissue compositions were compared between species. Trout could tolerate up to 14% CM diets without affecting the growth compared to the control, while salmon fed ≥8% CM gained less weight than the control (P = 0.008). The feed conversion ratio in trout fed 21% CM was higher than the control (P = 0.002), and feed intake in salmon fed ≥8% CM was lower than the control (P = 0.006). Trout fatty acid and amino acid composition showed minimal differences between CM‐fed and control‐fed fish, while salmon showed significant alterations after feeding CM diets. Multivariate analyses emphasized differences in tissue composition between species fed CM diets.     

High‐oil residue camelina meal,a viable source of protein at low levels in diets for juvenile salmonids

The oilseed Camelina sativa has been studied as a lipid source for farmed salmonids, but recommended inclusion as a protein source has not been determined. This study evaluated low inclusion of camelina high‐oil residue meal (HORM) at 20, 40 and 60 g/kg of the diet, to determine an adequate level for Atlantic salmon parr (Salmo salar) and rainbow trout (Oncorhynchus mykiss). Salmon and trout were fed experimental diets containing up to 60 g/kg HORM for 16 weeks. At 40 g/kg HORM, trout and salmon growth performance were similar to those fed a control diet. However, at 60 g/kg HORM, trout showed lower final weight, weight gain and feed intake than those fed the control diet. Rainbow trout fed 40 and 60 g/kg HORM showed significantly lower whole body ash (p = .005), slightly lower whole body protein levels and higher fat than the control. In salmon fed 60 g/kg HORM diets, whole body ash (p = .024), and the submucosal layer of the intestine was thicker than the control (p = .007). Current results indicate that up to 40 g/kg HORM can be included in diets for rainbow trout and salmon juveniles.     

An assessment of salmon farms and wild salmonids as sources of Lepeophtheirus salmonis (Krøyer) copepodids in the water column in Loch Torridon, Scotland

Wild salmonids and farmed salmon can both be sources of Lepeophtheirus salmonis (Krøyer, 1838) larvae . Farmed salmon smolts free of L. salmonis infections are stocked in sea cages and may subsequently contract L. salmonis infections, probably from wild fish. The contribution of gravid L. salmonis at Atlantic salmon, Salmo salar L., farms to populations of L. salmonis larvae in the water column has in the past been based on estimated parameters, such as louse fecundity. This present study augments these calculations by combining empirical data on densities of infective L. salmonis copepodids in the field with estimates of the number of gravid L. salmonis on farmed and wild salmonids in Loch Torridon. Data collected between 2002 and 2007 show a significant correlation between mean densities of L. salmonis copepodids recovered in the water column and the numbers of gravid L. salmonis at the local salmon farms. Generally, the farms with greatest numbers of salmon were observed to have stronger correlations with densities of copepodids in the water than the farms with fewer fish. The study suggests that louse management approaches, e.g. treatment trigger levels, need to take account of individual farm biomass, or numbers of fish. This study highlights the importance of control of L. salmonis on salmon farms for the co-existence of both wild salmonid populations and the aquaculture industry.     

Levels of Renibacterium salmoninarum antigens in resident and anadromous salmonids in the River Ellidaár system in Iceland

In relation to stock enhancement programmes, wild salmon broodfish have been routinely screened for the presence of Renibacterium salmoninarum antigens (Rs‐Ag) for decades. A sudden increase in the prevalence of Rs‐Ag experienced caused extensive problems to this industry as eggs from positive fish are discarded. The prevalence and level of Rs‐Ag were examined in resident and anadromous salmonids in the River Ellidaár system and the progress of Rs‐Ag in a cohort of salmon followed. Both prevalence and Rs‐Ag levels were high in resident salmonids and emigrating salmon smolts in the river system. When the smolts re‐entered their home river as adults the following summer, they were almost free of Rs‐Ag, but the longer they stayed in the river, the more Rs‐Ag they acquired; the majority being positive at spawning. This study demonstrates a high level of Rs‐Ag in salmonids in the River Ellidaár system which significantly reduces in the salmon during its seawater phase. Accordingly, it seems ideal to sample salmon broodfish as soon as possible after ascending the river and subsequently transfer to Rs‐free environment for storage until stripping, which could result in lower Rs‐prevalence and minimize the problems that stock enhancement programmes have faced due to Rs‐positive wild broodfish.     

Growth and feed utilization of large size rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) reared in freshwater: diet and species effects, and responses over time

Four diets differing in crude protein/crude lipid concentrations (CP/CL), 570/200, 510/220, 460/240, 430/260 (g kg^?1 dry diet) were fed to near‐satiety to rainbow trout (initial body weight, IBW = 268 g, domestic strain) and Atlantic salmon (IBW = 456 g, wild strain) for 308 days to determine the effect of diets, species and fish size on efficiency of feed, nitrogen (N) and energy utilization. Within species, weight gain, feed efficiency (FE), and energy retention efficiency (ERE, E gain/E intake) were not affected by diet (P < 0.05). N retention efficiency (NRE, N gain/N intake) increased linearly (P < 0.05) with decreasing CP/CL for both species. NRE and FE were higher for salmon compared with trout. Whole body CP of salmon was significantly (P < 0.05) higher, and lipid significantly (P < 0.05) lower, than those of trout. There was a significant (P < 0.05) linear decrease in FE as fish grew, regardless of diet and species. NRE linearly decreased (P < 0.0001) and lipid to protein deposition ratio (LD/PD) increased (P < 0.05) as trout grew. NRE linearly increased (P < 0.05) while ERE and LD/PD linearly decreased (P < 0.05) as salmon grew. Salmon appeared to rely to a greater extent on non‐protein energy‐yielding nutrients to support cost of growth than did trout.     

Pancreas disease in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in Norway

The present paper describes, for the first time, clinical signs and pathological findings of pancreas disease (PD) in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in sea water in Norway. Similarities and differences with reports of PD from Ireland and Scotland are discussed. Samples of 68 rainbow trout from disease outbreaks on 14 farms and from 155 Atlantic salmon from outbreaks on 20 farms collected from 1996 to 2004 were included in the present study. The histopathological findings of PD in Atlantic salmon and rainbow trout in sea water were similar. Acute PD, characterized by acute necrosis of exocrine pancreatic tissues, was detected in nine Atlantic salmon and three rainbow trout. Salmonid alphavirus (SAV) was identified in acute pancreatic necroses by immunohistochemistry. Most fish showed severe loss of exocrine pancreatic tissue combined with chronic myositis. Myocarditis was often but not consistently found. Kidneys from 40% and 64% of the rainbow trout and Atlantic salmon, respectively, had cells along the sinusoids that were packed with cytoplasmic eosinophilic granules. These cells resembled hypertrophied endothelial cells or elongated mast cell analogues. Histochemical staining properties and electron microscopy of these cells are presented. SAV was identified by RT-PCR and neutralizing antibodies against SAV were detected in blood samples.     

Plasma insulin,glucagon, glucagon-like peptide and glucose levels in response to feeding,starvation and life long restricted feed ration in salmonids

Plasma insulin, glucagon, glucagon-like peptide (GLP) and glucose were measured in samples taken from rainbow trout,Oncorhynchus mykiss. (1.5 years of age) before feeding and at selected times up to 9 days after feeding. The feed contained 21.7% carbohydrate (65% digestibe) in the dry matter. The fish responded to feeding with an elevated plasma insulin level (p<0.005) 0.5 h post-feeding, which may account for the unchanged plasma glucose levels. Twentyfour hours after feeding, plasma insulin level had returned to prefeeding levels, while 4–9 days after feeding, a significant reduction compared to pre-feeding levels was observed (p<0.001). During this period plasma glucose levels remained unchanged. The corresponding plasma glucagon or GLP levels showed no significant elevation in response to starvation, the plasma GLP concentration was even significantly reduced on days 4–9 post-feeding (p<0.01–0.001).Atlantic salmon,Salmo salar, (3.5 years of age) fed a calculated satiation ration (RL=100) throughout their lifetime had, in addition to a higher body weight, significantly higher plasma insulin (p<0.005) glucagon (p<0.0001) and GLP levels (p<0.0001) than fish fed half the satiation ration. The plasma glucose levels were, however, not significant different between the groups.     

Lake-use by juvenile Atlantic salmon (Salmo salar L.) and other salmonids in northern Norway

Abstract– The utilization of lakes, and inlet and outlet streams by juvenile Atlantic salmon ( Salmo salar L.), brown trout ( Salmo trutta L.) and Arctic charr ( Salvelinus alpinus (L.), were investigated in 16 watercourses northern Norway, all known to inhabit salmon stocks. In lakes, fish were caught by small mesh size gill nets, while in rivers fish were caught electrofishing. In the shallow littoral (0-3 m depth) there were juvenile salmon in 15 of 19 investigated lakes, juvenile trout in 17 and juvenile charr in seven. Trout dominated significantly in numbers in the shallow littoral of seven lakes, while salmon and charr dominated in three lakes each. When trout and salmon were frequent in the shallow littoral, charr was usually not present in this habitat, but were found in the profundal zone in most of the lakes. Atlantic salmon parr utilized both shallow and deep lakes, and used both stones and macrophytic vegetation as shelter. The utilization of lakes by salmon parr seemed to be closely related to utilization of small inlet streams for spawning. In most inlet and outlet streams salmon dominated over trout in numbers, while charr were absent. This is the first documentation of lake-use by naturally occurring salmon parr in Scandinavia.     

Energy and nutrient utilization of Atlantic cod, Atlantic salmon and rainbow trout fed diets differing in energy content

The growth and feed utilization of Atlantic cod (Gadus morhua) (437 g), Atlantic salmon (Salmo salar) (485 g) and rainbow trout (Oncorhynchus mykiss) (413 g) fed a diet (170 g kg⁻¹ fat, 600 g kg⁻¹ crude protein; LE) similar to that used in commercial cod production or one that was top dressed with additional fat (280 g kg⁻¹ fat, 530 g kg⁻¹ crude protein; HE), were compared in an 11‐week trial. In the cod, relative feed intake was 41–58% and thermal growth coefficient 63% of that in the salmonids, but the feed efficiency ratio (FER) was 38% better (P ≤ 0.05). In contrast to the cod where there was no effect of diet on feed intake, growth or FER, both the salmon and trout fed the HE diet had greater feed intake than those fed the LE diet, but the effect of this was only positive for growth in the salmon. The cod retained more of the digested nitrogen (44.9 ± 2.7%) than the salmon (39.4 ± 0.8%), and both of these species retained more than the trout (33.6 ± 1.1%) (P ≤ 0.05). The retention of digested energy was significantly higher in the salmon (52.2 ± 0.9%) than in the trout (44.8 ± 1.1%), with the cod (44.9 ± 4.9%) not different from either of the other species. There were no differences between the species in the retention of absorbed phosphorus (65.9 ± 3.6%). There were very few dietary effects on nutrient utilization in this trial and, for the cod, this indicates that higher energy diets may be feasible for use in production.     

Endocytosis via the scavenger- and the mannose-receptor in rainbow trout (Salmo gairdneri) pronephros is carried out by nonphagocytic cells

Endocytosis of modified human serum albumin (HSA) and mannose-terminated glycoproteins was studied in pronephros cells from rainbow trout (Salmo gairdneri). Blood clearance and tissue uptake of dinitrophenylated human serum albumin (DNP-HSA) was dependent on the number of DNP-groups conjugated to the albumin molecule. Uptake of DNP₃₅-HSA in isolated pronephros cells was saturable. Pronephros cells also internalized the mannose-terminated glycoprotein invertase by a receptor-mediated process. DNP-HSA and invertase were recovered in the same cell fractions when pronephros cells containing in vivo endocytosed ligands were separated by density gradient centrifugation. The cells endocytosing these ligands were apparently not macrophages. The macrophages were recovered in cell fractions with higher densities. They were identified by their ability to adhere to glass and to carry out phagocytosis. Cultured macrophages did not endocytose chemically modified albumin (DNP-HSA and formaldehyde-treated albumin) and mannose-terminated glycoproteins (ovalbumin) in vitro. These ligands were not recovered in glass adherent pronephros cells after in vivo endocytosis of the proteins. The present data suggest that macrophages in rainbow trout pronephros possess neither the receptor for chemically modified albumin, the scavenger receptor, nor the mannose-specific receptor.     

Stress-induced changes in the affinity and abundance of cytosolic cortisol-binding sites in the liver of rainbow trout,Oncorhynchus mykiss (Walbaum), are not accompanied by changes in measurable nuclear binding

Plasma cortisol levels and the number (N_max) and affinity (K_d) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml^–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml^–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml^–1). There was a 60% reduction in cytosolic N_max in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg^–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the K_d of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in K_d was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in N_max in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.