Thermal habitat use and juvenile growth of Svalbard Arctic charr: evidence from otolith stable oxygen isotope analyses

Abstract – Stable oxygen isotopes (δ¹⁸O) derived from otoliths were used to estimate mean annual water temperatures experienced by individual Svalbard Arctic charr, Salvelinus alpinus (L.), during their first four growth seasons. The analysed Arctic charr experienced a high variety of temperatures, indicating the use of different thermal habitats. A higher proportion of the juveniles experienced warmer temperatures during their first summer compared with later summers, suggesting the selective use of the shallowest littoral areas of the lake. Although the estimated temperatures were consistent with water temperatures found in High Arctic rivers and lakes during summer, they did not represent the annual variation in air temperature registered over the 20 years of otolith measurement. Furthermore, summer otolith increment width did not correlate with the experienced temperature. However, after the second year, otolith increment width was highly dependent on increment width during the previous summer. This study estimated mean summer water temperatures experienced by individual Arctic charr during the first four growth seasons providing additional evidence that stable oxygen isotope analysis can be used to provide insight into the thermal habitat use by juvenile Arctic charr.     

The acute stress response in fed and food deprived Atlantic cod, Gadus morhua L.

The aim of the present study was to evaluate the effect of stress and nutritional state (fed vs food deprived) on the generalized stress response and intestinal integrity in Atlantic cod (Gadus morhua L.). Cod in feeding or food deprived states were subjected to 15 min of acute stress (exhaustive exercise). Blood was collected at 9 intervals from before stress (t = 0), to t = 48 h post stress and analysed for blood haematocrit and haemoglobin, and plasma cortisol, lactate, glucose, osmolality, chloride, as well as the tissue damage indicators glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and thiobarbituric acid reactive substances. Intestinal segments were prepared for histology with the same intervals, while assessment of intestinal integrity and microbiology was performed at t = 0, 4 and 48 h post stress.Subjecting cod to exhaustive stress initiated a standard stress response including increased blood Hct and plasma cortisol, glucose, chloride, osmolality and lactate. Food deprived fish did in general have reduced stress resistance compared to fed fish. For many parameters, cod returned slowly to basal levels. Cellular indicators of tissue damage and oxidative stress increased in a biphasic manner following stress. Stress did not affect gut histology but did transiently increase gut permeability. Furthermore, stress had no effect on the adherent bacterial population level in midgut, but did cause a small decrease in hindgut (non-significant) and hindgut chamber (p < 0.05). Isolates belonging to Carnobacterium were predominant but not affected by stress.In conclusion, food deprived cod are less resistant to stress than fed cod. The magnitude of the response is less than in salmonids, but the effects are persistent (including tissue damage indicators and oxidative stress) and may have negative long term consequences. The gut is relatively resistant to stress, there is however a transient increase in the intestinal permeability and alterations in microbiota that may indicate lower protection against invading pathogens.     

Effect of soybean oil and soybean lecithin on intestinal lipid composition and lipid droplet accumulation of rainbow trout, Oncorhynchus mykiss Walbaum

Rainbow trout (Oncorhynchus mykiss Walbaum) were fed purified diets containing fish oil for six weeks and then soybean lecithin or soybean oil for 25 days. The gastrointestinal tract segments, stomach, midgut and hindgut were then sampled for lipid and fatty acid composition and electron microscopy. Membrane lipid class composition was fairly similar in all three segments of trout fed fish oil. Hindgut contained slightly more phosphatidylserine than stomach and midgut, while midgut contained more phosphatidylcholine and less lysophospatidylcholine/sphingomyelin. Feeding soybean products appeared to marginally decrease free cholesterol. The fatty acid compositions of the main lipid classes showed significant regional differences. In control fish, stomach had higher levels of arachidonic acid (20:4n-6) and n-6 polyunsaturated fatty acids than midgut and hindgut, and lower content of docosahexaenoic acid (22:6n-3) and n-3 polyunsaturated fatty acids. Midgut phosphatidylethanolamine also had higher levels of saturated fatty acids and less n-3 polyunsaturated fatty acids than the other tissues. Feeding soybean products decreased the n-3/n-6 ratio mainly due to increases in linoleic (18:2n-6) and 20:4n-6 and decreases in 22:6n-3 and eicosapentaenoic acid (20:5n-3). Phosphatidylcholine and to a lesser extent phosphatidylethanolamine adapted more readily to dietary changes by major increases in 18:2n-6 and C₂₀₋₂₂ n-6 polyunsaturated fatty acids. The composition of phosphatidyl-serine and -inositol appeared to be under more strict metabolic control. Linoleic acid hardly increased at all while the increase in other n-6 polyunsaturated fatty acids was less pronounced than for the other lipid classes. Regardless of lipid class, stomach resisted dietary changes more strongly than midgut and hindgut. Increases in n-6 polyunsaturated fatty acids were minor as were the loss of n-3 polyunsaturated fatty acids. The dead-end product 20:2n-6 accumulated to a higher degree in hindgut phosphatidyl-ethanolamine and -coline compared to midgut and stomach, suggesting that the activity of Δ₆ desaturation is higher in the anterior part of the intestine where most of the lipid is absorbed. Feeding soybean oil caused massive accumulation of free lipid droplets in midgut enterocytes while little lipid droplets were observed in trout fed fish oil or soybean lecithin. Since both soybean products influenced intestinal composition to the same degree, altered fatty acid profiles in membranes is not responsible for the observed lipid accumulation. This supports previous observations in Arctic charr (Salvelinus alpinus L.), suggesting that fish may require exogenous phospholipids in order to sustain a sufficient rate of lipoprotein synthesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effect of dietary inulin on aerobic bacteria associated with hindgut of Arctic charr (Salvelinus alpinus L.)

The primary aim of the present study was to evaluate the population level of adherent (autochthonous) aerobic and facultative anaerobic bacteria in the hindgut of healthy Arctic charr (Salvelinus alpinus L.) fed dextrin or inulin. This was assessed by the dilution plate technique, and visualized using both transmission and scanning electron microscopy. A population level of 4.8 × 10⁵ adherent bacteria per gram wet mass was found in the hindgut of fish fed a casein‐based diet supplemented with 15% dextrin. However, substituting dextrin with 15% inulin reduced the bacterial population level in the hindgut (3.56 × 10⁴). A total of 217 bacterial isolates were identified by key phenotypical and biochemical characteristics. In addition, 22 strains were also identified by partial sequencing of the 16S rRNA gene. The composition of bacteria colonizing the hindgut of Arctic charr fed dextrin was dominated by the genera Staphylococcus, Pseudomonas, Micrococcus, Psychrobacter glacincola and Streptococcus. However, bacteria colonizing the hindgut of fish fed inulin were dominated by Gram‐positive bacteria of the genera Staphylococcus, Streptococcus, Carnobacterium and Bacillus. While Carnobacterium divergens‐like strains were isolated from charr fed dextrin, Carnobacterium maltaromicus‐like strains were isolated from the hindgut of fish fed inulin. Electron microscopical analysis of hindgut regions confirmed traditional culture‐based microbial analysis as fewer bacterial cells were observed between microvilli and associated with the surfaces of enterocytes of fish fed inulin rather than dextrin.     

Phospholipids in formulated start-feeds – Effect on turbot (Scophthalmus maximus L.) larval growth and mitochondrial alteration in enterocytes

Turbot Scophthalmus maximus, larvae were start‐fed with formulated feeds containing marine phospholipids (MP) or soya phospholipids (SP). The experiment was performed with six experimental groups, four groups were given formulated feeds, one group was given live feed and one group was starved. Phospholipid (PL) contents of the formulated feeds were respectively 5% MP, 15% MP, 5% SP and 15% SP. Larvae were offered feed from day 3 post‐hatch. There was no significant size difference on day 5 between the group fed 15% MP and the group given rotifers. Electron microscopical examination of enterocytes was performed on larval intestine on day 5. Larvae fed 5% and 15% MP and larvae fed rotifers had normal looking enterocytes with numerous normal looking mitochondria. In the enterocytes of larvae fed 5% SP and 15% SP the mitochondria appeared swollen with a translucent matrix and fragmented cristae. Thus, MP and not SP seemed suitable as a lipid and PL source for start‐feeding turbot larvae.     

The effects of soybean, linseed and marine oils on aerobic gut microbiota of Arctic charr Salvelinus alpinus L. before and after challenge with Aeromonas salmonicida ssp. salmonicida

Populations of heterotrophic bacteria present in the hindgut region of Arctic charr Salvelinus alpinus L. fed dietary soybean, linseed and marine oils before challenge with Aeromonas salmonicida ssp. salmonicida and marine oil after challenge were estimated using the dilution plate technique. There were differences in bacterial composition between the rearing groups before and after challenge, as well as interindividual variations. For example, carnobacteria were only isolated from the hindgut region of fish fed soybean oil and linseed oil before challenge, whereas Carnobacterium spp. and Carnobacterium funditum‐like species were isolated from fish fed the same oils after challenge. Three non‐motile Aeromonas spp. were isolated from infected fish fed marine oil. One of these isolates was identified as identical to A. salmonicida ssp. salmonicida used in&the challenge test by microbial fingerprinting (amplified fragment length polymorphism). Electron microscopic examinations of hindgut regions demonstrated substantial numbers of bacterial cells associated with enterocytes, but bacterial colonization of the enterocyte surface varied between different rearing groups. The potential of bacteria found associated with the hindgut region to inhibit the fish pathogens A. salmonicida, Vibrio salmonicida and Vibrio anguillarum differed between rearing groups.     

A Method for Collection of Bile in the Conscious Sheep

A method to collect bile directly from the hepatic duct is described for use in the sheep. The technique is a combination of the intestinal re-entrant cannulae and a catheter from the duodenal lumen to the hepatic duct. The cystic duct is ligated near its junction with the common bile duct. The catheter is fixed in the proximal visible end of the hepatic duct. One plastic cannula is fixed to the duodenum opposite to the opening of the common bile duct and the other is fixed in the same way about 15 cm posterior to the first one. The two plastic cannulae fixed together with a plastic tube serve as an extra-abdominal anastomosis. During the collection periods the bile duct catheter is passed through an opening in the wall of the connection tube into a collection bag that is fixed to the plastic cannulae. Between the collection periods the catheter ends in the lumen of the anastomosis.     

The influences of dietary linseed oil and saturated fatty acids on caecal enterocytes in Arctic char (Salvelinus alpinus L.): a quantitative ultrastructural study

Arctic char (Salvelinus alpinus L.) were fed over a three-week period with a commercial diet or one of seven casein-based diets. The latter were either lipid-free or supplemented with 50–200 g linseed oil kg^–1 diet by dry weight, 160 g linseed oil and 40 g 14:0 or 160 g linseed oil and 40 g 16:0. Three fish having filled guts were sampled from each dietary group and analysed for ultrastructural changes in the pyloric caecum.Increasing the dietary lipid level increased the accumulation of lipid droplets in columnar absorptive enterocytes from about 9% of epithelial volume in fish fed a diet of 50 g linseed oil, to almost 61% in fish fed a diet of 200 g linseed oil. Replacing linseed oil in the diet with 14:0 (160 g linseed oil + 40 g 14:0 kg^–1 diet) appeared to produce a smaller lipid loading (roughly 53%) but the difference was not statistically significant. However, replacing 40 g linseed oil with 40 g of 16:0 in the diet decreased lipid loading significantly to just under 10%. Epithelial damage to enterocytes was assessed using a ranking scale based on ultrastructural signs of cell and organelle swelling and degeneration. The extent of damage closely followed the level of lipid loading, being lowest in fish fed the lipid-free or low-lipid (damage index 0.07–0.13) diets, and highest in char maintained on a diet containing 200 g linseed oil (index 1.41). Replacing linseed oil with 14:0 (160 g linseed oil + 40 g 14:0) appeared to reduce the damage index to 0.77 but this was not significant. However, a significant reduction of the damage index to 0.27 was observed when linseed oil was replaced by 16:0.We conclude that higher dietary linseed oil promotes lipid droplet accumulation in enterocytes. The droplets are probably related to the amount of polyunsaturated fatty acids in the lipid. Intracellular droplet formation and cellular damage are both reduced by adding saturated fatty acids to the diet. This could be related to disruptions in the lipoprotein assembly rate. The cellular damages observed with high lipid diets are likely to be pathological and may lead to intestinal malfunction and represent a major infection route for pathogenic bacteria.     

Absorption of acoustic energy in dense herring schools studied by the attenuation in the bottom echo signal

The absorption of acoustic energy in herring schools was studied by echo integration of the bottom signal under various densities of herring. The investigations were carried out in a fjord in northern Norway, where large schools of adult herring overwinter. The intensity of the bottom signal, expressed by the integrator value (reflected area in square metres per square nautical mile (m² NM⁻²)) of the bottom decreased significantly as the density of herring increased. The bottom integrator value decreased exponentially from 1.39 to 0.36 × 10⁶ as the area density of herring increased from 0 to 550 × 10³, corresponding to an increase in number of herring per square metre of 0–180. A method for correction of this effect is also presented.