Effects of dietary soybean meal and photoperiod cycle on osmoregulation following seawater exposure in Atlantic salmon smolts

Atlantic salmon Salmo salar juveniles were fed either fishmeal-based diets (FM) or diets in which soybean meal (SBM) partly replaced the FM from first feeding on. The fish were kept at continuous daylight during the juvenile stage. During the last 3 weeks before reaching 100 g body mass, all fish were subjected to 12L:12D. Starting at 100 g body mass, groups of 60 fish from each feeding background were subjected to continuous light for 12 weeks (short winter), or a square-wave photoperiod cycle to stimulate parr to smolt transformation with 8L:16D during the first 6 weeks, and then continuous light during the last 6 weeks (long winter). After the 12 weeks, 20 fish from each treatment were subjected to 0, 24 or 96 h seawater exposure at a water salinity of 34. Hypo-osmoregulatory ability at seawater exposure was assessed by mortality, intestinal pathology, plasma ion concentrations and osmolality, gill Na⁺/K⁺-ATPase activity and element concentrations in the cytoplasm of distal intestinal enterocytes using X-ray microanalysis. The hypo-osmoregulatory capacity was higher in fish kept at short winter than at long winter, apparently due to more rapid development of gill Na⁺/K⁺-ATPase activity. Fish fed SBM suffered typical soybean meal-induced histological alterations of the distal intestine and apparent reductions in digestive function in the more proximal gastrointestinal regions. The net osmoregulatory capacity of these fish was maintained, as indicated by higher gill Na⁺/K⁺-ATPase activity and lower plasma Na⁺, Ca²⁺ and osmolality compared to the FM-fed fish. Thus, feeding SBM did not impair the hypo-osmoregulatory ability of the Atlantic salmon following seawater exposure.     

The stress and metabolic responses of juvenile Atlantic cod Gadus morhua L. to an acute thermal challenge

Survival, oxygen consumption (     ), total plasma cortisol and glucose levels and gill heat-shock protein 70 (hsp70) expression were measured in 10 and 50 g juvenile Atlantic cod Gadus morhua during an acute temperature increase (2° C h⁻¹) to their critical thermal maximum. Ninety three per cent of the fish in both size classes survived to 24° C; however, mortality was 100% within 15 min of reaching this temperature. The     for both size classes increased significantly with temperature, reaching peak values at 22° C that were c. 2·8-fold those of control (10° C) fish. Resting plasma cortisol and glucose levels were lower in 10 g as compared to 50 g fish. Plasma glucose levels were highly variable in both size classes, and significant increases were only seen at >22° C for the 10 g fish. In contrast, plasma cortisol showed an exponential increase with temperature starting at 16° C in both size classes, and reached maximum levels at 22° C that were 19-fold (10 g fish) and 35-fold (50 g fish) higher than their respective control groups. Both the constitutive (73 kDa) and inducible (72 kDa) isoforms of hsp70 were detected in both size classes using the widely utilized mouse monoclonal antibody. Expression of these isoforms, however, did not change when Atlantic cod were exposed to elevated temperature, and the 72 kDa isoform was not detected using salmonid-specific antibodies. These results indicate that juvenile Atlantic cod are very sensitive to acute increases in water temperature. In addition, they (1) show that     and plasma cortisol, but not plasma glucose or gill hsp 70 levels, are sensitive indicators of thermal stress in Atlantic cod and (2) support previous reports that the upper critical temperature for this species is 16° C.     

The interaction of temperature and fish size on growth of juvenile halibut

Growth rate of individually tagged juvenile halibut was influenced significantly by the interaction of temperature and fish size. The results suggest an optimum temperature for growth of juvenile halibut in the size range 5–70 g between 12 and 15° C. Overall growth rate was highest at 13° C (1·62% day ⁻¹). At c. 5 g at the beginning of the experiment, fish at 16° C had the highest growth rate (3·2% day ⁻¹), but reduced this rate as they grew bigger. At 9 and 11°p C, growth rates were equal or only slightly lower during the later stages of the experiment, while the fish at 6° C showed significantly lower overall growth rate (0·87% day⁻¹). Optimal temperature for growth decreased rapidly with increasing size, indicating an ontogenetic reduction in optimum temperature for growth. Moreover, a more flattened parabolic regression curve between growth and temperature as size increased indicated reduced temperature dependence with size. Although individual growth rates varied significantly at all times within the experimental temperatures, significant size rank correlations were maintained during the experiment. This indicated an early establishment of a stable size hierarchy within the fish groups. Haematocrit was highest at the highest temperature while Na⁺/K⁺-ATPase activity was inversely related to temperature. There was no difference in plasma Na⁺, Cl⁻ and K⁺ concentrations among the temperature groups.     

Short-term thermal acclimation induces adaptive changes in the inner mitochondrial membranes of fish skeletal muscle

In the oxidative muscles (musculi laterales superficiales) of crucian carp Carassius carassius acclimated for 6 weeks to either 5 or 25° C, the volume density and the surface density of fibres per tissue did not differ significantly between the control and experimental groups. The correlation ratio (μ²) for these values was below 50, 39·3 and 43·9 respectively. After acclimation to 5° C, the surface density of outer mitochondrial membrane per fibre increased significantly from 0·93 to 1·23m² cm⁻³ in the summer population but dropped from 0·94 to 0·67 m² cm⁻³ in the winter population. The surface density of outer mitochondrial membrane per mitochondrion increased from 3·24 to 4·52 m² cm⁻³ in summer fish. After acclimation to 25° C, the surface density of inner mitochondrial membranes per muscle fibre decreased from 4·04 to 1·79 m² cm⁻³ in summer fish and from 3·86 to 1·07 m² cm⁻³ in winter fish. The surface density of inner mitochondrial membranes per mitochondrion increased from 14·17 to 15·60 m²cm⁻³ in summer fish but dropped from 13·91 to 10·67 m² cm⁻³ in winter fish. Correlation matrices demonstrate a negative correlation of the surface density of outer mitochondrial membrane per mitochondrion with the volume density of mitochondria per fibre and temperature, suggesting cold-induced proliferation of small mitochondria. It was concluded that short-term cold acclimation increased surface area of the inner mitochondrial membranes in summer fish.     

Cortisol stimulates hypo-osmoregulatory ability in Atlantic salmon, Salmo salar L.

Hypo-osmoregulatory ability in juvenile Atlantic salmon, Sulrno salur L., was improved by cortisol treatment. Implantation of a vegetable shortening pellet containing cortisol (50 mg kg⁻¹) resulted in elevated plasma cortisol titres. Maximum cortisol levels (160–170ngml⁻¹) were observed at days 6 and 12 after the implantation and dropped significantly by day 55. Cortisol-implanted fish in fresh water developed a twofold increase in gill Na⁺/K⁺-ATPase activity at days 6 and 12, and a threefold increase by day 55. Intestinal mucosa Na⁺/K⁺-ATPase activity was not affected by cortisol. Cortisol-implanted fish exposed to 28 ppt sea water for 48 h tended to show an improved ability to regulate their plasma osmolarity and reduce their ionic load. The osmo-regulatory ability attained at days 12 and 55 was further evaluated by exposing fish to 37 ppt sea water for 96 h. While all the control fish died relatively early in these tests, cortisol-implanted fish showed a clear reduction in their mortality rate. These results indicate that cortisol can induce biochemical and organismal changes during winter months that typify preadaptive events normally occurring in the spring.     

Inter‐individual differences in rates of routine energy loss and growth in young‐of‐the‐year juvenile Atlantic cod

Inter‐individual differences in rates of routine (non‐feeding) metabolism and growth were evaluated in young‐of‐the‐year (YOY) juvenile Atlantic cod Gadus morhua . Rates of O₂ consumption, CO₂ production and ammonia (TAN) excretion were measured in 64, 25–43 mm standard length ( L _S) YOY growing at different rates (0·27–0·47 mm day⁻¹) in a common rearing tank. Parameter rates ( y ) increased allometrically ( y = a·M^b ) with increasing body mass ( M ) with b ‐values for O₂ production, CO₂ consumption and TAN excretion equal to 0·81, 0·89 and 0·56, respectively. In some cases, residuals from these regressions were significantly negatively correlated to fish growth rate. In no cases did residuals of parameter rates increase with increasing growth rate. These data suggest that, during unfed periods, relatively fast‐growing fish were more metabolically efficient than slower‐growing fish from the same cohort. The fish condition factor, derived from     , also significantly decreased with increasing growth rate. Results indicated differences in both the rates of routine energy loss and the patterns of growth allocation among YOY Atlantic cod. Since these physiological attributes were positively correlated with growth rate, they may be indicative of 'survivors' in field populations.     

Effect of increased flow on the behaviour of Atlantic salmon parr in winter

The effect of increased flow on movement and microhabitat use of Atlantic salmon Salmo salar parr in winter was investigated using radiotelemetry. To simulate hydropeaking operations, flow was increased four‐fold from 1·3 m³ s⁻¹ to 5·2 m³ s⁻¹ for 24 h periods. Flow did not affect fish habitat use or displacement and had little effect on fish activity within diel periods. During high flow periods in late winter, fish reduced night‐time activity. Stranding rates during flow reduction were also very low (only one fish).     

Temperature effects on metabolic rate, swimming performance and condition of Pacific cod Gadus macrocephalus Tilesius

Critical swimming speed ( U _crit) and rate of oxygen consumption of Pacific cod Gadus macrocephalus acclimated to 4 and 11° C were determined to assess the influence of water temperature on performance. The physiological effect of exercise trials on fish held at two temperatures was also assessed by comparing haematocrit and plasma concentrations of cortisol, metabolites and ions collected from fish before and after testing. The U _crit of fish acclimated and exercised at 4° C did not differ from those acclimated and exercised at 11° C [1·07 body lengths (total length) s⁻¹]. While the standard metabolic rate of 11° C acclimated fish was 28% higher than that of 4° C fish, no significant difference was observed between fish acclimated at the two temperatures. Plasma concentrations of cortisol, glucose and lactate increased significantly from pre- to post-swim in both groups, yet only concentrations of cortisol differed significantly between temperature treatments. Higher concentrations of cortisol in association with greater osmoregulatory disturbance in animals acclimated at the lower temperature indicate that the lower water temperature acted as an environmental stressor. Lack of significant differences in U _crit between temperature treatments, however, suggests that Pacific cod have robust physiological resilience with respect to swimming performance within temperature changes from 4 to 11° C.     

Haematology of juvenile Acipenser oxyrinchus and Acipenser brevirostrum at rest and following forced activity

In vivo experiments were conducted to examine the haematology of juveniles from two relic bony fishes, Atlantic sturgeon Acipenser oxyrhinchus and shortnose sturgeon Acipenser brevirostrum . Oxygen transport characteristics (haematocrit, haemoglobin and mean erythrocytic haemoglobin concentration), ionic composition (Na⁺, Cl⁻, K⁺ and osmolality), metabolite concentration (lactate, cortisol and glucose) and protein content in blood were measured or calculated at rest and during recovery from forced activity. Under resting conditions, plasma osmolality and concentrations of Na⁺, Cl⁻, lactate, cortisol and total protein were significantly different between Atlantic and shortnose sturgeon. All other resting variables were not different between species. Following forced activity, plasma lactate levels were significantly higher in both species than at rest. Plasma cortisol levels in both species were only significantly higher 1 h following forced activity compared to resting values. Plasma lactate levels were significantly higher in Atlantic sturgeon than in shortnose sturgeon, but these levels returned to resting levels by 1 h in both species. Cortisol increases were greater in shortnose sturgeon than in Atlantic sturgeon. In general, oxygen transport characteristics, blood glucose, plasma protein and plasma osmolality were not altered by forced activity in either sturgeon species. Overall, both species had reduced responses ( i.e . the magnitude of changes in measured variables) to forced activity compared with teleosts.     

A method to probe the cytoplasmic osmolality and osmotic water and solute fluxes across the cell membrane of cyanobacteria with chlorophyll a fluorescence: Experiments with Synechococcus sp. PCC7942

We present a method with which osmotic properties of the cytoplasm of cyanobacterial cells and the osmotic permeability of plasma membranes to water and solutes can be assessed from measurements of chlorophyll a fluorescence. When the electron transport of photosystem II is inhibited, the quantum yield of chlorophyll a fluorescence in cyanobacterial cells varied between a low yield limit that was attained after acclimation to darkness (state 2) and a high yield limit that was attained after acclimation to light (state 1). It was shown recently that the difference between chlorophyll a fluorescence of light‐acclimated and of dark‐acclimated cells relates quantitatively to the internal osmolality of cyanobacteria (G. C. Papageorgiou and A. Alygizaki‐Zorba. 1997. Biochim. Biophys. Acta 1335: 1‐4). In the present work we employed rapid mixing of Synechococcus sp. PCC7942 (strain PAMCOD) suspensions with solutions of defined osmolality in order to measure cell osmolality and turgor threshold, as well as water and solute fluxes across cell membranes. Concentration upshocks with sorbitol, glycine betaine, Na⁺ and K⁺ salts caused rapid (t_1/2 < 10 ms) depression of fluorescence that was correlated to osmotic water outflow from the cells. The fluorescence remained depressed in all cases except for NaCl. With NaCl, the depression was transient and fluorescence recovered with an apparent time constant of 200 ms. The fluorescence rise correlates to inflows of NaCl and water.     

Changes in movement, range and habitat preferences of adult grayling from late summer to early winter

Radio‐tagged adult grayling Thymallus thymallus ( n  = 22), monitored from mid August to mid December 1999 in the River Kuusinkijoki, Finland, shifted by the end of September (water temperature 10·0–14·5° C) from riffle sites to deeper and slower pool sites 0·7–1·6 km up‐ or downstream. In early winter ( c . 0° C water temperature), eight of 13 fish still under study made a further shift into new pool sites, possibly triggered by ice formation. The summer range of grayling in the riffles was smaller (mean ±  s . d . length: 75 ± 146 m) than the autumn range (99 ± 46 m) in the pools, but gross daily movements were equally short in both the seasons (18 ± 34 m and 15 ± 7 m, respectively). In late summer, adult grayling preferred water depths 80–120 cm and mean velocities >40 cm s⁻¹. In autumn, the preferred ranges were 100–240 cm and <30 cm s⁻¹, respectively. Substratum was mainly boulders in the summer sites, and gravel or pebbles in the autumn sites.     

The effect of temperature and fish size on growth and feed efficiency ratio of juvenile spotted wolffish Anarhichas minor

The growth properties of juvenile spotted wolffish Anarhichas minor reared at 4, 6, 8 and 12° C, and a group reared under 'temperature steps', (T‐step) i.e . with temperature reduced successively from 12 to 9 and 6° C were investigated. Growth rate and feed efficiency ration was significantly influenced by temperature and fish size. Overall growth rate was highest at 6° C (0·68% day⁻¹) and lowest at 12° C (0·48% day⁻¹), while the 4 and 8° C, and the T‐step groups had similar overall growth rates, i.e . 0·59, 0·62 and 0·51% day⁻¹ respectively. Optimal temperature for growth ( T _{opt G} ) and feed efficiency ratio (T_{opt FCE}) decreased as fish size increased, indicating an ontogenetic reduction in T _{opt G} and T _{opt FCE}. The results suggest a T _{opt G} of juvenile spotted wolffish in the size range 135–380 g, dropping from 7·9° C for 130–135 g to 6·6° C for 360–380 g juveniles. The T _{opt FCE} dropped from 7·4° C for 120–150 g to 6·5° C for 300–380 g juveniles. A wider parabolic regression curve between growth, feed efficiency ratio and temperature as fish size increased, may indicate increased temperature tolerance with size. Individual growth rates varied greatly at all time periods within the experimental temperatures, but at the same time significant size rank correlations were maintained and this may indicate stable size hierarchies in juvenile spotted wolffish.     

Coho salmon haematological, metabolic and acid‐base changes during exercise and recovery in sea water

Cannulated, seawater‐acclimated coho salmon Oncorhynchus kisutch were swum to exhaustion in a seawater tunnel (10° C, mean U _crit 50 cm s⁻¹), resulting in metabolic acidosis and increased plasma electrolyte and cortisol concentrations, which were corrected during a 4 h recovery. Because the swimming and physiological performance data were similar to those of other salmonids, it was concluded that life‐history limitations, besides their exercise capabilities in upwelling zones, probably explain declining coho salmon populations.     

Seasonal growth of Atlantic cod: effects of temperature, feeding and reproduction

Growth of 2659 Atlantic cod Gadus morhua aged 4 to 9 years examined in Placentia Bay, Newfoundland, peaked in most cases in June and was at a minimum in October or November. Water temperature, partial fullness index ( I _P) and gonado‐somatic index ( I _G) explained between 31 and 52% of the monthly variability in growth. Temperature and I _P of capelin Mallotus villosus had significant effects on growth of all age groups and explained most of the variance for ages 6–8 and 4–5 years, respectively. The I _P of large invertebrates (ages 4 to 7 years), sandlance ( Ammodytes sp. age 6 years) and demersal fishes (age 9 years) had age‐specific effects in the model. Overall, amphipods, decapods and echinoderms dominated the Atlantic cod diet in most seasons, but fish consumption by Atlantic cod was high in June and July, particularly on capelin. The rapid increase in somatic mass during June and July occurred despite cold water temperatures ( < 3° C at 50 m) and moderate to high gonado‐somatic index. The findings of this study suggest that when food was not a limiting factor, growth tended to increase even when Atlantic cod occupied colder waters, but when food was limiting, the opposite may have occured.     

Allometric growth and condition factor of Atlantic cod (Gadus morhua) fed to satiation: effects of temperature and body weight

The effect of thermal environment on condition factor was examined for six different size-classes of Atlantic cod Gadus morhua fed to satiation. A weight–length relationship for 8 to 1303 g fish reared at 4–20°C indicated an allometric growth ( W  =  aL ^b , a  = 0.0045, b  = 3.257) of cod. Changes in relative condition factor ( K _rel) with temperature were described with a second order polynomial. The most pronounced effect of temperature on body condition was found in the smallest size-classes, but the curves flattened with increased size. Temperature had size-dependent effects on the relative condition factor obtained from an overall weight–length relationship for all fish in the experiment, i.e. K _rel increased with weight at 4°C, but decreased with weight at 16 and 20°C. K _rel remained high for most size-classes at 8 and 12°C. The slopes ( b -values) of the weight–length relationships decreased linearly with temperatures from 4 to 16°C.     

The response of trout red cells to adrenaline during seasonal acclimation and changes in temperature

Adult rainbow trout were acclimated to three different temperature and photoperiod regimes: 17°C, 14 h light: 10 h dark (summer); 7° C, 14 h light: 10 h dark; and 5° C, 8 h light: 16 h dark (winter). Blood was collected from these fish after 40 days acclimation, and the response of red blood cells to in vitro adrenergic stimulation was assessed. To examine potential seasonal variations in endogenous levels of circulating catecholamines, plasma levels of adrenaline (Ad) and noradrenaline (NAd) were measured at rest and after exercise. At rest, there were no differences between groups in plasma levels of either Ad or NAd, but, after exercise, the pattern of catecholamine elevation differed. In fish acclimated to 17 and 7° C in summer, Ad and NAd increased by about the same amount (10–15 times). In fish acclimated to 5° C in winter, NAd increased about three-fold, compared to the near 50-fold increase in Ad levels. Whether this difference between groups can be attributed to seasonal influences is unclear. At both low (0·5%) and high (2%) PCO ₂, adrenergic stimulation (2 × 10^-7 M Ad) of trout red cells in vitro led to a significant reduction in MCHC (mean cell [haemoglobin]), compared to non-stimulated cells. However, only at the high PCO ₂ were pHe and red cell pHi significantly different from those in the non-stimulated cells: the latter was higher and the former lower in the stimulated cells. There were no differences in the response of red cells to adrenergic stimulation between groups of fish. Under the conditions of the present study no influence of season and/or temperature on the in vitro response of trout red cells to adrenergic stimulation was shown.     

The behavioural and physiological response of Atlantic cod Gadus morhua L. to short-term acute hypoxia

The average rate of swimming speed and the physiological status or stress of individual Atlantic cod Gadus morhua was monitored in response to short-term acute (STA) hypoxia ( i.e. partial pressure of oxygen,     , reduced from 20·9 to 4·3 kPa within 1 h at 10° C). The STA hypoxic response of Atlantic cod was associated with a large primary increase (+29%) and a large secondary decrease (−54%) in swimming speed as well as major physiological stress ( e.g. plasma cortisol = 214·7 ng ml⁻¹ and blood lactate = 2·41 mmol l⁻¹).     

Salinity effects on oxygen consumption, gill Na+, K+-ATPase and ion regulation in juvenile coho salmon

The metabolic response of juvenile coho salmon Oncorhynchus kisutch to different salinities was examined, using whole-animal oxygen consumption rates and gill Na⁺, K⁺-ATPase activities as indicators of osmoregulatory energetics. Coho salmon smolts were acclimated to fresh water (FW), isosmotic salinity (ISO, 10‰) and sea water (SW, 28‰) and were sampled for up to 6 weeks for plasma levels of cortisol, glucose and ions (Na⁺, K⁺, Cl⁻), gill Na⁺, K⁺-ATPase activity and oxygen consumption rates. Following an initial adjustment period, plasma constituents in SW fish returned to near-FW values, indicating that the fish were acclimated to SW by day 21. Gill Na⁺, K⁺-ATPase activities on days 21 and 42 were lowest in ISO, higher in FW and highest in SW. This result is consistent with the idea that less energy would be required to maintain ion balance in an isosmotic environment, where the ionic gradients between extracellular fluid and water would be minimal. Oxygen consumption rates of swimming fish (1 body length s⁻¹), however, did not differ significantly between the three test salinities after 6 weeks. The results of this study suggest that the metabolic response of juvenile salmonids to changes in salinity is dependent on life-history stage (e.g. fry v . smolt), and that oxygen consumption rates do not necessarily reflect osmoregulatory costs.     

Hsp70 is not a sensitive indicator of thermal limitation in Gadus morhua

The levels of heat‐shock proteins of the 70 kDa family (Hsp70s) were measured in different soft tissues of Atlantic cod Gadus morhua from different locations and after exposure to various thermal conditions: acute temperature increments (1° C day⁻¹), mid‐term (73 days at 4–15° C) and long‐term thermal acclimation (278 days at 8–15° C), and seasonal and latitudinal temperature variations (field samples). Tissue specific distribution patterns of Hsp70s were observed: liver > gills > red blood cells > brain > white muscle. Thus, different tissues may have required different levels of protection by Hsp70s, and possibly this was related to the rate of protein synthesis. There were no differences in tissue Hsp70s between Arctic cod populations (Arctic, i.e . Barents and White Seas, Norwegian coast, and North or Baltic Seas). No changes in Hsp70s levels were observed in response to temperature variation of any intensity (acute fluctuation or seasonal and latitudinal) within the range of physiological temperatures (4–15° C) in wild and laboratory Atlantic cod. This confirms previous observations that changes in Hsp70 caused by such temperature variation are often small in fishes. Probably, the constitutive level of Hsp70s in Atlantic cod was high enough to overcome potentially harmful effects of temperature variations within the physiological range. A suppressing effect of high temperature (15° C) has already been observed at a systematic level (as reduced rate of somatic growth), whereas it is not reflected in modified Hsp70s. Therefore, Hsp70s apparently played a secondary role in defining thermal tolerance limits in Atlantic cod. These conclusions are in line with a recent concept of thermal tolerance which indicated that the first line of thermal limitation in the cold and warm is a loss in aerobic scope.     

Body temperature and seawater adaptation in farmed Atlantic salmon and rainbow trout during prolonged chilling

The core temperature of the rainbow trout Oncorhynchus mykiss (3·5 kg) dropped to 1·0° C during the first 6 h of chilling at 0·5° C, remained stable until 24 h, and dropped significantly to 0·7° C after 39 h. Blood plasma osmolality increased and muscle moisture content decreased gradually with increasing chilling time. After 39 h of chilling, the rainbow trout experienced 40 mosmol l^-1 higher blood plasma osmolality and 2·8% less muscle moisture content compared with initial values. In the Atlantic salmon Salmo salar (5·3 kg), core temperature dropped to 1·3° C and blood plasma osmolality increased significantly during the first 6 h of chilling at 0·5° C, but remained relatively stable throughout the rest of the experimental period. After 39 h of chilling, the salmon experienced 20 mosmol l^-1 higher blood plasma osmolality and 0·5% less muscle moisture content compared with initial values. In rainbow trout muscle moisture content was inversely related to blood plasma osmolality indicating reduced seawater adaptation with increasing hours of chilling. No such relationship was observed in the Atlantic salmon. Hence, changes in plasma osmolality and muscle moisture in the Atlantic salmon do not indicate osmoregulatory failure since the new levels, once established, were maintained throughout the chilling time.