Production performance of Atlantic salmon (Salmo salar L.) postsmolts in cyclic hypoxia,and following compensatory growth

This study investigated the production performance of the Atlantic salmon postsmolt (Salmo salar L.) subjected to cyclic oxygen reductions (hypoxia) of varying severity. Triplicate groups (N = 955) were kept at constant 80% O₂ (control) or subjected to 1 h and 45 min of hypoxia (50, 60 or 70% O₂, termed 80:70, 80:60 and 80:50 groups) every 6 h at 16°C for 69 days. Feed was provided in normoxia. One third of the fish were kept further for 30 days in normoxia to study possible compensatory growth. Cyclic hypoxia did not alter the oxygen uptake rates of fish, measured in night‐time. Fish subjected to 50% and 60% O₂ reduced feeding by 13% and 6% compared with the controls, respectively, with corresponding reductions in specific growth rates. Feed utilization was not reduced. Compensatory growth was observed in fish from the 80:50 group, but full compensation was not achieved. The main conclusions were that feeding in normoxia does not fully alleviate negative effects of cyclic hypoxia on feeding and growth, when oxygen is reduced to 60% or below in hypoxic periods, that feed utilization is maintained, and that compensatory growth may lessen negative effects.     

Environmental conditions alter the effect of organic acid salts on digestibility and intestinal morphology in Nile tilapia (Oreochromis niloticus)

The impact of two dietary organic acids (OAs) on nutrient digestibility and intestinal morphology was determined in Nile tilapia under conditions of dissolved oxygen in the water: normoxia and hypoxia. Four diets designated as control (0 g/kg organic acid salt), KDF (2 g/kg potassium diformate), CAB (2 g/kg calcium butyrate) and their combination (4 g/kg of a mixture of KDF and CAB, ration 1:1) were formulated with 520 g/kg of soybean meal in order to produce soybean meal enteritis‐like symptoms. The four diets were tested first under normoxic conditions (6 mg/L) for a period of 5 weeks, followed by a test period under hypoxic conditions (3 mg/L). The results showed that OAs were unable to significantly improve growth and nutrient digestibility under normoxic conditions but under hypoxic conditions, there was a significant enhancement of the growth and nutrient digestibility. Fish fed OA‐supplemented diets showed improvements in the intestinal morphology under the normoxic conditions, and these effects were more pronounced under the hypoxic conditions. Experimental findings suggest that OAs can improve the nutrient digestibility and intestinal morphology under hypoxic conditions. A synergistic effect by the combination of formic and butyric acid on growth, digestibility and intestinal morphology was not found.     

Temperature and dissolved oxygen influence growth and digestive enzyme activities of yellowtail kingfish Seriola lalandi (Valenciennes, 1833)

A 5 week experiment was carried out with juvenile yellowtail kingfish Seriola lalandi to investigate the interactive effects of water temperature (21, 24, or 27°C) and dissolved oxygen regime (normoxic vs. hypoxic) on the growth rate, feed intake and digestive enzyme activity of this species. Specific growth rate (SGR) was highest at 24°C, regardless of oxygen regime, but the SGRs of the fish exposed to hypoxia at 21, 24 and 27°C were 13%, 20% and 17% lower, respectively, than the SGRs recorded for the fish reared under normoxic conditions. The digestive enzyme activities (i.e. trypsin, lipase and α‐amylase) were influenced by temperature but did not appear to be affected by dissolved oxygen concentration. Information about the effects of water temperature and dissolved oxygen on feeding, growth and digestive capacity of juvenile yellowtail kingfish could contribute to improving feed management decisions for production of this fish species under different environmental conditions.     

Effect of Sublethal Hypoxia on the Immune Response and Susceptibility of Channel Catfish, Ictalurus punctatus, to Enteric Septicemia

The effect of sublethal hypoxia exposure on stress and immune responses and susceptibility to Edwardsiella ictaluri infection in juvenile channel catfish, Ictalurus punctatus, was investigated. Fish were monitored for temporal changes in glucose and cortisol concentrations before, during, and after 2 h exposure to sublethal hypoxia (<2 mg/L dissolved oxygen [DO]) and when maintained under normoxic conditions (6.0 ± 0.3 mg/L DO). Both blood glucose and plasma cortisol increased significantly in response to hypoxic conditions. Fish exposed to hypoxic or normoxic conditions were challenged with a high dose (1.3 × 10⁷ colony‐forming units [CFU]/mL) or a low dose (1.3 × 10⁵ CFU/mL) of E. ictaluri or sterile culture broth by 30‐min immersion bath. Approximately 1% of fish in both the normoxic and the hypoxic groups died when challenged with the low dose of E. ictaluri. However, when challenged with the high dose of E. ictaluri, catfish exposed to hypoxic conditions had significantly higher cumulative mortality (36 ± 12.1%) than those maintained under normoxic conditions (12 ± 1.1%). Total hemolytic complement and bactericidal activities and antibody response were lower in hypoxia‐exposed channel catfish, indicating that increased susceptibility of channel catfish to E. ictaluri may be the result of the immunosuppressive effects of the stress response to hypoxia.     

Air-breathing behavior and physiological responses to hypoxia and air exposure in the air-breathing loricariid fish, Pterygoplichthys anisitsi

Hypoxic water and episodic air exposure are potentially life-threatening conditions that fish in tropical regions can face during the dry season. This study investigated the air-breathing behavior, oxygen consumption, and respiratory responses of the air-breathing (AB) armored catfish Pterygoplichthys anisitsi. The hematological parameters and oxygen-binding characteristics of whole blood and stripped hemoglobin and the intermediate metabolism of selected tissue in normoxia, different hypoxic conditions, and after air exposure were also examined. In normoxia, this species exhibited high activity at night and AB behavior (2–5 AB h^?1). The exposure to acute severe hypoxia elicited the AB behavior (4 AB h^?1) during the day. Under progressive hypoxia without access to the water surface, the fish were oxyregulators with a critical O₂ tension, calculated as the inspired water O₂ pressure, as 47 ± 2 mmHg. At water O₂ tensions lower than 40 mmHg, the fish exhibited continuous apnea behavior. The blood exhibited high capacity for transporting O₂, having a cathodic hemoglobin component with a high Hb–O₂ affinity. Under severe hypoxia, the fish used anaerobic metabolism to maintain metabolic rate. Air exposure revealed physiological and biochemical traits similar to those observed under normoxic conditions.     

Hypoxia effects on gill surface area and blood oxygen-carrying capacity of the Atlantic stingray,Dasyatis sabina

Atlantic stingrays, Dasyatis sabina, are common residents of shallow-water seagrass habitats that experience natural cycles of severe hypoxia during summer months. We hypothesized that stingrays exposed to hypoxic episodes would improve their hypoxia tolerance by increasing branchial surface area and altering blood oxygen-carrying capacity. To this end, we compared critical oxygen minimum, gill morphology, and hemoglobin/hematocrit levels in a control group of Atlantic stingrays held at continuous oxygen saturations of 80–90 % (≥5.5 mg/l), to treatment groups exposed to a 7-h hypoxic interval at 55 % (~4.0 mg/l), or 30 % oxygen saturation (~2.0 mg/l). Stingrays in hypoxic treatment groups significantly improved their hypoxia tolerance. Critical oxygen minimum values fell from 0.7 ± 0.11 mg/l in control fish to 0.4 ± 0.05 and 0.4 ± 0.06 mg/l in the 55 and 30 % saturation treatment groups, respectively. Mass-specific gill surface area between control fish and the 30 % saturation treatment group increased by 1.7-fold, from 85 to 142 mm²/g. Although stingrays did not show an increase in hematocrit or hemoglobin levels, production of more efficient hemoglobin isoforms could not be ruled out. An increase in hypoxia tolerance allows Atlantic stingrays to forage for longer times and across a wide range of hypoxic habitats that are less accessible to predators and competitors.     

Effects of repeated hypoxic shocks on growth and metabolism of turbot juveniles

Turbot juveniles (45 g) were exposed for 41 d (17 °C, 34‰ salinity) to constant normoxic (100–100% air saturation, 100–100) or moderate hypoxic (75–75% air saturation, 75–75) conditions and to repeated hypoxic shocks (20% saturation for 1 h, 5 d per week) from normoxic (100–20% air saturation, 100–20) or moderate hypoxic (75–20% air saturation, 75–20) conditions. A normoxic group was feed restricted (100-FR). Mass increase of 100–100 and 75–75 groups fed to satiation was not significantly different. In comparison, it was significantly lower in the 100–20 and 75–20 groups (NS between the two hypoxic shocks groups). Intermediate results were obtained in the 100–100-FR group. The lowest mass increase under hypoxic shocks was explained by a significant decrease in both feed intake and food conversion efficiency (FCE). FCE was lower in the two hypoxic groups, but only the 75–20 group was significantly different from all the other groups. There was no sign of stress and no change in the physiological status of fish in any group. When challenged, pre-conditioning of turbot to regular hypoxic shocks extended survival time, slightly but significantly, for 50% of the population. It was 8 h longer in starved than in fed fish. When reared for 1 year in normoxic water, the growth rate of post-challenged survivors was dependent on pre-conditioning: day 0–375 specific growth rate was significantly higher in the two groups acclimated to repeated hypoxic shocks. In the second experiment, it was shown that exposure to 20% air saturation for 12 h led to major physiological changes within 4 h: a significant decrease in plasma total CO₂ and increase in plasma lactate contributing in maintaining blood pH stable, and a significant increase in osmolarity and chloride concentration. When returned to normoxic water, the recovery capacity of the fish was high: plasma osmolarity and total CO₂ returned to pre-exposure levels within 1 h. The results are discussed in terms of turbot capacity to cope with repeated hypoxic shocks and to acclimate.     

Effect of Oxygen Saturation in Water on Reproductive Performances of Pacu Piaractus brachypomus

Broodstock pacu Piaractus brachypomus as well as their eggs during their embryonic development were exposed to either normoxia (5.5–7.5 mg O₂/L) or hypoxia (2.0–4.5 mg O²/L) conditions. The plasma concentrations of 11-ketotestosterone in males and estradiol-17β in females, as well as that of their precursor testosterone (T) were significantly ( P < 0.01) higher in fish maintained under normoxic conditions than in fish exposed to hypoxia. After ovulation and spermiation induced by hormonal treatments, the plasma concentrations of T and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) significantly ( P 0.05) increased in both sexes under both normoxic and hypoxic conditions. The plasma levels of T and 17,20βP achieved under normoxic conditions were higher than the ones recorded under hypoxia, except for those of 17,20βP in males. Males responded positively to the hormonal treatments, and the concentration of spermatozoa was 10.5 ± 0.8 10⁹/mL under both oxygen conditions. Hypoxia resulted in significantly lower survival of embryos (17.3 ± 28%) in comparison to normoxic conditions (68.5 ± 25%). Moreover, larval deformities were found when exposed to hypoxia (91.6 ± 6%). During embryonic development of this species 4 mg/L of oxygen is tolerated at 26–27 C without negative impact. We conclude that despite the highly adaptable nature of adult pacu to environmental hypoxia, oxygen concentrations below 4 mg/L severely impacted survival of embryos.     

Oxygen consumption and ventilatory reflex responses are influenced by dietary lipids in sturgeon

The effects of one year's feeding with diets enriched either in highly unsaturated fatty acids of the w3 series (w3 HUFA) (fish oil-supplemented diet, FOD) or in saturated fatty acids (SFA) (coconut oil-supplemented diet, COD) on fatty acid composition of tissue lipids, on patterns of resting oxygen consumption and on responses to hypoxia and hypercapnia were investigated in the Adriatic sturgeon (Acipenser naccarii). FOD sturgeon had higher levels of w3 HUFA in liver and muscle lipids than COD fish, which had higher levels of SFA. A frequency distribution of instantaneous oxygen uptake rates (M]O2), as measured every 10 min for 8 h, revealed a different pattern of resting oxygen consumption between the two groups. The FOD sturgeon consumed oxygen in a narrow range of low rates, with a lower mean M]O2 than COD sturgeon, which showed a wide range of more elevated rates. FOD sturgeon had a lower opercular pressure amplitude than COD fish in normoxia. Exposure to three levels of hypoxia PO2s = 10.8±0.2; 6.6±0.2 and 4.6±0.2 kPa) or mild hypercapnia (PCO2 = 1.0±0.2 kPa) did not affect ventilation in FOD fish but elicited hyperventilation in COD animals. Mild hypoxia (PO2 = 10.8±0.2 kPa) and hypercapnia caused less reduction in blood oxygen content in FOD as compared with COD sturgeon. The effects of adding vitamin E supplements to the diets was investigated; groups fed vitamin E supplements had elevated M]O2 and hyperventilated in hypoxia. The data indicate that dietary fatty acid composition influences resting M]O2 in sturgeon and that this influences the regulation of ventilation and blood O2 levels in hypoxia and hypercapnia. The low resting M]O2 of fish fed w3 HUFA supplements (the FOD group) obviated the need for hyperventilation in hypoxia or hypercapnia, thereby making them less sensitive to these stresses than sturgeon fed SFA (COD group) or sturgeon fed either diet supplemented with vitamin E. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of temperature and Artemia enriched with ω‐3 PUFAs on the early ontogenesis of Atlantic sturgeon,Acipenser oxyrinchus Mitchill, 1815

Successful breeding of fish species in aquaculture depends on several factors, among which the temperature and feed are the most significant ones. This study was designed to evaluate the effect of temperature in the range of 14–18°C on the rate of embryogenesis and duration of larva period and to estimate the efficiency of Artemia nauplii enriched with PUFAs in growing sturgeon larvae. The temperature of 16°C is the most suitable for both egg incubation and Atlantic sturgeon prelarvae maintenance under aquaculture conditions. Even minor temperature fluctuations of 1 degree up or down leads to increased loss both of eggs and prelarvae. Increased temperature shortens the incubation period but has a lesser impact on the duration of prelarvae onset of external feeding. The technology of Artemia nauplii bioencapsulation with a PUFA‐containing supplement in A. oxyrinchus rearing increases sturgeon larvae weight by 1.5 times at a constant survival dynamic.     

The effects of temperature on respiration of Amur sturgeon,Acipenser schrenckii,at two acclimation temperatures

In order to clarify the respiratory responses strategy of Amur sturgeon Acipenser schrenckii exposed to water temperature changes, respiratory parameters of the fish were studied under two temperature regimes: fish acclimated at 13°C for Group I, temperature was increased to 16°C, 19°C, 22°C and 25°C and then returned stepwise to 22°C, 19°C, 16°C and 13°C; and fish acclimated at 25°C for Group II, the water temperature was reduced in steps to 22°C, 19°C, 16°C and 13°C, subsequently, returned to 16°C, 19°C, 22°C and 25°C. The results showed that the respiratory frequency (f_R), oxygen consumption rate (VO₂) and gill ventilation (V_G) of the fish were directly dependent on the acute temperature in both acclimation groups (p < .05). The initial 25°C VO₂ in Group II was significantly higher than the initial 13°C VO₂ in Group I (p < .05), but was significantly lower than that at 25°C in Group I (p < .05). In Group I, respiratory stroke volume (V_S.R) of fish significantly increased or decreased with the acute temperature increases or decreases, respectively (p < .05); oxygen consumption efficiencies (EO₂) of fish did not significantly show differences when temperature increased to 25°C from 13°C (p > .05), but the EO₂ significantly declined while returning to acclimation temperature (p < .05). In Group II, the V_S.R of the fish did not significantly change with acute temperature fluctuations between 25 and 13°C (p > .05), while the EO₂ increased with acute temperature increases (p < .05). The Q₁₀ values for f_R, VO₂, V_S.R, V_G and EO₂ were 1.53–1.72, 1.92–2.06, 1.07–1.60, 1.78–2.44 and 1.11–1.65 at 13–25°C of temperature interval respectively. Amur sturgeon showed partial metabolic compensation to temperature changes. The study results suggest that the ability of Amur sturgeon to regulate metabolism in response to acute temperature changes makes this species good adaptability in the aquaculture rearing.     

Lack of arterial PO<Subscript>2</Subscript> downregulation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) during long-term normoxia and hyperoxia

Regulation of arterial partial pressure of O₂ (P_aO₂) in Atlantic salmon (Salmo salar) was investigated during resting conditions in normoxic and hyperoxic water. Dorsal aorta cannulated adult Atlantic salmon (1.2–1.6 kg, n = 8) were exposed to 2 week sequential periods of normoxia [16.7 ± 1.1 kPa (mean ± SD)] and hyperoxia (34.1 ± 4.9 kPa) in individual tanks containing seawater (33.7 ± 0.2 ppt) at stable temperature conditions (8.7 ± 0.7°C) and a light regime of L:D = 12:12. Tank design and sampling procedures were optimized to provide suitable shelter and current for the fish, and to allow repeated, undisturbed sampling of blood from free-swimming fish. Fish were sampled regularly through the experimental period. P_wO₂, P_aO₂, blood ion composition (Na⁺, K⁺, Cl⁻), acid–base status (pH, PCO₂, HCO₃ ⁻), haematocrit and glucose were measured. The most frequently observed P_aO₂ values were in the range of 60–80% of P_wO₂, both during normoxia and hyperoxia, and P_aO₂ values were significantly lower during normoxia than during hyperoxia. Blood pH, PCO₂ and HCO₃ ⁻ were significantly elevated during hyperoxia, while, Na⁺, Cl⁻ and Hct were significantly lower. K⁺ and glucose showed no significant differences. This study demonstrates a lack P_aO₂ regulation in Atlantic salmon to low partial pressures, in contrast to previous reports for many aquatic gill breathing animals. Both during normoxia and hyperoxia, P_aO₂ reflects P_wO₂, and alterations in external PO₂ consequently result in proportional arterial PO₂ changes. Physiological adaptation to hyperoxia, as illustrated by changes in several blood parameters, does not include down-regulation of P_aO₂ in Atlantic salmon. The lack of P_aO₂ regulation may make Atlantic salmon vulnerable to the oxidative stress caused by increased free radical formation in hyperoxic conditions.     

Compensatory growth of Chinese shrimp, <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis> following hypoxic exposure

Compensatory growth following stress is a strategy aquatic animals use to adjust themselves to a variable environment. Studies on the recovery growth of aquatic animals are not only of theoretical value in ecophysiology and evolution, but also important to applications in aquaculture and fisheries resource management. In this experiment, juvenile Fenneropenaeus chinensis with an initial average body weight of about 3.72 g were exposed to hypoxic water (about 2.08 and 3.11 mg/l of dissolved oxygen (DO) content) for 10 days and then switched to normoxic water (about 5.63 and 5.59 mg/l DO). Compared with juveniles in normoxia, juveniles in the hypoxia period allocated a greater proportion of energy to metabolism and exuviations but allocated less energy to daily metabolism per gram shrimp weight (J/g/day). This reduced feed conversion efficiency and feeding rate. Finally, F. chinensis suffered growth depression. The juveniles completely compensated for hypoxia-induced growth depression in 30 days after being switched into normoxic water and the compensation was achieved mainly by hyperphagia and slightly by improvement of feed conversion efficiency. During the recovery period the hypoxic-stressed shrimp showed higher daily metabolic energy (J/g/day) than controls (P < 0.05). Which means the stressed shrimp had more energy for feeding-related activities. So hyperphagia was observed. Energy analysis indicated that F. chinensis improved feed-conversion efficiency mainly by reducing the percentage of energy lost in feces and exuviations. The results showed that short-term non-lethal hypoxia would not affect the growth of juvenile F. chinensis if there were enough time for the stressed shrimp to recover. This suggested F. chinensis was capable of adapting to DO fluctuation to some extent and short-term non-lethal hypoxia would not have an obvious effect on natural, released, and cultured shrimp stock.     

Skeletal anomaly assessment in diploid and triploid juvenile Atlantic salmon (Salmo salar L.) and the effect of temperature in freshwater

Triploid Atlantic salmon tend to develop a higher prevalence of skeletal anomalies. This tendency may be exacerbated by an inadequate rearing temperature. Early juvenile all‐female diploid and triploid Atlantic salmon were screened for skeletal anomalies in consecutive experiments to include two size ranges: the first tested the effect of ploidy (0.2–8 g) and the second the effect of ploidy, temperature (14 °C and 18 °C) and their interaction (8–60 g). The first experiment showed that ploidy had no effect on skeletal anomaly prevalence. A high prevalence of opercular shortening was observed (average prevalence in both ploidies 85.8%) and short lower jaws were common (highest prevalence observed 11.3%). In the second experiment, ploidy, but not temperature, affected the prevalence of short lower jaw (diploids > triploids) and lower jaw deformity (triploids > diploids, highest prevalence observed 11.1% triploids and 2.7% diploids) with a trend indicating a possible developmental link between the two jaw anomalies in triploids. A radiological assessment (n = 240 individuals) showed that at both temperatures triploids had a significantly (P < 0.05) lower number of vertebrae and higher prevalence of deformed individuals. These findings (second experiment) suggest ploidy was more influential than temperature in this study.     

Characteristics of Blood in Common Carp,Cyprinus carpio,Exposed to Low Temperatures

Changes in blood hemoglobin, hematocrit, leucocrit, osmolarity, lactate, glucose and cortisol concentrations were monitored in common carp, Cyprinus carpio, exposed to decreasing temperatures (from 24°C to 4°C). Two experiments [rapid change (acute) and slow change (chronic)] were conducted. In the acute experiment, it took 3 days for the decrease of water temperatures from 24°C to 4°C. In the chronic experiment, it took 33 days for the decrease of water temperatures from 24°C to 4°C. Blood lactate and hemoglobin concentrations, hematocrit, and leucocrit significantly (P < 0.05) decreased in the cold-treated common carp in both experiments. Hemoglobin concentrations did not decrease until the temperature reached 4°C. Plasma glucose and cortisol levels increased in common carp in acute exposure to 4°C. The levels of plasma chloride and osmolarity did not change significantly (P > 0.05) in the cold-treated common carp in both acute and chronic exposure. Data indicate that levels of blood lactate, hematocrit, hemoglobin, and leucocrit were consistently responsive to the stress of low temperatures in common carp.     

Effects of different step‐wise temperature increment regimes during egg incubation of Atlantic cod (Gadus morhua L.) on egg viability and newly hatched larval quality

We carried out an experiment to determine how rapidly the early incubation temperature of Atlantic cod eggs can be increased without affecting normal embryonic development and hatching. Atlantic cod eggs were incubated at a constant low temperature (4.5 ± 0.5°C; T5 – control) and four temperature increment treatments where the temperatures were increased stepwise from 4.5°C at zygote stage to 9.5 ± 05°C (T1‐8 h, T2‐32 h, T3‐64 h and T4‐96 h). Embryonic cell symmetry, embryonic mortality, hatching success and larval skeletal abnormalities, length and yolk sac volume were recorded. Larval samples were also taken at hatch for histological analysis. Except for higher egg mortality and lower hatching success in the T1, the differences among experimental groups were minor. Cell asymmetries and embryo mortalities were not significantly different between the control and T2–T4 treatment groups. Control larvae were significantly longer and had smaller yolk reserves at hatch than T1–T4 larvae and larvae from T2 had the largest yolk reserves. Tissue and organ histology of hatched larvae were similar. Considering embryonic cleavage pattern, hatching success and larval morphology and histology, a gradual increment of temperature in 32 h seems to be the better choice for future developmental programming studies in Atlantic cod.     

Effects of epinephrine exposure on contractile performance of compact and spongy myocardium from rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) during hypoxia

Hypoxia results in elevated circulating epinephrine for many fish species, and this is likely important for maintaining cardiac function. The aims of this study were to assess how hypoxia impacts contractile responses of ventricular compact and spongy myocardium from rainbow trout (Oncorhynchus mykiss) and to assess how and if epinephrine may protect myocardial performance from a depressive effect of hypoxia. Work output and maximum contraction rate of isolated preparations of spongy and compact ventricular myocardium from rainbow trout were measured. Tissues were exposed to the blood PO₂ that they experience in vivo during environmental normoxia and hypoxia and also to low (5 nM) and high (500 nM) levels of epinephrine in 100% air saturation (PO₂ 20.2 kPa) and during hypoxia (PO₂ 2 kPa, 10% air saturation). It was hypothesized that hypoxia would result in a decrease in work output and maximum contraction rate in both tissue types, but that epinephrine exposure would mitigate the effect. Hypoxia resulted in a decline in net work output of both tissue types, but a decline in maximum contraction rate of only compact myocardium. Epinephrine restored the maximum contraction rate of compact myocardium in hypoxia, appeared to slightly enhance work output of only compact myocardium in air saturation but surprisingly not during hypoxia, and restored net work of hypoxic spongy myocardium toward normoxic levels. These results indicate hypoxia has a similar depressive effect on both layers of ventricular myocardium, but that high epinephrine may be important for maintaining inotropy in spongy myocardium and chronotropy in compact myocardium during hypoxia.     

Cardiac activity,ventilation rate and acid-base regulation in rainbow trout exposed to hypoxia and combined hypoxia and hypercapnia

A computerised system for non-invasive monitoring of heart and ventilation rates and the time intervals between heart beats and between breaths was developed and used to investigate cardio-respiratory changes in rainbow trout exposed to hypoxia and to combined hypoxia and hypercapnia. Upon exposure to hypoxia and hypoxia-hypercapnia the arterial O₂ tension decreased from about 90 mmHg to about 30 mmHg. Acid-base changes were small in hypoxia whereas exposure to combined hypoxia-hypercapnia caused a large extracellular respiratory acidosis. This acidosis was completely compensated within 24h by accumulation of bicarbonate in plasma to concentrations twice the normoxic values. The ventilation rate was increased to higher values in hypoxic-hypercapnic trout than in hypoxic trout. In contrast to previous reports, the heart rate increased in hypoxia. On top of the tachycardia response to hypoxia, the heart rate was governed by circadian rhythms, with higher heart rates during the day than during the night. The time interval between heart beats varied considerably in normoxic fish. Hypoxia strongly reduced this variability, which may originate in a reduced cholinergic tone to the heart. The width of the frequency distribution of the time intervals between breaths was not affected by hypoxia. The degree of cardio-respiratory synchronization was low in both normoxic and in hypoxic and hypoxic-hypercapnic trout.     

Biochemical changes in oyster tissues and hemolymph during long-term air exposure

The Pacific oyster Crassostrea gigas is a sessile bivalve that inhabits the intertidal zone and therefore frequently exposed to air during the tidal cycle. It is highly adaptive to hypoxic conditions. We have studied the physiological state of oysters during long-term exposure to air. The oysters became hypoxic when exposed to air or hypoxic seawater. The 50% lethal time of oysters exposed to air at 4, 15 and 20°C was 47.8, 15.9 and 12.2 days, respectively. The hemolymph pH decreased by day 3; however, it showed a slight increase by day 5 at both 4 and 20°C. The adenylate energy charge (AEC) values decreased rapidly on the first day of air exposure in the adductor muscle, mantle, gill and body trunk, and these decreases were accompanied by decreases in ATP concentrations and increases in AMP concentrations. The AEC values in all of the tissues had fallen to below 30% by day 50 of air exposure at 4°C. These data suggest that the energy state of oysters deteriorates rapidly with air exposure. Consequently, AEC values may be useful indices of the physiological state of the oyster during long-term exposure to air.     

Vaccination of Atlantic lumpfish (Cyclopterus lumpus L.) at a low temperature leads to a low antibody response against Aeromonas salmonicida

We present a study on the effect of water temperature on immunization of Atlantic lumpfish. In total, 360 fish were vaccinated with either 50 μl of an oil‐based injection vaccine (VAX), with Aeromonas salmonicida and Vibrio salmonicida antigens, or PBS. Fish were vaccinated at three different water temperatures, 5°C, 10°C and 15°C, and sorted into six groups (N = 60). Lumpfish were weighed every 3 weeks after vaccination, sampled at 3, 6, 9 and 18 weeks post‐immunization (wpi) and evaluated by modified Speilberg score, ELISA and immunoblotting. Vaccinated fish showed low antibody response against V. salmonicida. Fish vaccinated at 5°C showed significantly lower antibody response against A. salmonicida throughout the study. At higher temperatures, vaccinated fish showed significantly increased antibody responses, at 18 wpi for 10°C and at 6 and 18 wpi for 15°C. Immunoblotting demonstrated specific response against the LPS antigen of A. salmonicida in the 10°C and 15°C VAX groups. Mean body weight increased in all groups throughout the study. Vaccinated fish had low Speilberg scores with no melanization of abdominal tissue. Our results show that vaccinating lumpfish at a lower water temperature may lead to a low antibody response against A. salmonicida.