Impact of a pathogenic haemoflagellate, Cryptobia salmositica Katz, on the metabolism and swimming performance of rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. Oxygen consumption of juvenile rainbow trout (5 g at 13°C) at moderate swimming speeds did not change significantly when infected with Cryptobia salmositica. However, significant reductions of as much as 44% of the maximum aerobic scope for activity and 24% of the critical swimming speed were observed when the parasitaemia reached a maximum of 57.6 × 10⁶ ml⁻¹ fish blood at 3 weeks post- infection. Blood haematocrit was significantly reduced from the initial 34.1 to 19.7% at 4 weeks post- infection, probably as a result of haemolysis by the parasite. The destruction of red blood cells clearly led to lower oxygen carrying capacity, and reduced respiratory and swimming performance.     

Effects of prolonged epinephrine infusion on blood respiratory and acid-base states in the rainbow trout: Alpha and beta effects

Rainbow trout were infused continuously for 12h with epinephrine in the presence or absence of alpha-and/or beta-adrenergic blockade to characterize the specific adrenergic mechanisms involved in the control of blood acid-base status and oxygen transport capacity. Infusion of epinephrine, alone, produced a transient respiratory acidosis, as indicated by an increase in carbon dioxide tension and a decrease in whole blood pH, yet arterial oxygen tension was elevated. Red blood cell pH increased by approximately 0.2 pH units during epinephrine infusion and this increase as well as the increase in oxygen tension were prevented by pretreatment with propranolol (a beta-adrenergic antagonist). Epinephrine infusion during alpha-adrenergic blockade caused a prolonged elevation of blood carbon dioxide tension and abolished the increases in hematocrit and hemoglobin concentrations observed during epinephrine infusion alone. Infusion of the alpha-adrenergic agonists phenylephrine (an alpha₁ agonist) or clonidine (an alpha₂ agonist) caused respiratory acidosis (decreased pH, increased CO₂ tension) and a reduction in oxygen tension. Infusion of isoprenaline (a non-specific beta agonist) caused delayed increases in carbon dioxide and oxygen tensions. We speculate that the increased carbon dioxide tension observed during epinephrine infusion is a result of beta-adrenoceptor mediated inhibition of red blood cell bicarbonate dehydration and not branchial convective or diffusive adjustments. The effects of epinephrine on blood O₂ tension, content and carrying capacity are discussed with reference to the participation of alpha- and beta-adrenergic mechanisms at the gill, spleen and red blood cell.     

Effects of hypoxia on respiratory physiology of turbot, Scophthalmus maximus

Variations in respiratory and acid-base status were studied in turbot (Scophthalmus maximus) during progressive severe hypoxia followed by recovery under normoxic conditions. The first behavioural strategy of turbot under hypoxia was an increase in amplitude and frequency of ventilation. Consequently, standard O₂ consumption remained unchanged over a broad range of O₂ tensions, until a low critical level of 30 mmHg. The hyperventilation induced a moderate blood alkalosis, compensated by a lactic acidosis. The fact that blood pH did not decrease below control values could be explained by the retention in white muscle of most of the lactate produced and by a high capacity for H⁺ excretion. During the recovery period, the marked increase in O₂ uptake corresponding to an oxygen debt repayment, was partly related to the lactate elimination. When total energy contributions of aerobic and anaerobic processes were assessed in terms of ATP, the anaerobic contribution, estimated at the deepest hypoxia level, was higher than 20% of the total energy budget and appeared to totally compensate for the decline in aerobic metabolism. Moreover, the high value of O₂ tension in arterial blood in normoxia and during recovery from hypoxia showed high diffusing capacity of gills in turbot. Our results explain the high tolerance of turbot for O₂ deficient waters.     

Isometric twitch tension of lateral muscle in carp, Cyprinus carpio L., with sekoke disease

Abstract. Changes in blood properties and the chemical components, histology and isometric twitch tension of lateral muscle were studied in carp, Cyprinus carpio L., showing clinical signs of sekoke disease induced by a diet containing oxidized oil. When compared to normal fish, sekoke fish were found to have significantly lower mean blood pH, haematocrit, haemoglobin concentration and muscle water content. However, plasma lactate, lipoperoxide and total lipid concentrations were significantly higher in sekoke fish. It was found that in sekoke fish the maximum tension and the number of isometric twitches leading to complete fatigue of lateral muscle were greatly reduced. Histological studies confirmed that these reductions were related to the degree of structural damage within this tissue.     

The effects of experimental anaemia on CO2 excretionin vitro in rainbow trout,Oncorhynchus mykiss

The effects of severe experimental anaemia on red blood cell HCO₃ ^– dehydrationin vitro were examined in rainbow trout,Oncorhynchus mykiss. After 5 days of anaemia (haematocrit=4.9±1.1%) induced by intraperitoneal injection of phenylhydrazine hydrochloride, fish displayed elevated arterial CO₂ tensions (anaemic PaCO₂=3.19±0.42 torrvs. control PaCO₂=1.35±0.17 torr) and a significant acidosis (anaemic pHa=7.73±0.04vs. control pHa=7.99±0.04). However, after 15–20 days of anaemia (hct=6.6±0.8%) induced by blood withdrawal, the arterial CO₂ tension was significantly lower than the control value, suggesting that physiological adjustments occurred within this time period to compensate for the lowered haematocrit. Compensation probably did not involve alterations in ventilation, which was unaffected by 5 days of anaemia (anaemic ;w=786±187 ml min^–1 kg^–1 vs. control ;w=945±175 min^–1 kg^–1), based on indirect Fick principle measurements.Potential adaptations to longer term anaemia at the level of the red blood cells were investigated using a radioisotopic HCO₃ ^– dehydration assay. Owing to the difference in haematocrits, the HCO₃ ^– dehydration rate for blood from anaemic fish was significantly lower than that for control fish following equilibration at the same CO₂ tension. This difference was eliminated when HCO₃ ^– dehydration rates were measured on blood samples adjusted to the same haematocrit, a result which implies that the intrinsic rate of CO₂ excretion at the level of the red blood cell was not up-regulated during anaemia. The difference was also eliminated by equilibrating the blood samples with CO₂ tensions appropriate for the group from which the sample was obtained,i.e., PCO₂=1.4 torr for control samples and PCO₂=3.2 torr for anaemic samples; each at the appropriate haematocrit. It is concluded that the elevated PaCO₂ helps to reset CO₂ excretion to the control level, but that some additional physiological adjustment occurs to lower the PaCO₂ after 15–20 days of anaemia.     

Influence of salinity and dietary composition on blood parameter values of hybrid red tilapia, Oreochromis niloticus (Linnaeus) ×O. mossambicus (Peters)

The influence of salinity and dietary composition on blood parameter values (haematocrit, leucocrit, immature lymphocytes, mature lymphocytes, granulocytes, plasma osmolarity and total plasma protein) of red hybrid tilapia, Oreochromis niloticus (Linnaeus) x O. mossambicus (Peters), was studied. Two groups of tilapia were fed a high-protein diet while kept in fresh or brackish water, respectively, and compared with two groups fed a low-protein diet under similar environmental conditions. Treatments were executed in duplo. Results show that salinity influenced all cellular blood parameters except the haematocrit. Dietary composition influenced the total plasma protein and haematocrit, while all parameters changed as time progressed during the experiment. It was concluded that the environmental parameters investigated in the present study should be considered when estimating fish health based on blood parameter values.     

Influence of sampling methods and anaesthetization on various haematological parameters of several teleosts

Effects of blood sampling procedure on various haematological parameters were investigated in four Baltic teleost species. In pike, parameters in samples from permanently cannulated and thus minimally disturbed specimens were used as a standard. Blood lactate and plasma K⁺ concentrations were the most labile. Because of the reduced muscular activity and shorter sampling time, blood from fish immobilized by stunning gave a better picture of the resting state than samples from anaesthetized fish. Incubation of venous blood under falling O₂ tension (sample ageing) caused a rapid change in plasma K⁺ concentration. For reliable plasma K⁺ values immediate centrifugation of blood samples is necessary.     

Cardiac remodelling,blood chemistry,haematology and oxygen consumption of Atlantic cod, <Emphasis Type="Italic">Gadus morhua</Emphasis> L., induced by experimental haemolytic anaemia with phenylhydrazine

Anaemia is a common pathology associated with many infectious and non-infectious diseases. The effects of haemolytic anaemia induced by i.p. injection of phenylhydrazine (PHZ) were studied in Atlantic cod. Phenylhydrazine injection (0.3 mg kg⁻¹) in a DMSO and saline vehicle induced a reproducible and stable anaemia reducing haematocrit, (Hct) by 62% over 3 weeks. Controls consisted of fish injected with saline and DMSO/saline vehicle with minimal effects on Hct or whole blood haemoglobin (Hb). Although anaemia resulted in reduced blood lactate and glucose in PHZ injected fish, there were no effects of anaemia on blood, sodium, chloride or potassium. Similarly, there were no changes in the relative proportions of leucocytes in the blood although an increase in the number of immature erythrocytes was observed in the anaemic fish. Anaemic fish showed a 29 and 22% increase in cardiac somatic index (CSI) relative to saline and vehicle controls, respectively, although there were no significant differences in the linear dimensions of the ventricle. Changes in cardiac somatic and ventricular somatic index correlated positively and significantly with Hct but not with whole blood Hb concentration. Anaemic fish had significantly reduced resting routine oxygen consumption compared with vehicle controls but were not able to increase oxygen consumption following a bout of exhaustive exercise. Plasma lactate concentrations increased significantly after exercise to a greater extent in anaemic fish compared with vehicle control fish. Phenylhydrazine is a useful model for studying haemolytic anaemia in Atlantic cod with minimal effects on blood biochemistry and haematology and clearly reduces the aerobic capacity in Atlantic cod.     

Metabolic responses of Nephrops norvegicus to progressive hypoxia

This work studied some of the metabolic responses of Nephrops norvegicus to a progressive reduction in water oxygen tension (PwO₂) at 12 °C. Experiments were designed to simulate water quality conditions that may occur during the trade of live crustaceans. Oxygen consumption rates and ammonia efflux rates were found to be constant over a wide range of PwO₂ values (20.4–5.9 kPa). A similar result was found for the difference between post-branchial and pre-branchial oxygen concentrations (20.4–2.6 kPa), obtained from a separate experiment. Anaerobic pathways, however, were activated after PwO₂ reached 6.3 kPa, as blood lactate and glucose concentrations increased from 1.24 ± 0.08 and 1.17 ± 0.19 (T₀ values) to 10.55 ± 8.99 and 3.63 ± 0.89 mg · 100 mL⁻¹ respectively. N. norvegicus was able to maintain blood pH levels at relatively constant values despite a drop in water pH levels and the accumulation of lactate observed at low PwO₂. Heart rates also remained stable during PwO₂ reductions, but scaphognathite beat rate increased considerably, probably as an attempt to maintain steady weight-specific oxygen consumption rates. N. norvegicus appeared to be well adapted to cope with progressive hypoxia as may occur during holding and transportation procedures.     

Plasma lysozyme levels in sheatfish Silurus glanis (L.) subjected to stress and experimental infection with Edwardsiella tarda

Two experiments were conducted to assess plasma lysozyme levels, leucocrit and haematocrit in juvenile sheatfish (90–140 g) under experimentally induced stress and infection. Stress was provoked by random modification of environmental light for 15 min, 16 times a day with a halogen lamp (500 W) and by holding fish in a net for 3 min twice a day. In the first trial, repeated measuring of marked sheatfish showed a progressive significant decrease in plasma lysozyme level, a significant increase in leucocrit and a significant decrease in haematocrit. After a rest period of 1 week, only the leucocrit value was back to the prestress value. In the second trial, light‐induced stress did not lead to any variation in lysozyme, leucocrit or haematocrit values. However, after intraperitoneal injection of Edwardsiella tarda (7 × 10⁸ cfu), a strong increase in plasmatic lysozyme in both stressed and unstressed groups of sheatfish was observed. The increase was significant vs. prechallenge levels 3 and 6 days after contamination, but no significant difference was detected between stressed and unstressed fish, despite the strongest stress (light+handling stress) applied to the fish at the beginning of contamination. Decreases in haematocrit values were observed in both groups of fish. A clear increase in leucocrit appeared in unstressed fish, whereas stressed fish showed a significant decrease in leucocrit values. Condition factor was significantly decreased by all kinds of stress. There were no difference in mortality between stressed and unstressed groups.     

Anaesthetic Effects on the Hepatic Portal Vein and on the Vascular Resistance of the Tail of the Chinook Salmon (<Emphasis Type="Italic">Oncorhynchus tshawytscha</Emphasis>)

We used blood vessel myography and a perfused tail preparation from the Chinook salmon (Oncorhynchus tshawytscha) to investigate the effect of 2 commercially used fish anaesthetics, AQUI-S^TM and MS222, on vascular tone. Hepatic portal vein rings were exposed to 1 × 10⁻⁷ M adrenaline in the presence or absence of either AQUI-S^TM or MS222 and changes in vessel tension measured. Tail preparations were perfused with saline containing increasing concentrations of either anaesthetic. Exposure to either anaesthetic did not alter the response of vessel rings to adrenaline in vitro. However, both anaesthetics caused a significant (P ≤ 0.05) decrease in vessel tension when compared to controls. In tail preparations, perfusion with either anaesthetic caused a significant (P ≤ 0.05) yet reversible, dose dependent decrease in vascular resistance. Further to this, it was found that for both anaesthetics there was a significant effect of fish holding time on the response of preparations, with fish that were acclimated for less than 1 week exhibiting less vasodilation in response to anaesthetic exposure. We conclude that commonly used concentrations of AQUI-S^TM and MS222 can have a direct effect on vascular tone of salmon vessels, causing a significant but reversible vasodilation of vessels. This vasodilation may offset increases in haematocrit seen during anaesthesia in the whole animal, and in recovery prevent large rises in peripheral resistance associated with catecholamine release. A reduced vascular resistance may allow the heart to pump greater volumes of blood during recovery.     

Effects of Sangrovit‐supplemented diet on growth performance,blood biochemical parameters,survival and stress resistance to salinity in the Caspian roach (Rutilus rutilus) fry

This study was carried out to show the effects of Sangrovit on growth, blood biochemical parameters, survival and resistance to salinity stress in the Caspian roach. Fish (1.24 ± 0.13 g) were divided into four groups fed diets containing Sangrovit at different levels: 0 (control), 0.05, 0.1 and 0.15% for 45 days. The results showed that there was a significant increase in final weight, weight gain and specific growth rate in those fish fed 0.15% Sangrovit diets compared with the control (P < 0.05). Food conversion rate of fish fed 0.05% Sangrovit diet was significantly (P < 0.05) lower than the control group. Condition factor was not significantly affected by Sangrovit dosage (P > 0.05). The blood glucose and cholesterol levels were significantly lower in treated groups compared with the control (P < 0.05). The highest total protein was obtained in the 0.15% group and it was significantly higher than in all other groups (P < 0.05). Challenges were carried out after 45 days of feeding, to determine the effect of Sangrovit on resistance to salinity stress. Blood samples were obtained four times after challenge (day 1, 3, 5 and 7) for evaluate haematocrit levels. On the first day, haematocrit increased significantly in all groups compared to day 5 and 7 (P < 0.05). The haematocrit levels decreased gradually in all groups from the third day. Before and after stress, no differences were observed for survival rate among the experimental diets (P > 0.05). These results indicate that the inclusion of 0.15% Sangrovit in the diet of the Caspian roach enhances feed efficiency, fish growth and blood biochemical parameters.     

Oxygen consumption and ammonia-N excretion in fed and starved tiger prawns,Penaeus esculentus Haswell

Prawns, 17.7 ± 0.26 g wet weight, were held at 25°C (full moulting cycle at this size, 18–20 days) until 2 days after moulting. Groups of seven, eight or nine were then starved for 5, 10 or 15 days, respectively. Control groups of seven prawns each were fed ad lib for the same periods. The oxygen consumption and ammonia-N excretion over 24 h of each prawn was measured, using antibiotics to minimise bacterial growth during the excretion measurements. Oxygen consumption fell sharply by 24–29% during the first 5 days of starvation and then levelled out. Ammonia-N excretion increased 46–73% with starvation. O:N ratios fell to about 7:1 after the first 5 days of starvation and remained at this level up to 15 days, indicating that protein is virtually the only energy substrate under these conditions. These results support previous research that showed that the carnivorous P. esculentus uses muscle protein preferentially as a source of energy during starvation. In fed prawns, O:N ratios were also low (13.6–16.4:1), indicating that protein is normally a major source of energy.     

Simple field methods for monitoring stress and general condition of fish

While physiological stress in fish can be assessed by several laboratory-based methods, many of those methods use very sensitive and expensive instruments, which make them impracticable under field conditions. The objective of this study was to determine the feasibility of detecting stressed states in fish, under Held conditions, using simple but reliable methods. The first set of experiments were conducted in the laboratory, where a standard handling stressor was applied to juvenile coho salmon, Oncorhynchus kisutch (Walbaum). Results of blood samples analysed by both laboratory-based and portable methods (e.g. glucometers for diabetic patients) were in good agreement. The second set of experiments focused on field-testing the most promising methods based on the laboratatory tests. Production lots of Atlantic salmon. Salmo salar L., at commercial fish farms were sampled to assess stress and health status with the selected field techniques, from the swim-up to smolting stages. The measured variables included: blood glucose, protein, haemoglobin concentrations: haematocrit: erythrocyte and lymphocyte numbers: and an autopsy-based health profile, which involved the internal and external examination of the body for texture and colour. In addition to establishing resting, or normal, values for each variable under field conditions, blood glucose levels were observed to increase significantly at 4 h after a stressful event (e.g. grading, handling) using these field methods. Some of these field techniques were found to be reliable indicators of stress in fish and have potential for use in aquaculture facilities and field monitoring programmes.     

Some haematological observations on carp, Cyprinus carpio L., experimentally infected with Trypanoplasma borreli Laveran & Mesnil, 1901 (Protozoa: Kinetoplastida)

Abstract. Common carp, Cyprinus carpio L., were experimentally infected with cloned Trypanoplasma borreli and some haematological data were determined. The haematocrit and total erythrocyte counts were reduced and were lowest at day 29 post-inoculation during the peak of infection. Differential erythrocyte number showed a reduction in mature erythrocytes and an increase in immature erythrocytes. The total leucocyte number was markedly increased and peaked at day 44 post-inoculation. Differential blood cell counts showed an increased abundance of leucocytes, especially granuloblasts and granulocytes.     

The test of a centrifugal hematology system for use in clinical practice

The QBC is a centrifugal haematology system. Modified haematocrit capillaries are measured optically. The parameters haematocrit, total leukocyte count, relative and absolute values of granulocyte and lympho-/monocyte fractions, and total thrombocyte count are stated. The microhaematocrit method, the counting chamber, and the differential blood count are reference methods. Blood samples of the dog, cat and horse were used for the study. As a screening method the QBC analysis meets all requirements for the veterinary practice.     

The use of additional lighting and artificial photoperiods to recondition early maturing Atlantic salmon (Salmo salar) in Tasmania

Salmonid culture in Tasmania experiences high rates of early maturation due to the rapid growth rates achieved under favorable environmental conditions. The present study investigates the use of artificial photoperiod to recondition previously mature Atlantic salmon to provide a larger fish with higher market value. Individual groups of previously mature female Atlantic salmon (Salmo salar) were put under 24 hour artificial light from either May, June, July or maintained under ambient conditions during the austral winter. Fish maintained under lights attained a reversion from dark skin pigmentation, typical of mature individuals, to a silver sea-going coloration more rapidly than the control group. In addition flesh pigmentation, ovarian absorption and body wall thickness was significantly greater than that of the control fish resulting in a higher market price. The study demonstrates that artificial lights can facilitate the reconditioning of previously mature Atlantic salmon under southern hemisphere conditions.     

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.     

Effect of stocking density,oxygen level,light regime and swimming velocity on the incidence of sexual maturation in adult Atlantic salmon (Salmo salar)

The effects of various environmental parameters on sexual maturation of two sea-winter Atlantic salmon (Salmo salar) were tested in two separate experiments. In the first experiment Atlantic salmon with initial mean individual weight 1.5 kg (smolt 13 months before) were reared for 8 months from June to February at different oxygen levels and stocking densities using continuous light. Oxygen levels of 5–7, 7.5–9.5 and 10–12 mg O₂ l⁻¹ and stocking densities starting at about 20, 30 and 40 kg m⁻³ and increasing as the fish grew to 80–90 kg m⁻³ for the highest densities were tested in a factorial design. Only male fish matured, and incidence of maturation among males varied from 4.1% to 25% between tanks. The highest percentage of mature males was found in the tanks with low stocking density. No clear effect on oxygen level was found.The second experiment lasted 20 months from seawater transfer in May until the fish weighed 3.3–3.5 kg. Two water current speeds (14–16 and 20–24 cm s⁻¹) and two photoperiod regimes (LD 20:4) and continuous light (LL) were tested in a factorial design. Neither swimming velocity nor photoperiod affected growth rate. Continuous light reduced the incidence of sexual maturation. The average proportion of maturation among males was 8% and 25% under the LL and LD 20:4 regimes respectively. The fish reared under the LD 20:4 light regime had a significant lower condition factor and significant larger hearts than the fish reared under continuous light. Swimming velocity had no significant effect on the incidence of maturation. The results indicate that the swimming velocity must be higher than 0.5 BL s⁻¹ in order to influence the energy stores. An important finding in this study is that light cues are not required for gonadal growth. The results also indicate that environmental factors can affect maturation even after the first sea-winter.     

单养条件下雄性中华绒螯蟹摄食行为及其与螯肢关系的研究

为了解摄食行为对中华绒螯蟹(Eriocheir sinensis)的个体生长发育以及群体生活的影响,选取12只附肢健全的雄蟹,使用高清摄像头连续6 d记录其在1 h内的摄食行为。结果显示:每天摄食的河蟹比例约50%,第2天的摄食量显著高于另外5 d,第1天摄食反应时间显著高于另外5 d;河蟹摄食连续天数以连续摄食2 d的河蟹数量最多,各连续天数间蟹的摄食量无明显差异,连续摄食6 d的河蟹对饵料反应时间显著高于连续摄食3、4和5 d。河蟹摄食期间使用右螯、左螯和双螯的频率分别为55%、41%和4%。右螯比左螯长的河蟹比例为91.67%。螯长与摄食量无明显相关性,但螯长与体长存在正相关性(R^2=0.968 3,P<0.01)。结论:每天摄食的河蟹比例约占一半,平均摄食量约占体重的1.6%;摄食反应时间第1天最高,连续摄食可造成蟹对饵料的反应时间升高。雄性河蟹右螯较长,是摄食期间最常使用的螯肢。