Ontogeny of the cortisol stress response in yellow perch (Perca flavescens)

The ontogeny of the cortisol stress response in yellow perch (Perca flavescens) was documented by measuring resting and stress-induced whole-body cortisol levels in fish at several early development stages. In newly fertilized eggs resting cortisol levels averaged 41.0 ± 3.0 pg/embryo, and decreased to 8.0 ± 3.0 pg/embryo by day 6 where they remained constant until hatch on day 10. Acute stress challenge tests (30 sec cold shock) conducted on embryonic (days 6 and 8 post-fertilization) and larval (hatch and 2 days post-hatch) yellow perch revealed no differences in whole-body cortisol levels between stressed and unstressed individuals. In acute stress challenge tests (30 sec net confinement) conducted on post-hatch yellow perch, differences in cortisol levels between stressed (30 min post-stress) and unstressed fish were detected one week after hatching (12.3 ± 1.6 ng/g and 7.0 ± 1.3 ng/g, respectively). The magnitude of the stress response was greater in 2- to 5-week-old fish than in one-week-old fish. Changes in post-stress cortisol levels were closely associated with changes in interrenal morphology. Cumulatively, these results indicate that cortisol biosynthesis, integration and maturation of the hypothalamic-pituitary-interrenal (HPI) axis can be observed in yellow perch as early as one week after hatching. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of butaphosphan and cyanocobalamin combination on plasma immune and biochemical parameters of Olive flounder (Paralichthys olivaceus) subjected to crowding stress

The study evaluated the effects of intramuscular injection of butaphosphan and cyanocobalamin combination (BPC) on the plasma levels of cortisol, cytokines, and the activities of selected enzymes in Oliveflounder exposed to crowding stress. Three groups of fish (n = 12 per group) were kept in different glass tanks with stocking densities of 46 kg/m³(group BP and SCH) and 15 kg/m³ (group SCL). Group BP was treated with 0.5 ml of BPC. While groupSCH and SCL were treated with 0.5 ml of saline. Blood was collected a day after injection (T1), and weekly for two consecutive weeks (T2 and T3). The plasma levels of interleukin (IL) ‐1β, IL‐6, cortisol, lysozyme, and the activities of aspartate aminotransferase (AST) and alanine transaminase (ALT)were determined using an ELISA kit. The statistical significance of differences was assessed using ANOVAfollowed by Tukey‐HSD test (P < 0.05 considered significant).A significantbody weight gain was measured in the BP and SCL groupscompared with SCH at T3. The activities of AST (at T1, T2, and T3) and ALT (at T1 and T2) were declined significantly (P < 0.05) following treatment of overcrowded fish with BPC than saline. Similarly, BPCcaused a significant reduction in the levels of cortisol and IL‐1β at T2 and T3.This is the first report of the effect of the combination of butaphosphan and cyanocobalamin on plasma immune and biochemical parametersin fish. Therefore, the combination could be beneficial in preventing overcrowding‐induced immune suppression and tissue damage in Olive flounder.     

Physiological stress responses in spotted wolffish (Anarhichas minor) subjected to acute disturbance and progressive hypoxia

The spotted wolffish (Anarhichas minor) is considered a promising species for coldwater aquaculture. It is a sedentary, bottom-dwelling fish which exhibits a calm and “non-stressed” behaviour in captivity. There are, however, no reports on the physiological responses to stressors in this species. In the present study we investigated primary (cortisol secretion) and secondary (glucose mobilization) responses to common aquaculture stressors like disturbance and hypoxia. Pre-stress plasma cortisol levels were within those generally considered representative for unstressed fish (~ 10 ng ml^− 1), whereas basal glucose levels were unusually low (0.3–0.4 mM). After exposure to a short, but severe disturbance challenge (emptying the tank of water for 10 min), the increase in plasma cortisol level was slow and relatively weak, reaching a peak level of 25 ng ml^− 1 4 to 8 h after disturbance. When the fish were exposed to a gradually decreasing oxygen level in the tank, a significantly elevated plasma cortisol level (35 ng ml^− 1) was seen in the fish that remained in the tank until oxygen saturation had decreased to 20% oxygen saturation after 2.5 h. However, a two-fold higher plasma cortisol level (~ 70 ng ml^− 1) was seen in all fish exposed to reduced oxygen levels (60, 40 and 20% oxygen saturation) after 3.5 h recovery in normoxic water. Plasma glucose levels showed only moderate increases (~ 70%) following disturbance and hypoxia challenges. An in vivo injection of ACTH caused a strong elevation of plasma cortisol (peak level ~ 170 ng ml^− 1), demonstrating a high capacity for interrenal steroidogenesis in the spotted wolffish. The slow and relatively weak cortisol response to stressors, and low plasma glucose levels, may relate to the sedentary lifestyle of the spotted wolffish. The stress-response is characterised by a passive (reactive) coping style, which is considered adaptive for farming of this species.     

Oral administration of Escherichia coli lipopolysaccharide enhances the immune system of striped catfish,Pangasianodon hypophthalmus (Sauvage)

Lipopolysaccharide (LPS), a component of Gram negative bacteria, was reported as important immunostimulant for fish. In this study, striped catfish were fed diets containing different Escherichia coli LPS concentrations (0%, 0.01% and 0.05%) for 2 weeks and then fed control feed (0% LPS) for 4 weeks. Plasma cortisol and glucose were rather low and did not differ significantly among treatments (P > 0.05). The respiratory burst activity, lysozyme, complement, total of antibody as well as mortality in fish challenged with Edwardsiella ictaluri were recorded every 2 weeks (W2, W4 and W6). The lysozyme activity significantly increased in fish treated with LPS (P < 0.05) in W2, W4 and W6. The highest values of respiratory burst activity were observed at week 4 in fish fed 0.01% LPS. There were significant differences in total of antibody between fish fed LPS (0.01%) and control in W2, W4. The challenge test with Edwardsiella ictaluri showed that fish fed 0.01% LPS had lower cumulative mortality (40%, 33% and 42%) compared with the fish fed 0.05% LPS (50%, 40% and 47%) and control fish (40%, 57% and 53%) in the three difference sampling times respectively. These results suggest that feed supplemented with 0.01% LPS could enhance immunity of striped catfish after 2 weeks of oral administration and fish could be protected against bacterial infection during the following 4 weeks.     

Cortisol and liver metabolism of immature American eels,Anguilla rostrata (LeSueur)

Plasma and liver composition, liver enzyme activities, and metabolite flux in isolated hepatocytes have been studied in immature American eels,Anguilla rostrata, injected daily IP with saline or cortisol (0.35 mg/kg eel). Plasma cortisol values were significantly increased above saline controls in those eels receiving cortisol at 3h and 6h following the final (tenth) injection. On day 6 and 10 of injection plasma cortisol levels were significantly below saline controls 24h following cortisol injection. Plasma glucose values were significantly depressed in the cortisol-injected eels at both 6 and 24h following the final (tenth) injection. At the 24h sampling time, plasma protein had significantly increased, but there was no change in either plasma amino acid or fatty acid levels. An increased hepatosomatic index was attributed to a major increase in total lipids, as both protein and glycogen contents were decreased. Of the liver enzymes assayed, significant activity changes occurred only for lactate dehydrogenase (decreased), mitochondrial citrate synthase (increased) and phosphoenolpyruvate carboxykinase (increased) 24h following the final (tenth) cortisol injection. Although enzyme activity changes implied increased liver gluconeogenesis, the absolute rate of lactate, alanine, and aspartate incorporation into glucose declined in viable hepatocytes isolated from cortisol-injected eels compared to the saline controls. Relative changes in metabolite flux did support a preferential increase in gluconeogenesis from amino acids. These results are consistent with the increase in amino acid gluconeogenesis as a result of cortisol administration implied in previous studies, but failed to show a definitive cortisol effect on this pathway in the eel liver. It is suggested that other hormones (e.g. thyroxine, catecholamines, glucagon) may interact in a complex way with cortisolin vivo to bring about the biochemical changes observed in this study. The rapid clearance of exogenously injected cortisol noted in this study makes causal relationships between the injected hormone and any observed metabolic effect in the intact animal difficult.     

甘草次酸对斑点叉尾（鱼回）免疫应激下血浆抗氧化酶和免疫指标的影响

为探讨斑点叉尾(鮰)免疫应激状态下血浆抗氧化酶和免疫指标的变化,并研究甘草次酸对斑点叉尾(鮰)免疫应激的调控机制,实验设计两组,每组6个重复:1)对照组,饲喂基础饲料;2)GA组,饲喂含0.15 g/kg甘草次酸饲料,养殖8周后,腹腔注射大肠杆菌脂多糖(LPS),分别在注射后0、3、6、12、24、48 h采集血液和肝脏组织,进行红细胞、白细胞计数,分离血浆后检测血浆皮质醇、溶菌酶、ACH50、谷草转氨酶、谷丙转氨酶、碱性磷酸酶活性,肝脏匀浆液检测SOD、MDA和CAT抗氧化酶活性.结果显示,在腹腔注射LPS后,血浆溶菌酶、皮质醇、谷草转氨酶、谷丙转氨酶、ACH50、SOD均呈现先升高后下降的趋势,白细胞计数、血浆CAT呈现先下降后升高的趋势;饲料中添加0.15 g/kg甘草次酸显著抑制了各指标变化,缩短了由LPS诱导产生各指标变化的恢复时间.研究表明,腹腔注射LPS使斑点叉尾(鮰)产生免疫应激,并在注射后6～12h达到顶峰;饲料中添加0.15 g/kg甘草次酸可以有效抑制由LPS诱导的免疫应激反应.     

Effect of L‐tryptophan supplemented diets on serotonergic system and plasma cortisol in Totoaba macdonaldi (Gilbert, 1890) juvenile exposed to acute stress by handling and hypoxia

Brain serotonin and cortisol play a central role in integrating the neuroendocrine response to stress. Previous studies in aquatic species show that dietary supplementation with the 5‐HT precursor L‐tryptophan amino acid is able to modulate this response. The aim of this study was to evaluate the effect of dietary tryptophan (Trp) on the serotonergic system and acute stress response in Totoaba macdonaldi. Four groups of fish were fed with one of four isonitrogenous and isocaloric diets formulated with different Trp content: 0.5% (1X TRP, control diet), 1.1% (2X TRP), 1.7% (3X TRP) and 2.3% (4X TRP). After a feeding period of 21 days, groups were exposed to acute stress by handling with a net or hypoxia. Feeding with Trp‐enriched diets resulted in increased 5‐HT content in telencephalon of undisturbed fish. Handling increased the 5‐hydroxyindole acetic acid (5‐HIAA) and serotonin turnover (5‐HIAA/5‐HT) in telencephalon of fish fed with 1X TRP and 2X TRP but was not significant for the fish fed 3X TRP and 4X TRP, and no significant increase was observed by hypoxia in both telencephalon and hypothalamus in any of the diets. An inverted U‐shaped response was observed in plasma cortisol increasing with 2X TRP after both handling and hypoxia and decreasing with diets rich in Trp, 3X TRP and 4X TRP. A higher increase was observed in haematocrit, plasma glucose and lactate in hypoxia than the handling groups. Our results indicate that Trp supplementation modulated cortisol response and mediated negative feedback of 5‐HT system in telencephalon after acute stress.     

Effects of repeated crowding on the stress response and growth performance in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

A 64-day growth experiment was conducted in which two groups of Atlantic salmon parr were grown under either control conditions or subjected to a weekly crowding stressor. Subjecting fish to the stressor resulted in a 7.7% reduction in wet weight after 29 days, which was maintained at 7.9% by day 64. This reduction in weight was reflected in a 44% reduction in specific growth rate and 38% increase in feed conversion ratio over the first 29 days of the experiment. Elevation in plasma cortisol was observed in crowded fish on days 1, 29 and 64. Similarly, on days 1 and 29 an increase in both plasma glucose and lactate was detected. On day 64, however, no differences in plasma glucose and lactate were observed, with the magnitude of the cortisol response also significantly reduced. Overall, the relatively moderate impact on growth performance and reduction in magnitude of measured stress parameters at the end of the experiment suggests possible habituation to the applied stressor.     

Effect of blood glucose level on acute stress response of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Stress has a considerable impact on welfare and productivity of fish, and blood glucose level of fish may be a factor modulating stress response. This study evaluated the effect of blood glucose level and handling on acute stress response of grass carp Ctenopharyngodon idella. Fish were intraperitoneally injected with glucose at 0, 0.2, 0.5, and 1.0 mg g^?1 body mass (BM) and then were exposed to handling for 5 min. Glucose injection resulted in increase of plasma glucose level and liver glycogen content and decrease of plasma lactate level. Handling resulted in increase of plasma levels of cortisol, glucose, and lactate and plasma lactic dehydrogenase (LDH) activity and decrease of liver glycogen content. At 1 h post-stress, the plasma cortisol level was lower in the stressed fish injected with glucose at 0.5 mg g^?1 BM than the stressed fish injected with glucose at 0, 0.2, and 1.0 mg g^?1 BM. No significant differences were found in the activities of phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate kinase (PK) in the liver between the stressed and unstressed fish, regardless of the dose of glucose injection. At 1 h post-stress, the liver glucose-6-phosphatase (G6Pase) activity was higher in the fish without glucose injection than in the fish injected with glucose. This study reveals that blood glucose level can affect stress response of grass carp by modulating cortisol release and glucose homeostasis through glycogen metabolism and gluconeogenesis in the liver.     

Species‐specific effects of subdaily temperature fluctuations on consumption,growth and stress responses in two physiologically similar fish species

Fluctuations in water temperature can have important physiological consequences for fishes. Effects of daily thermal cycles are well studied and can be beneficial, increasing prey consumption and growth rates when mean and maximum temperatures of the fluctuations are at or below the species’ optimum temperature. While less studied, subdaily temperature fluctuations are also common in many aquatic habitats and can be caused by both natural and anthropogenic processes. We performed laboratory experiments to examine how two fish species (yellow perch, Perca flavescens, and walleye, Sander vitreus) with similar thermal preferences respond to chronic exposure to subdaily temperature variability. We selected temperature treatments that reflected observed thermal variation after examining water temperature data from multiple aquatic systems. We then separately exposed yellow perch and walleye to a stable 23 °C treatment and 12‐h cycles of 23 ± 2 °C or 23 ± 4 °C for 45 days. Adult yellow perch exposed to fluctuations of 23 ± 4 °C over 12 h expressed higher consumption, growth and food conversion efficiency than fish experiencing stable 23 °C. Temperature fluctuations, though, resulted in mortalities and the development of skin ulcers in yellow perch that did not occur under stable temperatures. In contrast, the same 12‐h temperature fluctuations did not result in mortalities or stress responses in juvenile walleye. Moreover, unlike yellow perch, growth rates of walleye were lower under 12‐h temperature fluctuations compared with the stable 23 °C treatment. Our results indicate that species with similar thermal preferences can respond differently to the same subdaily temperature fluctuations.     

Effects of hormonal manipulation on stress responses in male and female broodstocks of pikeperch Sander lucioperca

This study was designed to determine the effects of hormonal manipulation on stress responses in female and male pikeperch. Two-year-old cultured female and male broodstocks with an average weight of 337.4 ± 20.1 (mean ± SE; n = 16) and 318.7 ± 15.1 g (n = 16), respectively, were randomly allocated into four hormonal treatments each containing 4 fish. Two sexual groups of 16 fish for each gender were considered. Sexually mature male and female pikeperch were injected with either physiological saline solution (as control group), common carp pituitary extract (CPE), human chorionic gonadotropin (hCG) and or luteinizing hormone-releasing hormone analog (LHRHa₂). The blood samples were taken before hormonal injection and after ovulation and spermiation. Then the plasma levels of stress indices (cortisol, glucose, and lactate) were determined. The results showed that all CPE-, HCG-, and LHRHa_2- injected males produced sperm. In females treated with CPE and hCG, three of four ovulated, but none of LHRHa_2- and saline-injected fish spawned. Significant changes in cortisol, glucose, and lactate levels were observed among the females injected with different hormones. Plasma cortisol and glucose levels increased significantly in males injected with CPE and females injected with hCG, but no significant change was observed in lactate levels before and after hormonal induction. Comparison of two sexes revealed significant differences in glucose levels for females in some groups before injection, while CPE-injected sexes showed significant changes in cortisol and lactate concentrations. The results indicated that the induction of ovulation or spermiation stimulated stress responses especially in female pikeperch, and therefore, all the procedures should be made to minimize the disturbance during the artificial spawning.     

Leukocyte response and phagocytic activity in Nile tilapia experimentally infected with <Emphasis Type="Italic">Enterococcus</Emphasis> sp.

This study evaluated the total and differential leukocyte counting and the phagocytic activity in Nile tilapia Oreochromis niloticus experimentally injected with Enterococcus sp. in the swim bladder. Fish were distributed in four treatments in triplicates of non-injected fish, fish injected with 1 ml of sterile saline solution 0.65%, and fish injected with 1 × 10³ and 1 × 10⁶ colony-forming units (CFU) of Enterococcus diluted in 1 ml sterile saline. Twenty-four hours after injection, the fish were anesthetized and the blood collected for white blood cell (WBC) counts, differential counting of WBC, and phagocytic activity of blood leukocytes. The increased numbers of WBC and lymphocytes were followed by decreased number of monocyte after infection. The percentages of phagocytic activities in the blood were 55.3 and 55.9%, respectively, in tilapia injected with 1 × 10³ and 1 × 10⁶ CFU/ml.     

Capture and handling stress affects the endocrine and ovulatory response to exogenous hormone treatment in snapper, Pagrus auratus (Bloch & Schneider)

Sexually mature female hatchery‐reared snapper, Pagrus auratus (Bloch & Schneider) were captured from sea cages by handline and injected at first capture (control) or 24 h after capture, transport and subsequent confinement (delayed injection) with either saline, luteinizing hormone releasing hormone analogue, human chorionic gonadotropin, or 17α‐hydroxyprogesterone. Blood was sampled before hormone treatment and again after 168 h, and fish were checked daily for ovulation. Plasma levels of 17β‐estradiol (E2), testosterone (T), 17α, 20β dihydroxy‐4‐pregnen‐3‐one (17, 20βP) and cortisol were determined by radioimmunoassay. The ovulatory response was assessed from the proportion of fish ovulating, ovulation volume, egg quality and fertility. A delay in injection resulted in significantly lower plasma E2 and T levels in response to hormone treatment, smaller ovulation volumes, and poorer egg quality than in control fish. The results are consistent with the generally inhibitory effects of stress on reproduction in fish, and confirm the requirement to treat fish with hormones designed to induce ovulation, as soon as possible after capture and disturbance.     

Carbohydrate utilization by juvenile silver perch, Bidyanus bidyanus (Mitchell). I. Uptake and clearance of monosaccharides following intraperitoneal injection

Abstract Intraperitoneal carbohydrate tolerance tests were done to assess the ability of silver perch, Bidyanus bidyanus, to utilize the predominant monosaccharides in plant ingredients currently being used in the formulation of aquaculture feeds for this species. Preliminary experiments carried out to assess baseline plasma glucose concentrations indicated that blood glucose levels were elevated within 2 min of handling and silver perch required a period of 48 h without feeding before plasma glucose levels remained constant. In the first carbohydrate test, either glucose, galactose or xylose were administered by injection into the intraperitoneal cavity at a dose rate of 1 g carbohydrate kg^?1 body weight (BW). In the second carbohydrate test, glucose was administered at a dose rate of either 2 or 4 g glucose kg BW^?1. Following injection, uptake and clearance rate of the carbohydrates from the blood stream was monitored over a 24‐h period. Silver perch were significantly more efficient at the uptake and clearance of glucose from the blood stream than xylose or galactose. Maximum plasma glucose concentrations (22.2 mmol L ^?1) were recorded at 1 h following injection and basal levels (3.44 mm ) were attained between 6 and 12 h following injection. For both galactose and xylose, maximum concentrations were recorded at 1 and 3 h, respectively, and concentrations of both monosaccharides remained significantly elevated 24 h after the administration. Plasma glucose concentrations of silver perch administered with either 2 or 4 g glucose kg BW^?1 were significantly elevated and peaked at similar levels (30.2 mmol L ^?1 and 30.7 mmol L ^?1 respectively) 3 h after injection. Basal plasma glucose concentrations were attained in silver perch injected with 2 g glucose kg BW^?1 at 24 h following administration. Plasma glucose concentrations remained significantly elevated in fish injected with 4 g glucose kg BW^?1 after 24 h. These findings indicate that silver perch are more efficient at utilizing glucose than either xylose or galactose, and that there are also differing maximum threshold for the inclusion of ingredients rich in glucose, galactose and xylose into the diets of silver perch.     

Determinism of the induction of the reproductive cycle in female Eurasian perch, Perca fluviatilis: Identification of environmental cues and permissive factors

In Eurasian perch, Perca fluviatilis, gametogenesis is induced by decrease of both temperature and photoperiod. However, a multiplicity of other factors can display non-permissive or modulating effects on the induction of the reproductive cycle. Consequently, a 9-week study was carried out to determine the environmental cues and permissive factors controlling the induction of the reproductive cycle in this species. A two-level fractional factorial design 2^8-4 was used in order to identify the influent factors from 8 environmental and nutritional ones and their 28 interactions. Tested factors were: (1) amplitude of temperature decrease, (2) time, (3) kinetics and (4) amplitude of photoperiod decrease, (5) initial nutritional state, (6) handling, (7) feeding rate and (8) light spectrum. Increase of gonadosomatic index (GSIi), proportion of female in exogenous vitellogenesis (EVP), plasma 17 β-estradiol (E₂) and cortisol levels, fat consumption and food intake were evaluated.This study showed that the amplitude of the decrease of temperature and the time of photoperiod decrease are the two main environmental cues controlling the induction of the reproductive cycle in Eurasian perch. GSIi was the highest (3.8%) when females were exposed to both low amplitude of temperature decrease and precocious decrease of photophase. It was the lowest (1.1%) when high amplitude of temperature decrease and late decrease of photophase were applied. Handling in association with temperature modulated the broodstock response. 100% of the unhandled fish held at 14 °C were at the exogenous vitellogenesis stage with mean E₂ and basal cortisol levels of 1.6 ng mL^− 1 and 9 ng mL^− 1 respectively. Handled fish and fish held at 6 °C exhibited lower vitellogenesis (40-73%) associated with lower E₂ (0.6-1.1 ng mL^− 1) and higher basal cortisol levels (37-89 ng mL^− 1). No other factor nor interaction displayed a cueing or non permissive effect on the induction of the reproductive cycle in female Eurasian perch. A first schematic model is proposed to explain the factorial determinism of the induction of the reproductive cycle.     

Validation of a whole-body cortisol extraction procedure for channel catfish (<Emphasis Type="Italic">Ictalurus punctatus</Emphasis>) fry

We validated a whole-body cortisol extraction technique for channel catfish, Ictalurus punctatus, fry. Three volume enhancement methods were tested: CAL method (zero calibrator A diluent added to lipid extract), PBS method (phosphate buffered saline added to lipid extract), and VO method (food grade vegetable oil added to lipid extract). The volume enhancement extracts were evaluated using a commercial radioimmunoassay kit. Sensitivity, accuracy, precision, reproducibility, and parallelism could not be determined for the PBS method as cortisol levels were not detected in any of the extracted samples. Intra-assay coefficient of variation (CV) for the CAL and VO methods were 7.3 and 8.3%, respectively, while inter-assay CV were 9.6 and 10.6%, respectively. Based on the sensitivity, accuracy, precision, reproducibility, and parallelism results, we conclude that the CAL method is the most appropriate method for volume enhancement of catfish fry lipid extract. Using the CAL method to detect cortisol in catfish fry, fish were stressed daily for 2 weeks. Fry weights were similar throughout the study while whole-body cortisol levels were higher (P < 0.01) in stressed fish after 1 day of stress. These data show the CAL method can effectively measure whole-body cortisol in catfish fry.     

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.     

Stress responses of Atlantic salmon (Salmo salar L.) elicited by water level reduction in rearing tanks

The Atlantic salmon (Salmo salar) stress response was examined by measuring plasma cortisol, glucose and chloride in fish after water level reduction within rearing tanks. Maximum plasma cortisol levels (366±43 (SD) nM and 534±280 nM for Groups 1 and 2, respectively) were observed 20 min after application of the stressor. Cortisol levels were down to control levels 24 h later. The pattern for cortisol changes observed within these two groups were comparable. Plasma glucose and chloride concentrations did not change significantly in either of the experimental groups. The fish in group 2 were then repeatedly subjected to the same stressor every third day. After the fifth exposure to the stressor, blood was obtained from fish at times corresponding to sampling after the first exposure. Their maximum plasma cortisol level only reached 223 (±96) nM, and was down to prestress levels within 2 h. Plasma glucose and chloride concentrations did not change significantly in this second experiment. This challenge test revealed an acute primary response in Atlantic salmon without any apparent harmful secondary responses, that may thus serve as a standardized reference stressor using other fish groups under comparable conditions.     

Carbohydrate tolerance in the fruit‐eating fish Piaractus mesopotamicus (Holmberg, 1887)

Some fish species have a limited ability to metabolize dietary carbohydrates. An important tool for understanding carbohydrate metabolism is the application of the glucose tolerance test, which can be performed orally or intraperitoneally. To evaluate carbohydrate tolerance in the fruit‐eating fish pacu, two experiments were performed, one with oral administration by gavage of three carbohydrate types (glucose, fructose and starch, 2.0 g/kg body weight (BW)) and the other with intraperitoneal injection (IP) of glucose (500 mg/kg BW). Oral glucose resulted in an increase in plasma glucose 2 hr later with the peak at 4 hr (8.30 mmol/L), and return to baseline between 6 and 12 hr; starch administration promoted a peak after 4 hr (7.70 mmol/L), returning to the baseline at 6 hr. The administration of fructose promoted a moderate peak after 2 hr (5.71 mmol/L), and return to baseline for the time points that followed. Elevated serum cholesterol levels were observed 2 and 24 hr after administration of glucose and starch. Hepatic glycogen levels increased within 24 hr, regardless of the type of carbohydrate administered. IP glucose load resulted in a peak of plasma glucose 3 hr post injection (6.91 mmol/L), returning to baseline 6 hr later. There was a reduction in the concentration of triglycerides at 24 hr. The results demonstrate that pacu metabolize both oral (glucose or starch) and intraperitoneal (glucose) carbohydrate loads after 6 hr, suggesting good ability to deal with dietary carbohydrates.