Egg cortisol response to stress at early stages of development in Persian sturgeon Acipenser persicus

One of the most important periods in artificial breeding of sturgeon is incubation. In general, little is known about ontogeny of the stress response in early developmental stages of sturgeon. In this study, cortisol content was measured by radioimmunoassay for the first time in eggs of Persian sturgeon Acipenser persicus to elicit at which times during early developmental stages, that they were particularly sensitive to stressful events. In this study, fertilized eggs of mature Persian sturgeons were placed in special boxes (as replicates) that were put in incubation units of Yushchenko. To assess pre-stress samples (resting cortisol levels), 5 eggs were selected randomly from each box in 7 important stages of incubation (2-cell division, blastula, end of blastula, mid of gastrula, neurula, eyed stage and heart beaten). In another experiment, eggs were exposed to an acute stress (10 min out of water) in 7 developmental stages. For determination of post-stress cortisol, eggs were collected 2 and 6 h after the stress. The results indicated significant difference in various stages of incubation but cortisol levels revealed no differences in whole-body levels between stressed samples (2 and 6 h post-stress) and unstressed samples (P > 0.05). The highest percentage of hatching rate for post-stressed eggs was recorded in 2-cell division stage while this percentage declined steadily in the heart beaten stage when a statistically significant difference was revealed with regard to other stages (P = 0.000). These findings suggested that sturgeon eggs were not so much sensitive in their early developmental stages and relatively resistant to stress. However, significant decrease in survival rate in heart beaten stage showed a sensitivity of this stage to stress. Lack of an increase in cortisol after acute stress indicated that hypothalamus-pituitary-interrenal (HPI) axis for cortisol synthesis was not yet functional. However, further studies seem necessary to clarify when HPI axis is activated in this species.     

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.     

Ontogeny of the stress response in chinook salmon, Oncorhynchus tshawytscha *

Whole body concentrations of cortisol were determined via radioimmunoassay in chinook salmon, Onchorynchus tshawytscha, during early development in both stressed and non-stressed fish to determine when the corticosteroidogenic stress response first appeared. Progeny from both pooled and individual females were examined to determine if differences existed in offspring from different females. Levels of cortisol were low in eyed eggs, increased at hatch, decreased 2 weeks later and then remained constant thereafter. Differences in cortisol between stressed and control fish were found 1 week after hatch and persisted for the remainder of the study. The magnitude of the stress response, or relative amount of cortisol produced, generally increased from the time when it was first detected, but a decrease in the ability to elicit cortisol was seen 4 weeks after hatching. Cortisol content of separate progeny from two individual females showed a similar pattern to that seen in pooled eggs. Our results indicate that chinook salmon are capable of producing cortisol following a stressful event approximately 1 week after the time of hatching. The decrease in endogenous cortisol content seen 2 weeks after hatching, and the decrease in the magnitude of the stress response seen 4 weeks after hatching may be comparable to developmental events documented in mammals where corticosteroid synthesis is inhibited to neutralize possible detrimental effects of these hormones during critical periods of development. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Plasma cortisol and hypothalamic monoamine responses in yellow perch <Emphasis Type="Italic">Perca flavescens</Emphasis> after intraperitoneal injection of lipopolysaccharide

The concentrations of monoamines in the hypothalamus were determined in yellow perch Perca flavescens before and after injection with lipopolysaccharide (LPS; 3 mg kg⁻¹ fish weight) or saline to test for the presence of neurochemical changes potentially associated with changes in plasma cortisol characteristic of intraperitoneal (ip) challenge with LPS. In the first experiment, yellow perch were injected with saline or LPS and the hypothalamus removed and plasma sampled before and at 0.5, 1.5, 3.0, and 6 h after injection. Plasma cortisol was elevated in both saline- and LPS-injected fish through 1.5 h after injection and returned to levels resembling pre-injection by 3 h after injection. Significantly higher amounts of cortisol in plasma from LPS-injected relative to saline-injected fish were observed 6 h following injection. A significant decrease relative to levels observed 0.5–3 h after handling was observed in serotonin concentrations at 6 h following LPS and saline injection with a concomitant increase in the ratio of 5-hydroxyindoleacetic acid:serotonin. In the second experiment, hypothalamic monoamines were sampled before and at 3, 6, 9, 12, and 24 h after injection with LPS or saline. Significant increases from pre-injection levels were observed in the ratio 5-hydroxyindoleacetic acid:serotonin at 9, 12, and 24 h after injection, but no differences were detected between LPS- and saline-injected fish. These results support a model linking serotonergic system activation following handling stress, but no correlations with the sustained elevations of plasma cortisol associated with inflammatory challenge were observed.     

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 dietary nucleotides supplementation on rainbow trout (Oncorhynchus mykiss) performance and acute stress response

This experiment was conducted to examine the effect of dietary nucleotides (NT) on fish performance and acute stress response on fingerling rainbow trout (23 g ± 0.01, mean weight ± SEM). Five experimental diets according to different levels of supplemented nucleotides (0, 0.05, 0.1, 0.15, and 0.2%) were assayed on experimental fish for 8 weeks. Growth, hematological parameters (hematocrit, hemoglobin, erythrocyte, lymphocyte, and neutrophil count), serum proteins (globulin, albumin), and plasma enzymatic activity (alkaline phosphatase, ALP; aspartate transaminase, AST; lactate dehydrogenase, LDH; alanine transaminase, ALT) were assayed. At the end of feeding trial, fish fed the control and 0.2% diets were subjected to handling and crowding stress. Modulatory effects of nucleotides on acute stress response (cortisol and glucose) and plasma electrolytes (Na⁺, Cl⁻, K⁺, and Ca²⁺) were studied. The percentage of body weight gain (WG) and feed efficiency (FE) of fish were better when the fish were fed 0.15–0.2% diets. Fish fed the nucleotide-supplemented diets tended to have lower levels of serum enzymes including ALP, AST, LDH, and ALT. Plasma cortisol levels of fish fed on 0.2% diet under handling and crowding stress were significantly lower than fish fed the control diet at all post-stress time intervals. In our study, fish fed nucleotide-supplemented diet had significantly lower concentrations of glucose compared to those fed the basal diet. The concentrations of sodium, chloride, calcium, and potassium of fish fed the control diet were significantly lower than in fish fed nucleotide-supplemented diet. Dietary nucleotides administration seems to promote growth and to enhance resistance against handling and crowding stress in fingerling rainbow trout.     

Stress-induced changes in the affinity and abundance of cytosolic cortisol-binding sites in the liver of rainbow trout,Oncorhynchus mykiss (Walbaum), are not accompanied by changes in measurable nuclear binding

Plasma cortisol levels and the number (N_max) and affinity (K_d) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml^–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml^–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml^–1). There was a 60% reduction in cytosolic N_max in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg^–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the K_d of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in K_d was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in N_max in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.     

Emergence of larval yellow perch, Perca flavescens, in South Dakota lakes: potential implications for recruitment

Abstract  Temporal patterns in length frequency distributions and hatch dates were described for larval yellow perch, Perca flavescens (Mitchill), captured in surface ichthyoplankton trawls from late April to mid-June 2000 to 2002 in six South Dakota, USA lakes. Fewer than 15 larval yellow perch were collected in four of six lakes during 2002, suggesting that in some cases factors prior to, during or immediately after hatching likely play a critical role in the perch recruitment process. When larval yellow perch were encountered in larger numbers, temporal trends in total length (TL) frequencies indicated that only a single cohort was produced annually in each lake. Most yellow perch in these lakes hatched between 29 April and 17 May, and most hatching occurred during 5–11 days each year. Larval TL was not related to hatch date. The apparent prevalence of relatively short hatch periods in these yellow perch populations probably increases the risk of catastrophic losses resulting from periods of poor environmental conditions.     

Cortisol response to air exposure in <Emphasis Type="Italic">Solea senegalensis</Emphasis> post-larvae is affected by dietary arachidonic acid-to-eicosapentaenoic acid ratio

An experiment was conducted in order to evaluate the effects of feeding frozen Artemia diets differing in arachidonic acid-to-eicosapentaenoic acid ratios (ARA/EPA) on growth, survival and stress coping ability of Senegalese sole post-larvae (19–31 days after hatch). Two experimental diets presenting high (‘High’; 3.0) or low (‘Low’; 0.7) ARA/EPA ratios were tested under two rearing conditions: undisturbed (C) and stressed by a 2-min air exposure every two days (S). Growth, survival and basal cortisol levels were similar between groups indicating that independently of dietary ARA/EPA ratios, fish were able to cope with the repeated stress imposed. Also, cortisol levels at 3 h past air exposure were determined in all groups at the end of the experiment. Among fish fed the ‘Low’ diet, C groups seemed to present a quicker recovery from the acute stress (basal-like levels) than S groups. Repeated stress effects were not apparent in fish fed the ‘High’ diet and, relative to basal levels, twofold higher cortisol concentrations were detected at 3 h, in both C and S groups. This study suggests the importance of ARA in steroidogenesis regulation and the modulatory role of EPA in this process. Despite the tolerance to a wide range of dietary ARA/EPA as indicated by growth and survival results, acute stress coping response may be more efficient in Senegalese sole post-larvae fed low ARA/EPA ratios and, under these particular conditions, a faster recovery of cortisol to basal values could be indicative of rearing conditions (undisturbed vs. repeatedly stressed).     

Effects of Temperature on Performance Characteristics and the Cortisol Stress Response of Surubim Pseudoplatystoma sp.

Three experiments were conducted in flow‐through tanks at 24, 27 and 30 C to evaluate growth, food conversion, morbidity, survival, and the cortisol stress response of surubim Pseudoplatystoma sp. at different temperatures. In Experiment 1, fish (mean initial weight, 33.3 ± 7.2 g) reared at 27 and 30 C for 60 d grew significantly faster than fish reared at 24 C (P < 0.05). Fish at the lowest temperature showed the poorest feed conversion ratio (FCR, 5:1), while fish at 27 and 30 C had FCRs of 2.5:1 and 3.5:1, respectively. Mortality and morbidity rates were inversely proportional to the temperatures tested. In Experiment 2, serum cortisol levels following an acute handling stressor peaked at 30 min after stress, returned to near‐resting levels at 1 h after stress, and completely returned to resting levels at 3 h after stress in fish at all temperatures tested. Peak serum cortisol levels were higher in fish at 30 C (321.1 ± 38.8 ng/mL) than those in fish at 27 and 24 C (143.3 ± 37.4 ng/mL and 104.2 ± 37.2 ng/mL, respectively). In Experiment 3, fish (mean initial weight, 67 ± 8.58 g) reared at 27 C for 137 d grew significantly faster than fish at 30 and 24 C. Virtually, all fish reared in 30 C had some sign of morbidity (skin lesions and fin abrasion) compared to only a small percentage of fish in the other two temperatures. Again, the highest cortisol peak level was measured in fish reared at 30 C (117.6 ± 16.7 ng/mL), as compared to fish at 24 and 27 C (99.2 ± 15.5 ng/mL and 80.2 ± 12.8 ng/mL, respectively). Our findings indicate that the optimum temperature for rearing surubim in tanks is 27 C. At 30 C, these fish may suffer from stress, as suggested by the elevated peak of cortisol and morbidity and mortality rates. The relatively rapid cortisol stress response of surubim suggests that they may be more tolerant to handling stressors than other fish species.     

Cortisol and Behavioral Response to Handling (Acute) and Confinement (Chronic) Stressors in Juvenile Spotted Wolffish,Anarhichas minor

The purpose of this study was to determine the cortisol response of spotted wolffish (Anarhichas minor) after exposure to air immersion and netting (acute stress) under a range of increasing densities (chronic stressor). In addition, the presence of a cumulative negative impact from chronic stress was assessed by the application of an additional acute stressor following rearing at various densities, and proxies of behavior were evaluated (feeding and aggression data). The stress response to air immersion and netting led to increasing plasma cortisol values at 0.5, 13, and 37 h post-stress, with the highest value achieved after 13 h. At 168 h, cortisol concentrations returned to similar pre-stress value. Changes in cortisol concentrations were relatively low after acute stress (five to six fold increases) and recovery time long lasting (>37 h). Prolonged rearing at 20, 30, and 40 kg m^?2 showed a non-significant trend of increasing cortisol values with increasing density, and the cortisol response after the application of an additional acute stressor was significantly altered, i.e., higher cortisol levels were measured in unstressed (chronic stress only) compared to stressed fish (chronic stress + acute stress). Our results suggest that (1) spotted wolffish is a low responder that additionally displays a long-lasting cortisol elevation following an acute stressor; (2) fixed densities did not lead to significant differences of plasma cortisol concentration or aggression levels, although involved marked changes in daily feed intake and food conversion efficiency were observed; and (3) cortisol levels reached acute stress values after prolonged chronic stress. Although this species displays indications of being relatively resistant to handling disturbances during aquaculture practices, it is nevertheless responsive to chronic stress factors that could alter the physiological response to subsequent acute stressor.     

Diel changes in plasma cortisol and effects of size and stress duration on the cortisol response in European sea bass (Dicentrarchus labrax)

European sea bass (Dicentrarchus labrax), one of the most economically important fish in Mediterranean mariculture, shows high basal cortisol concentrations compared with other teleosts. The present study aims (a) to identify cortisol diel variation in fish held under a 12L:12D cycle and minimum handling stress, and (b) to establish the effect of fish size and stressor duration on the cortisol response. The results indicate high intrapopulation variability in plasma cortisol and a significant diel fluctuation with a peak value at dusk (18 h). Stressors of different intensity and/or duration affected the cortisol stress response in a differential manner according to fish size (and/or age). Maximum cortisol values in small-size fish were found at 1 and 2 h post-stress, depending on the duration of the stressor, while at 0.5 h post-stress in large fish regardless stress duration.     

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.     

Whole-body concentrations of cortisol and sex steroids in white sturgeon (Acipenser transmontanus, Richardson 1836) during early development and stress response

In general little is known about hormones and the ontogeny of the stress response in the early developmental stages of chondrostean fishes and in particular of white sturgeon (Acipenser transmontanus, Richardson 1836). In this study, we measured for the first time cortisol and sex steroids (testosterone and estradiol) in eggs, larvae, post-larvae, and fry of white sturgeon by radioimmunoassay (RIA), to elucidate some endocrine aspects of its development. The cortisol, testosterone, and 17β-estradiol of maternal origin found in unfertilized eggs of white sturgeon probably regulate both growth and development of the embryo. Cortisol decreased after fertilization, whereas testosterone and 17β-estradiol did not significantly change. During the late stages of embryo development and immediately after hatching, endogenous production of cortisol and sexual steroids, respectively, occurred. Sex steroids may be physiological inducers of gonad sex differentiation in sturgeon. All steroids showed an increase 10 days post-hatch (dph), near the transition from an endogenous to an exogenous energy source. Cortisol maintained the same basal levels even after metamorphosis, whereas testosterone and 17β-estradiol declined significantly in post-larvae at 35 and 45 days post-hatch. In addition, to evaluate the ontogeny of a functional hypothalamic-pituitary-interrenal (HPI) axis, larvae and fry were submitted to acute stress. The HPI axis did not seem to be functional on the first day post-hatch, but became so from the third day post-hatch onward.     

Effects of Lighting Spectrum and Disturbance Level on the Growth and Stress Responses of Yellow Perch Perca flavescens

Abstract.— We evaluated the effects of different lighting spectra and routine disturbances on the growth and stress responses of yellow perch Perca, flavescens. Fish were reared in tanks under one of three different lighting spectra: blue (405–530 nm), red (600–750 nm). or full spectrum (380–750 nm). For each lighting spectrum, fish were reared under one of two disturbance levels: one in which the fish were exposed to 2–4 daily routine disturbances which typify intensive tank culture conditions (e.g., casting shadows over the tank, sham tank siphoning, and dissolved oxygen measurement: ‘disturbed’). or a second treatment in which such disturbances were minimized (‘undisturbed’). The fish were reared for 87 d under temperature, photoperiod. and feeding conditions near optimal for the growth of yellow perch. The cortisol stress response of the different fish groups was evaluated by measuring baseline serum cortisol levels on days 0, 1,3, and 21, and by measuring cortisol levels 30 min after the application of an acute handling stressor on days 0, 1, and 3. Treatment effects on growth were measured from days 21–86 of the study. On day 87, various organs and parameters were evaluated to compare the health and condition of fish reared under the different treatments. Disturbed fish gained less weight and length, and had a significantly lower condition factor than undisturbed fish. Additionally, the disturbed fish showed no signs of acclimation to the frequent disturbances. The fish under red and full spectrum light gained more length than those under blue light. Frequent disturbance had a greater negative effect on length gain in fish reared under red light than in those reared under the other light treatments. On day 21, but not on other days, baseline cortisol levels were higher in disturbed than in undisturbed fish. Undisturbed fish had higher levels of mesentery fat than disturbed fish. Our results demonstrate that yellow perch growth can be significantly affected by disturbance level and lighting conditions. These factors should be taken into consideration when developing culture strategies to maximize the growth of yellow perch reared under intensive culture conditions.     

Fin damage and growth parameters relative to stocking density and feeding method in intensively cultured European perch (Perca fluviatilis L.)

We evaluated the relationship of stocking density to survival, growth performance and fin condition of European perch Perca fluviatilis with hand feeding and self-feeders. Hand-fed perch (body weight 19.1 ± 5.1 g and total length 107 ± 9 mm) were reared at 0.5, 1.0, 1.5 and 2.0 fish/L. Self-feeding perch (body weight 25.4 ± 3.9 g and total length 128 ± 7 mm) were reared at stocking densities of 0.6, 1.0 and 1.4 fish/L. Pond-reared perch served as a comparison group for fin damage assessment. We found no differences in survival rate among stocking densities with either feeding method. Hand-fed fish displayed the highest weight gain and SGR at stocking density of 0.5 fish/L. The self-feeding fish showed a non-linear association of weight gain with stocking density with the highest growth at 1.0 fish/L. Fin length was noticeably greater in pond-reared fish compared with RAS-reared fish regardless of feeding method. In both experiments, fin length relative to standard length showed a negative relationship with stocking density, with pectoral fins showing the greatest effect. Fin condition deteriorated with increasing stocking density, and growth was highest at 0.5 and 1.0 fish/L in hand-fed and self-feeding fish, respectively.     

Cortisol is responsible for positive and negative effects in the ovarian maturation induced by the exposure to acute stressors in Nile tilapia, Oreochromis niloticus

The aim of the present study was to evaluate the effect of acute stress and cortisol injection on oocyte final maturation process in female Nile tilapia (Oreochromis niloticus). Handling followed by a prophylactic treatment (0.3?mL?L^?1 H₂O₂, 5?g?L^?1 NaCl solution during 30?min) and an environmental change (transfer from a 2?m³ fibreglass square tank to 50?L aquaria) were used as acute stressors and compared to a single cortisol injection (0.5 or 5?mg?kg^?1 body weight). For both acute stress and cortisol injection (0.5?mg?kg^?1 body weight), serum cortisol level was significantly increased from 2.3 to 134.1?ng?mL^?1 1?h post-stress/injection and returned to a resting basal value 24?h after the stress/injection. In fish injected with 5?mg?kg^?1 body weight cortisol, mean serum cortisol level reached a peak up to 2500?ng?mL^?1 1?h after injection. 63?% of the females (mean body weight: 242?±?4?g) submitted to the acute stress ovulated within 72?h after the stress. In the same way, cortisol injection (5?mg?kg^?1 body weight) at the 10th day of the maturation cycle led to a twofold reduction of the time before ovulation compared to vehicle injected control fish. Relative and total fecundity were significantly decreased in females submitted to an acute stress or cortisol injected at 5?mg?kg^?1 body weight, but not fertilization or hatching rates. In conclusion, acute stress and cortisol induction exert both positive and negative effects on the final reproductive process in O. niloticus, and cortisol is the endocrine mediator causing these changes.     

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.     

Effects of commonly used disinfectants on bacterial load,hatchability and survival of Bluefin Sea bream (Sparidentex hasta) eggs

The efficacy of four chemical reagents, iodophor, formalin, hydrogen peroxide and bronopol as fish egg surface disinfectants were evaluated in bluefin sea bream (Sparidentex hasta). Fertilized eggs were counted and subjected to a static bath dip treatment in different concentrations of the above chemicals for 4 min before being incubated at 20 ± 0.5°C for 40 h. Treatment efficacy of the different disinfectants was evaluated by assessing the bactericidal activity, egg hatch percentage and survival of larvae up to 3 days post hatch. Results showed that iodophor at medium concentrations (75 and 100 ppm) was the best of all tested disinfectants in bacterial killing ability (12% reduction in the bacterial counts), egg hatching per cent (99.8% and 99.6% respectively) and larval survival up to 3 days post hatch (50.8% and 54.8% respectively). Formalin was the second best disinfectant at levels of 100 and 150 ppm. Hydrogen peroxide gave good results compared with the control while, bronopol showed discouraging results. In conclusion, iodophor appeared to be suitable for bluefin sea bream eggs disinfection with a 4 min exposure to 75–100 ppm when applied 14–16 h after egg fertilization.