Effect of a nitric oxide releasing compound isosorbide dinitrate on development and cardiovascular physiology of rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) eggs and alevins were exposed for up to 4 weeks to the nitricoxide (NO) donor isosorbide dinitrate (ISDN) at concentrations up to 100 mol l^–1. Hatching success was over 95%, but increasing concentrations of ISDN significantly delayed hatching, without significant affects on survival. Alevins exposed to ISDN concentrations of 50 or 100 mol l^–1 showed significant cardiovascular and developmental changes compared to unexposed alevins. These included lowered heart rate and ventilation rate, doubling of cardiac output, dilation of the vitelline vein and increased in alevin weight. The results indicate a vasodilatory role of NO in trout alevins, and their cardiovascular responses to NO, with reference to the role of the vitelline vein, are discussed.     

The role of açaí (Euterpe oleracea Mart. 1824) as a chemoprotective agent in the evaluation of antioxidant defence,oxidative damage and histology of juvenile shrimp Litopenaeus vannamei (BOONE, 1931) exposed to ammonia

This study investigated the effects of the use of the inclusion of açaí on the diet of shrimp Litopenaeus vannamei on antioxidant and histopathological responses after exposure to ammonia. The shrimps were fed two experimental diets: control and with 10% of açaí inclusion (W/W), for 35 days. Afterwards, the organisms were exposed at four concentrations of ammonia (0.01‐control; 0.26; 0.48 and 0.91 mg NH₃‐N L^?1) for 96 hr. The total antioxidant capacity (ACAP) of the gills decreased significantly in both diets when exposed to ammonia, whereas in the muscle, the açaí promoted an increase in ACAP. Concomitantly, lipid peroxidation levels increased in the gills and decreased in muscle. After exposure to ammonia, glutathione‐S‐transferase activity increased in hepatopancreas in shrimps fed with açaí facilitating the detoxification of lipid peroxidation by‐products, and the concentration of protein sulfhydryl groups decreased in the gills and muscle of the shrimp of the control diet, evidencing protein damage, an unobserved response in shrimps that received the açaí diet. Histopathological changes decreased in açaí‐fed shrimps about the control diet after exposure to ammonia. It is concluded that açaí mitigated ammonia‐induced histopathological changes, improved the antioxidant defence system (gills and muscle) and attenuated the lipid damage in the muscle.     

Bioaccumulation of cadmium and its biochemical effect on selected tissues of the catfish (<Emphasis Type="Italic">Clarias gariepinus</Emphasis>)

The present study examines the pattern of accumulation of cadmium (Cd) and its biochemical effects on selected tissues of a variety of African catfish (Clarias gariepinus), after exposure to various doses of Cd. The results obtained indicate that at the end of 21 days of exposure, the total tissue organ cadmium concentration followed the pattern kidney > gill > liver > muscle for each of the exposure concentrations. The levels of Cd in these organs were higher than those in ambient water. Moreover, while the rate of uptake of Cd increased with time in the kidney, liver and muscle, it decreased in the gill. Superoxide dismutase (SOD) was significantly elevated only in the kidney of catfish treated with 0.2 and 0.4 ppm of Cd for 7 days compared with the control. Conversely, gill SOD was significantly decreased in the same concentrations of Cd-treated catfish relative to the control. Statistically similar levels of SOD were observed in the liver, brain and muscle with all the treatments after the same duration of treatment. In the fish exposed for 21 days, SOD activity was significantly decreased in the kidney with a corresponding increase in lipid peroxidation (LPO), but it manifested only with the 0.2- and 0.4-ppm Cd treatment relative to the control. In the liver, however, Cd exposure significantly increased SOD in the 0.2- and 0.4-ppm treatments of the same duration. In conclusion, the present study indicates that the accumulation of Cd and its effect on SOD and LPO in C. gariepinus is dependent on concentration, tissue and time     

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.     

Effect of phenanthrene on haematological parameters in olive flounder, Paralichthys olivaceus (Temminch et Schlegel)

Effects of phenanthrene on haematological parameters in olive flounder, Paralichthys olivaceus, were studied by exposing fish to water‐borne phenanthrene (0.5, 1 and 2 μM) for 4 weeks. Red blood cell count, haemoglobin concentration, haematocrit levels decreased with increasing exposure time of phenanthrene to the fish. Plasma potassium level increased significantly in the experimental group but the levels of sodium and chloride were significantly decreased compared with control. There was no significant change in plasma magnesium, calcium and phosphate in fish of all treated groups compared with the control. The most profound physiological disturbance in phenanthrene‐exposed fish was a decrease in blood osmolality. Malondialdehyde level in the plasma in the phenanthrene group (2 μM) was 1.4‐fold higher following 4‐week exposure compared with the control group. Acetylcholinesterase activity decreased as the concentration of phenanthrene increased, reaching 65.9% of the control value at the highest dose at the 4th week. A key finding from this study is that olive flounder exposed to phenanthrene at concentrations more than 1.0 μM are likely to experience negative impacts on haematological parameters.     

Effect of glyphosate-based herbicide on hematological and hemopoietic parameters in common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L)

The effects of Roundup (glyphosate-based herbicide) on peripheral blood parameters and hematopoietic tissue cellular composition and activity in common carp juveniles were evaluated. The fish were exposed for 7 days at 0.0, 0.1, 0.5, or 5.0 mg/L of glyphosate. In fish exposed to Roundup, hematological alterations were observed; however, most of them were not directly related to the herbicide concentration. An increase in Ht and MCV, and decrease in Hb, MCH, and MCHC compared to the control were observed. Fish exposed to Roundup showed also a reduction in WBC and oxidative metabolic activity of phagocytes (NBT) compared to the control. The fish exposed to 0.1 and 5.0 mg/L showed increased glucose values, whereas in those subjected to 0.5 mg/L blood glucose concentration declined compared to the control. Cholesterol significantly increased at 0.1 mg/L and decreased at 5.0 mg/L. Analysis of head kidney hematopoietic tissue revealed that Roundup at concentrations 0.5 and 5.0 mg/L caused a significant increase in the rate of cell proliferation accompanied by an increase in frequency of early blast cells. No significant differences occurred in percentages of most cell lineages but the frequency of monocytoid, eosinophilic, and basophilic lineage cells significantly increased in the herbicide-exposed fish compared to the control. The obtained results revealed that sublethal concentrations of Roundup that may occur in polluted natural waters caused a slight anemic and significant immunosuppressive response in common carp juveniles. On the other hand, they indicate a considerable compensatory potential of carp hematopoietic system.     

A comparative study on the effects of a pesticide (cypermethrin) and two metals (copper, lead) to serum biochemistry of Nile tilapia, Oreochromis niloticus

The present study was designed to compare the responses in freshwater fish Oreochromis niloticus exposed to a synthetic pyrethroid, cypermethrin (CYP); an essential metal, copper (Cu); and a nonessential metal, lead (Pb). Fish were exposed to 0.05 μg/l CYP, 0.05 mg/l Cu, and 0.05 mg/l Pb for 4 and 21 days, and the alterations in serum enzyme activities, metabolite, and ion levels were determined. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities increased in response to CYP, Cu, and Pb exposures at both exposure periods. While elevations in alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activities and in cholesterol level were observed in pesticide-exposed fish at 4 and 21 days, they increased in Cu- and Pb-exposed fish at 21 days. Although metal-exposed fish showed increases in cortisol and glucose levels at 4 days followed by a return to control levels at the end of the exposure period, their levels elevated in pesticide-exposed fish at both exposure periods. Total protein levels decreased in Pb- and pesticide-exposed fish at 21 days. Na⁺ and Cl⁻ levels decreased in pesticide-exposed fish at both exposure periods and in Cu- and Pb-exposed fish at 21 days. The exposures of pesticide and metals caused an elevation in K⁺ level at the end of the exposure period. The present study showed that observed alterations in all serum biochemical parameters of fish-treated pesticide were higher than those in fish exposed to metals.     

Intrafollicular thyroid hormone staining in whole-mount zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) embryos for the detection of thyroid hormone synthesis disruption

Endocrine-disrupting chemicals are known to impact multiple hormonal axes of vertebrates, among which the thyroid system is crucial for multiple developmental and physiological processes. Thus, the present study focused on the semi-quantitative visualization of intrafollicular triiodothyronine (T3) and thyroxin (T4) in zebrafish embryos as a potential test system for the detection of disrupted thyroid hormone synthesis. To this end, an antibody-based fluorescence double-staining protocol for whole-mount zebrafish embryos and larvae was adapted to simultaneously detect intrafollicular T3 and T4. During normal development until 10 days post-fertilization (dpf), the number of thyroid follicles increased along the ventral aorta. Concentrations of T4 and T3, measured by fluorescence intensity, increased until 6 dpf, but decreased thereafter. Exposure of zebrafish embryos to propylthiouracil (PTU), a known inhibitor of TH synthesis, resulted in a significant decrease in the number of follicles that stained for T3, whereas a trend for increase in follicles that stained for T4 was observed. In contrast, fluorescence intensity for both thyroid hormones decreased significantly after exposure to PTU. Overall, the zebrafish embryo appears to be suitable for the simultaneous visualization and detection of changing intrafollicular TH contents during normal development and after PTU treatment.     

Plasma proteins,hepatic enzymes,thyroid hormones and liver histopathology of Cyprinus carpio (Linnaeus, 1758) exposed to an oxadiazin pesticide,indoxacarb

Indoxacarb is a relatively new pesticide from oxadiazin class, which is used near carp ponds for agricultural purposes. Thus, the aim of this study was to determine indoxacarb effects on common carp (Cyprinus carpio). The fish were exposed to 0 (control), 0.75, 1.5 and 3 ppm of indoxacarb over 21 days and plasma biochemical characteristics and liver histopathology were examined. Exposure to indoxacarb induced fall in total protein after 21 days. Globulin increased after 7 days and then decreased after 14‐ and 21‐day exposure. Plasma alanine transaminase and aspartate transaminase activities increased in 1.5 and 3 ppm treatments after 7‐day exposure. Indoxacarb exposure significantly decreased plasma alkaline phosphatase after 7 days with no change at the second and third samplings. After 7 days, plasma T₃ levels had no significant change; however, it decreased after 14 days in the 1.5 and 3 ppm treatments and also reduced after 21‐day exposure to the 3 ppm treatment compared to the control group. Plasma T₄ level only decreased significantly in the 3 ppm treatment compared to the control group after 21 days. Different histopathological symptoms such as necrosis, hyperaemia, sinusoidal space extension, pyknotic nuclei, leucocyte infiltration and melanomacrophage aggregates were observed after 21‐day exposure to indoxacarb. The symptoms intensity was dependent on indoxacarb concentration. In conclusion, the present results show that indoxacarb exposure adversely affects common carp health and welfare, which consequently may induce serious problems in this species aquaculture.     

Supplementation of maca (Lepidium meyenii) tuber meal in diets improves growth rate and survival of rainbow trout Oncorhynchus mykiss (Walbaum) alevins and juveniles

Maca tuber meal is used in fish diet formulations in Andean trout culture and knowledge of its effects on fish growth is paramount to healthy human food production. In the first experiment with rainbow trout alevins (0.096±0.002 g), starter diets were offered from first feeding until 15 weeks. We formulated high protein content (～60%) semi‐purified starter diets supplemented with 0%, 5%, 10%, or 15% maca tuber meal (control, M‐5, M‐10, and M‐15 respectively). The second feeding trial was conducted with juveniles (1.56±0.02 g) fed one of three diets (control, M‐15, and commercial) for 8 weeks. In the first experiment, fish fed M‐10 and M‐15 diets exhibited significantly higher growth rates than the other dietary groups. Survival was significantly improved in the groups fed diets supplemented with maca tuber meal (60.0–69.2%) in comparison with the group fed a control diet (21.7%). The second experiment showed a higher growth rate in the M‐15 group compared with the control and a commercial diet fed group. Leucocyte numbers were increased by dietary supplementation of maca tuber meal. The findings of the present study suggest that a maca tuber meal inclusion at least 5% improves growth rate, feed utilization, immunity by increased leucocyte number, and survival of rainbow trout alevins and juveniles.     

Modulation of calcium uptake in cadmium-pretreated tilapia (Oreochromis mossambicus) larvae

Tilapia larvae were exposed to 0 (control), 50 (50-Cd) or 100 (100-Cd) μg l^-1 cadmium for 4 days and then transferred to cadmium-free fresh water for 3 days of detoxification. Total length and weight, calcium influx and total body calcium and cadmium content were examined at various times during detoxification. All the groups grew normally with regards to total length and body weight. Within the first 12h of detoxification the 50- and 100-Cd exposed groups released cadmium at the similar rate of about 24 ng mg^-1 h^-1 (or 140 ng larva^-1 h^-1). Later, however, this rate declined to only 4–16% of the initial level. Calcium influx in the control group showed a 10–26% increase during the detoxification period. Calcium influx in the 50-Cd group increased by about 280% and reached it peak at 12h. Calcium influx in the 100-Cd group increased by 440% and did not peak until 24h after transfer. After peaking, the influxes in both 50- and 100-Cd groups declined to the level of control at the end of the experiment. Calcium contents in 50- and 100-Cd groups increased more rapidly than that in control group within first 24h of the detoxification period. However the rate of increase in calcium content in three groups was the same after 24h. The changes in calcium influx appeared to be correlated with those in calcium content, and these suggested that tilapia larvae regulate the mechanism of calcium balance to compensate for the reduced calcium level in the body. This revised version was published online in July 2006 with corrections to the Cover Date.     

Physiological short-term response to sudden salinity change in the Senegalese sole (Solea senegalensis)

The physiological responses of Senegalese sole to a sudden salinity change were investigated. The fish were first acclimated to an initial salinity of 37.5?ppt for 4?h. Then, one group was subjected to increased salinity (55?ppt) while another group was subjected to decreased salinity (5?ppt). The third group (control group) remained at 37.5?ppt. We measured the oxygen consumption rate, osmoregulatory (plasma osmolality, gill and kidney Na⁺,K⁺-ATPase activities) and stress (plasma cortisol and metabolites) parameters 0.5 and 3?h after transfer. Oxygen consumption at both salinities was higher than for the control at both sampling times. Gill Na⁺,K⁺-ATPase activity was significantly higher for the 55?ppt salinity at 0.5?h. Plasma osmolality decreased in the fish exposed to 5?ppt at the two sampling times but no changes were detected for high salinities. Plasma cortisol levels significantly increased at both salinities, although these values declined in the low-salinity group 3?h after transfer. Plasma glucose at 5?ppt salinity did not vary significantly at 0.5?h but decreased at 3?h, while lactate increased for both treatments at the first sampling time and returned to the control levels at 3?h. Overall, the physiological response of S. senegalensis was immediate and involved a rise in oxygen consumption and plasma cortisol values as well as greater metabolite mobilization at both salinities.     

长江口中华鲟幼鱼血液水分、渗透压及离子浓度的变化规律

为了研究长江口中华鲟幼鱼盐度适应过程及其调节规律,将7月龄幼鱼直接转入0(淡水对照),5,10,15等4个盐度组中养殖32天,分别在0.5,1,2,4,8,16,和32 d 检测幼鱼血液水分、血清渗透压和血清Na 、Cl-、K 的浓度,结果显示：试验过程中,淡水和盐度5组中华鲟幼鱼血液各项指标始终保持一致,未呈现显著性差异。盐度10和15组,中华鲟幼鱼血液水分含量呈先下降后上升趋势,下降程度与盐度呈正相关,16 d时各组幼鱼血液水分无显著性差异。中华鲟幼鱼转入盐度10和15条件下,其血清渗透压与Na 和Cl-浓度的变化趋势一致,表现为先上升后下降,最后达到新的平衡;12 h是其上升和下降的拐点。而血清K 浓度的变化趋势与血清渗透压和Na 、Cl-离子不同,呈现先下降后趋于平稳的趋势。从结果可以看出,中华鲟幼鱼与其他广盐性鱼类一样,其盐度适应过程可分为2个阶段,即临界期和调整期。     

杂色鲍血细胞中一氧化氮合酶活性的鉴别

一氧化氮合酶(Nitric oxide synthase,NOS)是催化L-精氨酸与O_2产生一氧化氮(Nitric ox- ide,NO)的一种合成酶,它广泛存在于生物体的各个器官和组织中,其活力大小可通过测定NO合成量的多少来决定。NO作为一种新型生物信使分子、效应分子和免疫调节分子,参与机体多种重要的生理病理活动,可以通过非特异性地杀伤细菌、真菌、寄生虫及病毒等。增强机体的非特异性免疫。本文采用生物化学法,以副溶血弧菌和脂多糖为刺激因子.并采用硝酸盐还原酶法测定了NO的水平,对杂色鲍血细胞中的NOS活性进行了初步鉴别。结果表明,加入副溶血弧菌组或LPS组NO水平显著高于对照组(P<0.05),孵育4h后分别为对照组的1.84和1.92倍,孵育8h后分别为对照组的2.38和3.05倍,而且这一反应能被NOS抑制剂L-NAME所阻断,说明杂色鲍血细胞中存在NOS活性。     

Toxic effects of indoxacarb on gill and kidney histopathology and biochemical indicators in common carp (Cyprinus carpio)

The aim of this study was to determine the biochemical and histopathological effects of indoxacarb on an economic fish species Cyprinus carpio. The fish were divided into four triplicated groups exposed to either of 0% (control), 5% (0.75 mg/L), 10% (1.5 mg/L) or 20% (3 mg/L) of the pesticide LC₅₀. Blood sampling was performed after 7, 14 and 21 days exposure. There were no changes in blood calcium levels at any times. In the indoxacarb‐treated fish, chloride levels decreased at the first and increased at the second sampling compared to the control. In 3 mg/L treatment, phosphorus increased significantly after 7 days compared to the control. Plasma glucose levels decreased significantly at the 7th and 14th days sampling; however, it increased at the 21st day. After 7 days, creatinine level in 3 mg/L treatment was significantly lower than the control; however, the creatinine levels of indoxacarb treatments were higher than the control at the 21st day. The indoxacarb‐treated fish had higher plasma urea levels compared to the control at the 7th day. At the 21st day, plasma urea level at 3 mg/L was significantly lower than the control treatment. Edema, Lamellar curling, hyperplasia, lamellar fusion and hyperaemia were observed in the indoxacarb‐treated fish gill. Tubular necrosis, hematopoietic necrosis, melanomacrophage aggregates, Bowman's capsule edema, glomerulus degeneration and hyperaemia were observed in the indoxacarb‐treated fish kidney. Generally, sublethal concentrations of indoxacarb cause stress, hydromineral imbalance, metabolism alteration and gill and kidney damages in common carp.     

Effects of purified microcystin-LR and cell extracts of Microcystis strains PCC 7813 and CYA 43 on cardiac function in brown trout (Salmo trutta) alevins

Sub-lethal cardiac responses of brown trout alevins (Salmo trutta L.) were determined in response to aqueous extracts of the cyanobacterium Microcystis strains PCC 7813 (microcystins detectable by HPLC) and CYA 43 (no microcystins detectable by HPLC) and to the purified cyanobacterial hepatotoxin, microcystin-LR (MC-LR) at concentrations of 5, 50 and 500 g microcystin-LR equivalents l^–1. Responses were determined using a flow chamber and video camera attached to a low power microscope. Heart rate in brown trout alevins was acutely sensitive to cyanobacterial extracts and significant increases were observed within 15–60 sec of exposure to aqueous extracts, although no change was observed on exposure to purified MC-LR. Stroke volume increased in all treatments at 50 and 500 g MC-LR equivalents l^–1, which may, at least in part, have been due to vasodilation of the yolk-sac blood vessels. Cardiac output increased significantly at all three concentrations of cyanobacterial cell extracts but not at the lowest concentration of MC-LR, although the rate increased at levels at/or above 50 g l^–1. Increased heart rate, stroke volume and cardiac output occurred at environmentally relevant microcystin concentrations of Microcystis PCC 7813 and CYA 43 aqueous extracts.     

Amelioration of LPS‐induced inflammatory response and oxidative stress by astaxanthin in Channa argus lymphocyte via activating glucocorticoid receptor signalling pathways

The present study was conducted to evaluate the effects of astaxanthin (AST) against lipopolysaccharide (LPS)‐induced lymphocyte viability, ultrastructural lesions, apoptosis, oxidative stress and inflammatory responses in Channa argus. Lymphocytes exposed to more than 10 μg/ml LPS alone for 24 hr showed significantly decreased cell viability, elevated nitric oxide (NO) and malondialdehyde (MDA), lactate dehydrogenase (LDH) contents, and increased nuclear factor κB p65 (NF‐κB p65), myeloid differential protein‐88 (MyD88), tumour necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), interleukin‐8 (IL‐8), caspase‐3, caspase‐8 and caspase‐9 gene expression. LPS at a concentration of 10 μg/ml could induce oxidative stress and inflammatory responses in lymphocytes. The activities of antioxidant enzymes (catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD)) were significantly decreased after exposure to 10 μg/ml LPS. Besides, AST strikingly antagonized the LPS‐induced negative effects. AST significantly increased the expression of HSP70, HSP90, IκB‐α, and glucocorticoid receptor (GR) and decreased inflammatory responses. Further study showed that AST can activate GR signalling pathway and inhibit p65 phosphorylation. In addition, AST attenuated LPS‐induced apoptosis, mitochondrial swelling, degeneration and vacuolization. Collectively, these findings suggest that AST has protective roles in LPS‐induced cell damage via modulating GR activation in C. argus lymphocytes.     

Changes in plasma prolactin and growth hormone concentrations during freshwater adaptation of juvenile chum salmon (Oncorhynchus keta) reared in seawater for a prolonged period

Freshwater adaptability of chum salmon was examined in juvenile fish reared in seawater for 4 months. The fish, weighing about 40g, were transferred directly to fresh water in October, when their cohorts are migrating in the North Pacific Ocean. Plasma sodium concentration decreased from 167 mM in seawater to about 130 mM during the first 24h, and increased gradually during 2–7 days after the transfer. No immunoreactive prolactin (PRL) was detected in the plasma of the seawater-adapted fish nor during the first 24h in fresh water. Significant levels of PRL were detected after 2–3 days. The maximal level (2.6 ng/ml) was observed after 5 days and became undetectable again after 7 days; no significant correlation was seen between the changes in plasma sodium and PRL levels during the transfer. Plasma growth hormone levels were relatively constant, except for a significant decrease 12h after the transfer. Although plasma thyroxine levels were highly variable during the experiment, a significant decrease and an increase were observed 12h and 5 days after the transfer, respectively. The present study indicates that juvenile chum salmon retain hyperosmoregulatory ability even after prolonged rearing in seawater. Examination of turnover rates, rather than changes in plasma levels, seems to be essential to clarify the osmoregulatory roles of the hormones.     

Effects of night‐time light intensity on the survival rate and stress responses in juvenile Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel)

Land‐based cultured juvenile Pacific bluefin tuna Thunnus orientalis (PBT) have high mortality rates due to collisions or contacts with tank walls after about 30 days of hatching. To determine the effect of night‐time lighting on their survival, juvenile PBT were reared under four different night‐time light intensities (0, 5, 15 and 150 lx) for 9 days, followed by a 3‐day observation period. High‐intensity, night‐time lighting (150 lx) significantly improved the survival rate (75.8%; P < 0.001) compared with the unlit control group (0 lx, 64.3%). The survival rate in the high‐intensity group decreased after the end of the lighting period. Lighting did not influence whole‐body cortisol levels, glucose levels, or diel changes in plasma cortisol levels. In contrast, the survival rates of fish exposed to light intensities between 5 and 15 lx were slightly lower than that of the unlit control group. These results suggest that providing night‐time lighting of 150 lx or higher is an effective method for reducing the mortality of cultured PBT.     

Physiological effects of stocking density on the fish Piaractus mesopotamicus fed with red seaweed (Pyropia columbina) and β‐carotene‐supplemented diets

This study aimed at investigating the physiological responses of Piaractus mesopotamicus exposed to high stocking density and the potential protective role of supplemented diets. Fish were fed with basal, red seaweed (Pyropia columbina) or β‐carotene‐supplemented diets for 90 days. Then, fish were distributed at low (1.5 g/L) and high (22 g/L) stocking densities for 15 days. Fish exposed to the high density showed increased hepatosomatic index, haemoglobin content and mean corpuscular haemoglobin concentration (all diets); decreased haematocrit, mean corpuscular volume (basal and seaweed) and white blood cells count (all diets) were observed. High density‐exposed fish showed decreased plasmatic metabolites as well as the hepatic lipids content in basal and seaweed diets. Regarding oxidative stress, increased activity of glutathione S‐transferase in high density‐exposed fish muscle (all diets), and lower lipid peroxidation in liver (basal and β‐carotene) and intestine (basal and seaweed) were evidenced. Interactions between diet and stocking density were recorded regarding the triglycerides (decrease in fish exposed to high density fed with basal and seaweed) and hepatic lipids (decrease in fish exposed to high density fed with basal). The major changes occurred in haematologic and metabolic parameters as strategies to cope with overcrowding stress. Fish response to stocking density was not affected by diets.