Acute and chronic effects of aqueous ammonia on marbled spinefoot rabbitfish,Siganus rivulatus (Forsskål 1775)

Ammonia is a metabolite of aquatic organisms which might reach deleterious levels in intensive fish farms. The aim of the present study was to determine median lethal concentrations (96‐h LC₅₀) of total ammonia nitrogen (TA‐N) on marbled spinefoot rabbitfish (Siganus rivulatus) and chronic effects of TA‐N on survival, growth and behaviour of juvenile rabbitfish over a 50 day period. In the first experiment, fish were exposed to 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mg L^?1 TA‐N for 96 h and survival evaluated. In the second experiment, 12 fish were stocked per 50‐L tank and treated with one of 0, 2, 4, 6, 8, 10 and 12 mg L^?1 TA‐N with three replicate tanks per treatment. Survival and growth were determined and histopathological alterations of gills due to chronic ammonia exposure were studied by light and electron microscopy. The 96‐h LC₅₀ values were 16–18 mg L^?1 TA‐N. In the chronic exposure experiment, fish reared in water with 0 mg L^?1 TA‐N had 100% survival and had 50% weight increase in 50 days. Fish at 2 and 4 mg L^?1 TA‐N all died whilst fish in 6, 8, 10 and 12 mg L^?1 TA‐N survived and grew albeit less than in treatment 0 mg L^?1. Gills from ammonia treated fish displayed severe histological and ultrastructural alterations including hyperplasia, hypertrophy and fusion of secondary lamellae, aneurysms and presence of pleomorphic altered cells. Chronic exposure to ammonia is deleterious to marbled spinefoot rabbitfish and low concentrations of ammonia appear to kill the fish in <50 days whilst fish can survive for more than 50 days at concentrations between 6 and 12 mg L^?1 TA‐N.     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

Effects of dietary α‐ketoglutarate supplementation on the antioxidant defense system and HSP 70 and HSP 90 gene expression of hybrid sturgeon Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress

This study was conducted to determine the effects of dietary α‐ketoglutarate (AKG) supplementation on the antioxidant defense system and gene expression of heat shock protein (HSP) 70 and HSP 90 in hybrid sturgeons Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress. A 2 × 3 factorial experiment was arranged, in which each diet (0%, 1% AKG) was randomly assigned to 0.25 (control) 5 and 10 mg L^?1 ammonia‐N groups with three replicate aquaria for each 72 h. The 10 mg L^?1 ammonia‐N significantly increased serum ammonia concentrations and intestinal Gln concentrations and GS activity compared with the 0.25 or 5 mg L^?1 ammonia‐N groups. The intestinal Gln concentration and GS activity increased, and the serum ammonia concentration decreased, in fish given dietary supplementation of 1.0% AKG compared with fish given diets without AKG. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in serum, gills and intestines decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N, and their activity increased in fish given diets with 1% AKG. Catalase in the serum and gills decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N and increased in fish given diets with 1% AKG. The 10 mg L^?1 ammonia‐N or 1% AKG supplementation increased HSP 70 and HSP 90 gene expression in the liver. The increased activity of antioxidant enzymes, and increased HSP 70 and HSP 90 gene expression in fish fed diets containing 1% AKG suggested higher tolerance to ammonia‐N stress.     

Acute ammonia toxicity and gill morphological changes of Japanese flounder Paralichthys olivaceus in normal versus supersaturated oxygen

Ammonia toxicity and morphological changes in gills of juvenile Japanese flounder Paralichthys olivaceus (5.76 ± 0.12 g) were investigated when fish were separately exposed to normal dissolved oxygen (DO) at 6.5 ± 0.5 mg L^?1 and supersaturated oxygen at 16.0 ± 2.0 mg L^?1 at different ammonia concentrations. Under normal oxygen, ammonia concentrations were tested from 0.04 (control) to 93.3 mg L^?1 total ammonia nitrogen (TAN), whereas under oxygen supersaturation, ammonia concentrations ranged from 0.04 (control) to 226.7 mg L^?1 TAN in the trial. After exposure to ammonia for 96 h, the ammonia LC₅₀ for fish was 62.48 mg L^?1 TAN (0.50 mg L^?1 NH₃–N) at normal oxygen and 160.71 mg L^?1 TAN (0.65 mg L^?1 NH₃–N) at oxygen supersaturation. Light microscopic observations confirmed that gill damage in normal oxygen was more profound than in oxygen supersaturation when fish were exposed to the same level of TAN (93.3 mg L^?1). Furthermore, electron microscopic scanning also showed more crimple, retraction and fibrosis on the secondary lamella surface in fish exposed to normal oxygen than those in fish exposed to supersaturated oxygen at the same TAN (93.3 mg L^?1). This study suggests that supersaturated oxygen can increase ammonia tolerance in Japanese flounder through reducing gill damage by ammonia, which partially explains the merit of using pure oxygen injection in intensive fish farming.     

The efficacy of MS‐222 as anaesthetic agent in four freshwater aquarium fish species

The efficacy of anaesthetic tricaine methanesulfonate (MS‐222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L^?1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio – 75, 100 and 125 mg L^?1, P. reticulata – 125, 150 and 200 mg L^?1, S. discus – 75 and 100 mg L^?1 and X. helleri – 125 and 150 mg L^?1. The induction time generally decreased significantly with increasing concentration of MS‐222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS‐222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS‐222 concentration. MS‐222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Redox dysregulation and bioaccumulation of copper in tilapia Oreocromis niloticus

The objective of this study was to evaluate the effect of two concentrations of copper (0.13 and 0.5 mg L^?1) on juveniles of tilapia, Oreocromis niloticus (Linnaeus), using two toxicological assays (I‐ copper exposure and II‐ copper exposure followed by recovery). Doses were taken from the liver of the test organisms to determine the concentrations of copper, reduced glutathione, activities of glutathione S‐transferase and catalase, and lipid peroxidation. Exposure to copper in both assays resulted in significant accumulation of this metal in the analysed tissue and in decreased concentration of reduced glutathione. There was no alteration in the activities of catalase and glutathione S‐transferase and in lipid peroxidation.     

Dietary Lactobacillus plantarum supplementation decreases tissue lead accumulation and alleviates lead toxicity in Nile tilapia (Oreochromis niloticus)

Pollution by the heavy metal lead (Pb) has become a threat to both aquaculture and food safety. In this study, the protective effects of a selected probiotic with good Pb binding capacity (Lactobacillus plantarumCCFM8661) against waterborne Pb exposure were evaluated in Nile tilapia (Oreochromis niloticus). One hundred and eighty fish were divided into four groups as control, Pb‐only, Pb‐plus‐CCFM8661 and CCFM8661‐only. Fish were exposed to a dose of waterborne Pb level at 1 mg L^?1 for 4 weeks, and the probiotic was administered at 10⁸ CFU g^?1 in fish diet twice daily. The results showed that dietary supplementation of CCFM8661 ameliorated the growth performance and prevented the death of Pb‐exposed fish. Cotreatment with Pb and CCFM8661 also effectively decreased Pb accumulation in kidney, liver, gonad, brain, gills and muscle. Moreover, the administration of this probiotic alleviated Pb‐induced oxidative stress, recovered digestive enzyme activities and blood δ‐aminolevulinic acid dehydratase activity, reversed alterations in innate immune status and decreased the frequencies of the nuclear abnormalities in peripheral blood erythrocytes of fish. These results suggested that CCFM8661 may be a novel dietary supplement against Pb toxicity at least in tilapia.     

Effectiveness of eugenol sedation to reduce the metabolic rates of cool and warm water fish at high loading densities

Effects of eugenol (AQUI‐S^®20E, 10% active eugenol) sedation on cool water, yellow perch Perca flavescens (Mitchill), and warm water, Nile tilapia Oreochromis niloticus L. fish metabolic rates were assessed. Both species were exposed to 0, 10, 20 and 30 mg L^?1 eugenol using static respirometry. In 17°C water and loading densities of 60, 120 and 240 g L^?1, yellow perch controls (0 mg L^?1 eugenol) had metabolic rates of 329.6–400.0 mg O₂ kg^?1 h^?1, while yellow perch exposed to 20 and 30 mg L^?1 eugenol had significantly reduced metabolic rates of 258.4–325.6 and 189.1–271.0 mg O₂ kg^?1 h^?1 respectively. Nile tilapia exposed to 30 mg L^?1 eugenol had a significantly reduced metabolic rate (424.5 ± 42.3 mg O₂ kg^?1 h^?1) relative to the 0 mg L^?1 eugenol control (546.6 ± 53.5 mg O₂ kg^?1 h^?1) at a loading density of 120 g L^?1 in 22°C water. No significant differences in metabolic rates for Nile tilapia were found at 240 or 360 g L^?1 loading densities when exposed to eugenol. Results suggest that eugenol sedation may benefit yellow perch welfare at high densities (e.g. live transport) due to a reduction in metabolic rates, while further research is needed to assess the benefits of eugenol sedation on Nile tilapia at high loading densities.     

Bioassay‐guided isolation and identification of active compounds from Macleaya microcarpa (Maxim) Fedde against fish pathogenic bacteria

Four alkaloids (Sanguinarine, 6‐Methoxyl‐dihydro‐chelerythrine, Cryptopine and β‐Allocryptopine) were isolated from aerial parts of Macleaya microcarpa (Maxim) Fedde using bioassay‐guided isolation method, and the inhibitory activity of ethanolic extract, various fractions and these four alkaloids against four fish pathogenic bacteria (Aeromonas hydrophila, Aeromonas salmonicida, Vibrio anguillarum and Vibrio harveyi) was assessed in vitro using the agar dilution method and the microdilution assay method respectively. A. hydrophila was the most sensitive strain to all the tested compounds. Minimum inhibitory concentration (MIC) values were lower for sanguinarine against all tested Gram‐negative strains than other three alkaloids, with MIC values of 12.5 mg L^?1 for A. hydrophila and 50 mg L^?1 to other pathogenic bacteria. Followed by 6‐methoxyl‐dihydro‐chelerythrine, which showed considerable antibacterial activity with MIC values of 80 mg L^?1 for A. hydrophila, 100 mg L^?1 for V. harveyi, and 125 mg L^?1 for both V. anguillarum and A. salmonicida. Cryptopine and β‐allocryptopine revealed similar inhibitory activity with MIC values of 100 mg L^?1 for A. hydrophila and 200 mg L^?1 for other three bacterial species. These finding provided evidence that extract, as well as isolated compounds from M. microcarpa might be potential sources novel antibacterial agents for the treatment of fish infectious diseases.     

Application of low‐frequency sonophoresis and reduction of antibiotics in the aquatic systems

A major concern in aquaculture is the use of chemical therapeutics, such as antibiotics, because of their impact on the environment as well as on the fish product. As a potential tool for reducing antibiotic use, we tested the application of low‐frequency ultrasound as a method for enhancing antibiotic uptake. Rainbow trout juveniles (Oncorhynchus mykiss) were exposed to two different concentrations of oxytetracycline (OTC), flumequine (FLU) and florfenicol (FLO), administered by bath after the application of ultrasound. After exposure, concentrations of these substances were measured in the liver and blood of treated fish. Results showed that the ultrasound treatment can significantly increase the uptake for all three antibiotics. The uptake of OTC for example, in fish exposed to an OTC concentration of 20 mg L^?1 after prior treatment with ultrasound, was similar to the OTC concentrations in their liver and blood to fish exposed to 100 mg L^?1 without sonication. For FLU and FLO, the use of ultrasound caused significant differences of uptake in the liver at high antibiotic concentrations. This suggests that the use of ultrasound as a technique to deliver antibiotics to fish can ultimately reduce the amount of antibiotics discharged into the aquatic environment.     

Anaesthetic efficacy of eugenol on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus)

Anaesthetic efficacy of eugenol was investigated on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus). A total of 104 fish with average weights of 12 ± 2.5, 28 ± 5 and 53 ±5.1 g were subjected to 25–200 mg L^?1 eugenol and behavioural responses as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 49.9–127.3 s after exposure to 50–200 mg L^?1 eugenol, fish reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 57.3–140.4 s post exposure to such concentrations. Recovery time was 91.7–312 s in all weight classes for all eugenol concentrations. Mortality (23%) was only observed in 12‐g fish when were subjected to 200 mg L^?1 eugenol. This study showed the behavioural response of Flowerhorn to anaesthesia and eugenol efficacy as an anaesthetic in this important ornamental species. The general quadratic equation revealed that concentrations of eugenol and fish size along with their interactive effects have significantly contributed to the model, with concentration recording the highest beta value in all models (β = ?0.809, ?0.818 and ?0.909, P = 0.000). According to the results, minimum eugenol concentration to induce anaesthesia in less than 3 min was 50 mg L^?1.     

Effects of different dietary vitamin C supplementations on growth performance,mucus immune responses and antioxidant status of loach (Misgurnus anguillicaudatus Cantor) juveniles

A 60‐day feeding trial was conducted to determine the effects of different dietary vitamin C levels on growth performance, immune response and antioxidant capacity of loach juveniles. Six isonitrogenous (58.6% of crude protein), isoenergetic (17.5 kJ g^?1) practical diets supplemented with 0 (VC0), 100 (VC100), 200 (VC200), 500 (VC500), 1000 (VC1000) and 5000 mg kg^?1 (VC5000) of VC (35% ascorbic acid equivalent) were fed to fish (mean initial weight 0.11 ± 0.02 g) in triplicate. Results showed that fish fed VC0 diet had significantly lower body weight gain (BWG) and survival rate (SR). However, BWG and SR improved significantly in fish fed VC100 and VC200 diet respectively. Whole body ascorbic acid concentration increased with incremental dietary VC levels from 0 to 100 mg kg^?1. The activity of mucus alkaline phophatase was significantly increased by the dietary VC level. Incremental levels of VC significantly reduced activities of glutathione peroxidase (GPx) and catalase. Moreover, fish fed diets containing more than 100 mg kg^?1 VC significantly down‐regulated the superoxide dismutase and GPx mRNA expression in liver. Meanwhile, the expressions of liver heat shock protein (HSP70) and nuclear factor‐erythroid 2‐related‐2 (Nrf2) were affected by fish fed diets containing VC from 100 to 5000 mg kg^?1. In conclusion, VC requirement of loach juveniles for optimum growth and functionally preventing lipid peroxidation was more than 200 mg kg^?1 of diet. Moreover, high dose of VC supplementation did not show any detrimental effects on loach growth performance.     

Impacts of chloramine‐T treatment on antioxidant enzyme activities and genotoxicity in rainbow trout,Oncorhynchus mykiss (Walbaum)

Juvenile rainbow trout Oncorhynchus mykiss (Walbaum) were exposed to therapeutic, and higher concentrations of chloramine‐T (Cl‐T) to assess the effects of this chemical on the antioxidant enzyme system and genetic structure. Red blood cells acetylcholinesterase, ?‐aminolevulinic acid dehydratase, paraoxonase and liver glutathione S‐transferase activity were increased at 10 and 20 mg L^?1 Cl‐T‐exposed fish, while they were decreased at 30 mg L^?1 Cl‐T‐exposed fish. On the other hand, liver catalase activity and liver protein levels increased at 10 mg L^?1 and decreased at 20 and 30 mg L^?1 concentrations of Cl‐T. Liver super‐oxide dismutase activity decreased at 10 mg L^?1 and 20 mg L^?1 Cl‐T and increased at 30 mg L^?1 of Cl‐T. Compared to control, comet assay indicated that Cl‐T did not cause significant DNA damage to red blood cells of the fish. Results indicate that 10 or 20 mg L^?1 Cl‐T can be safely used to prevent or treat external parasitic and bacterial infection of rainbow trout.     

Gill damage to juvenile orange‐spotted grouper Epinephelus coioides (Hamilton, 1822) following exposure to suspended sediments

Juvenile Epinephelus coioides were exposed to three nominal concentrations (8, 32, 128 mg wet sediments L^?1) of suspended sediments (SS) from Port Shelter (PS), Mirs Bay (MB) and Victoria Harbour (VH) for 10 and 30 days using semi‐static system. Sediments from VH contained higher concentrations of Cu and polycyclic aromatic hydrocarbons (PAHs) than sediments from PS and MB. Gill damages including lamellar blood sinus dilation and vascular congestion were prevalent after just 10 days of exposure to SS. Fish exposed to SS at the highest concentration of 128 mg L^?1 showed higher incidence of lamellar aneurism. Hyperplasia in the base of lamellae was recorded in fish that had been exposed to contaminated SS from VH. Significant increases in the density of chloride cells and mucous cells were found on the gills of fish that had been exposed to 128 mg L^?1 of SS from PS. Clogging of gills by SS produced hypoxic‐like responses in fish. Polluted sediments from VH produced addictive or synergistic effects between SS and chemical contaminants on fish.     

Efficacy of formalin,iodine and sodium chloride in improvement of egg hatching rate and fry survival prior to the onset of exogenous feeding in yellow perch

Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L^?1) and 25 mg L^?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L^?1), iodine at three concentrations (50, 100 and 250 mg L^?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L^?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L^?1 formalin for 30 min, 50 mg L^?1 iodine for 15 min and 500 mg L^?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L^?1), iodine (250 mg L^?1) and sodium chloride (1000 and 3000 mg L^?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L^?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.     

Acute toxicity of nitrite on white shrimp Litopenaeus vannamei (Boone) juveniles in low‐salinity water

The nitrite toxicity was estimated in juveniles of L. vannamei. The 24, 48, 72 and 96 h LC₅₀ of nitrite‐N on juveniles were 8.1, 7.9, 6.8 and 5.7 mg L^?1 at 0.6 g L^?1; 14.4, 9.6 8.3 and 7.0 mg L^?1 at 1.0 g L^?1; 19.4, 15.4, 13.4 and 12.4 mg L^?1 at 2.0 g L^?1 of salinity respectively. The tolerance of juveniles to nitrite decreased at 96 h of exposure by 18.6% and 54.0%, when salinity declined from 1.0 to 0.6 g L^?1 and from 2.0 to 0.6 g L^?1 respectively. The safe concentrations at salinities of 0.6, 1.0 and 2.0 g L^?1 were 0.28, 0.35 and 0.62 mg L^?1 nitrite‐N respectively. The relationship between LC₅₀ (mg L^?1), salinity (S) (g L^?1) and exposure time (T) (h) was LC₅₀ = 8.4688 + 5.6764S – 0.0762T for salinities from 0.6 to 2.0 g L^?1 and for exposure times from 24 to 96 h; the relationship between survival (%) and nitrite‐N concentration (C) for salinity of 0.6–2.0 g L^?1, nitrite‐N concentrations of 0–40 mg L^?1 and exposure times from 0 to 96 h was as follows: survival (%) = 0.8442 + 0.1909S – 0.0038T – 0.0277C + 0.0008ST + 0.0001CT–0.0029SC, and the tentative equation for predicting the 96‐h LC₅₀ to salinities from 0.6 to 35 g L^?1 in L. vannamei juveniles (3.9–4.4 g) was 96‐h LC₅₀ = 0.2127 S² + 1.558S + 5.9868. For nitrite toxicity, it is shown that a small change in salinity of waters from 2.0 to 0.6 g L^?1 is more critical for L. vannamei than when wider differences in salinity occur in brackish and marine waters (15–35 g L^?1).     

Menthol as anaesthetic for lambari Astyanax altiparanae (Garutti & Britski 2000): attenuation of stress responses

The anaesthetic potential of menthol was evaluated in lambari Astyanax altiparanae by exposing fingerlings to concentrations 50, 100, 150, 200, 250 and 300 mg L^?1 and measuring the induction and recovery times to deep anaesthesia, the mortality rates during and 96 h after procedure and after 6 min of continuous exposure. The effect of menthol on stress responses were evaluated by comparing glucose and cortisol levels of juveniles subjected to anaesthesia (50 mg L^?1), stress (air exposure) or pre‐anaesthesia associated to stress. All concentrations induced deep anaesthesia within 0.5 to 1 min, with recovery between 1.83 and 4.16 min, without mortality during the induction or up to 96 h after exposure. Induction time decreased and recovery time increased linearly as the menthol concentration increased. Continuous exposure to 50, 100 and 150 mg L^?1 concentrations resulted in mortality rates of 0%, 20% and 80% respectively. Anaesthesia or air exposure increase blood glucose but prior anaesthesia with menthol suppressed the elevation of cortisol caused by stress. Menthol has an anaesthetic effect and attenuates the stress response in lambari and 50 mg L^?1 is the most effective concentration for inducing deep anaesthesia in 1.0 min, safe for up to 6 min exposure.     

Survival and behavioural responses of cool and warm water fish sedated with AQUI‐S®20E (10% eugenol) at high loading densities

This study evaluated the effects of AQUI‐S^®20E (10% eugenol) sedation on the survival and behaviour of yellow perch Perca flavescens (Mitchill) and Nile tilapia Oreochromis niloticus L. held in high loading densities. Fish were held in 0–300 mg L^?1 AQUI‐S^®20E (0–30 mg L^?1 eugenol) for up to 10 h in static tanks. At 17°C, yellow perch held in 200 and 300 mg L^?1 AQUI‐S^®20E were lightly sedated for up to 7 h. Yellow perch at 200 and 300 mg L^?1 AQUI‐S^®20E also had >95% mean survival 7‐days post exposure using loading densities up to 360 g L^?1. Nile tilapia were only sedated for ≤3 h in concentrations up to 300 mg L^?1 at 22°C and had >90% mean survival at loading densities ≤480 g L^?1. Ammonia in tanks increased significantly as loading density increased, but treatment with AQUI–S^®20E did not reduce ammonia accumulation. Results suggest that AQUI–S^®20E was effective to sedate yellow perch and Nile tilapia at high loading densities, but sedation varied with loading density and species.     

Performance of mono‐sex tilapia (Oreochromis niloticus) in rice field with different ditch size

The effects of ditch size on growth and production of mono‐sex tilapia, Oreochromis niloticus in rain fed concurrent rice–fish system were technically and economically evaluated for a period of 4 months. Three different ditch sizes were tried: 5%, 10% and 15% of the total cultivable rice field. The fish fingerlings stocking density (5000 ha^?1) and management practices were similar in all plots. The water temperature, dissolve oxygen, pH, Secchi disc visibility, ortho‐phosphate, ammonia nitrogen and alkalinity were found to be in the range of 31.5–31.9°C, 5.10–5.27 mg L^?1, 7.56–7.62, 34.81–35.41 cm, 0.88–0.96 mg L^?1, 0.014–0.021 mg L^?1 and 66.23–69.76 mg L^?1, respectively, and all the values were found suitable for tilapia fish farming. Sediment organic matter, available nitrogen and phosphorous were increased significantly (P < 0.05) in all plots after cultivating the fish. Larger ditch size (15% of the total land area) resulted in significantly (P < 0.05) higher specific growth rate, survival and fish production compared with the smaller (5%) and medium (10%) ditch sizes. However, significantly (P < 0.05) higher net income and benefit–cost ratio were obtained in medium ditch size with the values of 2.6% and 3.6% higher than the smaller and larger ditch sizes respectively. It is concluded that rice–fish farming in a rain fed ecosystem of Bangladesh with medium ditch size and stocking density of 5000 ha^?1 mono‐sex tilapia can achieve better economic return.