Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Pharmacokinetic–pharmacodynamic modelling for the determination of optimal dosing regimen of florfenicol in Nile tilapia (Oreochromis niloticus) at different water temperatures and antimicrobial susceptibility levels

Optimized dosing regimen is key to the effective use of antibacterials and to minimizing drug‐related side effects. The current study established a pharmacokinetic–pharmacodynamic (PK‐PD) model for the determination of optimal antibacterial dosing regimen in fish taken into consideration the temperature‐dependent PK and the pathogen‐dependent antimicrobial susceptibility, using florfenicol (FF) in Nile tilapia as an example. The calculated optimal dosages significantly varied by temperature and target MIC levels, ranging from 2.23 (MIC 1 µg/ml at 24°C) to 34.88 mg kg^?1 day^?1 (MIC 4 µg/ml at 32°C). The appropriateness of the calculated dosages was successfully verified by the in vivo studies. After 5 days of oral administration of the calculated optimal dosage at 24°C, the predicted plasma drug values were in line with the mean observed C_min(ss) while at 28 and 32°C underestimation of the C_min(ss) in a dose‐dependent manner was observed and likely due to the occurrence of non‐linear PK at high dosages. The averaged serum protein binding of FF was 19.1%. Our results demonstrated the appropriateness and clinical applicability of the developed PK‐PD approach for the determination of optimal dosing regimens at given temperatures and MICs. Saturation metabolism and PK non‐linearity of FF in tilapia warrant further study.     

Comparative effects of dietary supplementation with maggot meal and soybean meal in gibel carp (Carassius auratus gibelio) and darkbarbel catfish (Pelteobagrus vachelli): growth performance and antioxidant responses

A 6‐week growth trial was conducted to investigate the effect of dietary supplementation with maggot meal (MGM) and soybean meal (SBM) on the growth performance and antioxidant responses of gibel carp (GC) and darkbarbel catfish (DC). The basal diet was formulated to contain 114 g kg⁻¹ fish meal (FM) and 200 g kg⁻¹ SBM. The basal diet was supplemented with either 280 g kg⁻¹ FM (Control), 390 g kg⁻¹ MGM or 450 g kg⁻¹ SBM to obtain three isonitrogenous (crude protein: 380 g kg⁻¹) and isocaloric (gross energy: 16 kJ g⁻¹) diets. For GC, a significant decrease in specific growth rate (SGR) was only observed in fish fed the SBM diet compared with the control (P < 0.05). Principal components analysis (PCA) of GC showed a higher similarity in antioxidant response to dietary supplementation with MGM and SBM proteins between liver and intestine, but the DC did not. The present results suggest that supplementing 390 g kg⁻¹ MGM protein to basal diet cause an enhancement of the antioxidant capacity in GC, but supplementing 390 g kg⁻¹ MGM and 450 g kg⁻¹ SBM proteins to basal diets resulted in a significant attenuation of the antioxidant capacity in DC.     

Water flow and diurnal variation in metabolite production rates of Atlantic salmon (Salmo salar L.) post‐smolt

Diurnal variation of carbon dioxide (CO₂) and total ammonia production in Atlantic salmon post‐smolt were studied at different water flow rates. The experiment comprised four groups each with two replicates representing specific flow levels of 0.5, 0.4, 0.3 and 0.2 L kg^?1 min^?1. During the first diurnal cycle, the seawater samples were collected eight times during 21 hr. In the second diurnal cycle, six samplings were performed during a prolonged sampling period of 35 hr. The highest CO₂ concentrations were observed in the lowest water flow group (0.2 L kg^?1 min^?1) between 4 and 10 p.m. for the first sampling period and at about 2 p.m. for the second sampling period. The overall real CO₂ production rates were in the range 1.7–5.5 mg kg^?1 min^?1 including diurnal variation in all groups for both sampling periods. In general, a second‐order polynomial model describes the relationship between specific water flow and real mean CO₂ production rate (p < 0.001). Maximum concentrations of total ammonia nitrogen (TAN) occurred around 7 and 8 p.m. in all groups and was highest for the 0.2 L kg^?1 min^?1 group for both sampling periods. Note that maximum concentration of TAN and CO₂ both occurred in the afternoon. The TAN production rates were in the range 0.06–0.44 mg kg^?1 min^?1 including both sampling cycles.     

Effect of binder type and concentration on prepared feed stability,feed ingestion and digestibility of Litopenaeus vannamei broodstock diets

The effect of six different binding agents (agar, sodium alginate, cassava starch, gelatin, wheat gluten and kelp meal) in two concentrations (30 g kg⁻¹ and 50 g kg⁻¹) were evaluated with respect to physical quality of Litopenaeus vannamei broodstock pelleted feed, after 15, 30, 60, 90, 120, 150 and 180 min of water immersion. The best treatments in terms of water stability, water absorption and protein leaching were obtained with sodium alginate and wheat gluten at 50 g kg⁻¹. In a second experiment, the feed ingestion and diet digestibility with these two binders and their combination (1 : 1) were compared against a control diet containing 50 g kg⁻¹ wheat flour. There were no significant differences (P > 0.05) in the daily feed ingestion rate 2.39–3.33% of the biomass. The most representative values of apparent digestibility of protein (ADP) and apparent dry matter digestibility (ADMD) were achieved with diets containing wheat gluten and alginate + gluten mixture as binder. Based on these results, combinations with 50 g kg⁻¹ wheat gluten is recommended as binder in pelleted feed for broodstock L. vannamei.     

Determination of the muscle residue levels and withdrawal time of florfenicol in healthy and experimentally Lactococcus garvieae infected rainbow trout (Oncorhynchus mykiss,Walbaum 1792) grown in the Black Sea water

The aim of this study was to determine, muscle residue level (MRL) and withdrawal time (WT) of florfenicol (FF), in healthy and infected rainbow trout (Oncorhynchus mykiss) with Lactococcus garvieae. Fish were divided into the four groups, Group I (Infected fish fed with FF free diet, n = 10), Group II (Healthy fish fed with FF free diet, n = 10), Group III (Infected fish fed with FF containing diet, n = 80) and Group IV (Healthy fish fed with FF containing diet, n = 80). Fish in Group I and Group III were infected with 1.8×10⁶ cfu/ml Lactococcus garvieae by immersion. FF containing diet was applied fish (10 mg kg^‐1 day^‐1) in Group III and IV for 10 days. FF residues in fish muscles were analysed by validated HPLC method. The limit of detection (LOD) and limit of quantification (LOQ) values of the method were determined to be 0.31 μg/g and 0.64 μg/g, respectively, and the recovery was 92.61%. As a result of the experiments, there was no significant difference between the infected and healthy group in term of MRLs. However, in both groups, it was determined that third‐day values were below MRLs (1,000 μg/kg). Also, the WT values were lower than the 500°C‐day values reported in the EC commission decision in both groups and were determined as 14.5 and 14.12 (water temperature mean 18°C ± 1) per day, respectively.     

Pharmacokinetic/pharmacodynamic properties and P‐glycoprotein expression in pacific white shrimp,Litopenaeus vannamei after oral administration at three dosages of oxolinic acid

The experiments explored the pharmacokinetics (PK) properties of oxolinic acid (OXA) after oral administration at three dosages (10, 30 and 80 mg/kg) via medicated feed in the shrimp. The results showed that the C_max values of 4.31, 14.93 and 16.62 mg/L and AUC_0–∞ values of 92.61, 252.30 and 364.27 mg hr^?1 L^?1 were observed at three OXA dosage groups in the haemolymph respectively. In the hepatopancreas, C_max values of 7.90, 27.23 and 60.51 mg/kg and AUC_0–∞ values of 42.01, 133.06 and 219.06 mg hr^?1 L^?1 were observed at 0.5 hr post administration respectively. In the muscle, C_max values of 1.62, 5.80 and 7.36 mg/kg and the AUC_0–∞ values of 25.64, 98.10 and 134.24 mg hr^?1 L^?1 were observed at 2 hr post administration respectively. In the gills, C_max values of 2.87, 8.08 and 12.12 mg/kg and the AUC_0–∞ values of 51.38, 118.65 and 206.48 mg hr^?1 L^?1 were observed at 4 hr post administration respectively. In addition, the in vitro MIC values of OXA at three dosages against 132 strains of Vibrio were examined and showed that the minimum inhibitory concentration (MIC) values for OXA primarily ranged from 0.15–1.25 µg/ml, including eight strains of Vibrio showing MIC values ≥5 µg/ml. The MIC₅₀ and MIC₉₀ values of 132 strains were 0.62 and 1.25 μg/ml respectively. The AUC_0–24/MIC₉₀ ratios of Vibrio were 140.4 in 30 mg/kg group. Furthermore, the P‐glycoprotein (P‐gp) expression was determined in shrimp tissues after administration to three dosage groups (10, 30 and 80 mg/kg). The results showed that P‐gp expression was up‐regulated in the hepatopancreas (5.36‐, 13.68‐ and 31.06‐fold respectively) compared with the control group.     

Effects of dissolved oxygen and dietary lysine levels on growth performance,feed conversion ratio and body composition of grass carp,Ctenopharyngodon idella

In order to investigate the effects of lysine and dissolved oxygen on grass carp, the grass carp were fed with 13, 15 and 17 g kg ⁻¹ lysine diet at about 6 mg L⁻¹ (high dissolved oxygen, HO group) or 3.5 mg L⁻¹ (low dissolved oxygen, LO group) dissolved oxygen level, for 8 weeks. The fish were fed to apparent satiation by hand. The results showed that apparent digestibility of protein, energy and dry matter were decreased significantly when grass carp were fed at 3.5 mg L⁻¹ dissolved oxygen, and feed intake (FI) was also inhibited by low dissolved oxygen (P < 0.05). Weight gain, protein retention, protein efficiency, feed conversion ratio and amino acid retention of fish at 6 mg L⁻¹ dissolved oxygen level were significantly improved at 3.5 mg L⁻¹ dissolved oxygen level (P < 0.05). Weight gain, protein and amino acid retention, and feed efficiency of grass carp at the two dissolved oxygen levels were significantly improved by lysine supplementation (P < 0.05). The dietary lysine level and dissolved oxygen of water had an interaction effect on feed conversion ratios (P < 0.05). Grass carp fed at low dissolved oxygen level showed lower liver protein and fat contents. Plasma aspartate aminotransferase (AST) activity of grass carp fed at 3.5 mg L⁻¹ dissolved oxygen level was significantly increased compared to 6 mg L⁻¹ dissolved oxygen level (P < 0.05). Our results show that low dissolved oxygen level of water is harmful to the liver of grass carp.     

Pharmacokinetics,optimal dosages and withdrawal time of florfenicol in Asian seabass (Lates calcarifer) after oral administration via medicated feed

Asian seabass (Lates calcarifer) is an economically important fish in Asian and Australian markets, but few pharmacokinetic (PK) data of antimicrobial drugs in this species is available. The present study investigated the PK behaviour of florfenicol (FF) through medicated feed in Asian seabass cultured at 25°C. The serum and muscle/skin concentrations of FF and its metabolite florfenicol amine (FFA) were determined by the HPLC-FLD method and analysed by one-compartmental model. The optimal dosages were determined by pharmacokinetic-pharmacodynamic (PK-PD) approach and the linear regression analysis was used to determine the withdrawal time (WDT). The PK study following a single oral administration of 15 mg/kg FF via medicated feed revealed that the absorption half-life (t_1/2Ka), elimination half-life (t_1/2K), peak concentration (C_max), area under the concentration-time curve (AUC), volume of distribution (Vd/F) and clearance (CL/F) were 1.47 h, 8.07 h, 8.61 μg/ml, 146.41 h·μg/ml, 1.19 L/kg and 0.102 L/kg/h, respectively. The muscle/skin concentration-time profile was similar to that of the serum, suggesting well distribution but only a small fraction of FF was metabolized to FFA. The optimal dosage for a minimum inhibitory concentration of 2 μg/ml was calculated as 13.38 mg/kg/day. The appropriate WDT after multiple oral medications with 15 mg/kg FF once daily for 7 days was determined as 8 days. Information obtained from the current study can potentially be applied for the treatment of bacterial diseases in farming Asian seabass.     

Immunostimulatory effect of water soluble fraction of Nyctanthes arbortristis leaves on the immune response in Oreochromis mossambicus (Peters)

Use of immunostimulants as a prophylactic measure against diseases in fish is considered as an effective alternative to antibiotic use. Plant‐derived immunostimulants have recently received more attention, as most of them are cost‐effective and eco‐friendly. This study was carried out with the objective of evaluating the possible immunostimulatory activity of intraperitoneally injected water soluble fraction of Nyctanthes arbortristis (an Indian medicinal plant) leaves on (i) specific immunity (antibody response), (ii) nonspecific immunity (Lysozyme activity, ROS production) and (iii) functional immunity in terms of disease resistance against Aeromonas hydrophila in Oreochromis mossambicus. The results of the study showed that all the tested doses (i.e. 3.2, 16, 80 and 400 mg kg^?1) of water soluble fraction of Nyctanthes arbortristis leaves significantly enhanced both primary and secondary antibody responses to heat killed A. hydrophila. Significant enhancement in serum lysozyme activity by all the doses of water soluble fraction was observed on day 10. The lower doses of 3.2 and 16 mg kg^?1 of water soluble fraction alone enhanced intracellular ROS production. Fish treated with both single and double dose of water soluble fraction showed maximum disease resistance. The highest dose (400 mg kg^?1) of water soluble fraction exhibited highest relative per cent survival in both single dose and double dose groups.     

Pharmacokinetic disposition of enrofloxacin in brown trout (Salmo trutta fario) after oral and intravenous administrations

In this study, the pharmacokinetic profile of enrofloxacin (EF) and its major metabolite, ciprofloxacin (CF), were investigated in brown trout (Salmo trutta fario) (n = 150) after intravenous (i.v.) and oral (p.o.) administrations of a single dose of 10 mg kg^− 1 body weight (b.w.) at 10 °C. The plasma concentrations of the drugs were determined by high-performance liquid chromatography (HPLC-UV) from 0.08 to 120 h. Pharmacokinetic parameters were described by the two-compartment open model for intravenous and oral administrations, respectively. After intravenous administration, the elimination half-life (t_1/2β), apparent volume of distribution at steady-state (V_ss) and total body clearance (Cl_tot) of enrofloxacin were 19.14 ± 1.51 h, 3.40 ± 0.18 L kg^− 1 and 0.14 ± 0.01 L kg h^− 1, respectively. After oral administration, the maximum plasma concentration (C_max), time of maximum concentration (t_max) and bioavailability (F%) were 2.30 ± 0.08 µg mL^− 1, 8 h and 78 ± 4%, respectively. Ciprofloxacin was not detected in the present study. The elimination half-life for enrofloxacin following oral administration was longer than values calculated for other animals. After oral administration, the mean plasma concentration was well above the minimum inhibitory concentrations (MICs)—that is, > 0.5 µg mL^− 1 at 36 h—for most gram-negative fish pathogens. It is possible and practical to obtain therapeutic blood concentrations of enrofloxacin in brown trout (S. trutta fario) using oral administration of 10 mg kg^− 1 body weight; therefore, it may be effective in the therapy for brown trout diseases.     

Pharmacokinetics and tissue residues of vancomycin after oral administration in crucian carp

The pharmacokinetics and tissue residues of vancomycin in crucian carp (Carassius auratus) were studied after oral administration of a single dosage of 1 mg/kg body weight at water temperature of 18°C. The drug concentration‐time data were fitted using a two‐compartment open model with first‐order absorption. The vancomycin concentrations in plasma and tissues including muscle, liver and kidney were analyzed by means of liquid chromatography‐tandem mass spectrometry. As a result, the plasma absorption rate constant of vancomycin was 3.75/hr and the time point of maximum concentration in plasma was 3.0 hr respectively. The maximum concentration of vancomycin in plasma was 216.58 μg/L. The distribution half‐life and the elimination half‐life in plasma were 6.69 hr and 190 hr, as well as the absorption half‐life of plasma was 0.19/hr. The area under the plasma concentration‐time curve of plasma was 15.12 mg hr/L. The apparent volume of distribution of plasma was estimated to be 4.45 L/kg and the total body clearance was computed as 0.061 L hr^?1 kg^?1. In addition, the maximum tissue concentrations of vancomycin were in the order of liver > kidney > muscle. To guarantee the safety of tissue samples for consumption, it was suggested that the withdrawal time should not be less than 30 days at 18°C. All these results may provide guidance for future vancomycin treatment in aquaculture.     

Erythromycin and florfenicol treatment of rainbow trout Oncorhynchus mykiss (Walbaum) experimentally infected with Flavobacterium psychrophilum

Flavobacterium psychrophilum is responsible for significant economic losses in rainbow trout aquaculture. Antimicrobial treatment remains the primary means of control; however, there are limited choices available for use. The objectives of the study were therefore to determine the minimum inhibitory concentrations for erythromycin and florfenicol in selected F. psychrophilum isolates and to evaluate their clinical treatment efficacy in experimentally infected rainbow trout. All isolates tested had moderate susceptibility to florfenicol and erythromycin except one isolate, which had low susceptibility to erythromycin. Two isolates (one with moderate and one with low susceptibility to erythromycin) were used in an experimental infection trial. Rainbow trout juveniles were injected intraperitoneally with 10⁸ cfu/fish and after mortality had begun, fish were given erythromycin‐ and florfenicol‐medicated feed at a rate of 75 mg kg^?1 day^?1 and 10 mg kg^?1 day^?1 fish body weight, respectively, for 10 consecutive days. The splenic F. psychrophilum load was determined using an rpoC quantitative PCR throughout the 30‐day trial. Relative to antibiotic‐free controls, erythromycin treatment significantly (p < 0.05) reduced mortality of rainbow trout juveniles infected with FPG101, even when treatment was initiated after clinical signs developed.     

Deposition of conjugated linoleic acid in market size sea bass (Dicentrarchus labrax) and its effects on performance,composition and fillet sensory and texture attributes

The effect conjugated linoleic acid (CLA) accumulation in fish fillet on sensory and texture attributes has been evaluated. Fish were fed diets with graded levels of CLA, which were administered for different durations prior to harvest. Fish were sampled for fillet proximate composition and fatty acid profile for sensory and texture parameters at the end of the trial. Growth performance, feed conversion ratio and muscle proximate composition were not affected by dietary treatments. Muscle fatty acid profile was not affected by dietary treatments despite the increase in saturated fatty acids in fish fed diet with 10 g kg⁻¹ of CLA for 16 weeks before harvest. Besides, the deposition levels of CLA isomers increased gradually depending on the CLA treatment, reaching its maximum level in fish fed 10 g kg⁻¹ CLA diet for 20 weeks before harvest. Fish fed 10 g kg⁻¹ were juicier (P < 0.05) compared with control group, whereas fish fed 5 g kg⁻¹ CLA diet were less (P < 0.05) chewy than fish fed control diet regarding sensory attributes. Texture parameters were not significantly affected by dietary CLA. These results suggest that market size European sea bass could successfully incorporate CLA isomers with some minor positive effects on sensory analyses.     

Evaluation of Calanus finmarchicus copepod meal in practical diets for juvenile Atlantic halibut (Hippoglossus hippoglossus)

The copepod, Calanus finmarchicus, has potential as a new ingredient in practical feeds for marine fish. Two experiments were conducted to evaluate the nutritional value of C. finmarchicus meals when fed to juvenile Atlantic halibut. The first study determined protein, lipid and energy digestibility coefficients of four C. finmarchicus meals prepared under different processing conditions. The second study evaluated growth and nutrient utilization efficiency of juveniles fed diets containing varying proportions of fish meal, plant meals and C. finmarchicus meal. Moisture, ash, protein, lipid and gross energy contents of the C. finmarchicus meals were 28–93, 74–138, 505–648, 123–269 g kg⁻¹, and 21–26 MJ kg⁻¹, respectively. Protein, lipid and energy digestibilities of C. finmarchicus meals were 91–99%, 90–95% and 90–99%. The digestibilities significantly decreased for the Calanus meals processed at higher temperatures. During the growth study, halibut fed a diet containing 240 g kg⁻¹ C. finmarchicus meal had significantly higher weight gain and growth rate than all other groups. Nitrogen and energy retention efficiencies ranged between 35–45% and 33–43%, and were significantly higher for fish fed 160–240 g kg⁻¹ C. finmarchicus meal than for fish fed plant protein and control diets. The results indicate that growth and nutrient utilization efficiency are improved in Atlantic halibut fed diets supplemented with C. finmarchicus meal.     

Wheat gluten in extruded fish feed: effects on morphology and on physical and functional properties

This article focuses on understanding the role of vital wheat gluten on the structural parameters of extruded fish feed and its correlation to the physical and functional properties. Gluten–soy protein concentrate blends with five gluten concentrations (0–200 g kg⁻¹) were produced. An abrupt reduction in oil uptake was observed with the 200 g gluten kg⁻¹ blend. Inclusion of gluten from 100 to 200 g kg⁻¹ resulted in unacceptable product properties. Sinking of feed pellets with 0 and 50 g gluten kg⁻¹ was 100%, whereas only 36% of pellets with 200 g gluten kg⁻¹ sank. We suspect that this is due to a relationship between morphological structure and oil impregnation during coating of feeds. The addition of gluten at 200 g kg⁻¹ gave a smoother and non‐porous outer surface. Pellets without gluten had a larger number of cells that were smaller than 200 μm (P < 0.05) compared with pellets with 100 and 200 g gluten kg⁻¹. More spherical cell shapes (P < 0.01) and a compact structure were favoured in the presence of gluten. The closed porosity increased (P < 0.05), whereas interconnectivity between pores decreased (P < 0.01), with increasing gluten content from 0 to 200 g kg⁻¹. The effects of the addition of gluten are probably related to the film‐forming properties of gluten.     

Growth,feed utilization,body composition and swimming performance of giant croaker,Nibea japonica Temminck and Schlegel,fed at different dietary protein and lipid levels

A 50‐day feeding trial was conducted to examine the effects of dietary protein and lipid levels on growth, feed utilization, body composition and swimming performance of giant croaker, Nibea japonica. Fish (initial body weight 44.6 g ind⁻¹) were fed ten test diets which were formulated at 5 crude protein levels (360, 400, 440, 480 and 520 g kg⁻¹) and 2 crude lipid levels (90 and 150 g kg⁻¹). In addition, a raw fish diet (fillet of small yellow croaker) served as the reference. The weight gain (WG) increased, whereas the feed intake (FI) and feed conversion ratio (FCR) decreased, with increasing dietary protein level from 360 to 520 g kg⁻¹. At the same dietary protein level, no significant difference was found in the WG between fish fed the diets containing 90 or 150 g kg⁻¹ crude lipid. Fish fed the diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid exhibited higher WG, nitrogen retention efficiency (NRE) and energy retention efficiency (ERE) but lower nitrogen wastes output (TNW). At the end of the feeding trial, the hepatosomatic index (HSI) and viscerosomatic index (VSI) decreased, whereas the body protein content increased, with increase in dietary protein level. The body lipid content was higher in fish fed at the 150 g kg⁻¹ lipid level than in fish fed at the 90 g kg⁻¹ lipid level. No significant difference was found in the maximum sustained swimming speed (MSS) between fish fed at different dietary protein and lipid levels. The WG, NRE, ERE and condition factor (CF) were higher, whereas the FI, FCR, HSI, VSI and TNW were lower, in fish fed the raw fish diet than in fish fed the diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid. No significant difference was detected in the MSS between fish fed the raw fish diet and diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid. The results of this study suggest that the suitable dietary crude protein and crude lipid levels are 480 g kg⁻¹ and 90 g kg⁻¹ for giant croaker reared in net pens.     

Nitrite toxicity to Litopenaeus schmitti (Burkenroad, 1936, Crustacea) at different salinity levels

Litopenaeus schmitti juveniles (total length 15 ± 0.4 cm) were exposed to different concentrations of nitrite using the static renewal method at different salinity levels (5‰, 20‰ and 35‰) at pH 8.0 and 20 °C. The 24, 48, 72 and 96 h LC50 values of nitrite in L. schmitti juveniles were 40.72, 32.63, 24.63, and 19.12 mg L⁻¹ at 5‰; 53.52, 38.60, 27.76, 25.55 mg L⁻¹ at 20‰; 54.32, 47.87, 41.67 and 38.88 mg L⁻¹ at 35‰ salinity. As the salinity decreased from 35‰ to 5‰, susceptibility to nitrite increased by 33.4%, 46.7%, 69.2% and 103.3%, after 24, 48, 72 and 96 h of exposure respectively. Furthermore, we found that exposure of shrimp to nitrite caused an increase in oxygen consumption by 137.3%, 99.2% and 81.4% and an increase in the ammonia excretion level by 112.5%, 87% and 64.3% with respect to the control with decreasing salinity levels.     

Molecular cloning of cDNA of mammalian and chicken II gonadotropin-releasing hormones (mGnRHs and cGnRH-II) in the beluga (<Emphasis Type="Italic">Huso huso</Emphasis>) and the disruptive effect of methylmercury on gene expression

Two gonadotropin-releasing hormone (GnRH) isoforms were identified in the beluga (Huso huso) brain by cDNA sequencing: prepro-mammalian GnRH (mGnRH) and prepro-chicken GnRH-II (cGnRH-II). The nucleotide sequences of the beluga mGnRH and cGnRH-II precursors are 273 and 258 base pairs (bp) long, encoding peptides of 91 and 86 amino acids, respectively. To investigate the effect of methylmercury (MeHg) on GnRH gene expression, animals were fed with four diets containing increasing levels of MeHg (0 mg kg⁻¹ [control]; 0.76 mg kg⁻¹ [low]; 7.8 mg kg⁻¹ [medium]; 16.22 mg kg⁻¹ [high]) for 32 days. The effects of MeHg on brain GnRH mRNA levels were evaluated by real-time PCR. A significant decrease in brain mGnRH and cGnRH-II mRNA levels were detected in fish receiving high dietary MeHg dose compared to controls on day 11 (P < 0.05). On day 18 and 32, all treatment groups had significantly lower brain mGnRH and cGnRH-II mRNA levels compared to the control group (P < 0.05). These findings demonstrate a disruptive role of MeHg on the level of brain mGnRH and cGnRH-II mRNAs in immature beluga.