Acute effects of ammonia exposure on the plasma and haematological parameters and histological structure of the juvenile blunt snout bream,Megalobrama amblycephala,and post‐exposure recovery

In this study, we investigated the toxic effects of ammonia‐N on the plasma and haematological parameters and histological structure of blunt snout bream (Megalobrama amblycephala) juveniles. The fish (initial weight, 14.79 ± 0.01 g) were randomly sorted into six tanks (200 L), and each tank was stocked with 40 fish for culture. The juveniles were exposed to two ammonia‐N levels—0 mg/L (control group) and 25 mg/L (experimental group)—and sampled at 0, 6, 12, 24, 48 and 72 hr, and then they underwent 96 hr of post‐exposure recovery. The results showed that ammonia‐N had significant effects on the plasma and haematological parameters. The treatment group showed increased cortisol, plasma ammonia and haematocrit levels and white blood cell count with increasing exposure time, up to 24 hr, and then the levels and count decreased. A significantly higher plasma glucose level was observed in the treatment group at 12 hr. After 96 hr of post‐exposure recovery, all parameter levels decreased to the control levels. The fish displayed histopathological alterations in the gills, liver and kidney. The results indicate that the severity of the lesions clearly differed among the organs, with the liver showing the most extensive damage, followed by the gills and kidney. Adverse effects to physiological indicators and histological structure increased with increasing exposure time before 24 hr. The fish showed self‐regulation; however, the histological structure could not recover fully, the gill tissue showed irreversible changes and the kidney tissue exhibited the worst recovery ability.     

Pharmacokinetic model of florfenicol in turbot (Scophthalmus maximus): establishment of optimal dosage and administration in medicated feed

The pharmacokinetics of florfenicol (FF) in turbot (Scophthalmus maximus) was studied after single intravenous (10 mg kg⁻¹) and oral (100 mg kg⁻¹) administration. The plasma concentration–time data of florfenicol were described by an open one‐compartment model. The elimination half‐life (t_1/2) was estimated to be 21.0 h, and the total body clearance, Cl, was determined as 0.028 L kg h⁻¹. The apparent volume distribution (V_d) was calculated to be 0.86 L kg⁻¹ and the mean residence time (MRT_iv) was 30.2 h. Following oral administration, the maximum plasma concentration (C_max) of 55.4 μg mL⁻¹ was reached at 12 h (T_max). The absorption constant (k_a) was 0.158 h⁻¹. The bioavailability was estimated to be 57.1%. The low bioavailability observed at higher doses was explained by the saturation of the mechanisms of absorption. The drug absorption process was limited by its inherent low solubility, which limited the amount of available FF absorbed in the gastrointestinal tract. Based on the pharmacokinetic data, an optimal dosing schedule for FF administration is hereby provided. Based on the minimum inhibitory concentration found for susceptible strains of Aeromonas salmonicida, oral FF administration of first, an initial dose of 30 mg FF kg⁻¹, followed by 6 maintenance doses at 18 mg kg⁻¹/daily could be effective against furunculosis in turbot.     

Single Intravascular and Oral Dose Pharmacokinetics of Mebendazole in Blunt Snout Bream,Megalobrama amblycephala

Mebendazole (MBZ) is a broad‐spectrum benzimidazole methylcarbamate anthelmintic used widely in animal husbandry and aquaculture. However, there is no information available on the pharmacokinetic behavior of MBZ in blunt snout bream, Megalobrama amblycephala. In this study, pharmacokinetic parameters of MBZ were estimated in blunt snout bream after intravascular (3 mg/kg body weight [BW]) and oral (20 mg/kg BW) administration. The analyses of plasma samples were performed using ultra performance liquid chromatography with ultraviolet detector. After intravascular administration, plasma concentration–time curves were best described by a two‐compartment open model. The distribution half‐life (t_1/2α), elimination half‐life (t_1/2β), and area under the concentration–time curve (AUC) of blunt snout bream were 0.1 h, 27.9 h, and 56666.0 h.µg/L, respectively. After oral administration, a one‐compartment open model with first‐order absorption best fit the plasma data. The absorption half‐life (t_1/2Ka), elimination half‐life (t_1/2Ke), peak concentration (C_max), time‐to‐peak concentration (T_max), and AUC of blunt snout bream were estimated to be 1.9 h, 34.6 h, 918.1 µg/L, 8.4 h, and 54201.4 h.µg/L, respectively. The oral bioavailability (F) was 14.3 %. The pharmacokinetics of MBZ in blunt snout bream displayed low bioavailability, relatively rapid absorption, and relatively rapid elimination.     

Pharmacokinetics of minocycline in crucian carp (Carassius auratus) after intravenous and oral administration

The pharmacokinetic of minocycline was studied after a single intravenous as well as oral dose (5 mg/kg body weight) in crucian carp (Carassius auratus) reared in freshwater at 10°C. Plasma samples were randomly collected from six fish at each sampling time. Plasma concentrations were determined by high‐performance liquid chromatography and further subjected to noncompartmental analysis. Initial concentration of minocycline just after intravenous administration was calculated as 7.320 μg/ml, while the other parameters after intravenous injection were determined as flows: apparent elimination rate constant (λ_z) of 0.064 per hr, apparent elimination half‐life () of 10.82 hr, total body clearance (Cl) of 142.72 ml/hr/kg, volume of distribution (Vz) of 2,227.38 ml/kg and volume of distribution at steady‐state (Vss) of 1,937.08 ml/kg. While after oral administration, the λ_z, , mean absorption time (MAT), absorption half‐life (t_1/2ka) and bioavailability were determined as 0.059 per hr, 11.74, 5.55, 3.84 hr, and 81.98%, respectively, and the peak concentration was observed as 1.474 ± 0.362 μg/ml at 8 hr. It was shown that minocycline was slowly but relatively completely absorbed, extensively distributed, and slowly eliminated in crucian carp. Based on the ratios of AUC_0–24 hr/MIC90, a minocycline dosage of 5 mg/kg body weight administered intravenously or orally would be only effective to successfully treat crucian carp infected by bacterium with MIC values ≤0.25 μg/ml.     

Pharmacokinetics and tissue distribution of bronopol in grass carp,Ctenopharyngodon Idella,at 15 and 20°C

This study explored the pharmacokinetic characteristics of bronopol in grass carp (Ctenopharyngodon idella) under different temperatures. The concentrations of bronopol in the plasma and tissues (kidney, liver and muscle) of grass carp after soaking in bronopol (6.75 µg/ml) at 15 and 20°C were determined using liquid chromatography–mass spectrometry. The concentrations of bronopol in both plasma and tissue first increased and then decreased with soaking time at both tested temperatures. The peak bronopol concentrations at 15°C (plasma: 8.934 µg/ml; kidney: 9.23 µg/g; liver: 9.47 µg/g; and muscle: 7.98 µg/g) were slightly lower than those at 20°C (plasma: 9.654 µg/ml; kidney: 10.83 µg/g; liver: 13.40 µg/g; and muscle: 8.68 µg/g). In an analysis of pharmacokinetic parameters, the bronopol concentration–time profiles for plasma were best described by a two‐compartment, open pharmacokinetic model with first‐order absorption. The t_1/2β for plasma at 15 and 20°C was 82.933 and 92.041 hr, respectively, indicating that the elimination of bronopol was faster at 15°C than at 20°C. In addition, AUC_0‐t and AUC_0‐∞ values of bronopol were 477.892 and 495.809 µg/L hr at 15°C and 495.809 and 589.859 µg/L hr at 20°C, indicating that the content of bronopol in the organism was higher at higher temperature.     

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

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

Does Ralstonia eutropha,rich in poly‐β hydroxybutyrate (PHB), protect blue mussel larvae against pathogenic vibrios?

The natural amorphous polymer poly‐β‐hydroxybutyrate (PHB‐A: lyophilized Ralstonia eutropha containing 75% PHB) was used as a biological agent to control bacterial pathogens of blue mussel (Mytilus edulis) larvae. The larvae were supplied with PHB‐A at a concentration of 1 or 10 mg/L for 6 or 24 hr, followed by exposure to either the rifampicin‐resistant pathogen Vibrio splendidus or Vibrio coralliilyticus at a concentration of 10⁵ CFU/ml. Larvae pretreated 6 hr with PHB‐A (1 mg/L) survived a Vibrio challenge better relative to 24 hr pretreatment. After 96 hr of pathogen exposure, the survival of PHB‐A‐treated mussel larvae was 1.41‐ and 1.76‐fold higher than the non‐treated larvae when challenged with V. splendidus and V. coralliilyticus, respectively. Growth inhibition of the two pathogens at four concentrations of the monomer β‐HB (1, 5, 25 and 125 mM) was tested in vitro in LB₃₅ medium, buffered at two different pH values (pH 7 and pH 8). The highest concentration of 125 mM significantly inhibited the pathogen growth in comparison to the lower levels. The effect of β‐HB on the production of virulence factors in the tested pathogenic Vibrios revealed a variable pattern of responses.     

Pharmacokinetics and physio‐metabolic response of single and multiple dose of fenbendazole in Labeo rohita (Hamilton, 1822) fingerlings

An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (C_max) 1.85 and 3.09 µg/ml in plasma at 4 hr (T_max) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.     

Efficacy and effects of clove oil and MS‐222 on the immune‐biochemical responses of juvenile rohu Labeo rohita

The present study determined the effective concentrations of clove oil and MS‐222 in juvenile rohu Labeo rohita for quick induction and recovery. The immune‐biochemical responses due to 0, 1 and 24 hr exposure to those anaesthetics were also evaluated. Of four concentrations of the anaesthetics examined, the lowest effective concentration of clove oil and MS‐222 were 50 µl/L and 125 mg/L respectively. Clove oil and MS‐222 significantly increased the myeloperoxidase, total protein and alkaline phosphatase activity at some of the holding durations. However, superoxide anion production (after 0 and 1 hr) and antiprotease activity (after 24 hr) were significantly reduced in fish exposed to clove oil. Serum glucose content was significantly elevated in the MS‐222‐treated group. Furthermore, the clove oil‐treated group showed significantly higher levels of serum Na⁺ and K⁺, while the aspartate and alanine aminotransferase activities were significantly enhanced in the MS‐222 group. The use of both clove oil and MS‐222 is advised as an anaesthetic agent for rohu with a bias towards clove oil, considering its economic and operational feasibility.     

Change in blood stress and antioxidant markers and hydromineral balance of common carp (Cyprinus carpio) anaesthetized with citronellal and linalool: Comparison with eugenol

The present study aimed to investigate the effects of anaesthesia with citronellal and linalool on stress, antioxidant and hydromineral responses of common carp. Eugenol was used as control anaesthetic, as it is a common anaesthetic in aquaculture. The fish (110 ± 5.65 g) were exposed to 406 mg/L citronellal, 982 mg/L linalool or 43 mg/L eugenol within 5 min (short‐term anaesthesia) or 10 and 20 mg/L of each anaesthetic for 3 hr (long‐term exposure), before blood sampling. The results showed that the short‐term anaesthesia had no significant effects on plasma lactate, sodium, chloride and phosphorus levels and blood catalase (CAT) and superoxide dismutase activities. The short‐term anaesthesia with linalool led to higher stress responses (increased cortisol and glucose levels) and oxidative stress (increased malondialdehyde [MDA] level and decreased total antioxidant capacity [TAC] and glutathione peroxidase [GPx] activity) compared to the eugenol and citronellal. Citronellal group had significantly higher cortisol and lower TAC level and GPx activity compared to eugenol group. Under the long‐term exposure, increase in anaesthetic concentration led to significant increase in plasma cortisol, glucose and phosphorus levels. The linalool‐treated fish had significantly higher cortisol and MDA levels compared to the other groups. The eugenol group had significantly higher lactate and phosphorus and lower TAC levels compared to the other groups. Although citronellal causes slight increase in plasma cortisol level, it induces lower tissue damages compared to eugenol. Thus, citronellal might be an alternative anaesthetic for carp anaesthesia based on the present tested parameters.     

Pharmacokinetics and tissue residues of marbofloxacin in crucian carp (Carassius auratus) after oral administration

Pharmacokinetics and residue elimination of marbofloxacin (MBF) were studied in crucian carp (Carassius auratus, 250±30 g) kept at two water temperatures of 15 and 25 °C. Marbofloxacin concentrations in plasma and tissues were analysed by means of high‐performance liquid chromatography using an ultraviolet detector. The limits of detection were 0.02 μg mL^?1, 0.02 μg g^?1, 0.025 μg g^?1, 0.02 μg g^?1 and 0.025 μg g^?1 in plasma and muscle, skin, liver and kidney respectively. Fish were administered orally at a single dosage of 10 mg kg^?1 body weight in the PK group. The data were fitted to two‐compartment open models at both temperatures. At 15 °C, the absorption half‐life () and distribution half‐life (t_1/2α) of the drug were 0.36 and 4.48 h respectively. The corresponding values at 25 °C were 0.23 and 0.87 h respectively. The elimination half‐life (t_1/2β) was 50.75 h at 15 °C and 25.05 h at 25 °C. The maximum MBF concentration (C_max) differed little between 15 (6.43 μg mL^?1) and 25 °C (8.36 μg mL^?1). The time to peak concentration was 1.74 h at 15 °C and 0.78 h at 25 °C. The apparent volume of distribution (V_d/F) of MBF was estimated to be 1.36 and 0.87 L kg^?1 at 15 and 25 °C respectively. The area under the concentration–time curve (AUC) was 301.80 μg mL^?1 h at 15 °C and 182.80 μg mL^?1 h at 25 °C. The total clearance of MBF was computed as 0.03 and 0.05 L h^?1 kg^?1 at 15 and 25 °C respectively. After repeated oral administration at a dosage of 10 mg kg^?1 body weight per day for 3 days, the results showed that the elimination half‐lives () of MBF from all tissues at 15 °C were longer than that at 25 °C. Therefore, water temperature is an important factor to be considered when deciding a reasonable withdrawal time.     

Effects of various dietary factors on astaxanthin absorption in Atlantic salmon (Salmo salar)

The experiment was designed to investigate the dietary factors that might enhance or interfere with astaxanthin (Ax) absorption in salmon including potentially interfering factors such as certain carotenoids (zeaxanthin and lutein), plant sterols, fibre and enhancing compounds such as cholesterol and vitamin E. Two hundred and eighty‐eight salmon (778 ± 78 g) were reared in sea water under controlled conditions and fed practical experimental diets. The experimental diets were supplemented with 40 mg Ax kg^?1, in addition to various dietary factors, including cholesterol (2%), vitamin E (450 IU kg^?1), wheat bran (5%), lutein (40 mg kg^?1), zeaxanthin (40 mg kg^?1) and phytosterol (2%). After 26 days of feeding, blood was collected and plasma was separated to determine the plasma Ax concentration. Ax was not detected in the plasma of fish fed the non‐pigmented diet. Fish fed diet containing 2% cholesterol significantly improved Ax absorption, which was reflected in the higher Ax concentration in plasma of Atlantic salmon. Other supplements including vitamin E, wheat bran, lutein, zeaxanthin and phytosterols in diet had no significant effect on plasma Ax concentration . Fish fed diet containing 2% cholesterol significantly increased cholesterol concentration in fish plasma. Phytosterol had no benefit to lower cholesterol plasma level in fish fed 2% phytosterol‐supplemented diet.     

A single‐dose pharmacokinetic study of emamectin benzoate in cod,Gadus morhua L., held in sea water at 9 °C

The pharmacokinetic profile of the antiparasitic agent emamectin benzoate was studied in plasma after intravenous (i.v.) injection and in plasma, muscle and skin following oral (p.o.) administration to cod, Gadus morhua, held in sea water at 9 °C and weighing 100–200 g. Following i.v. injection, the plasma drug concentration‐time profile showed two distinct phases. The plasma distribution half‐life (t_1/2α) was estimated as 2.5 h, the elimination half‐life (t_1/2β) as 216 h, the total body clearance (Cl_T) as 0.0059 L kg^?1 h^?1 and mean residence time (MRT) as 385 h. The volume of distribution at steady state, V_d(ss), was calculated to be 1.839 L kg^?1. Following p.o. administration the peak plasma concentration (C_max) was 15 ng mL^?1, the time to peak plasma concentration (T_max) was 89 h and t_1/2β was 180 h. The highest concentration in muscle (21 ng g^?1) was measured after 7 days and t_1/2β was calculated to be 247 h. For skin, a peak concentration of 28 ng g^?1 at 3 days was observed and a t_1/2β of 235 h was determined. The bioavailability following p.o. administration was calculated to be 38%.     

Anaesthetic efficacy and biochemical effects of 1,8‐cineole in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792)

The aim of the present study was to investigate the anaesthetic efficacy and biochemical effects of 1,8‐cineole (cineole) in rainbow trout (Oncorhynchus mykiss). The fish were exposed to 200, 300, 400, 500, 600 and 800 μl/L cineole and time of induction of anaesthesia and recovery from anaesthesia were recorded. Thereafter, the fish haematological and biochemical responses to anaesthesia with different concentrations of cineole were studies. Moreover, the haematological and biochemical response of fish anaesthetized for 300 s with either cineole (283 μl/L) or eugenol (25 μl/L) were compared. Cineole at the concentrations of 200–800 μl/L induced stages 2, 3 and 4 anaesthesia within 109‐29.3, 226‐59 and 418‐117 s respectively. Increase in anaesthesia induction time led to higher stress responses and enzymes’ activity characterized by elevation in red blood cell (RBC), white blood cell (WBC), blood haematocrit and haemoglobin, and plasma cortisol, glucose and lactate levels, and aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase and creatine phosphokinase (CPK) activities. However, cineole concentrations had no significant effects on plasma alanine aminotransferase (ALT) activity and ions levels. Comparison of cineole with eugenol showed that cineole showed less change in blood RBC and plasma AST, ALP, CPK, cortisol and glucose levels compared to eugenol. In conclusion, cineole is efficient to anaesthetize trout at concentrations of 200–800 μl/L. Increase in cineole concentration shortens anaesthesia induction time, stress response and probably tissue damage. The concentrations of 600–800 μl/L cineol is recommended for rapid sampling as it causes the least stress and enzymatic responses. The present results suggest that cineole causes slightly lower side effects in trout compared to eugenol.     

Acute and chronic toxicity of nitrate to fat greenling (Hexagrammos otakii) juveniles

We evaluated the acute and chronic toxicity of nitrate to juvenile fat greenling Hexagrammos otakii. The 24‐, 48‐, 72‐, and 96‐hr LC₅₀s of nitrate to 1.91 ± 0.7 g greenlings were 2,741, 2,413.5, 2,357.6, or 2,339.2 mg/L nitrate‐N, respectively. Greenlings (6.55 ± 1.83 g) were exposed to 5 mg/L (control) and 157 mg/L for 4 weeks in a recirculating aquaculture system. After 4 weeks, length, weight, feed conversion ratio, and specific growth rate were significantly (p < 0.05) lower for nitrate‐exposed fish than for control fish. Elevated nitrate exposure was associated with decreased plasma hemoglobin concentration and red blood cell count. Our results demonstrate that nitrate poses a threat to greenlings and provide information that is useful for establishing water quality criteria for early life stages of this cultured fish. The sensitivity of greenlings to elevated NO₃^? should be evaluated at other life stages to determine how chronic exposure might impact survival, growth, health, reproductive success, and harvest quality.     

Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

A high amylase‐producing bacterial strain exhibiting the phosphorus‐dissolving ability,isolated from freshwater aquaculture pond

Amylase‐producing bacteria could improve water quality contaminated by waste from feed residue and fish metabolism, thereby increasing the efficiency of aquaculture systems. The objective of this research was to screen and optimize fermentation conditions of a high amylase‐producing strain. Four amylase‐producing bacterial strains (named S458‐1, G05, H38 and B09) were isolated from a grass carp pond, and the strain S458‐1 showed the highest amylase‐producing ability, with 19.58 ± 0.38 mm hydrolysis circle diameter. The strain S458‐1 was identified as Bacillus cereus based on morphological identification, biochemical identification and 16S rDNA sequence analysis. The optimal culture medium formula included (in g/L) Ca²⁺ 0.8, Mg²⁺ 0.2, Mn²⁺ 0.4, Fe²⁺ 0.6, Al³⁺ 0.2, 1% soluble starch and 1% peptone. The optimal fermentation conditions were determined as initial pH 9, culture temperature 37°C, fermentation time of 60 hr and 2% inoculum. Under the optimal formula and condition, its enzyme activity increased from 32 U/ml to 173.01 U/ml, a 5.41‐fold increase. Surprisingly, our research found that the strain S458‐1 also had phosphorus degradation capabilities. Its phosphorus‐dissolving ability was both time‐ and concentration‐dependent. Thus, this study will make a contribution to the bacterial amylase based on the fermentation process and provide a theoretical basis for further research of aquatic probiotics.     

Colonization of Aeromonas salmonicida subsp. masoucida strains in Atlantic salmon (Salmo salar L.) during infection

Aeromonas salmonicida subsp. masoucida (ASM) is classified as atypical A. salmonicida and brought huge economic damages to the local salmonid aquaculture in China. An ASM strain named AS‐C4 was used to investigate the colonization of ASM in Atlantic salmon (Salmo salar L.) by an immersion challenge with the control group (T0, no AS‐C4), group T1 (2.67 × 10⁴ CFU/ml AS‐C4) and group T2 (2.67 × 10⁷ CFU/ml AS‐C4). The numbers of AS‐C4 copies in different fish tissues (gill, intestine, skin, blood, muscle, spleen, liver and kidney) were determined at different time points post challenge using the quantitative real‐time PCR (qRT‐PCR). AS‐C4 were detected in the gill and intestine as early as 0 hr after the challenge both in T1 and T2 groups, suggesting that the gill and intestine were probably the portals of entry of AS‐C4 into salmon. Although AS‐C4 could not be detected in the skin until 24 hr after the challenge in T1 group, it could be detected in the skin as early as 0 hr after the challenge in T2 group, indicating that the skin may also be a portal of entry of AS‐C4 into salmon. AS‐C4 was immediately detected in the blood within 3 hr after it entered the host, suggesting that AS‐C4 successfully invaded the bloodstream of fish. After AS‐C4 colonized the host, it colonized the internal tissues, such as the spleen, liver, kidney and muscle. The results of this study will contribute to the understanding of the pathogenesis of the ASM strains and give a broader understanding of the infection route of ASM in it's host, providing more information for the development of new therapeutic strategies to protect against this pathogen in aquaculture.     

Effects of dietary turmeric administration on stress,immune, antioxidant and inflammatory responses of common carp (Cyprinus carpio) during copper exposure

The present study assessed the effects of dietary turmeric on Cyprinus carpio resistance and responses to copper exposure. First, the fish were assigned to four treatments received diets supplemented with 0 (control), 5, 10 and 20 g/kg turmeric for 3 weeks. Thereafter, the fish were exposed to lethal concentration (3.5 mg/L) of ambient copper for 24 hr and mortality was 65.3%, 41.8%, 22.7% and 20.6%, respectively. In the second experiment, the fish were fed with the aforementioned diets and simultaneously exposed to sub‐lethal concentration (0.25 mg/L) of ambient copper for 3 weeks. Copper exposure led to increases in plasma cortisol, glucose, malondialdehyde (MDA), alanine transaminase (ALT) and aspartate transaminase (AST), and decrease in plasma T₄, T₃, lysozyme, alternative complement haemolytic (ACH50), bactericidal activities, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and blood red blood cell count (RBC) and haemoglobin. Moreover, copper exposure led to significant upregulation of tumour necrosis factor‐alpha (TNF‐a) and interleukin 1‐beta (IL1‐b), and significant downregulation of interleukin 10 (IL10) gene expressions in the fish liver. Turmeric administration at 10 g/kg significantly mitigated/inhibited the copper‐induced negative effects, which seems to be due to the augmenting of the antioxidant defence.     

Growth and survival of winter flounder (Pseudopleuronectes americanus) larvae reared on different photoperiod regimes from hatch to metamorphosis

The aim of this study was to determine the effect of photoperiod on growth and survival of winter flounder, Pseudopleuronectes americanus, from hatch until metamorphosis. Sixteen 100 L upwelling tanks were stocked with larvae at an initial stocking density of 20 larvae/L. Larvae in eight tanks were provided with continuous light (24 hr light/0 hr dark), while those in the remaining eight tanks were grown under a photoperiod of 14 hr light/10 hr dark. Tanks were sampled for larval standard lengh from 6 to 28 days post‐hatch, and the number of live larvae, live metamorphosed and dead metamorphosed fish were recorded at the termination of the experiment (starting 46 dph). Results indicated that larvae provided with continuous light were larger than those sampled from 14 hr light/10 hr dark treatment. No effect of photoperiod on the number of non‐metamorphosed fish recovered at the end of the experiment was detected. On the contrary, over three times as many fish survived and reached metamorphosis under continuous light. An ANCOVA model showed a significant interaction between light and age. Both regressions were significant and provided the following growth equations: SL_{24 hr light/0 hr dark} = 0.101 × age + 2.55 (r² = 0.96) and SL_{14 hr light/10 hr dark} = 0.094 × age +2.45 (r² = 0.97). Using 6.36 ± 0.059 mm SL as the mean size at metamorphosis, larvae experiencing continuous and 14 hr light/10 hr dark photoperiods would reach metamorphosis in 38 and 42 days respectively.