Ca. Branchiomonas cysticola,Ca. Piscichlamydia salmonis and Salmon Gill Pox Virus transmit horizontally in Atlantic salmon held in fresh water

Elucidation of the role of infectious agents putatively involved in gill disease is commonly hampered by the lack of culture systems for these organisms. In this study, a farmed population of Atlantic salmon pre‐smolts, displaying proliferative gill disease with associated Candidatus Branchiomonas cysticola, Ca. Piscichlamydia salmonis and Atlantic salmon gill pox virus (SGPV) infections, was identified. A subpopulation of the diseased fish was used as a source of waterborne infection towards a population of naïve Atlantic salmon pre‐smolts. Ca. B. cysticola infection became established in exposed naïve fish at high prevalence within the first month of exposure and the bacterial load increased over the study period. Ca. P. salmonis and SGPV infections were identified only at low prevalence in exposed fish during the trial. Although clinically healthy, at termination of the trial the exposed, naïve fish displayed histologically visible pathological changes typified by epithelial hyperplasia and subepithelial inflammation with associated bacterial inclusions, confirmed by fluorescent in situ hybridization to contain Ca. B. cysticola. The results strongly suggest that Ca. B. cysticola infections transmit directly from fish to fish and that the bacterium is directly associated with the pathological changes observed in the exposed, previously naïve fish.     

Wound‐healing potential of curcumin in the carp,Labeo rohita

In this investigation, the effect of dietary administration of curcumin on the healing of skin wound in fish, Labeo rohita, has been reported. Fish were divided into three groups: control group (fish without skin wound), sham group (fish with skin wound without curcumin treatment) and curcumin‐treated group (fish with skin wound and subjected to dietary administration of 1% curcumin). Experiments were conducted for 30 days to assess the healing of skin wounds at different time intervals using scanning electron microscopy, histology, and mucopolysaccharide and enzyme histochemistry. In the curcumin‐treated group, healing of skin wounds was found to be enhanced than in the sham group as indicated by early restoration of morphology of the surface layer of epithelial cells; the density of the mucous goblet cells; the density of club cells in epidermal layer; and early granular tissue formation, collagen deposition and tissue remodelling in dermal layer. Furthermore, peroxidase and catalase enzyme activity showed increased endogenous defence system in the curcumin‐treated group compared with the sham group. It could be concluded that dietary administration of curcumin is beneficial in rapid healing of skin wounds in fish. Early healing of wounds could be considered to prevent the invasion of pathogens and to maintain the integrity of the surrounding tissue.     

Improvement of growth performance,digestive enzymes and body composition of Persian sturgeon (Acipenser persicus) following feeding on probiotics: Bacillus licheniformis,Bacillus subtilis and Saccharomyces cerevisiae

The individual and combinational effects of probiotics on Persian sturgeon (Acipenser persicus) were evaluated through assessing the growth performance, proximate body composition, digestive enzymes and intestinal morphology. On the basis of feeding with experimental diets, the fish were assigned into four groups denoted as follows: control (basal diet), D‐Pro (basal diet + Bacillus subtilis and Bacillus licheniformis), D‐Pro/Yeast (basal diet + B. subtilis and B. licheniformis + Saccharomyces cerevisiae), Yeast (basal diet + S. cerevisiae). Maximum mean weight and length were measured for the fish fed with D‐pro regime. weight gain (WG), body weight gain (BWI%),  feed efficiency (FE%), protein efficiency rate (PER), lipid efficiency rate (LER), and condition factor (CF) demonstrated the highest values for the fish fed with the diets containing D‐Pro and/or D‐Pro/yeast. LER and food conversion ratio (FCR) showed the highest performance for the fish received the D‐Pro diet. The diets containing D‐Pro exhibited higher performance in enhancing the activity of protease and lipase than the one supplemented only with the yeast, whereas the regimes treated with the yeast exhibited higher amylase activity. The fish received D‐Pro exhibited the highest protein and fat contents, yet did the lowest contents of moisture and total ash. Taken together, the findings revealed that the dietary administration of the D‐Pro and D‐Pro/Yeast could improve the growth performance of A. persicus.     

Effects of dietary fishmeal replacement with Spirulina platensis on the growth,feed utilization,digestion and physiological parameters in juvenile gibel carp (Carassis auratus gibelio var. CAS III)

A 60‐day growth experiment was conducted to investigate the effect of dietary replacement of fishmeal by Spirulina platensis on growth performance, digestion and physiological parameters in juvenile gibel carp (5.0 ± 1.0 g). Four diets were formulated to replace 0 (SP0), 50% (SP50), 75% (SP75) and 100% (SP100) of dietary fishmeal protein by S. platensis respectively. Other two diets based on SP100 were supplemented with 2% dicalcium phosphate (SP100 + P) or 0.28% coated lysine (SP100 + Lys). The results showed that no significant differences of specific growth rate (SGR), feed efficiency (FE), feeding rate (FR) and protein efficiency ratio (PER) were observed between SP0 group and the replacement groups (SP50, SP75 and SP100). FE and PER of fish‐fed SP100 + Lys diet were significantly higher than the fish‐fed SP100 diet (p < .05). Apparent digestibility coefficients (ADCs) of dry matter, protein, energy and phosphorus increased significantly with the increasing S. platensis inclusion. Meanwhile, fish from SP100 + P or SP100 + Lys group had higher ADCs of nutrients compared to the SP100 group (p < .05). Compared with SP0 diet, fish‐fed S. platensis diets had higher activities of plasma superoxide dismutase, hepatopancreas total antioxidant capacity, plasma alkaline phosphatase and plasma lysozyme, as well as the lower content of plasma malondialdehyde. The present results indicated that dietary fishmeal could be fully replaced by S. platensis without any negative effects on growth performance of gibel carp. Supplementation of lysine in the S. platensis replacement diet could further improve the feed utilization of gibel carp.     

Dietary combination of pawpaw seed and onion peel powder: Impact on growth,haematology, biochemical and antioxidant status of Clarias gariepinus

A feeding trial was conducted to evaluate the effect of pawpaw–onion powder (POP) mixture on the growth, and haemato‐biochemical and antioxidant responses of Clarias gariepinus (4.02 ± 0.01g/fish) for 60 days. Five trial diets were formulated as control (without POP or antimicrobial growth promoter (AGP)), AGP (basal diet + 10ml AGP/kg diet), POP 2.5 (basal diet + 2.5g POP/kg diet), POP 5.0 (basal diet + 5.0g POP/kg diet) and POP 10 (basal diet + 10g POP/kg diet). Two hundred and twenty‐five fish were equally distributed into five groups in triplicate and fed twice daily. The results indicate that AGP or POP supplementation exerted no effects on the growth and blood profile among the various groups, but a significantly higher lymphocyte count was observed in POP 10g/kg. The highest whole‐body protein and lipid contents were noticed in fish fed the control diet (p < .05), whereas POP 5.0g/kg group recorded the highest hepatosomatic value. Furthermore, the cholesterol level was found to be lower in the AGP‐ and POP‐fed fish compared with the higher level recorded in the control. The glucose concentration and superoxide dismutase enzyme activity were found to be higher in POP 2.5‐fed fish, whereas POP 10‐fed fish showed higher catalase activities compared with other groups (p < .05). Based on the result obtained, this study showed that dietary POP had no significant impact on the growth performance but has direct effects on the whole‐body lipid content, lymphocyte count, cholesterol level, alanine aminotransferase, alkaline phosphatase and antioxidant response of Clarias gariepinus.     

Growth performance,lipid requirement and antioxidant capacity of juvenile Russian sturgeon Acipenser gueldenstaedti fed various levels of linoleic and linolenic acids

To evaluate the effects of dietary linoleic (LA) and linolenic acids (LN) on growth, enzyme activities to lipid addition and antioxidant capacity of the Russian sturgeon, Acipenser gueldenstaedti, 10 diets with different sources of lipid were formulated. Coconut oil in the basal diet was replaced by sunflower oil to make three diets (LA0.5 + LN0, LA1.0 + LN0 and LA2.0 + LN0) with dietary LA at 0.5%, 1.0% and 2%, by perilla oil to obtain three diets (LA0 + LN0.5, LA0 + LN1.0 and LA0 + LN2.0) with dietary LN at 0.5%, 1.0% and 2%, and by the combination of sunflower oil and perilla oil to provide three diets (LA0.25 +LN0.25, LA0.5 + LN0.5 and LA1.0 + LN1.0) containing LA and LN at 0.25, 0.50 and 1.00%. Coconut oil was used as the control diet at 10%, but without LA and LN (LA0 + LN0). After feeding for 56 days, the fish whole body, the serum and the liver of sturgeon were sampled. Fish fed LA0 +  LN0 and LA0 + LN2.0 gained lowest weight but was not significantly different from the fish fed LA0 +  LN1.0. Fish fed LA1.0 + LN1.0 gained highest weight among all groups. Feed conversion ratio was the best in fish fed LA1.0 +  LN1.0. Survival, condition factor and viscerosomatic index did not differ among treatments. Total body n‐6 fatty acids increased with the levels of n‐6. Total body n‐3 fatty acids also increased with dietary n‐3, and fish fed 2.0% LN had the highest content of total body n‐3 fatty acids among all groups. Triglyceride and cholesterol in the serum of fish fed LA1.0 +  LN1.0 were lowest. Fish fed LA1.0 +  LN1.0 showed the highest lipoprotein lipase and lipase activities, but the lowest malate dehydrogenase activity. Fish fed LA1.0 +  LN1.0 showed higher catalase, superoxide dismutase activity and total antioxidant capacity than fish fed other diets. This study indicates that diets containing both LA and LN are best for the growth of Russian sturgeon. The level of LA1.0 + LN1.0 in the diet is most beneficial for growth performance and antioxidant capacity in juvenile Russian sturgeon.     

Effects of dietary supplementation of fructooligosaccharide on growth performance,body composition,intestinal enzymes activities and histology of blunt snout bream (Megalobrama amblycephala) fingerlings

A feeding trial was conducted to investigate the effects of dietary fructooligosaccharide (FOS) on growth performance, body composition, intestinal enzymes activities and histology of fingerling Megalobrama amblycephala. A total of 1200 fish (1.42 ± 0.01 g) were fed diets containing graded levels of FOS (0, 0.5, 1, 2, 4 and 8 g kg⁻¹ diet) for 8 weeks in a recirculating system indoor. The weight gain, specific growth rate (SGR) and survival rate were all improved in dietary supplementation of FOS fed fish. Increasing FOS levels resulted in both higher whole‐body lipid and lower moisture contents, whereas ash and protein contents showed no significant differences among all the treatments. Intestinal amylase, protease, Na⁺, K⁺‐ATPase, alkaline phosphatase, γ‐glutamyl transpeptidase and creatine kinase activities all increased with dietary FOS levels up to 4 g kg⁻¹ (P < 0.05). Transmission electron microscopy analysis indicated that microvilli length in the mid‐intestine was significantly increased with increased dietary FOS levels (P < 0.05). In conclusion, dietary supplementation of FOS could confer benefits on growth performance, intestinal digestive and absorptive ability, histology of fingerling Megalobrama amblycephala.     

Non‐fish meal,non‐fish oil diet development for red sea bream,Pagrus major,with plant protein and graded levels of Schizochytrium sp.: Effect on growth and fatty acid composition

A feeding experiment was conducted to determine the optimal formulation level of algae meal, which is rich in docosahexaenoic acid (DHA), in a non‐fish meal diet. Six iso‐nitrogenous (450 g/kg) and iso‐lipidic (130 g/kg) experimental diets were prepared. The control diet was formulated with fish meal (400 g/kg), fish oil (60 g/kg), plant protein sources (220 g/kg) and rapeseed oil (50 g/kg). Plant protein sources (soy protein concentrate, soybean meal and corn gluten meal), rapeseed and fish oil were formulated in the second diet (NFM + FO). In the third diet, fish oil of the NFM + FO diet was replaced by rapeseed oil (NFM + NFO) and designated as the negative control. In the other three diets, rapeseed oil in the NFM + NFO diet was replaced with algae meal (Schizochytrium sp. powder) at 50 g/kg, 100 g/kg and 150 g/kg (AM5, AM10 and AM15, respectively). Triplicate groups of juvenile red sea bream (8.8 g) were fed the experimental diets for 12 weeks near satiation. The growth was lowest in the fish fed NFM + NFO diet. This was improved by the formulation of algae meal, which reached the growth level of the NFM + FO group in the AM10 group. The lipid content of the whole fish body in the NFM + NFO group was significantly lower than those of other groups. The fatty acid profile showed significant differences among dietary treatments. DHA content in total and polar lipids of the whole body and liver was highest in the AM10 and AM15 groups. These results reconfirm that microalgae are a suitable lipid source for the replacement of dietary fish oil for marine fish, and the optimal level was estimated as 50 g/kg?100 g/kg in diet.     

Effects of dietary inclusion of soybean meal and cholesterol on the growth,cholesterol status and metabolism of the giant grouper (Epinephelus lanceolatus)

The study evaluated effects of cholesterol supplementation in a diet with high soybean meal (SBM) on the growth and cholesterol metabolism of giant grouper (Epinephelus lanceolatus). All‐fish‐meal diet was used as control. The diet including SBM (replaced 50% of the fish meal protein, SBM diet) and the SBM diet supplemented with 10 g/kg cholesterol (SBM + cholesterol) were used as experimental diets. Three diets were each fed to triplicate groups of juvenile grouper (initial body weight: 12.39 ± 0.36 g) in a recirculating aquaculture system for 8 weeks. Grouper fed the control diet showed higher (p < .05) weight gain, feed intake, feed efficiency and protein efficiency ratio than the other two dietary treatments. Hepatic cholesterol concentrations and 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase gene expressions were higher in fish fed the control diet than fish fed the control diet and SBM + cholesterol diet. Hepatic cholesterol 7α‐hydroxylase gene expression was higher in fish fed the SBM + cholesterol diet than that in fish fed the control diet. Results indicate that giant grouper on a diet low in cholesterol can regulate cholesterol synthesis, suggesting that the reduced dietary cholesterol intake in the fish fed diet containing SBM is sufficiently compensated by increased cholesterol synthesis.     

Evaluating the protective efficacy of antigen combinations against Photobacterium damselae ssp. piscicida infections in cobia,Rachycentron canadum L.

Cobia, Rachycentron canadum L., is a very important aquatic fish that faces the risk of infection with the bacterial pathogen Photobacterium damselae ssp. piscicida, and there are few protective approaches available that use multiple antigens. In the present study, potent bivalent antigens from P. damselae ssp. piscicida showed more efficient protection than did single antigens used in isolation. In preparations of three antigens that included recombinant heat shock protein 60 (rHSP60), recombinant α‐enolase (rENOLASE) and recombinant glyceraldehyde‐3‐phosphate dehydrogenase (rGAPDH), we analysed the doses that elicited the best immune responses and found that this occurred at a total of 30 μg of antigen per fish. Subsequently, vaccination of fish with rHSP60, rENOLASE and rGAPDH achieved 46.9, 52 and 25% relative per cent survival (RPS), respectively. In addition, bivalent subunit vaccines – combination I (rHSP60 + rENOLASE), combination II (rENOLASE + rGAPDH) and combination III (rHSP60 + rGAPDH) – were administered and the RPS in these groups (65.6, 64.0 and 48.4%, respectively), was higher than that achieved with single‐antigen administration. Finally, in combination IV, the trivalent vaccine rHSP60 + rENOLASE + rGAPDH, the RPS was 1.6%. Taken together, our results suggest that combinations of two antigens may achieve a better efficiency than monovalent or trivalent antigens, and this may provide new insights into pathogen prevention strategies.     

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.     

Effects of two dietary probiotics (Bacillus subtilis or licheniformis) with two prebiotics (mannan or fructo oligosaccharide) in Japanese eel,Anguilla japonica

A 12‐week feeding trial was conducted to investigate the effects of two dietary probiotics; Bacillus subtilis KCTC 2217 or Bacillus licheniformis KCCM 11775 with two prebiotics; mannan oligosaccharide (MOS) or fructooligosaccharide (FOS) in Japanese eel. Fish averaging 12.8 ± 0.47 g (mean ± SD) were randomly distributed into five treatments with triplicate tanks. A basal control diet (CON) and four synbiotic diets supplementing B. subtilis + MOS (BSM), B. subtilis + FOS (BSF), B. licheniformis + MOS (BLM), and B. licheniformis + FOS (BLF). Weight gain and specific growth rate of fish fed all synbiotic diets were higher than those of fish fed CON. Immune‐related gene expression of heat shock protein 70 and immunoglobulin M of fish fed BSF and BLM were significantly higher than those of fish fed CON. Fish fed BSF and BLM had significantly higher intestinal villi length than those of fish fed BLF and CON (p < .05). Disease resistance against Aeromonas hydrophila of fish fed all synbiotic diets were significantly higher than those of fish fed CON (p < .05). Therefore, these results indicated that dietary B. subtilis with FOS (BSF) and B. licheniformis with MOS (BLM) could have beneficial effects on intestinal morphology, and immune‐related gene expression in Japanese eel.     

Combined or individual effects of dietary magnesium and selenium nanoparticles on growth performance,immunity, blood biochemistry and antioxidant status of Asian seabass (Lates calcarifer) reared in freshwater

This study was conducted to investigate effects of and interactions between nanoselenium (NanoSe) and nanomagnesium (NanoMg) on growth, humoral immunity, serum biochemistry and antioxidant capacity of juvenile Asian seabass Lates calcarifer reared in freshwater. Four groups of fish with an average weight of 32.78 ± 2.23 g were fed one of the experimental diets for 6 weeks: (a) control (basal diet); (b) NanoSe (basal diet + 4 mg NanoSe/kg diet); (c) NanoMg (basal diet + 500 mg NanoMg/kg diet); and (d) combination (basal diet + 4 mg NanoSe/kg diet + 500 mg NanoMg/kg diet). Fish fed with NanoSe‐supplemented diets (NanoSe and combination) showed higher weight gain, specific growth rate and feed intake. The combination of NanoSe and NanoMg enhanced the immune response. Also, fish fed on combination diet showed higher serum alanine aminotransferase and aspartate transaminase levels but lower glucose concentration. The activities of liver superoxide dismutase, glutathione peroxidase and catalase were not different among all groups, but liver malondialdehyde level was lower in fish fed diets supplemented with NanoSe and/or NanoMg. It could be concluded that NanoSe improved growth performance; the combination of both NanoSe and NanoMg enhanced humoral immunity; and NanoSe and/or NanoMg improved antioxidant capacity in Asian seabass.     

Symbiotic supplementation on the hemato‐immunological parameters and survival of the hybrid surubim after challenge with Aeromonas hydrophila

This study evaluated the effects of dietary supplementation with two symbiotics in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum) prior to and after challenge with Aeromonas hydrophila. The fish were divided into unsupplemented fish, supplemented fish with Weissella cibaria and inulin, and supplemented fish with Lactobacillus plantarum and inulin. Twenty days after being fed symbiotics, the fish were sampled for immunological, haematological, and microbiological analysis pre‐challenge. After this period, they were challenged with Aeromonas hydrophila, and after 96 h the fish were sampled for blood collection and survival. Fish fed W. cibaria and inulin from the pre‐challenge showed the highest values of lysozyme activity. Fish fed L. plantarum and inulin presented a higher number of thrombocytes and granular leukocyte PAS⁺ than the other fish. After challenge, fish fed both symbiotics presented an increased red blood cells number. Fish fed L. plantarum and inulin showed an increased total leucocyte count (WBC), neutrophils, and monocytes number as well as total immunoglobulin. The W. cibaria‐ and inulin‐supplemented fish showed improved haematological and immunological parameters and reduced cumulative mortality after the challenge.     

Responses of antioxidant enzymes,lipid peroxidation,and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase in liver of the fish <Emphasis Type="Italic">Goodea atripinnis</Emphasis> exposed to Lake Yuriria water

Lake Yuriria, located in the heavily populated and polluted Mexican Central Plateau, receives domestic sewage, industrial effluents, and municipal wastewaters that are still directly discharged without treatment into the tributaries and the lake. Pollutants in water and sediments include heavy metals, aromatic hydrocarbons, and organochlorine pesticides. Activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), as well as Na⁺/K⁺-adenosine triphosphatase (Na⁺/K⁺-ATPase) activity, and lipid peroxidation (LPO) were evaluated in the livers of the fish Goodea atripinnis after 96 h of exposure to water collected in March and June 2005 from three sites: Y (limnetic zone), L (Lerma tributary), and C (la Cinta tributary). Physical and chemical parameters of the lake water were also analyzed. Increases in CAT activity and LPO levels at all three sites were detected compared with control fish (P < 0.05), while GPx and SOD activities decreased significantly (P < 0.05). Na⁺/K⁺-ATPase activities were similar to the control in fish exposed to limnetic water from both March and June but were higher than control at the two tributary sites in March (P < 0.05); fish exposed to water from the Lerma tributary in June exhibited lower Na⁺/K⁺-ATPase than the control (P < 0.05). During March, CAT and Na⁺/K⁺-ATPase activities were increasing more than in June in Y and L, respectively, while in June, SOD and GPx were depleted more than March in L and Y and L, respectively. Despite the antioxidant defenses of the fish liver, exposure to all water samples from Lake Yuriria exerted alterations in hepatic LPO levels, antioxidant enzymes, and Na⁺/K⁺-ATPase activities that could substantially impair the mechanisms of fish defenses against oxidative stress.     

Effects of stocking density on the growth performance,digestive enzyme activities,antioxidant resistance,and intestinal microflora of blunt snout bream (Megalobrama amblycephala) juveniles

In this study, effects of stocking density on the growth performance and physiological responses of blunt snout bream, Megalobrama amblycephala juveniles were evaluated. The fish (average body weight, 25.76 ± 2.25 g) were randomly stocked at densities of 30F (30 fish/m³), 60F, 90F and 120F in 12 cages (1 m × 1 m × 1 m) in a concrete pond, with three cages for each density, for a period of 6 weeks. The higher stocking densities had a negative effect on individual growth performance. The results indicated that serum cortisol, triglyceride, alanine aminotransferase, aspartate transaminase, alkaline phosphatase and malondialdehyde activities; and Acinetobacter, Aeromonas, Pseudomonas and Vibrio numbers in the intestinal microflora increased significantly as the stocking density increased. In contrast, the viscerosomatic index, hepatosomatic index survival rate; serum glucose, total cholesterol, lipase, protease, glutathione peroxidase and superoxide dismutase activities; and Clostridium, Bacteroides, Lactococcus, Lactobacillus and Bacillus numbers in the intestinal microflora decreased significantly. The 90F and 120F groups showed obvious enlargement of the lamina propria and goblet cell damage, indicating that the gut showed inflammatory responses. The specific growth rate and weight gain rate increased significantly as the stocking density increased from 30 to 60 fish/m³, but decreased significantly when the stocking density was over 60 fish/m³.     

Effects of malic acid and citric acid on growth performance,antioxidant capacity,haematology and immune response of Carassius auratus gibelio

The aim of this study was to investigate the effects of citric acid (CA) and malic acid (MA) on growth performance, antioxidant capacity, haematology and immune response of Carassius auratus gibelio. Carassius auratus gibelio were randomly divided into six groups, with three replicates in each group and 20 fish in each replicate. The six groups were fed a basic diet with organic acid supplementation (control group, 0.2%MA, 0.2%MA + 0.1%CA, 0.2%MA + 0.2%CA, 0.2%MA + 0.4%CA and 0.2%MA + 0.6%CA) for 8 weeks respectively. The results showed that compared with the control group, the weight gain rate and specific growth rate significantly increased in organic acid‐fed groups (p < .05). The highest value was in the 0.2%MA + 0.2%CA group. However, there were no significant differences in survival rate, red blood cells or white blood cells between groups. The contents of ALT, AST and LDH in Carassius auratus gibelio fed with organic acids increased, especially in 0.2%MA + 0.4%CA and 0.2%MA + 0.6%CA groups. However, there was no significant difference between the 0.2%MA + 0.2%CA group and the control group. In addition, the highest values of GSH and T‐AOC were found in the 0.2%MA + 0.6%CA group. Compared with the control group, there were no significant differences in the expression of HSP70 or LZM (p > .05), but the expression of Il‐1β, IL‐10 and TNF‐α was up‐regulated in the 0.2%MA + 0.4%CA and 0.2%MA + 0.6%CA groups. The results showed that the growth performance, antioxidant capacity, haematological parameters and expression of immune‐related genes were significantly improved by adding organic acids to the feed of Carassius auratus gibelio. However, excessive addition of organic acids may cause liver injury, and the optimal addition amount in this experiment was 0.2%MA + 0.2%CA.     

Antioxidative role of selenium against the toxic effect of heavy metals (Cd<Superscript>+2</Superscript>, Cr<Superscript>+3</Superscript>) on liver of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> Walbaum 1792)

The main purpose of this study is to discuss the effect of Cd⁺², Cr⁺³ and Se metals on biochemical parameters in liver tissue of Oncorhynchus mykiss. The rainbow trout were exposed to heavy metal stress (Cd⁺², Cr⁺³) at 2 ppm dosage. The present study was undertaken to determine the protective effect of selenium treatment at the same dosage (2 ppm) on some biochemical parameters. The activity of catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and the changes in levels of malondialdehyde (MDA) from biochemical parameters were determined in liver tissue of the fish groups exposed to heavy metals, especially for the selenium-applied groups. Results of this study showed that the activities of CAT, GSH-Px and SOD in the tissues of fish exposed to the stress of Cd⁺² and Cr⁺³ were significantly lower than the control groups (P < 0.05). Meanwhile, the closer values to the control groups were obtained in selenium-added groups (Cr⁺³ + Se⁺⁴, Cd⁺² + Se⁺⁴). For the level of MDA, the last production of lipid peroxidation showed increases (P < 0.05) in the groups exposed to the metal stress, whereas significant decreases were obtained in selenium-applied groups. The result of the statistical evaluation showed that the negative effects occurring in the biochemical parameters of the applied groups exposed to the toxicity of heavy metal were significantly eliminated (P < 0.05) as a result of selenium treatment.     

Morphological diversity of Paramoeba perurans trophozoites and their interaction with Atlantic salmon,Salmo salar L., gills

Amoebic gill disease (AGD) caused by the ectoparasite Paramoeba perurans affects several cultured marine fish species worldwide. In this study, the morphology and ultrastructure of P. perurans in vitro and in vivo was investigated using scanning and transmission electron microscopy (SEM and TEM, respectively). Amoebae cultures contained several different morphologies ranging from a distinct rounded cell structure and polymorphic cells with pseudopodia of different lengths and shapes. SEM studies of the gills of AGD‐affected Atlantic salmon, Salmo salar L., revealed the presence of enlarged swellings in affected gill filaments and fusion of adjacent lamellae. Spherical amoebae appeared to embed within the epithelium, and subsequently leave hemispherical indentations with visible fenestrations in the basolateral surface following their departure. These fenestrated structures corresponded to the presence of pseudopodia which could be seen by TEM to penetrate into the epithelium. The membrane–membrane interface contained an amorphous and slightly fibrous matrix. This suggests the existence of cellular glycocalyces and a role for extracellular products in mediating pathological changes in amoebic gill disease.