The non-specific activation of rainbow trout, Salmo gairdneri Richardson, complement by Aeromonas salmonicida extracellular products and the correlation of complement activity with the inactivation of lethal toxicity products

Abstract The possible mechanism of inactivation of the toxicity of Aeromonas salmonicida extracellular products (ECP) by normal rainbow trout serum was investigated using juvenile rainbow trout. ECP was prepared from culture supernatant by an acetone precipitation method. The ECP was incubated with normal rainbow trout serum at 20°C for 2 h, and the interrelationship between ECP proteolytic activity and immune complex-initiating, haemolytic complement activity (CH₅₀) of normal serum against antibody-sensitized goldfish red blood cells was evaluated. When normal serum was incubated with increasing concentrations of ECP, the CH₅₀ activity of serum decreased. The CH₅₀ activity was completely abolished in serum treated with undiluted ECP. ECP treated with serum was administered to trout intraperitoneally to determine mortality. All the fish receiving untreated ECP (0.05 ml = 0.5 mg protein) alone died within 24 h. When ECP was treated with serum at 1:1 to 4:1 (serum: ECP) in volume a similar high mortality was produced. These inocula possessed high protease activity and no or low CH₅₀ activity. However, mortality decreased and finally no mortality was recorded as ECP was treated with large volumes of serum (9:1 to 19:1). These inocula had lower protease activity and considerably higher CH₅₀ activity. Fish receiving ECP treated with heat-inactivated serum at 19:1 showed 100% mortality. A serum: ECP inoculum derived from fish which had been administered lipopolysaccharide from Salmonella enteritidis and which possessed a low CH₅₀ activity also gave a high mortality when used at 19:1. These results suggest that rainbow trout complement is implicated in the inactivation of toxicity of A. salmonicida ECP.     

The role of the lethal extracellular cytolysin of Aeromonas salmonicida in the pathology of furunculosis

Abstract. The histopathological effects of intramuscular (i.m.) and intraperitoneal injections of the purified lethal cytolytic toxin of A. salmonicida in Atlantic salmon, Salmo salar L., were restricted to extensive degranulation of eosinophilic granular cells (EGCs) in the gills (both routes) and a limited coagulative necrosis of muscle fibres at the site of i.m. injection. The severity of these lesions did not apparently differ amongst fish that were moribund within 48 h and the surviving fish at 90 h. Previous studies indicate a maximum time to death caused by this toxin of 48 h. Thus, it is difficult to account for death of the fish from the toxin solely on the grounds of the histopathological features. It is suggested that a complex metabolic dysfunction results in death. The mechanism of 'furuncle' formation was demonstrated to result from a combined effect of the cytolysin and the purified 70-kDa protease present in the ECP. When i.m.-injected singly, the purified cytolysin produced a limited coagulative necrosis and the purified protease produced a limited liquefaction and haemorrhaging. In combination, they induced the extensive liquefactive haemorrhagic 'furuncle' characteristic of the whole ECP.     

Effects of cadmium on some haematological and biochemical characteristics of <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (Linnaeus, 1758) dietary supplemented with tomato paste and vitamin E

The present study investigates the potential protective effects of tomato paste (9 mg/kg-lycopene) in comparison with vitamin E (50 mg/kg) against the impacts of cadmium (Cd) toxicity (4.64 mg/l: ¼ of 96 h LC50) on fishes Cd exposed for 15 and 30 days. Cd impacts were evaluated in terms of biological, haematological and biochemical characteristics. Cd significantly induced free radicals in serum and liver. The activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly increased due to Cd. Treatment with Cd caused a significant increase in Lipid peroxidation and DNA fragmentation in liver tissue and serum glucose and total lipid. On the other hand, Cd significantly led to decline in serum total protein, blood haemoglobin, red blood cell count, haematocrit value, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Dietary supplementation with vitamin E and/or tomato paste to Cd-exposed fish declined significantly the increased lipid peroxidation and DNA fragmentation in liver tissue and the increased aspartate aminotransferase, alanine aminotransferase, glucose and total lipid in serum to the normal condition. This supplementation also significantly increased the declined serum total protein, blood haemoglobin, red blood cell count, haematocrit value, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration to the normal state. Cd impacts and tomato paste/or vitamin E supplementations did not reflected on the condition factor of the fish. These findings demonstrated the beneficial diet supplementation of tomato paste phytonutrients and vitamin E in counteracting the harmful effects of Cd on the characters investigated.     

Blood and haemoglobin meal as protein sources in diets for gilthead sea bream (Sparus aurata): effects on growth, nutritive efficiency and fillet sensory differences

Two parallel experiments were conducted to evaluate the effect of partial substitution of fish meal by two different animal protein sources, blood meal (B) and haemoglobin meal (H) at 0, 50 and 100 g kg⁻¹ of level inclusion in diets for gilthead sea bream, considering a long feeding period for juveniles (Trial 1) and a short feeding period (Trial 2) for on-growing fish. In Trial 1, 33 g juveniles were fed for 242 days and the fish fed with 5% and 10% of haemoglobin dietary inclusion obtained less growth, although feed efficiency, protein efficiency ratio and muscle composition were similar in all diets. In Trial 2, 179 g initial weight fish were fed for 164 days and growth of fish fed H10 showed the lowest growth, although nutrient efficiency and muscle composition were not affected significantly. The results of these experiments demonstrated that blood meal can substitute fish meal (up to 10%) with no effect on performance, but may lead to sensory differences compared with fish fed diet 0, while the inclusion of 5% blood meal had no effect on growth or sensory characteristics. Fish fed 10% haemoglobin inclusion had the poorest growth values.     

Effect of sublethal toxicity of some pesticides on growth parameters, haematological properties and total production of Nile tilapia (Oreochromis niloticus L.) and water quality of ponds

The effects of sublethal doses of dimethoate (20, 10 and 5 mg L^?1) and malathion (2.0, 1.0 and 0.5 mg L^?1) on growth parameters, haematological properties and total production of Nile tilapia (12.0 cm length and 40.0 g weight) and water quality were studied in fibreglass tanks (3 m²) for 24 weeks. Results showed that malathion compounds were more toxic than dimethoate. The sublethal toxicity of pesticides decreased plankton abundance and water quality in fish ponds. In comparison with controls, sublethal levels of these pesticides led to a significant decrease (P<0.05) in final body weight, specific growth rate and normalized biomass index of fish. The survival rate of Nile tilapia decreased with increasing concentrations of pesticides. Feed utilization (total food consumed, feed conversion ratio and protein efficiency ratio) varied with pesticide treatments. The erythrocyte count, haematocrit value and haemoglobin content of Nile tilapia were slightly decreased with increasing concentrations of pesticides. In addition, serum protein decreased, while serum glucose and lipid increased. Further, glycogen, protein and lipid in fish muscle gradually decreased with increased pesticide concentrations. Pesticide residues in the liver, gills and muscles of fish increased with increased pesticide concentrations in fish ponds. Their bioaccumulation in the liver was higher than in gill or muscle, which had the lowest residues for these pesticides. On the other hand, total production, net returns and profitability of reared fish decreased with increasing of concentrations of pesticides. Generally, sublethal concentrations of two pesticides in aquatic areas led to degradation of water quality and fish production in ponds.     

Some physiological studies on the effect of onion and garlic juices on the fish, Clarias lazera

Fish, Clarias Lazera were orally administered onion (Allium cepa) or garlic (Allium sativum) juices. Contents of glucose, free amino acids, total lipids, triglycerides, cholesterol, albumin, total protein, urea, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum as well as glycogen, free amino acids, total lipids, triglycerides and cholesterol contents in liver, white and red muscle were determined 5 h and 24 h after a single dose or after repeated doses (1 dose every 24 h for 5 days). Both onion and garlic treatments caused a temporary hypoglycemia as well as liver glycogen muscle depletion accompanied with a significant elevation in white muscle glycogen suggesting that dietary onion or garlic may enhance the process of glycogenesis in white muscle. Onion feeding caused hypolipidemia accompanied with significant decreases in liver content of total lipids in response to the repeated doses for 5 days and a severe rise in the lipid content of red muscle at all periods. Such results suggest that dietary onion may enhance the process of lipogenesis in red muscle of Clarias lazera. Probably, the main precursor of such lipogenesis in the red muscle was the serum fatty acids. Moreover, dietary onion led to hypocholesterolemia accompanied with a significant rise in cholesterol level of both liver and red muscle. On the other hand, dietary garlic significantly decreased serum free amino acids after 5 h and 5 days. Furthermore, significant elevations of liver free amino acids were dedected in the onion-treated fish group (24 h and 5 days) and in the garlic-treated fish group (5 h), probably due to oxidation of thiol components (sulfur attached to the proteins). This result was accompanied with either a marked increase in white muscle free amino acids at all periods of garlic fish group, or a noticeable drop in the onion group (repeated doses). Also, dietary onion and garlic treatment led to an elevation in hepatic ureagenesis in spite of the unchanged serum urea level. Significant decreases of both serum AST and ALT in garlic groups (all periods) and in onion groups (repeated doses) were observed. In liver, AST and ALT showed temporary rises after onion or garlic supplementation. In general, it could be concluded that both dietary onion and garlic have a transitory hypoglycemic action as well as a promoting effect on the process of glycogenesis in white muscle whereas dietary onion promotes lipogenesis in red muscle of fish. Also, dietary garlic may enhance the white muscle uptake of free amino acids which, in turn, may promote protein synthesis.     

Is the use of recombinant cGnRH may be a future alternative to control the fish spawning? Let us go with the goldfish example

The use of recombinant gonadotropin-releasing hormone (rGnRH) has very rarely been tested in fish to promote spawning. This study evaluated the impact of recombinant chicken gonadotropin-releasing hormone (rcGnRH) with metoclopramide on the release of sex steroids and final maturation induction in goldfish (Carassius auratus) broodstock. For this purpose, goldfish broodstock was divided into four groups and treated with 0.9% NaCl with 20 mg/kg metoclopramide (Met) (C); 10 μg/kg body weight (BW) rcGnRH with 20 mg/kg metoclopramide (rcGn10); 15 μg/kg BW rcGnRH with 20 mg/kg metoclopramide (rcGn15); and 20 μg/kg BW rcGnRH with 20 mg/kg metoclopramide (rcGn20). The capability of the rcGnRH for eliciting biological response was tested in vivo by evaluating the changes of 17β estradiol (E2), testosterone (T), and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) and the induced spawning. Blood samples were obtained at 0 h, 12 h, and 24 h after injection. The rcGn10, rcGn15, and rcGn20 treatments induced lower E2 concentration, especially 24 h post-injection. T levels were significantly higher in rcGn10, rcGn15, and rcGn20 treatments 12 h post-injection than at 0 h and then decreased at 24 h post-injection. Furthermore, the rcGnRH tested significantly enhanced DHP secretion in rcGn10, rcGn15, and rcGn20 treatments 12 h post-injection before a decline at 24 h post-injection. No significant difference between the sampling times was found in the C treatment for the 3 sex steroids tested. The results also displayed that rcGnRH at 10–20 µg/kg of body weight can trigger spawning with the highest speed and efficiency of spawning at 20 µg/kg. The obtained results represent a possible strategy for enhancing the artificial reproduction and ovulation of broodstock fish by rGnRH and further support the use of recombinant hormones to promote reproduction in aquaculture.

     

The production of a lymphokine (macrophage activating factor) by rainbow trout, Oncorhynchus mykiss (Walbaum), leucocytes stimulated with the extracellular products of Mycobacterium sp.

The production of macrophage activation factor (MAF) by rainbow trout, Oncorhynchus mykiss (Walbaum) , head kidney leucocytes was examined after culturing in vitro with extracellular products (ECP) collected from Mycobacterium sp. Cultures of leucocytes were prepared from naive fish, or fish previously vaccinated with either the ECP or with formalin killed whole cell preparations (WC) of the bacterium. The cells were then incubated with the ECP in vitro and the ability of their supernatants to activate macrophages assessed. Macrophages from control fish were incubated with the supernatants, and their ability to reduce nitroblue tetrazolium (NBT) measured as an indicator of macrophage activation. Incubation of head kidney macrophages from naive fish directly with 1, 10 or 100 μg mL^–1 of ECP for 48 h significantly enhanced macrophage activation compared with control macrophages. Vaccination of fish with either ECP or WC had no significant effect on the respiratory burst of control macrophages 4 weeks post-vaccination. By the eighth week, however, absorbance levels of respiratory burst reflecting both the primary (cells from vaccinated fish cultured in vitro with PBS) and the secondary (cells cultured in vitro with ECP) MAF responses of fish vaccinated with ECP and WC, had peaked and these were significantly different from the non-vaccinated controls. This activity had fallen to levels similar to control fish by week 12 for fish vaccinated with WC.     

Effect of infestation with the isopod, Alitropus typus M. Edwards (Crustacea: Flabellifera: Aegidae) on the haematological parameters of the host fish, Channa striatus (Bloch)

Detailed accounts supported by experimental data are furnished regarding the haematological parameters of the host fish, Channa striatus infested by the isopod, Alitropus typus. Results demonstrate that the infested fish become anaemic through loss of blood as shown by a reduction in the total erythrocyte count, haemoglobin content and haematocrit. Considering the mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and volume index, the anaemia can be designated as macrocytic and hypochromic. The total and differential counts of white blood corpuscles of the infested fish indicate leucopenia coupled with lymphocytosis. The percentages of monocytes, neutrophils and macrophages, which were 0.65%, 0.39% and 1.04%, respectively, after 24 h of infestation, had increased to 2.31%, 4.91% and 2.02%, respectively, after 96 h of infestation and declined thereafter.     

拟微绿球藻粉对大菱鲆幼鱼生理和脂肪酸组成的影响

为探讨饲料中添加拟微绿球藻粉替代鱼油对大菱鲆幼鱼血清生化指标、体组成和脂肪酸组成的影响,用拟微绿球藻粉替代基础饲料中0(对照组)、8%、16%、24%和32%的鱼油,配制5种等氮等能的饲料。选取初始体质量(82.64±0.32)g的大菱鲆幼鱼375尾,随机分为5组,每组3个重复,每个重复25尾,养殖37d。试验结果显示,试验组的全鱼粗蛋白呈逐渐上升的趋势并且显著高于对照组(P<0.05),试验组的全鱼粗脂肪呈逐渐下降的趋势并且显著低于对照组(P<0.05),试验组的肌肉粗蛋白和粗脂肪与全鱼具有相同的趋势;肌肉和肝脏的花生四烯酸含量随着饲料中拟微绿球藻粉含量的增加而上升,而n-3/n-6高不饱和脂肪酸呈下降的趋势;血清总超氧化物歧化酶和总抗氧化能力呈先升后降的趋势,在替代16%试验组达到最大值并且显著高于对照组(P<0.05),血清溶菌酶也呈先升后降的趋势,替代16%试验组和替代24%试验组显著高于对照组(P<0.05),血清丙二醛呈先降后升的趋势,在替代16%试验组达到最低值并且显著低于对照组(P<0.05),血清低密度脂蛋白胆固醇呈下降趋势,试验组显著低于对照组(P<0.05)。研究表明,在16%～24%替代水平上,可提高试验组的非特异性免疫力和抗氧化能力,降低其血脂水平,而大菱鲆幼鱼高不饱和脂肪酸的含量不变,保持了其营养价值。     

In vivo fish models for visualizing Aeromonas hydrophila invasion pathway using GFP as a biomarker

The invasion pathway of Aeromonas hydrophila in vivo was studied in Crucian carp (Carassius auratus gibelio) with a virulent strain A. hydrophila J-1 transformed with a plasmid encoding green fluorescent protein (pGFPuv) (A. h J-1^GFP), which had similar virulence characteristics (haemolysin, extracellular proteases production, toxicity for EPC cells, survival in fish serum and LD₅₀ value) as the parent strain. Fish were divided into four experimental groups: (1) normal fish; (2) bacteria bath challenged, unwounded fish; (3) skin artificially wounded by scalpel, bacterial bath challenged fish; and (4) skin mucus layer partially removed by paper towel, bacterial bath challenged fish. The number of bacteria from blood, gills, kidney, muscle, liver and intestine were detected at 2, 4, 8, 12, and 24 h post-challenge. High bacterial numbers were observed in the muscle of the artificial wound group, in the kidney of the mucus removed group and in the gills of all groups. In conclusion, the gills and damaged skin are likely to be the main routes of entry for A. hydrophila, and GFP can be used as a real-time biomarker to study intimate host–pathogen interaction in fish.     

Non-specific immune response of Nile tilapia,Oreochromis nilotica,to the extracellular products of Mycobacterium spp. and to various adjuvants

Nile tilapia were immunized by injecting extracellular products (ECP) of Mycobacterium spp. (strain TB40, TB267 or the type strain Mycobacterium marinum) into their swim bladders. A variety of adjuvants – Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA) and Titremax – were similarly injected into additional groups of tilapia. Phosphate-buffered saline (PBS) was used as a control. The number of nitroblue tetrazolium (NBT)-positive cells observed in the swim bladder of the immunized fish had significantly increased by the fourth day post-immunization. By day 8, the number of NBT-positive cells in fish immunized with ECP from mycobacteria strains TB40 or TB267 were fewer than in fish immunized with ECP from M. marinum or fish injected with FCA or FIA. The level of lysozyme activity detected in the serum of fish 4 days after being immunized with ECP from various Mycobacterium spp. was also significantly higher than that found in the serum of the control fish. Head kidney macrophages showed an enhanced reduction of NBT when cultured in vitro with 1 μg ml^–1 of ECP. Concentrations greater than this (10 or 100 μg ml^–1) were found to suppress the reduction of NBT by the macrophages.     

Pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in healthy and Vibrio alginolyticus-infected large yellow croaker (Pseudosciaena crocea)

The present study was designed to explore pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in healthy and Vibrio alginolyticus-infected large yellow croaker (Pseudosciaena crocea) after a single 10 mg/kg oral dose. Concentrations of enrofloxacin and ciprofloxacin in serum, liver, kidney, muscle and skin of fish were determined using high-performance liquid chromatography. Pharmacokinetic parameters were analysed based on classical compartmental model analysis. The overall changes in enrofloxacin concentration–time curves in serum and tissues of diseased fish were similar to those of healthy fish. However, the peak concentration and peak time of enrofloxacin in serum and tissues were different in healthy and diseased fish. A delay of enrofloxacin peak time in serum and all tissues appeared in the diseased fish. The peak concentrations in serum and tissues of the diseased fish were lower than those of healthy fish. In healthy fish, the area under the concentration–time curve (AUC) was in the order serum >liver > kidney >muscle > skin, while AUC was serum >live > muscle >kidney > skin in the diseased fish. The peak concentrations of ciprofloxacin in the liver, serum, kidney, muscle and skin of healthy fish were 0.93 μg/g, 0.55 μg/ml, 0.36 μg/g, 0.37 μg/g and 0.12 μg/g respectively. T_max of ciprofloxacin in the corresponding tissues was 8, 24, 12, 12 and 16 h respectively. In the diseased fish, the peak concentrations of ciprofloxacin in the corresponding tissues were 0.52 μg/g, 0.52 μg/ml, 0.41 μg/g, 0.27 μg/g and 0.13 μg/g respectively. T_max in the corresponding tissues were 0.5, 8, 12, 16 and 48 h respectively. These data indicate that the health status of fish affects drug absorption and metabolism.     

Studies on the utilization by rainbow trout (Salmo gairdneri Rich.) of feed mixtures containing soya bean meal and an addition of amino acids

This paper deals with the growth of fish and utilization of feed mixtures in which fish meal is partly substituted by powdered soya beans. In order to improve the quality of protein in the feeds, they were enriched with synthetic amino acids. The food coefficients of the feeds were calculated, as were the protein efficiency ratio and apparent net protein utilization, by chemical analysis.

The results of the experiment showed that an addition of amino acids improved the utilization of feeds. It was found that rainbow trout can utilize soya bean protein enriched with cystine (1%) and tryptophan (0.5%) almost as well as protein from the fish meal.

Chemical composition of the experimental fish is discussed, as well as the amino acid composition of fish muscles and selected physiological parameters: haematocrit, haemoglobin content in the fish blood, the hepasomato index, and the content of protein, amino-transferases and cholesterol in the fish blood serum.     

低鱼粉饲料中添加酶解豆粕对凡纳滨对虾生长性能和抗胁迫机能的影响

为研究在养殖过程中降低鱼粉用量的同时保持凡纳滨对虾良好的生长性能和抗逆能力,实验在含10%鱼粉的基础饲料中,分别添加酶解豆粕(PSM) 0%(A)、2.5%(B)、3.5%(C)、4.5%(D)、5.5%(E)制成5组等氮等能饲料,分别投喂初始体质量为(0.45±0.02) g的凡纳滨对虾幼虾8周,检测对虾生长性能及抗胁迫机能。结果显示,8周养殖实验结束后,各实验组对虾的终末体质量为14.65～15.38 g/尾,各组间对虾终末均重、成活率和饲料系数指标均无显著性差异;A组对虾肌肉粗蛋白质含量显著低于其他各实验组;C组、D组和E组对虾肌肉粗脂肪含量显著高于A组;各组间灰分和水分均无显著性差异;D组和E组对虾肝胰腺蛋白酶、淀粉酶、脂肪酶、血清溶菌酶和血清总超氧化物歧化酶活性(T-SOD)均显著高于A组;A组对虾血清丙二醛(MDA)含量显著高于D组和E组。人工急性感染高剂量副溶血性弧菌的胁迫实验中,A组对虾在弧菌感染48和60 h时的累积死亡率均显著高于D组对虾同期的累积死亡率;低剂量副溶血性弧菌人工急性感染后,在凡纳滨对虾鳃组织中检测Toll受体、免疫缺陷(IMD)和溶菌酶3种免疫相关基因的表达量,结果显示,对虾Toll受体、IMD和溶菌酶mRNA表达量最大峰值分别出现在添加酶解豆粕的C组、B组和D组,峰值出现时刻分别为感染后24、42和24 h。研究表明,含10%鱼粉的饲料中添加0%～5.5%酶解豆粕对凡纳滨对虾的生长性能改善效果不显著,酶解豆粕会显著提高凡纳滨对虾肌肉粗蛋白质含量和粗脂肪含量;显著降低对虾血清丙二醛含量;同时也会显著改变凡纳滨对虾对弧菌的抵抗力及其免疫相关基因的时空表达,酶解豆粕添加量达到4.5%时可使养殖的凡纳滨对虾获得最佳的抗弧菌能力。     

Thermal stress altered growth performance and metabolism and induced anaemia and liver disorder in Labeo rohita

Sudden fish deaths occurred during summer peaks in India and expected to be more in future. This study was conducted to explain the biochemical responses of Labeo rohita under extreme thermal condition (treated: 37–38°C against controlled: 28–30°C). Exposure of 14 days resulted in 30% of fish mortality. Glucose concentration was maintained in serum for both treated and controlled groups. However, triglycerides, protein, globulin, calcium, cholesterol and haemoglobin were declined and enzymes (glutamate‐pyruvate transaminase, GPT and glutamate‐oxalate transaminase, GOT) were elevated in serum in the treated group. Indicators of overall metabolism (biomolecules: glucose, triglycerides, protein, cholesterol; enzymes: GPT, GOT; and RNA: DNA) were diminished in liver, whereas these were less affected in muscle. Higher GPT, GOT and lower albumin in serum confirmed disturbance in hepatic panel. Simultaneously compromised growth (lower SGR, DNA content in liver) was observed in treated fish due to impaired metabolism. Extreme thermal stress induced by such catastrophic warmer water exposure caused liver problems and anaemia which ultimately caused fish death.     

Biochemical responses of matrinxãBrycon cephalus (Günther, 1869) after sustained swimming

Juvenile matrinxã, Brycon cephalus, were submitted to sustained swimming for 72 days at 1.0 body length s^?1. Exercised fish (EF) grew more than non‐EF and their feed conversion ratio (FCR) improved; haematological responses demonstrated a decrease in haemoglobin and mean cell haemoglobin contents and increase in the mean cell volume. In the plasma, sodium, ammonia and amino acid concentrations increased; plasma triglycerides decreased while free fatty acids increased. Liver glucose, free amino acids, ammonia, the rate protein per fish weight and total lipid content increased, while the glycogen per fish ratio declined. Glutamate dehydrogenase (GDH) activity increased while pyruvate kinase (PK) and lactate dehydrogenase (LDH) decreased. White muscle glucose, lactate, the glycogen per fish‐weight ratio and total lipid content exhibited a decrease in their values; ammonia, free amino acids and the protein per fish‐weight ratio increased. GDH and PK decreased their activities. In the red muscle glycogen store, the glycogen per fish‐weight ratio and glucose were reduced. Juvenile matrinxãs, under sustained swimming, were physiologically and biochemically adapted to exercise as indicated by improved blood flow, transport and oxygen uptake, FCR, amino acid and protein incorporation and growth. Continuous exercise is a good practice for B. cephalus cultivation.     

Effects of additive iron on growth,tissue distribution,haematology and immunology of gilthead sea bream, <Emphasis Type="Italic">Sparus aurata</Emphasis>

The comparative effects of iron-supplemented levels on growth, tissue distribution, haematology and immunology of gilthead sea bream, Sparus aurata (2 g) were investigated, using four organic (50, 100, 200, 300 mg ORG/kg diet) and one inorganic iron source (200 INOR mg/kg diet). Fish were treated for 12 weeks with the experimental diets and maintained at a water temperature of 19–22°C. Growth (final weight and specific growth rate), tissue distribution (spleen, liver and muscle), haematological parameters (red blood cells, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration) and non-specific immune indexes (respiratory burst activity and antibacterial activity of serum) were analysed. No significant differences were found in growth and iron tissue distribution among the tested groups. Red blood cell counting was statistically higher in fish given 50 ORG, 100 ORG, 200 ORG and 200 INOR feeds. However, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration were not significantly affected by increasing dietary iron. Fish receiving the 100 ORG diet had the best performance with respect to the respiratory burst activity and significantly higher values for antibacterial activity of serum were obtained in fish fed with the 300 ORG diet. The present findings provided no clear evidence of the optimum iron concentration. However, there was adequate indication that iron supplementation enhanced the performance of gilthead sea bream, mainly from a haematological and immunological point of view.     

Effects of salinity level on the survival,growth, feed utilization,carcass composition,haematological and serum biochemical changes of juvenile Meagre (Argyrosomus regius) (Asso, 1801) grown in ground saltwater

A study was performed to examine the effects of salinity on water quality, fish performance, carcass composition and haemato‐biochemical parameters in juvenile meagre, Argyrosomus regius. Fish (5.0 g) were stocked in fibreglass tanks at four salinity levels: 8‰, 16‰, 24‰ and 32‰, and fed a pelleted diet (47/17 protein/lipid) for 56 days. Results indicated that the growth, feed utilization, carcass composition and haemato‐biochemical parameters of meagre gradually improved with the increase in salinity up to 24‰ and then significantly (p ≤ .05) decreased at 32‰. The survival per cent showed a significant decrease when A. regius exposed to 8‰ salinity. An improvement with 32%, 47% and 34.1% of FCR, protein productive value and energy utilization was detected at 24‰ compared with 8‰ salinity respectively. The highest content of protein and the lowest of lipids were recorded in fish carcass at 24‰ compared with the opposite trend at 8‰ salinity. The 24‰ salinity treatment exhibited the highest value of haemoglobin (4.9 g/dl) and the lowest ratio (0.73) of albumin/globulin. The serum total protein, albumin and globulin were significantly higher at 24‰ and 32‰ salinity than those at 8‰ and 16‰ salinity groups. These findings indicate that 24‰ salinity level might be the best for meagre.     

Histopathology of snakehead, Channa striatus (Bloch), experimentally infected with the specific Aphanomyces fungus associated with epizootic ulcerative syndrome (EUS) at different temperatures

Abstract Snakeheads, Channa striatus (Bloch), were inoculated with a spore suspension of the specific pathogenic Aphanomyces , isolated from fish affected by epizootic ulcerative syndrome (EUS), in South East Asia. Fish were held at three different temperatures: 19, 26 and 31 °C. Histological changes induced by the infection are described. In the early stages of the disease, degenerative changes were observed in all samples, but inflammatory infiltrate was much more marked in fish kept at 26 and 31 °C. By 8 days post-injection, extensive mycotic granulomatosis was observed in the samples kept at 26 and 31 °C. The fish kept at 19 °C developed a severe invasive myonecrosis with limited macrophage response. From 14 to 28 days post-injection, healing became well established at 26 and 31 °C and surviving fish kept at these temperatures recovered completely by 28 days. The lesion was still progressing at 21 days post-injection in fish kept at 19 °C, and all such fish succumbed by this time. Thus, mortalities in the fish kept at 19 °C were considerably higher than in the groups of fish kept at 26 and 31 °C. The findings help to explain why mortalities from EUS occur when water temperatures are low.