Localization of calcium regulatory hormones in fish

Immunocytochemical localization of hypocalcin, a hypocalcemic factor in the corpuscles of Stannius (CS), in American eels was examined at the light (ABC method) and electron microscopic (protein A-gold technique) levels with the specific antiserum raised against purified rainbow trout hypocalcin. Only type 1 cells in the CS were immunoreactive in the light microscopic immunocytochemistry. At the electron microscopic level, however, hypocalcin immunoreactivity was observed in secretory granules of both type 1 and type 2 cells. Our findings may indicate that type 1 cells are the main source of hypocalcin, but that type 2 cells also produce it, suggesting that the presence of two cell types reflects different physiological conditions of a single cell type, rather than functionally different cell types. In addition, we summarize our recent data on the localization of other calcium regulatory, or putative calcium regulatory, hormones in fish: parathyroid hormone, calcitonin and calcitonin gene-related peptide.     

Protective Immunity of Goldfish Against Ichthyophthirius multifiliis Infection Induced by Different Trophont Vaccine Preparations

Ichthyophthirius multifiliis, commonly called “Ich,” is a protozoan parasite that infects the epidermis and gills of freshwater fish. Here, we used goldfish (Carassius auratus) to determine whether the four vaccine preparations (live trophonts, formalin‐fixed trophonts, freeze‐thawed trophont lysates, and trophont cilial and cell cortical fractions) of I. multifiliis (Fouquet) can elicit resistance of immunized fish against subsequent theront challenge, and whether a relationship exists between in vitro immobilization of theronts in sera or mucus from immunized fish and host protective immunity against the parasite. Experimental goldfish were randomly divided into five groups with five parallel controls, with each group or subgroup consisting of 20 individuals. Each test goldfish was immunized with one of the four Ich vaccine preparations administered by one of the three routes: live trophonts by gavage (Group 1), formalin‐fixed trophonts by injection (Group 2) or by immersion (Group 3), freeze‐thawed trophont lysates by injection (Group 4), and trophont cilial and cell cortical fractions by injection (Group 5). The fish were then challenged by a standard Ich theront challenge on Day 21 postimmunization. For every 10 d following immunization, we tested the in vitro immobilization of Ich theronts by sera or mucus from immunized and control fish. We found that although all control (nonimmunized) and Group 3‐immunized goldfish died following theront challenge, more than half (55–65%) of the immunized fish from Groups 1, 2, 4, and 5 survived. Furthermore, except for the fish from Groups 3 and 4, the number of mean days to death in each test group was significantly higher than that in the respective control. These results indicate that goldfish immunized by injection or by gavage with any of the four vaccine preparations gained resistance to Ich infection. Further analysis indicated that this protective immunity was closely associated with the in vitro immobilization of Ich theronts by fish sera or mucus.     

Protective responses of intestinal mucous cells in a range of fish–helminth systems

Histopathological, immunofluorescence and ultrastructural studies were conducted on the intestines of four fish species infected with different taxa of enteric helminths. Brown trout (Salmo trutta trutta), eel (Anguilla anguilla) and tench (Tinca tinca) obtained from Lake Piediluco (central Italy) were examined. Brown trout and eel were infected with two species of acanthocephalans, and tench was parasitized with a tapeworm species. In addition to the above site, specimens of chub (Squalius cephalus) and brown trout infected with an acanthocephalan were examined from the River Brenta (north Italy). Moreover, eels were examined from a brackish water, Comacchio lagoons (north Italy), where one digenean species was the predominant enteric worm. All the helminths species induced a similar response, the hyperplasia of the intestinal mucous cells, particularly of those secreting acid mucins. Local endocrine signals seemed to affect the production and secretion of mucus in the parasitized fish, as worms often were surrounded by an adherent mucus layer or blanket. This is the first quantitative report of enteric worm effects on the density of various mucous cell types and on the mucus composition in intestine of infected/uninfected conspecifics. We provide a global comparison between the several fish–helminth systems examined.     

Branchial calcium uptake: possible mechanisms of control by stanniocalcin

The branchial Ca²⁺ uptake by teleost fish is under inhibitory control by the hormone stanniocalcin (STC) which is generated by the corpuscles of Stannius (CS). Removal of the CS in North American eel, Anguilla rostrata LeSueur, induced a rapid rise in blood calcium levels. Branchial Ca²⁺ influx following the extirpation of the CS (stanniectomy, STX) increased during the first four days and stayed elevated thereafter (in agreement with previous studies). The transepithelial potential (TEP) across the gills did not change after STX and this means that the electrochemical gradient for Ca²⁺ is less favourable for passive influx of Ca²⁺ in STX eel. Therefore, the Ca²⁺ influx in STX eels is a transcellular flux, with Ca²⁺ crossing the apical and basolateral membrane barrier. The kinetics of ATP-driven Ca²⁺-transport across basolateral plasma membranes from eel gills did not change after STX. Thus, the increased Ca²⁺-influx after STX is not correlated with changes in ATP-dependent Ca²⁺-extrusion across the basolateral membrane, suggesting a regulation at the apical membrane. Moreover, STC did not affect ATP-driven Ca²⁺-transport in isolated basolateral membranes (in vitro). STC (0.1 nM) reduced cAMP levels in dispersed eel gill cells. It had no significant effect on the IP₃ levels in these cells. We postulate that STC controls the permeability to Ca²⁺ of the apical membranes of the Ca²⁺ transporting cells of fish gills by controlling second messenger operated Ca²⁺ channels in the apical membrane.

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Résumé L'entrée de calcium au niveau des branchies est sous le controle inhibiteur de la stanniocalcine (STC) qui est synthétisée au niveau des corpuscules de Stannius (CS). L'ablation des CS chez l'anguille d'Amérique du Nord, Anguilla rostrata LeSueur, induit une augmentation rapide des niveaux de calcium dans le sang. Le flux entrant branchial de calcium consécutif à l'ablation des CS (stanniectomie, STX) augmente pendant les 4 premiers jours et reste élevé au-delà (en accord avec des études antérieures). Le potentiel transépithélial (TEP) à travers les branchies ne change pas après STX, ceci indiquant que le gradient électrochimique du Ca²⁺ est moins favorable pour le flux entrant passif du Ca²⁺ chez l'anguille STX. En conséquence, le flux entrant de Ca²⁺ chez l'anguille STX est un flux transcellulaire, avec le Ca²⁺ traversant la barrière membranaire apicale et basolatérale. La cinétique du transport de Ca²⁺ conduit par l'ATP à travers les membranes plasmatiques basolatérales de branches d'anguille n'est pas modifiée après STX. Ainsi, l'augmentation du flux entrant de Ca²⁺ après STX n'est pas corrlée avec des modifications de l'excrétion de Ca²⁺ conduit par l'ATP à travers la membrane basolatérale, suggérant donc une régulation au niveau de la membrane apicale. De plus, la STC ne modifie pas le transport de Ca²⁺ conduit par l'ATP dans des membranes basolatérales isolées (in vitro). La STC (0.1 nM) réduit les niveaux d'AMPc dans des cellules dispersées de branchies d'anguille. Cette hormone n'a pas d'effet significatif sur les niveaux d'IP₃ dans ces cellules. Nous suggérons que la STC régule la perméabilité au Ca²⁺ des membranes apicales des cellules branchiales transporteuses de Ca²⁺ en controlant un second messager agissant sur les canaux calciques de la membrane apicale.

     

In vitro and in vivo effects of rabbit anti-thymocyte serum on circulating leucocytes and production of complement fixing antibodies in thymectomized Oncorhynchus mykiss (Walbaum) infected with Cryptobia salmositica Katz 1951

Rabbit anti-thymocyte serum (RATS) against thymocytes of rainbow trout was toxic to leucocytes from intact and thymectomized rainbow trout at 10 °C under in vitro conditions. The total number of leucocytes decreased significantly in 24 h after RATS was injected intraperitoneally into intact rainbow trout, but the number returned to pre-injection level within 1 week. RATS destroyed a lower percentage of leucocytes in thymectomized fish than in intact fish under both in vitro and in vivo conditions and the recovery in the number of leucocytes was slower in thymectomized fish. The parasitaemia, packed cell volume and production of complement fixing antibody in thymectomized and intact fish (injected with RATS before Cryptobia salmositica infection) were not significantly different from control fish (not injected with RATS), and they both acquired protective immunity against cryptobiosis on recovery. This indicates that RATS is not cytotoxic to B-like cells in the lymphoid tissue which produce complement fixing antibody against C. salmositica and that the protective antigen in C. salmositica seems to be thymus-independent.     

PARASITES AND PARASITOSIS IN FISH CULTURE IN PORTUGAL

This paper deals with research work covering the period from May 1979 up to May 1981 and presents results of parasites and parasitosis diagnosed in a total of 195 complete dissections of six fish species from cultured and feral populations in Portugal: European eel (Anguilla anguilla) (49); mirror carp (Cyprinus carpio specularis) (29); common grey mullet (Mugil cephalus) (18); European plaice (Pleuronectes platessa) (11); rainbow trout (Salmo gairdneri) (42); common sole (Solea solea) (46). A list of protozoans and helminths identified with respective incidence is given: A) Feral fish: 1) European eel (17)—Echinorhynchus clavula 11.8% and Rhabdochona anguilla 5.9%; 2) common grey mullet (18)—Haploporus benedenii 16.7%, Neoechinorhynchus sp. 5.6%, and Raphidascaris sp. 5.6%; 3) European plaice (11)—Bothriocephalus sp. larvae 9.1%, Contracaecum aduncum 9.1%, and Cryptocotyle lingua 9.1%; 4) common sole (46)—Cucullanellus minutus 8.7%, Gyrodactylus elegans 4.3%, and Cestodes larvae 2.2%. B) Cultured fish: 1) European eel (32)—Trichodina anguillae 3.1% and Ichthyophthirius multifiliis 18.8%; 2) rainbow trout (42)—Eimeria truttae 2.4%, Hexamita intestinalis 4.8%, Holophrya simplex 2.4%, and Trichodina truttae 4.8%. The results show that feral fish harbor predominantly indirect life cycle helminths, in contrast with cultured fish only affected with protozoans, due mainly to overcrowding and deficient management, which propitiate a significant background infection. It was in these specimens that some lesions characteristic of protozoosis were found.     

Development and validation of glycoprotein‐based native‐subunit vaccine for fish against Aeromonas hydrophila

Aeromonas hydrophila is known to be causative agent of an infection named as Bacterial haemorrhagic septicaemia or red pest in freshwater fish. The aim of this study was to develop and validate the glycoprotein‐based fish vaccine against Aeromonas hydrophila. For this aim, after identification and characterization of A. hydrophila isolates from fish farms, one A. hydrophila isolate was selected as vaccine strain. Antigenic glycoproteins of this vaccine strain were determined by Western blotting and glycan detection kit. The connection types of these glycoproteins were examined by glycoprotein differentiation kit. Two glycoproteins, molecular weights of 19 and 38 kDa, with SNA connection type were selected for use in vaccination trials. After their purification by SNA‐specific lectin and size‐exclusion chromatography, protection studies with purified proteins were performed. For challenge trials, four experimental fish groups were designated: Group I (with montanide), Group II (with montanide and ginseng), Group III [with Al(OH)₃] and Group IV [with Al(OH)₃ and ginseng]. The survival ratings of fish were determined, and protection was calculated as 21.56%, 29.41%, 69.83% and 78.88% in groups I, II, III and IV, respectively. In conclusion, A. hydrophila glycoproteins with Al(OH)3 and ginseng could be used as a safe and effective vaccine for fish.     

Effects of exercise-training on cardiac performance and muscle enzymes in rainbow trout,Oncorhynchus mykiss

Rainbow trout,Oncorhynchus mykiss, were exercise-trained for 18 hours per day over 28 days at water velocities up to 60% of their measured U_crit. Anin situ perfused heart preparation was used to compare maximum cardiac performance between control and trained fish. Trained fish had a larger stroke volume at a given filling pressure, as well as an 18% higher cardiac output and a 25% greater maximum power output. These observations indicate that exercise training in rainbow trout improved maximum cardiac performance. Adrenaline produced positive inotropic and chronotropic effects on the perfused heart, but exercise training did not alter these stimulatory effects. Maximal activities of citrate synthase (CS), B-hydroxyacyl CoA dehydrogenase (HOAD), glutamate dehydrogenase (GDH) and carnitine palmitoyl transferase (CPT) were measured in cardiac and skeletal muscles. CS, HOAD and GDH increased in red and white skeletal muscle as a result of training. Training also increased GDH activity in the endocardium and epicardium, and increased HOAD in the epicardium. While the training regime did not result in a statistically significant increase in U_crit and produced a decrease in the condition factor of the fish, other training effects were clearly evident. Furthermore, significant correlations were observed between U_crit and the maximal activities of GDH and HOAD.     

Effects of adrenaline on ionic equilibria in red blood cells of rainbow trout (Salmo gairdneri)

Effects of adrenaline on the equilibrium distributions of Na⁺ , K⁺ , H⁺ , Cl^– , and H₂O across the cell membrane of rainbow trout (Salmo gairdneri) erythrocytes were determinedin vitro, as a function of P CO₂ (1.76–7.77 torr). CO₂-carrying capacity of the blood was also examined. Plasma catecholamine concentrations inunanaesthetized, unrestrained trout were 3.1 nM adrenaline and 1.2 nM noradrenaline. Elevation of the plasma adrenaline concentrationin vitro to 4.6 × 10³ nM resulted in net gains of Na⁺ , Cl^– and H₂O by red cells, a net loss of H⁺ from red cells, and a pronounced red cell swelling. Adrenaline also reduced the CO₂-carrying capacity of trout bloodin vitro. The magnitudes of these effects increased with P_CO2 and, thus, were sensitive to blood HCO₃ ^– concentrations. The distribution of K⁺ between red cells and plasma was unaffected by adrenaline. Adrenergic-mediated ion movements and red cell swelling were sensitive to both propranolol and SITS. These results are consistent with the symport NaCl uptake model for adrenergic-mediated swelling of Baroinet al. (1984). The adrenergic response of fish erythrocytes may function to ameliorate the effects of blood acidoses on O₂-carrying capacity by maintaining red cell pH in the face of a decrease in plasma pH.     

Extranuclear Histones in Teleost Gills: An Evolutionary Study

Extranuclear histones and histone fragments (13–15 kDa) in gill microsomes of a phylogenetic series of teleostean fishes (tilapia, trout, carp, eel and elephant fish) were studied. Histones H2A and H2B are the first occurring of these antimicrobial agents. Halfway the evolution of teleosts (carp, trout) H3 fragments emerge, which disappear in later stages of development and are replaced by truncated H1 (tilapia). The hypothesis is forwarded that variation in composition of extranuclear histones is caused by changes in specificity of histone transporters.     

Influence of gossypol from dietary cottonseed meal on haematology, reproductive steroids and tissue gossypol enantiomer concentrations in male rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) males were fed, during a 9‐month period, five experimental diets where fishmeal proteins were gradually replaced with cottonseed meal (CS) proteins (0, 25, 50, 75 and 100%; diets 1–5, respectively). This study was carried out to evaluate the action and tissue concentrations of gossypol. Growth performance of fish was not affected with the increasing levels of CS in the diets. Haemoglobin concentration and haematocrit significantly decreased in fish fed 100% CS compared with other dietary treatments. Gonadosomatic index, plasma sex steroids (testosterone, 11‐ketotestosterone and 17,20β‐dihydroxy‐4‐pregnen‐3‐one) and sperm characteristics (concentration, motility, protein concentration and lactate dehydrogenase activity) were not negatively affected by increasing levels of CS. For the first time, a comprehensive study of gossypol concentrations, total and (+) and (?)‐enantiomers, in several tissues (liver, blood plasma, spermatozoa and seminal plasma) was performed. The concentrations of both (+) and (?)‐enantiomers significantly increased with increasing levels of CS in the diet. The highest concentrations were found in the liver of fish fed diet 5 (185 ± 18 μg g^?1). In blood plasma, the concentrations of total gossypol were 10 times lower than the one found in the liver, but 10 and 100 times higher than the concentrations in the spermatozoa and seminal plasma, respectively. In all tissues studied, the concentration of (+)‐enantiomer was higher than the (?)‐enantiomer. The ratio (?)‐enantiomer/total gossypol did not change significantly with the increasing levels of CS in the diet. The results of the present study indicate that CS can be used over a period of 9 months to replace fishmeal proteins completely without compromising growth and reproduction of rainbow trout males.     

Reevaluation of the Dietary Protein Requirements and Optimum Dietary Protein to Energy Ratios in Japanese Eel, Anguilla japonica

A 2 × 3 factorial design was used to reevaluate the dietary protein requirements and to determine the optimum dietary protein to energy (P/E) ratios in Japanese eel, Anguilla japonica, reared in the recirculating system. For each of two experiments, six experimental diets (₄₅P₁₆, ₄₅P₁₇, ₄₅P₁₉, ₅₀P₁₆, ₅₀P₁₇, and ₅₀P₁₉) were formulated and prepared to contain two protein levels (45 and 50%) and three energy levels (16, 17, and 19 kJ/g diet) at each protein level. In the first experiment, glass eel of initial weight 0.1 ± 0.02 g (mean ± SD) were used, while the second experiment was conducted with juvenile eel of initial weight 15.0 ± 3 g (mean ± SD). The first and second experimental periods were 6 and 16 wk for the glass and juvenile eel, respectively. At the end of the first experiment, there were no protein, energy, and their interaction effects. Also, there were no significant differences in weight gain (WG), specific growth rate (SGR), feed efficiency (FE), and protein efficiency ratio (PER) for glass Japanese eel fed all diets. Although there were no significant differences in growth parameters of glass eel fed all experimental diets, these parameters were higher for fish fed ₅₀P₁₆ than for fish fed the other diets. For the second experiment, there were significant protein effects on WG, SGR, and PER (P < 0.05). However, there were neither significant energy effects nor protein and energy interaction effects on WG, SGR, FE, and PER. Fish fed ₄₅P₁₉ had a higher WG, SGR, and PER than did fish fed ₄₅P₁₆, ₅₀P₁₆, and ₅₀P₁₉ (P < 0.05). However, there were no significant differences in growth parameters among fish fed ₄₅P₁₆, ₄₅P₁₇, ₅₀P₁₆, ₅₀P₁₇, and ₅₀P₁₉ and among those fed ₄₅P₁₇, ₄₅P₁₉, and ₅₀P₁₇. These results may indicate that the optimum dietary protein requirement and the P/E ratio could be 44.3% and 24.1 mg protein/kJ (₄₅P₁₉), respectively, in juvenile Japanese eel, based on WG, SGR, and PER.     

Purification and characterization of pepsinogens and pepsins from the stomach of rice field eel (<Emphasis Type="Italic">Monopterus albus</Emphasis> Zuiew)

Three pepsinogens (PG1, PG2, and PG3) were highly purified from the stomach of freshwater fish rice field eel (Monopterus albus Zuiew) by ammonium sulfate fractionation and chromatographies on DEAE-Sephacel, Sephacryl S-200 HR. The molecular masses of the three purified PGs were all estimated as 36 kDa using SDS–PAGE. Two-dimensional gel electrophoresis (2D-PAGE) showed that pI values of the three PGs were 5.1, 4.8, and 4.6, respectively. All the PGs converted into corresponding pepsins quickly at pH 2.0, and their activities could be specifically inhibited by aspartic proteinase inhibitor pepstatin A. Optimum pH and temperature of the enzymes for hydrolyzing hemoglobin were 3.0–3.5 and 40–45°C. The K _m values of them were 1.2 × 10⁻⁴ M, 8.7 × 10⁻⁵ M, and 6.9 × 10⁻⁵ M, respectively. The turnover numbers (k _cat) of them were 23.2, 24.0, and 42.6 s⁻¹. Purified pepsins were effective in the degradation of fish muscular proteins, suggesting their digestive functions physiologically.     

An Evaluation of the Suitability of Selected Waste Products in Feeds for Three Fish Species

ABSTRACT

Five types of aquatic food industry waste products (carp offal, carp roe, fish frames, trout offal and surimi processing waste) together with fish meal were evaluated for their suitability as potential fish meal replacements, partially or wholly, in diets for three species (rainbow trout, Murray cod and shortfin eel) cultured in Australia, using a number of criteria.

The proximate composition of the ingredients on a dry matter basis including protein content, lipid and ash, varied considerably. The essential amino acid (EAA) contents of the waste products and fish meal decreased in the order: carp roe > fish meal > carp offal > ‘surimi’ processing waste > fish frames > trout offal. The results of cluster analysis of A/E ratios of waste products and fish whole body fell within three clusters. The EAAI of whole body tissue of Murray cod, rainbow trout and Australian shortfin eel however, were closest to fish meal, followed by fish frame waste and/or ‘surimi’ waste. The results on A/E ratios and EAAI did not conform to the raw data on TAA and EAA. Therefore, the study emphasizes the need to have a multi-prong approach to determine the suitability of ingredients for incorporation into fish feeds.     

Comparative study of digestive enzymes in fish with different nutritional habits. Proteolytic and amylase activities

This work provides a comparative study of the proteolytic and amylase activities in six species of fish with different nutritional habits: rainbow trout (Oncorhynchus mykiss), gilthead seabream (Sparus aurata), European eel (Anguilla anguilla), common carp (Cyprinus carpio), goldfish (Carassius auratus), and tench (Tinca tinca). Trout and carp showed the highest digestive proteolytic activity. When proteolytic activity was determined in a wide range of pHs, the highest values in the digestive tract of all species were found at alkaline pHs, except in eel where activity could be detected only at acid pHs. Eel showed the lowest digestive proteolytic potential among all the species studied. With respect to amylase activity, the omnivorous species presented higher activity than did the carnivores. Among the carnivorous species, the lowest activity was found in trout. The ratio of total amylase:total proteolytic activity was higher in omnivorous fish species, the carp having the greatest value, whereas in trout this ratio was lower than one. Digestive enzyme activity declined as the incubation temperature decreased, but this trend varied depending on the fish species and the tissue analyzed.     

Rapid response of fish and aquatic habitat to removal of a tidal barrier

1. River barrier removal is used increasingly as a conservation tool to restore lotic habitat and river connectivity, but evidence of its efficacy is incomplete. This study used a before–after methodology to determine the effects of removing a tidal-limit barrier on the fishes, macroinvertebrates, and habitats of an English coastal stream.
2. Following barrier removal, habitat diversity increased immediately upstream and remained similar downstream. Mobilized silt altered the substrate composition immediately downstream, but this was temporary as silt was flushed out the following winter. Changes to macroinvertebrate communities occurred upstream and downstream of the former barrier but these were transient.
3. A dramatic and sustained increase in fish density occurred immediately upstream of the barrier after its removal, but effects downstream were minor. The fish community upstream changed, largely due to rapid recruitment and dispersal of endangered European eel (Anguilla anguilla). Eel density in the formerly impounded zone increased from 0.5 per 100 m² before barrier removal to 32.5 per 100 m² 5 months after removal. By 17 months after barrier removal there was no difference in eel density across the six sections sampled.
4. Although resident stream fishes such as bullhead (Cottus gobio species complex, protected under the European Habitats Directive) were abundant in middle and upper-stream sections, brown trout (Salmo trutta, a listed species for biodiversity conservation in England and Wales) density remained low during the study and recruitment was poor. This suggests that although colonization access for anadromous trout was available, habitat upstream may have been unsuitable for reproduction, indicating that wider catchment management is required to complement the restoration of connectivity.
5. These findings suggest that tidal barrier removal is an effective method of restoring lotic habitats and connectivity, and can be beneficial for resident and migratory fishes including those of conservation importance (e.g. European eel) in coastal streams.

     

Use of probiotics to enhance growth,stimulate immunity and confer disease resistance to Aeromonas salmonicida in rainbow trout (Oncorhynchus mykiss)

An 8‐week feeding trial was conducted to evaluate the effects of dietary probiotics on growth, non‐specific immune responses and disease resistance in juvenile rainbow trout, Oncorhynchus mykiss. Fish averaging 5.8 ± 0.8 g (mean ± SD) were fed one of the five experimental diets; one control (Cont), and four other diets were prepared by supplementing single probiotics 1 (Bacillus subtilis; SP₁, 0.5%), single probiotics 2 (Bacillus licheniformis; SP₂, 0.5%), multi‐probiotics (B. subtilis + B. licheniformis; MP, 0.5%) and oxytetracycline (OTC) at 5 g OTC kg^?1 diet. After 8 weeks of the feeding trial, weight gain and specific growth rate of fish fed SP₁, SP₂ and OTC diets were significantly higher than those of fish fed Cont diet (P < 0.05). Superoxide dismutase (SOD) and lysozyme activities of fish fed SP₁, SP₂ and MP diets were significantly higher than those of fish fed Cont diet (P < 0.05). There were no significant differences in SOD and lysozyme activities among fish fed SP₁, SP₂, MP and OTC diets. In challenge test with Aeromonas salmonicida for 15 days, fish fed SP₁, SP₂ and MP diets showed significantly higher cumulative survival rate than those of fish fed Cont diet (P < 0.05). However, there were no significant differences in cumulative survival rate among fish fed SP₁, SP₂, MP and OTC diets. Although there was a little advantage in fish fed MP diet in terms of non‐specific immune responses, single or multi‐probiotics are equally effective statistically. These results indicate that single or multi‐probiotics had equal beneficial effects as an antibiotic replacer on growth performance, non‐specific immune responses and disease resistance in juvenile rainbow trout.     

Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout

The feasibility of using rainbow trout Oncorhynchus mykiss embryos as an expression system for proteins was investigated. For model proteins, we selected two goldfish gonadotropins (GTHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). To produce single-chain goldfish FSH (scgfFSH) and LH (scgfLH), cDNAs encoding glycoprotein hormone (GP) α and FSHβ were fused in tandem, and cDNAs encoding GPα and LHβ were fused in tandem. The fused cDNAs were ligated with β-actin promoter, and microinjected into fertilized rainbow trout eggs. After 4-days incubation, the embryos were subjected to western blotting and in vitro bioassays. The recombinant proteins produced by the embryos were immunoreactive to antisera against goldfish GPα, N-glycosylated, and biologically active. We conclude that scgfFSH and scgfLH were successfully produced in transgenic rainbow trout.     

Response of Rainbow trout (Oncorhynchus mykiss) to unrefined peanut oil diets: Effect on growth performance,fish health and fillet fatty acid composition

A 60‐day feeding study with rainbow trout, Oncorhynchus mykiss, was conducted to determine the effects of replacement of fish oil (FO) by unrefined peanut oil (PO) on growth performance, feed utilization, body composition, fatty acid composition and serum biochemical and haematological parameters. Rainbow trouts (51.60 ± 0.75 g) were fed five experimental diets formulated by replacing dietary FO with PO at levels of level 0 (PO₀), 1/4 (PO₂₅), 1/2 (PO₅₀), 3/4 (PO₇₅) and 4/4 (PO₁₀₀), respectively. As a result, the best growth performance was observed in fish fed with PO₀ and PO₅₀ diet. No significant differences were detected among the groups in terms of body compositions. Fatty acid profiles of the fish fillets reflected the fatty acid profiles of the feeds that the fishes were fed with. In this study, the haematological parameters detected that there were no significant differences compared to the control group, whereas the serum biochemical parameters generally worsened as the ratio of peanut oil in the ration exceeded half of fish oil. As a conclusion, the results of the study suggested that the unrefined peanut oil could be used as a replacer of fish oil in diets for rainbow trout.     

The role of nutritional factors in the prevention of peroxidative damage to tissues

The multi-level defence system present in vertebrate cells to protect against chain reactions initiated by free radicals (mainly toxic metabolites of oxygen) is outlined. It comprises superoxide dismutases (Cu-Zn and Mn dependent), glutathione peroxidase (Se dependent), vitamin E and glutathione S-transferase.The protective role of superoxide dismutase (SOD) was demonstrated by studies on trout (Salmo gairdneri) retina (Desrochers and Hoffert 1983). This tissue is subject to very high O₂ tensions but has a high resistance to O₂ toxicity that is dependent on a high dismutating capacity. The activities of both the cytosolic Cu-Zn SOD and the mitochondrial Mn SOD in the liver of rainbow trout were significantly altered in response to changes in dietary intake of these minerals.Deficiency of Se did not affect the growth rate of rainbow trout but led to greatly reduced activities of hepatic and plasma glutathione (GSH) peroxidase. There was no evidence of a Se-independent GSH peroxidase activity. In rainbow trout depleted of Se there was a compensatory increase in hepatic GSH S-transferase activity. This enzyme in conjunction with GSH has been shown to inhibit lipid peroxidation in the Festimulated microsomal systemin vitro. A dietary synergism between vitamin E and Se in rainbow trout has been demonstrated.The vitamin E requirement of rainbow trout is related to the polyunsaturated fatty acid (PUFA) content of the diet. The molar ratio PUFA: vitamin E in rainbow trout tissues is lower than that in rats. Time course studies on the uptake of orally administered vitamin E showed rapid incorporation into biomembranes; few differences between normal and vitamin E deficient fish were evident. The ester form of vitamin E was more slowly assimilated than the alcohol.