Infectious pancreatic necrosis virus: experimental infection of goldsinny wrasse, Ctenolabrus rupestris L. (Labridae)

The use of wrasse as cleaner fish in farming of Atlantic salmon, Salmo salar L., is now widespread in Scotland, Ireland and Norway, but little is known about the susceptibility of these fish to the common pathogens of cage-cultured salmon. The susceptibility of goldsinny wrasse, Ctenolabrus rupestris L., to infectious pancreatic necrosis virus (IPNV) was investigated. Captive-bred C. rupestris were infected with IPNV-Sp (Shetland strain) using a bath challenge method. Two infection doses were used, low (4.1 × 10⁵ pfu mL⁻⁻¹) and high (2.4 × 10⁶ pfu mL⁻⁻¹), with two replicates for each experiment. Bathing times were 1 h for the low dose and 5 h for the high dose. A maximum prevalence of infection (30%) was seen 2 weeks post-infection in replicate 1 of the low dose experiment and was accompanied by low tissue titres. The tissue titres dropped to an undetectable level by 4 weeks post-infection. In the high dose experiment, a high prevalence of infection was seen along with moderate titres in tissues as well as a marked pathological response. Both prevalence and intensity declined rapidly and the fish recovered. In the high dose experiment, faecal titres followed the same pattern as those in the tissues. The implications for the use of wrasse in the farming of Atlantic salmon are discussed.     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Acute LD50 and kidney histopathology following injection of erythromycin (Erythro-200) and its carrier in spring chinook salmon, Oncorhynchus tshawytscha (Walbaum)

We evaluated the toxicity of commercial injectable erythromycin (Erythro-200) and the injectable drug carrier alone (PEG-400, ethyl alcohol and ethyl acetate) in sub-adult spring chinook salmon, Oncorhynchus tshawytscha (Walbaum). Both compounds were toxic to salmon when administered in high dosages. The 50% lethal dose (LD₅₀) of the erythromycin within its carrier (Erythro-200) at 12 °C was estimated at 429 mg erythromycin kg^–1 or 2.145 mL kg^–1 when measured by volume. The LD₅₀ of the drug carrier alone was 2.95 mL kg^–1 by volume or equivalent to 3.21 g kg^–1 of the carrier. We evaluated and scored the histopathology in kidney sections removed from fish 96 h after injection with Erythro-200, the drug carrier alone, or sterile phosphate buffered saline (PBS) that served as a reference control. Kidneys from fish injected with saline had lowest average pathology scores; those injected with the carrier had moderate scores; and the pathology score was highest in tissues from erythromycin-injected fish. In tests at 12 °C, vacuoles were more prominent in proximal and distal tubules of fish injected with higher dosages of 403 and 545 mg erythromycin kg^–1 than in tissues from fish injected with 299 mg kg^–1.     

Dietary vitamin E requirement of the red drum Sciaenops ocellatus

A 12-week feeding trial was conducted to establish the minimum dietary vitamin E requirement of juvenile red drum by broken-line regression analysis. The semi-purified basal diet was supplemented with 10, 20, 30, 40, 60 or 80 IU vitamin E kg⁻¹ as all-rac -α-tocopheryl acetate. Juvenile red drum were conditioned by feeding the basal diet for 8 weeks prior to the feeding trial to reduce whole-body vitamin E levels. Then, fish initially averaging 12.2 ± 0.4 g fish⁻¹ (mean ± SD) were fed the experimental diets at a rate approaching apparent satiation for 12 weeks. Weight gain and feed efficiency responses of fish fed diets were significantly ( P  < 0.01) altered by the level of vitamin E supplementation but not strictly in a dose-dependent manner. Vitamin E concentrations in liver and plasma also were significantly ( P  < 0.001) influenced by dietary vitamin E level. Plasma ascorbic acid in fish fed the basal diet tended ( P  = 0.066) to be lower than in fish fed diets containing the various levels of vitamin E. In addition, fish fed the basal diet showed edema in the heart, while fish fed all other diets were normal. Fish fed 60 or 80 IU all-rac -α-tocopheryl acetate kg⁻¹ diet had significantly higher respiratory burst of head kidney macrophages than fish fed all other diets, although dietary effects on hematocrit and neutrophil oxidative radical production were not significant. The minimum dietary vitamin E requirement of juvenile red drum was established based on broken-line regression of liver thiobarbituric acid reactive substances to be 31 mg all-rac -α-tocopheryl acetate kg⁻¹ diet.     

Dose-dependent influences of dietary β-1,3-glucan on innate immunity and disease resistance of hybrid striped bass Morone chrysops×Morone saxatilis

To investigate potential use of dietary β-1,3-glucan for health management of hybrid striped bass, juvenile fish were fed diets supplemented with yeast glucan (MacroGuard^®) at 0.05%, 0.1% or 0.2% of diet for 4 weeks, followed by immune response assays and a bath challenge with Streptococcus iniae . Dietary glucan significantly ( P <0.05) enhanced neutrophil oxidative radical production, and fish fed 0.1% glucan had a significant ( P <0.05) reduction in mortality (10%) after bacterial challenge compared with fish fed the control diet (46.7%). However, accumulative mortality of fish fed 0.2% glucan was not significantly different from that of fish fed the control diet. To further elucidate this observation, macrophages from sub-adult hybrid striped bass were isolated and cultured in L-15 medium with 10% foetal calf serum and penicillin/streptomycin supplemented with 0.2, 0.5, 1, 2, 5, 20 and 100 μg soluble glucan (MacroGuard^®) mL⁻¹ for 24 and 48 h. Intracellular superoxide anion production was significantly ( P <0.001) increased by 0.5 μg glucan mL⁻¹, but significantly ( P <0.001) suppressed by doses >5 μg glucan mL⁻¹. It is concluded that dietary yeast glucan has potential for use in diet formulations of hybrid striped bass to limit the adverse effects of S. iniae , but dosage should be an important consideration in administration.     

An experimental study of the susceptibility of Atlantic salmon fry, Salmo salar L., to viral haemorrhagic septicaemia

Abstract. Infection trials using two serotypes of VHS viruses (type 1 and 23/75) demonstrated that Atlantic salmon fry were susceptible to the disease when injected intraperitoneally (i.p.) with 10³ pfu of virus/fish but resistant to infection by a bath method when exposed for 3 h in water containing 5 × 10⁴ pfu of virus/ml. In the i.p.-infected fish, mortality reached 78 and 67% within 13 days with VHSV₁ nad 23/75 serotypes, respectively. High virus yields were recovered from infected fish and virus shedding was demonstrated by the onset of VHS in rainbow trout kept in the outflow water from the aquaria containing infected salmon. Neither mortality nor virus shedding occurred in salmon infected by the water route but virus multiplication was demonstrated in 2 of 60 fish with VHSV₁ and 3 of 60 fish with virus 23/75. On day 79 post-infection the sera from surviving salmon of both i.p. and bath infection trials exhibited good neutralizing titres (around 1000) against the homologous viruses.     

Protection against enteric redmouth disease in rainbow trout, Salmo gairdneri Richardson, after vaccination with Yersinia ruckeri bacterin

Abstract. Rainbow trout, Salmo gairdneri Richardson, (average weight 100g) were vaccinated intraperitoneally (i.p.) with Yersinia ruckeri bacterin in saline or in oily adjuvant. Agglutinating antibody kinetics were followed during 445 days before challenge (1.2×10⁷ bacteria i.p.). Fourteen days after challenge 88.5% of the control fish had died while few mortalities were observed with vaccinated fish regardless of their agglutinating titre. Protection against Y. ruckeri does not seem to be due to antibodies.     

An experimental investigation on aspects of infectious salmon anaemia virus (ISAV) infection dynamics in seawater Atlantic salmon, Salmo salar L.

This study investigated infection dynamics of infectious salmon anaemia virus (ISAV) by conducting two experiments to examine minimum infective dose and viral shedding of ISAV. In terms of minimum infective dose, the high variability between replicate tanks and the relatively slow spread of infection through the population at 1 × 10¹ TCID₅₀ mL⁻¹ indicated this dose is approaching the minimum infective dose for ISAV in seawater salmon populations. A novel qPCR assay incorporating an influenza virus control standard with each seawater sample was developed that enabled the quantity of ISAV shed from infected populations to be estimated in values equivalent to viral titres. Viral shedding was first detected at 7 days post-challenge (5.8 × 10⁻² TCID₅₀ mL⁻¹ kg⁻¹) and rose to levels above the minimum infective dose (4.2 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹) on day 11 post-challenge, 2 days before mortalities in ISAV inoculated fish started. These results clearly demonstrate that a large viral shedding event occurs before death. Viral titres peaked at 7.0 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹ 15 days post-infection. These data provide important information relevant to the management of ISA.     

Effect of dietary lipid level on muscle composition in Atlantic salmon Salmo salar

This study was conducted to determine the effects of feeding increasing lipid concentrations (310, 380 and 470 g kg^–1 lipid on dry weight) in diets based mainly on herring byproducts to Atlantic salmon Salmo salar . The diets were isonitrogenous, varying in dietary lipid content at the expense dietary starch. Average fish weight increased from 1.2 kg in April to 2.2–2.7 kg at the end of the feeding trial in September. Significantly greater growth was found in fish fed either the 380 g kg⁻¹ or the 470 g kg⁻¹ lipid diets compared with the 310 g kg⁻¹ lipid diet. Muscle lipid content increased in all dietary groups on a wet weight basis from 7.7 ± 1.4% to 12 ± 3% in salmon fed the 310 g kg⁻¹ lipid diet, and to 16 ± 2% in salmon fed the 380 g kg⁻¹ and 470 g kg⁻¹ lipid diets. In fish of similar weight there was a positive correlation between dietary lipid and muscle lipid concentrations. Low concentrations of muscle glycogen were detected in fish fed each of the diets, while muscle vitamin E concentrations slowly decreased as muscle lipid increased. Muscle fatty acid composition reflected dietary fatty acid profiles, containing similar percentages of total saturated, monoenic and n-3 fatty acids (20:5n-3 and 22:6n-3) in fish from all dietary treatment groups. However, a higher ratio of n-3/n-6 was found in muscle from fish fed the 470 g kg⁻¹ lipid diet compared with the other two groups. Blood chemistry values varied somewhat, but all values were within normal ranges for Atlantic salmon of these sizes.     

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L.

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L., was evaluated in a dose titration study and two dose confirmation studies. Replicated groups of salmon with induced infestations of sea lice were given emamectin benzoate on pelleted feed at doses of 0, 25, 50 and 100 μg kg⁻¹ biomass day⁻¹ for seven consecutive days. Sea lice were counted at 7, 14 and 21 days from the start of treatment, and comparisons made with control fish given the same diet without emamectin benzoate. Total numbers of sea lice were significantly reduced at all doses in all three studies when compared to control fish. There was no significant difference between doses of 50 and 100 μg kg⁻¹, but the 50 μg kg⁻¹ dose resulted in significantly fewer lice than the 25 μg kg⁻¹ dose. Emamectin benzoate was highly effective in reducing numbers of preadult and adult lice and prevented the maturation of chalimus to motile stages. The optimum therapeutic dose was selected as 50 μg kg⁻¹ day⁻¹ for seven days. Treatment reduced the incidence of epidermal damage by sea lice and, in one study, survival of treated fish was 48% higher than in control groups. No fish mortalities or adverse effects were attributed to treatment with emamectin benzoate at any of the doses tested.     

Urea in feeds for sea water farmed Atlantic salmon: effect on growth, carcass quality and outbreaks of winter ulcer

The main objective of the present study was to evaluate whether dietary supplementation of urea might reduce the incidence of winter ulcer in sea water-farmed Atlantic salmon. Salmon destined to be S0 smolt were fed with a urea-supplemented diet (0 or 20 g kg⁻¹ urea) over an 8-week period prior to sea water transfer and were then fed supplemented diet (0, 5, 10 or 20 g kg⁻¹ urea) during the first and second winters in the sea. During the first winter positive relationships between dietary urea and plasma urea and between plasma urea and plasma osmolality were observed. Further, plasma osmolality displayed a negative relationship to mortality. Of the salmon that died during the first winter in the sea 90% had one or more skin ulcers. Both during the first and second winter there were fewer salmon with ulcer among fish fed with the diets supplemented with urea. Salmon fed with 20 g kg⁻¹ urea tended to have higher percentage water in the muscle. Mortality and incidence of salmon with ulcer seemed to relate to plasma osmolality amongst fish fed on diets that differed in levels of urea supplementation, suggesting that an osmotic imbalance may contribute to the development of winter ulcer in farmed salmon. Salmon fed with 20 g kg⁻¹ urea showed significantly greater body weight during the second winter in sea. Fish killed without prior starvation had significantly higher level of muscle urea in the 20 g kg⁻¹ urea group compared with fish fed with the unsupplemented diet. However, a 13-day starvation period reduced urea content in the muscle to the level of the control. No effects of dietary urea supplementation on the sensory quality of market size Atlantic salmon were observed.     

Brook charr, Salvelinus fontinalis (Mitchill), and cryptobiosis: a potential salmonid reservoir host for Cryptobia salmositica Katz, 1951

Abstract. Laboratory-raised Cryptobia -susceptible brook charr, Salvelinus fontinalis (Mitchill), and rainbow trout, Oncorhynchus mykiss (Walbaum), were vaccinated intraperitoneally with a live Cryptobia salmositica vaccine (250000 parasites per fish), and 4 weeks later were challenged with the pathogen (250000 parasites per fish). Unvaccinated and infected brook charr had high parasitaemias but no clinical signs of disease, while unvaccinated and infected rainbow trout had anaemia and general oedema. Vaccinated and challenged fish had very low parasitaemias compared to unvaccinated and infected brook charr and rainbow trout. Complement fixing antibodies were detected in vaccinated and challenged fish 2 weeks after challenge. Unvaccinated and infected brook charr had consistently higher litres of complement fixing antibody than unvaccinated and infected rainbow trout. Parasitaemias were lower in all fish in which titres of complement fixing antibody were high. In a second experiment, brook charr inoculated intraperitoneally or intramuscularly with 100000 C. salmositica per fish had high parasitaemias but no anaemia or other clinical signs. The results show that susceptible brook charr do not suffer from cryptobiosis and may serve as reservoir hosts for C. salmositica in areas where the disease is prevalent. Vaccination to reduce the parasitaemia when fish become infected may be a control strategy in these areas.     

The infectivity by different routes of exposure and shedding rates of Aeromonas salmonicida subsp. salmonicida in Atlantic salmon, Salmo salar L., held in sea water

Abstract. The infectivity of the bacterial fish pathogen Aeromonas salmonicida subsp. salmonicida to Atlantic salmon, Salmo salar L., in sea water was investigated and found to be similar to that reported for fresh water. The minimal infective dose in short duration bath exposures (1–3 days) was 10⁴ colony-forming units (cfu) per ml, while prolonged exposure for three weeks, but not for 1 week, produced infection with 10² cfu/ml. Intragastric intubation of A. salmonicida established infection with doses of >10⁵ cfu. Release of bacteria from dead or morbid infected fish was monitored and found to be in the order of 10⁵–10⁸ cfu/fish/h. These results emphasize the importance of removing dead fish from farm sites.     

The synthetic antioxidant, ethoxyquin, adversely affects immunity in tilapia (Oreochromis niloticus)

Ethoxyquin (EQ) has been used as an antioxidant in livestock, aquaculture and pet foods. Animal food safety law has established the upper limit of EQ in animal feed at 150 mg kg⁻¹. However, the risk of EQ at the approved level for aquaculture feed (150 mg kg⁻¹) to fish health is unknown. Here, we examine the effect of EQ on the immunity of tilapia ( Oreochromis niloticus ). EQ concentration in the blood reached 0.16 mg L⁻¹ in fish fed EQ at the approved level. This level of EQ inhibited phagocytic activity of leucocytes in vitro and antibacterial activity of whole blood in vivo . Furthermore, pyknosis in the liver was observed throughout the duration of feeding. However, after 30 days of experimental challenge with feed containing 150 mg kg⁻¹ of EQ, no significant difference was observed in mortality. Although EQ at the approved level in feed causes immunosuppression in fish, the severity of immunosuppression does not lead to a lowering of disease resistance for short feeding periods.     

Dietary menadione nicotinamide bisulphite (vitamin K3) does not affect growth or bone health in first-feeding fry of Atlantic salmon (Salmo salar L.)

The aim of the present study was to elucidate if increasing levels of added vitamin K in the feed influenced fish growth, health or the incidences of bone deformities in Atlantic salmon ( Salmo salar L.) from onset of start feeding to ∼100 g in freshwater. Duplicate groups of Atlantic salmon fry (∼0.20 g) were fed eight levels (0, 2.5, 5, 7.5, 10, 15, 20 and 50 mg menadione kg⁻¹) of the vitamin K derivate menadione nicotinamide bisulphite (MNB) in a regression design for 28 weeks. All fish maintained high growth rates throughout the experiment, and showed no significant differences in specific growth rate, condition factor, whole body proximate analysis, blood coagulation time, vertebra morphology or mechanical properties of vertebrae. We found a dose-response between whole body vitamin K concentration and the dietary MNB supplementation level. Analysis of liver γ-glutamylcarboxylase activity revealed significant dose-dependent differences between groups given the 0, 10 and 50 mg MNB kg⁻¹ diets. In conclusion, Atlantic salmon seems to require low levels of dietary vitamin K, and the amount of vitamin K found naturally in the presently used feed ingredients may be enough to maintain optimal growth, health and bone strength in Atlantic salmon fry from start feeding.     

Supplementing copper to a fish meal based diet fed to Atlantic salmon parr affects liver copper and selenium concentrations

The present experiment was conducted to study growth and tissue responses in Atlantic salmon, Salmo salar L., fed a fish meal based diet supplemented with copper (Cu). The findings of the experiment were used to evaluate the need for dietary Cu supplementation. Atlantic salmon parr, initially weighing ≊ 7.5 g, were randomly distributed among 10 tanks, with 300 fish in each tank. Duplicate groups of fish were fed a fish meal based diet containing 3.5 mg Cu kg⁻¹, or this diet supplemented with 5, 10, 50 or 100 mg Cu kg⁻¹ (as CuSO₄*5 H₂O) for 12 weeks. Growth was recorded and blood haemoglobin measured. The Cu concentrations in whole body, liver, serum and selected muscle samples were measured, as was liver selenium (Se) concentration.
There were no difference in growth among the dietary treatments. There were, however, significant differences among the dietary groups in liver Cu and Se concentrations. The fish fed the diet supplemented with 5 mg Cu kg⁻¹ had increased liver Cu concentration compared with the other groups. Similar trends were found for serum Cu concentration and whole-body Cu concentration, but these effects were not significant. Liver Cu and Se concentrations were positively correlated and liver Se concentrations were inversely correlated to dietary Cu concentration, confirming an interaction between these two elements in salmon.
There may be a positive effect of a modest Cu supplementation level, and we suggest that a small amount of Cu (5–10 mg kg⁻¹) should be added to fish meal based diets.     

Use of salt to control ichthyophthiriosis and prevent saprolegniosis in silver perch, Bidyanus bidyanus

The ciliate protozoan, Ichthyophthirius multifiliis , and the fungus, Saprolegnia parasitica , cause the diseases ichthyophthiriosis and saprolegniosis respectively. Both diseases are difficult to control and can cause high mortalities of freshwater fish, including the Australian silver perch, Bidyanus bidyanus (Mitchell). The efficacy of salt (NaCl) in controlling and preventing these diseases in silver perch was evaluated in aquaria and tanks. Low pH levels were also evaluated as a control for ichthyophthiriosis. Concentrations of 2 or 3 g L⁻¹ salt controlled infestations of I. multifiliis , and fish were free of both theronts and trophonts by day 8 at temperatures of 17.3–21.3 °C and by day 6 at 19.2–23.5 °C. Fish treated with 1 g L⁻¹ salt remained infested and all fish in a control treatment (0 g L⁻¹ salt) died. Although the mean survival rates of infested fish at pH levels of 5 or 6 were only 13.9% and 7.6%, respectively, there were no theronts or trophonts on surviving fish after 12 days. Silver perch harvested from a pond and treated with 2 or 3 g L⁻¹ salt did not become infected with S. parasitica and survival was 100%, whereas 16.6% of untreated (0 g L⁻¹ salt) fish became infected and survival was only 66.7%. A concentration of 2 g L⁻¹ NaCl is recommended for the control of ichthyophthiriosis and the prevention of saprolegniosis in silver perch held in tanks, aquaria and re-circulating aquaculture systems.     

Effects of dietary protein to metabolizable energy ratios and total protein concentrations on the performance of yellow perch Perca flavescens

Juvenile yellow perch Perca flavescens were fed semipurified diets with varying protein to metabolizable energy ratios (PME, g protein MJ⁻¹ metabolizable energy) and nutrient densities in three experiments to determine recommended dietary protein and energy concentrations. Experiment 1 fish (18.6 g) were fed diets containing 450 g crude protein kg⁻¹ dry diet and 14.5–18.8 MJ ME kg⁻¹ dry diet for 10 weeks. No differences were found in the growth of experiment 1 fish fed the different diets. Experiment 2 fish (21.9 g) were fed diets containing 15.7 MJ ME kg⁻¹ dry diet and 210–420 g crude protein kg⁻¹ dry diet for 8 weeks. Fish fed the diet containing 340 g kg⁻¹ protein (diet PME = 22) exhibited the greatest weight gain. Experiment 3 fish (27.1 g) were fed diets with a PME of 22 and varying nutrient density (yielding 205–380 g crude protein kg⁻¹ dry diet) for 8 weeks. No differences were found in the growth of experiment 3 fish. Yellow perch fed the semipurified diets exhibited increased liver fat content, liver size and degree of liver discoloration compared with fish fed a commercial fish meal-based diet. Liver changes may have resulted from high dietary carbohydrate levels. We conclude that a protein level of 210–270 g kg⁻¹ dry diet is suitable for juvenile yellow perch provided that the dietary amino acid profile and carbohydrate content are appropriate for yellow perch.     

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.