Plasma insulin levels during the spawning migration of the pink salmon, Oncorhynchus gorbuscha

Plasma insulin concentration was measured by homologous radioimmunoassay in male and female pink salmon. Oncorhynchus gorbuscha , during spawning migration in the Fraser and Thompson Rivers, British Columbia. Although the fish ceased feeding prior to entering fresh water, plasma levels of insulin remained stable (males) or even elevated (females) during the final stages of oogenesis and spermatogenesis, decreasing thereafter. Mean concentrations ranged from 0–69 to 1.24 ng ml⁻¹ in males and from 0.33 to 0.88 ng ml⁻¹ in females. At all stages in the anadromous migration where a significant difference in plasma insulin levels between the sexes was observed, males had higher concentrations than females.     

Differential gene expression in infarct scar and viable myocardium from rat heart following coronary ligation

Post-myocardial infarction (MI) remodeling of cardiac myocytes and the myocardial interstitium results in alteration of gross ventricular geometry and ventricular dysfunction. To investigate the mechanisms of the remodeling process of the heart after large MI, the expression of various genes in viable left ventricle and infarct scar tissue were examined at 16 weeks post-MI. Steady-state expression of Na⁺-K⁺ATPase α-1 and −2, phospholamban (PLB), α-myosin heavy chain (α-MHC), ryanodine receptor (Rya) and Ca²⁺ ATPase (Serca2) mRNAs were decreased in the infarct scar vs noninfarcted sham-operated controls (P < 0.05). On the other hand, Giα2 and β-MHC mRNAs were upregulated (P < 0.05, respectively) in the infarct scar whereas Na⁺-K⁺ ATPase-β, Na⁺-Ca²⁺ exchanger and Gs mRNAs were not altered vs control values. In viable left ventricle, the a-1 subunit of Na⁺-K⁺ATPase, α-3, β-isoforms, Rya, β-MHC, Giα2, Gs and Na⁺-Ca²⁺ exchanger were significantly elevated while expression of the a-2 subunit of Na⁺-K⁺ ATPase, PLB and Serca2 were significantly decreased compared to controls. Expression of CK2α mRNA was elevated in noninfarcted heart (145 ± 15%) and diminished in the infarct scar (66 ± 13%) vs controls. Expression of β-MHC mRNA was elevated in both viable and infarct scar tissues of experimental hearts (140 ± 31% and 183 ± 30% vs. controls, respectively). These results suggest that cardiac genes in the infarcted tissue and viable left ventricle following MI are differentially regulated.     

Effects of sampling on blood parameters in the rainbow trout, Salmo gairdneri Richardson

The effects of sampling and short-term storage of blood samples on the blood parameters of rainbow trout were evaluated by comparing values obtained from resting fish sampled via cannulae and from fish sampled by cardiac puncture. Sampling stress causes an increase in haematocrit value and a decrease in mean cellular haemoglobin concentration, indicating red cell swelling. These changes are aggravated by consecutive storage of samples. Also, blood ionic balance is affected. The K⁺ concentration of plasma increases and plasma Cl⁻ concentration decreases owing to sampling. During storage the plasma K⁺ concentration decreases far below resting levels in samples taken by cardiac puncture, whereas plasma Cl⁻ level returns to pre-stress levels. Owing to the sampling- and storage-induced changes in blood parameters, care must be taken to ensure that similar sampling procedure is employed throughout to allow reliable comparisons between groups to be made.     

Swimming activity and behaviour of European Anguilla anguilla glass eels in response to photoperiod and flow reversal and the role of energy status

To better understand migratory divergences among Anguilla anguilla glass eels, the behaviour of individuals caught at the time of their estuary entrance was studied through their response to a light:dark cycle and then to both water current reversal and light:dark cycle. In a first experiment, fish moving with the flow in response to dusk (M⁺ fish) and fish that had not exhibited any movement (M⁻ fish) were distinguished. Anguilla anguilla from these two groups were then individually marked and their response to water current reversal compared. M⁺ individuals mainly exhibited negative rheotaxis with a tidal periodicity, whereas positive rheotaxis was mainly exhibited by M⁻ individuals. Thus, M⁺ A. anguilla glass eels showing negative rheotaxis appear to have the strongest propensity to migrate, the converse applies to M⁻ ones showing positive rheotaxis. A small percentage of individuals (5%) were hyperactive, alternately swimming with and against the current with almost no resting phase. These fish lost c . 2 mg wet mass day⁻¹, whereas individuals which were almost inactive lost c . 1 mg day⁻¹. Wet and dry mass changes in relation to activity levels were compared with previous experiments and it was concluded that A. anguilla glass eel energy status might be involved in differences in migratory tendencies but other factors that might be important are discussed. It is proposed that any decrease in A. anguilla glass eel energy stores associated with global warming might lead to an increase in the proportion of sedentary individuals and thus be involved in the decrease in the recruitment to freshwater habitats.     

Evaluation of biochemical methods for the non‐destructive identification of sex in upstream migrating salmon and sea trout

Female‐specific markers of reproductive activity [plasma 17β‐oestradiol (E2), vitellogenin (VTG) and alkali‐labile phosphoprotein phosphorous (ALP)] were measured over 12 months in a captive population of brown trout Salmo trutta . During the early months of the reproductive season (February to May) and using the concentration of plasma E2 or plasma ALP as a marker for females the proportion of fish in which sex was misidentified was high (15–50%). The misidentification rate was considerably lower (1–8%) using plasma VTG. Preliminary evaluation of a commercial immunochromatographic VTG test system as a screen for the presence or absence of VTG in plasma from brown trout provided results that were consistent with those obtained from direct measurement of plasma VTG levels by enzyme‐linked immunosorbent assay (ELISA). These preliminary conclusions were verified by sampling upstream‐migrating anadromous brown trout, sea trout, and Atlantic salmon Salmo salar trapped over a 6 month period. Plasma E2 levels did not satisfactorily discriminate between male and female sea trout and Atlantic salmon. Plasma VTG levels in both species, however, were bimodally distributed and it was assumed that this divergence corresponded to male (plasma VTG levels <10 μg ml⁻¹) and female (plasma VTG levels >800 μg ml⁻¹) fishes. Plasma ALP provided a more accurate indication of sex in the wild Atlantic salmon and sea trout than was suggested by the pilot study on captive brown trout. The commercial immunochromatographic VTG test system provided results that were wholly consistent with the data obtained from the trapped fishes by direct measurement of plasma VTG.     

Seasonal changes in androgen levels in stream- and hatchery-reared Atlantic salmon parr and their relationship to smolting

In stream-reared Atlantic salmon Salmo salar , plasma androgens were significantly greater in mature male parr than immature males and females in October, but had declined by January and did not differ significantly from immature fish throughout the spring. Immature fish in March were significantly larger and had greater gill Na⁺, K⁺-ATPase activity than their previously mature counterparts. Bimodal growth distribution was seen in hatchery-reared Atlantic salmon and a proportion of the male fish in the lower mode matured. Plasma testosterone (T) and 11-ketotestosterone (11-KT) were significantly elevated from September to December in mature male (1+ year) parr. In January, plasma androgens had declined in mature males and did not differ significantly from immature fish. By May all the hatchery fish were large enough to smolt and a proportion of the previously mature males had increased gill Na⁺, K⁺-ATPase activity. Therefore elevated androgens in the previous autumn do not prevent smolting. Parr with higher plasma T and 11-KT in April and May, that are presumably beginning to mature, had lower gill Na⁺, K⁺-ATPase activity, indicating that future maturation and associated increases in androgens may inhibit smolting.     

Effects of five years of intensive trawling on a previously unexploited smelt stock

In the years 1989–1993, over 1000 metric tons of fish (392 kg ha⁻¹) was removed by trawls from the Enonselkä basin in Lake Vesijärvi in order to enhance the water quality. Roach (52%) and smelt (28%) were the most abundant species in the catches. In this study, the effects of the mass removal on the previously unexploited smelt stock was assessed. The total smelt catch was c . 282 metric tons. Due to the mass removal, the density of the stock collapsed from c . 46 000 fish ha⁻¹ in 1989 to c . 14 000 fish ha⁻¹ in 1993 ( M =1.0). The biomass of the stock decreased from 75 kg ha⁻¹ in 1989 to 12 kg ha⁻¹ in 1993. The age group composition of the stock changed clearly towards the dominance of younger age groups. The male/female relationship in the samples changed from clear dominance of females to the dominance of males and the infection rate by the sporozoan parasite Glugea hertwigi declined. The growth rate of the smelt did not change considerably during the years of mass removal. The results suggest that in biomanipulated lakes several mechanisms may prevent the exploited stock from responding to the increased mortality.     

Sex differences in pectoral muscles but not in pectoral fins in the three‐spined stickleback Gasterosteus aculeatus

The pectoral muscle index ( I _PM)( I _PM = 100 M _PM M⁻¹, where M _PM and M are the pectoral muscle and body masses, respectively) fin‐area and fin ray length were studied over a year in male and female three‐spined stickleback Gasterosteus aculeatus from a marine population (Öresund, Sweden) kept under simulated natural light and temperature conditions. A castration‐replacement experiment was used to test androgen effects on the I _PM, fin‐area and fin ray length. Non‐breeding males were castrated or sham‐operated in winter ( i.e . the fish had low levels of androgens). Castrated control and sham‐operated fish were implanted with empty Silastic capsules and castrated groups with capsules containing the androgens testosterone or 11‐ketoandrostenedione into the abdominal cavity. The experiment was terminated after 41 days, when the controls had matured. No morphological differences were found in pectoral fins between sexes during the year, except during the peak breeding season (May), where females showed larger fin‐area and longer fin ray in length compared to males. No effects of androgens treatment or of castration on pectoral fin‐area or fin ray length was observed. Breeding and non‐breeding males showed higher I _PM compared to females. The lower I _PM in females than in males could not be explained by the larger gonads in the former alone, as a sex difference in I _PM was still present after deduction of the ovaries from the female body mass. The I _PM was higher in sham‐operated compared to castrated fish. No effects of androgens treatment on I _PM was observed.     

The accumulation of zinc by O-group plaice, Pleuronectes platessa (L.), from high concentrations in sea water and food

The uptake of zinc by young O-group plaice, Pleuronectes platessa was investigated at levels of high input from sea water and food ( Artemia nauplii). Zinc concentrations in the fish were strongly weight-specific and accumulation effects were distinguished by regression analysis. Sea water was estimated to contribute >10% of the total input up to levels corresponding to high environmental concentrations ( c. 0.1 μg Zn ml⁻¹) and food was the major source of zinc up to sea water concentrations of c. 0.6 μg Zn ml⁻¹. A high degree of regulation obtained along the food pathway to the fish although, in the system studied, most of this regulation was effected by the food organism. Regulation of food input explained the lack of variation in zinc levels previously observed in wild populations.     

Body composition and energy allocation in life-history stages of brown trout

Energy contents of immature parr and smolts, and mature resident and anadromous brown trout Salmo trutta sampled from a small stream in southern Norway were estimated from lipid, protein and carbohydrate concentrations. In immatures the lipid concentrations were highest in parr in the autumn. Mean lipid concentrations increased significantly with age in parr sampled in autumn (1·3% in age 0+ to 3·4% in age 3+), whereas they did not in smolts. The lipid concentrations of parr in spring were not significantly different from those of similarly aged smolts. By contrast, the relative water content (%) decreased with age in parr in the autumn and increased with age in smolts, mean values being slightly higher in smolts (78%) than in parr (77%). Protein and carbohydrate concentrations did not vary with age in the immature fish, mean protein concentrations being 18·0, 17·5 and 16·8% in parr in the autumn and spring, and in smolts, respectively. In residents, the concentrations of lipids were lower and of water higher, in age group 1 than in older fish, whereas there was no significant variation with age amongst anadromous trout. The energy concentration of 2+ smolts (349 kJ 100 g^-1) was similar to that of 0+ parr in the autumn. Mean somatic energy density in autumn was 1·1 times higher in freshwater residents than in parr at age 1+ (407 and 387 kJ 100 g^-1) and marginally different at age 2+ (462 and 426 kJ 100 g^-1, respectively). The energy contents per unit mass of residents were 1·3–1·6 times that of similar aged smolts. Mean somatic energy density of anadromous trout (504 kJ 100 g^-1) was higher than that of residents (455 kJ 100 g^-1). Somatic energy, lipid and protein concentrations were correlated highly with water contents of all life stages, age and sex groups.     

Short-term thermal acclimation induces adaptive changes in the inner mitochondrial membranes of fish skeletal muscle

In the oxidative muscles (musculi laterales superficiales) of crucian carp Carassius carassius acclimated for 6 weeks to either 5 or 25° C, the volume density and the surface density of fibres per tissue did not differ significantly between the control and experimental groups. The correlation ratio (μ²) for these values was below 50, 39·3 and 43·9 respectively. After acclimation to 5° C, the surface density of outer mitochondrial membrane per fibre increased significantly from 0·93 to 1·23m² cm⁻³ in the summer population but dropped from 0·94 to 0·67 m² cm⁻³ in the winter population. The surface density of outer mitochondrial membrane per mitochondrion increased from 3·24 to 4·52 m² cm⁻³ in summer fish. After acclimation to 25° C, the surface density of inner mitochondrial membranes per muscle fibre decreased from 4·04 to 1·79 m² cm⁻³ in summer fish and from 3·86 to 1·07 m² cm⁻³ in winter fish. The surface density of inner mitochondrial membranes per mitochondrion increased from 14·17 to 15·60 m²cm⁻³ in summer fish but dropped from 13·91 to 10·67 m² cm⁻³ in winter fish. Correlation matrices demonstrate a negative correlation of the surface density of outer mitochondrial membrane per mitochondrion with the volume density of mitochondria per fibre and temperature, suggesting cold-induced proliferation of small mitochondria. It was concluded that short-term cold acclimation increased surface area of the inner mitochondrial membranes in summer fish.     

Plasma catecholamines and accumulation of adrenaline in the atrial cardiac tissue of aquacultured Atlantic salmon (Salmo salar) during stress

The effects of stress on plasma catecholamines (CA) and capacity for tissue accumulation of CA were studied in cardiac and skeletal muscle of cultured Atlantic salmon ( Salmo salar L.). Adrenaline (A) and noradrenaline (NA) were quantified by high performance liquid chromatography.
Plasma A and NA levels were 56±10 nmoll⁻¹ and 77±17 nmoll⁻¹ (± s.e.m. ), respectively, in a control group living under normal rearing conditions in a fish farm. Following a ±3 h period of pre-slaughter crowding and handling in the fish farm, plasma A reached 221 ± 72 nmol1⁻¹ with no increase in plasma NA. An 0.5 h period of struggling out of water led to even higher level of plasma A (480 ± 89 nmol1⁻¹), without change in NA.
Skeletal muscle was low in CA (A, 0.07 ± 0.02 and NA, 0.06 ± 0.01 nmol g⁻¹ wet wt). Tissue CA was higher in the atrium (A, 0.47 ± 0.04 and NA 0.94 ± 0.10 nmol g⁻¹) than in the ventricle (A, 0.25 ± 0.03 and NA, 0.30 ± 0.02 nmol g⁻¹). The 0.5 h period out of water and the 0.5-3 h period of pre-slaughter crowding led to accumulation of A, but not NA, in the atrium.
These data show that A, released during stress, accumulates in an undegraded form in the atrial tissue of the Atlantic salmon. This suggests a potent uptake mechanism for A in the atria presumably in the sympathetic nerve terminals. The acumulation of A in the atrium appears to reflect the period of high plasma A during stress.     

Quantitative observations on the pulmonary anatomy of the domestic Muscovy duck (Cairina moschata)

The lungs of five female domestic Muscovy ducks, mean body weight 1.627 kg, total lung volume 48.07 cm³, were analysed by standard morphometric methods. Principal results obtained are: lung volume per unit body weight, 30.17 cm³/g; volume densities of exchange tissue relative to lung volume, 49.24%, blood capillaries relative to exchange tissue, 29.63%, tissue of the blood gas (tissue) barrier relative to exchange tissue, 5.88%; surface area of the blood-gas (tissue) barrier per unit body weight, 30.04 cm²/g; ratios of the surface area of the blood-gas (tissue) barrier per unit volume of the lung and per unit volume of exchange area, 979 cm²/cm³ and 200.06 mm²/mm³, respectively; harmonic and arithmetic mean thicknesses of the tissue barrier, 0.199 μm and 0.303 μm, respectively. The anatomical diffusing capacity of the tissue barrier for oxygen ( DtO₂ ) and the total pulmonary diffusing capacity ( DLO₂ ), 49.58 ml O₂/min/mmHg/kg and 4.55 ml O₂/min/mm Hg/kg, respectively. The lungs of the domestic Muscovy duck appear to be about as well adapted anatomically for gas exchange as the lungs of wild anatid species, and there is no clear evidence that domestication has been associated with any deterioration in the anatomical capacity for oxygen uptake. The weight-specific anatomical diffusing capacity of the lung for oxygen ( DLO₂/W ) was about 3.6 times greater than the weight-specific physiological value, a factor which falls within the expected range.     

Physiological characteristics of wild Atlantic salmon post-smolts during estuarine and coastal migration

Changes were measured in some of the major physiological variables associated with seawater adaptability, growth and energetics in wild Atlantic salmon Salmo salar smolts and post-smolts migrating from the river and through the estuary, fjord and coastal areas in the River Orkla and the Trondheimsfjord, Norway during late May to early June. Gill Na⁺,K⁺-ATPase activity increased to levels of 12–16 µmol ADP mg protein⁻¹ h⁻¹ in post-smolts caught in higher salinity zones, probably representing long-term levels of Atlantic salmon post-smolts in oceanic conditions. Muscle moisture was regulated within narrow limits (77·7–78·7%) in fish from all zones during both years, suggesting that post-smolts adapt to marine conditions without any long-term disturbance of hydro-mineral balance. Lipid and glycogen content showed a general trend towards depletion from the river, through the fjord and into the ocean. There was, however, no significant change in protein content. The present results confirm that smolts are naturally 'energy deficient' during downstream migration, and suggest that post-smolts also mobilize energy reserves during their early marine phase, while protein is allocated for somatic growth. Plasma growth hormone (GH) levels increased transiently during passage through the estuary and fjord, with lower levels observed in post-smolts caught off-shore, i.e. in fish which were feeding on marine prey and had adapted to the marine environment. These physiological changes may confer substantial selective advantages during the critical early marine phase of anadromous salmonids, and hence are adaptive for long-term survival in sea water.     

Physiological impact of sea lice on swimming performance of Atlantic salmon

Atlantic salmon Salmo salar were infected with two levels of sea lice Lepeophtheirus salmonis (0·13 ± 0·02 and 0·02 ± 0·00 sea lice g⁻¹). Once sea lice became adults, the ventral aorta of each fish was fitted with a Doppler cuff to measure cardiac output ( ̇ ), heart rate ( f _H) and stroke volume ( V _S) during swimming. Critical swimming speeds ( U _crit) of fish with higher sea lice numbers [2·1 ± 0·1 BL (body lengths) s⁻¹] were significantly lower ( P  < 0·05) than fish with lower numbers (2·4 ± 0·1 BL s⁻¹) and controls (sham infected, 2·6 ± 0·1 BL s⁻¹). After swimming, chloride levels in fish with higher sea lice numbers (184·4 ± 11·3 mmol l⁻¹) increased significantly (54%) from levels at rest and were significantly higher than fish with fewer lice (142·0 ± 3·7 mmol l⁻¹) or control fish (159·5 ± 3·5 mmol l⁻¹). The f _H of fish with more lice was 9% slower than the other two groups at U _crit. This decrease resulted in ̇ not increasing from resting levels. Sublethal infection by sea lice compromised the overall fitness of Atlantic salmon. The level of sea lice infection used in the present study was lower than has previously been reported to be detrimental to wild Atlantic salmon.     

Testicular Development Induced by a Recessive Mutation during Gonadal Differentiation of Female Common Carp (Cyprinus carpio, L.)

The phenotypic effects of a new recessive mutation mas ⁻¹, which in homozygous condition induces testicular development in XX animals of common carp ( Cyprinus carpio L.), are described. Sexual differentiation of XX; mas ⁻⁺/ mas ⁻¹ and XX; mas ⁻¹/ mas ⁻¹ animals was compared with the gonad development of XX wild type females and XY males. In XX females gonadal differentiation starts with the formation of an ovarian cavity and entry into meiosis of germ cells at around 80 days post hatching (ph). Male gonads remain quiescent until 120 days ph during which period they develop a network of loose connective tissue. Spermatogenesis starts with tubule formation and the differentiation of germ cells into spermatogonia type B. Heterozygous XX; mas ⁻⁺/ mas ⁻¹ animals developed as normal females, but in homozygous XX; mas ⁻¹/ mas ⁻¹ animals two types of gonad development were observed. In the first type, germ cells did not enter meiosis until 100 days ph when they differentiated as spermatogonia. An ovarian cavity was not formed but male specific connective tissue developed instead. These gonad developed as normal testes. In the second type, germ cells differentiated at 80 days ph as either oocytes or spermatocytes, which resulted in the gonads developing as ovotestes. The formation of an ovarian cavity was in most cases incomplete. The phenotypic effects of mas ⁻¹ are interpreted as a timing mismatch between mas activation and female sex differentiation.     

Morphology and fine structure of the heart of the burbot, a cold stenothermal fish

The ventricle of the burbot Lota lota heart comprised 0·148 ± 0·006% of the body mass which is nearly two-fold heavier than the relative ventricular mass ( M _V) of other similarly sized teleosts. The shape of the ventricle is pyramidal and the wall is exclusively composed of spongious muscle without a distinct compact layer. The atrium forms 0·017 ± 0·002% of the body mass. Length, width, sarcolemmal surface area and volume of enzymatically isolated myocytes from burbot ventricle were 147·2 ± 10·2 μm, 6·3 ± 0·4 μm, 2440·8 · 251·5 μm² and 2356·8 ± 316·6 μm³, respectively. The myofibrils were peripherally located and their volume density was remarkably high: 65 ± 2 and 68 ± 3% in ventricle and atrium, respectively ( P >0·05). Although not particularly conspicuous, some nonjunctional and junctional sarcoplasmic reticulum (SR) was present in both atrial and ventricular myocytes. The SR formed peripheral couplings with the sarcolemma and the junctional clefts were frequently occupied by foot processes. These findings suggest that cold-adaptation is achieved by cardiac enlargement, high volume density of myofibrils and well-developed peripheral couplings in the SR in the heart of stenothermal burbot.     

Reproductive ecology of cultured fish in the wild

Domestication has been shown to have an effect on morphology and behaviour of Atlantic salmon ( Salmo salar ). We compared swimming costs of three groups of juvenile Atlantic salmon subject to different levels of domestication: (1) wild fish; (2) first generation farmed fish origination from wild genitors; and (2) seventh generation farmed fish originating from Norwegian aquaculture stocks. We assessed swimming costs under two types of turbulent flow (one mean flow velocity of 23 cm s⁻¹ and two standard deviations of flow velocity of 5 and 8 cm s⁻¹). Respirometry experiments were conducted with fish in a mass range of 5–15 g wet at a water temperature of 15° C. Our results confirm (1) that net swimming costs are affected by different levels of turbulence such that, for a given mean flow velocity, fish spent 1·5‐times more energy as turbulence increased, (2) that domesticated fish differed in their morphology (having deeper bodies and smaller fins) and in their net swimming costs (being up to 30·3% higher than for wild fish) and (3) that swimming cost models developed for farmed fish may be also be applied to wild fish in turbulent environments.     

Changes in serum cortisol, metabolites, osmotic pressure and electrolytes in response to different blood sampling procedures in cultured sea bass (Dicentrarchus labrax L.)

This study investigated the effect of five sampling procedures on serum cortisol, glucose, total protein, osmolality, Na⁺, Cl^–, K⁺ and Ca⁺⁺ concentrations in 2-year-old cultured sea bass ( Dicentrarchus labrax L).
Mild disturbance caused by rapid removal of fish and brief handling did not induce significant variation in any of the blood parameters investigated. Confinement and crowding elicited a high and significant increase in serum cortisol, glucose, osmolality, Na⁺, Cl^–, and Ca⁺⁺ concentrations. Exposure to MS 222 (140 mg L^–1) significantly increased osmolality, but not ionic concentration. Site of blood withdrawal (cardiac sinuses/caudal vein) had no effect on the concentration of analysed blood constituents, except for K⁺ levels. Scattered literature of sea bass blood chemistry is reviewed and compared with 'normal' ranges of blood constituents measured in this study. We conclude that it is necessary to select and rigorously execute an opportune blood sampling procedure whenever blood constituents are used as indicators of fish functional state.     

Muscle buffering capacity of yellowtail fed diets supplemented with crystalline histidine

Juvenile yellowtail Seriola quinqueradiata (initial body mass of 22 g) were fed either a commercial diet (control, diet 1) or diets supplemented with histidine (diet 2), histidine+β-alanine (diet 3), or histidine+β-alanine+thyroxine (diet 4), for 6 weeks. The dietary treatment did not affect the final body mass. Free histidine levels of white muscle in the fish fed the diets supplemented with histidine (diets 2-4) were significantly higher (>62 mmol kg⁻¹ of wet tissue) than that of control group (42 mmol kg⁻¹ of wet tissue). Dietary supplementation of β-alanine (diet 3) or β-alanine+thyroxine (diet 4) failed to increase muscle anserine (β-alanyl-π-L.-histidine) level. Muscle buffering capacity of the range from pH 6·0 to 7·5 of the fish fed the diets 2-4 (41·6-42·7 mmol NaOH pH⁻¹ kg muscle⁻¹) reflected the increase of muscle histidine level, having slightly but significantly intensified compared to control fish (36·6 mmol NaOH pH⁻¹ kg muscle⁻¹). Most of the free amino acids other than histidine were significantly lower in the fish fed the diets 2-4 than in control fish. Thus, crystalline histidine supplemented to diets appears to be deposited in muscular tissue, and consequently enhance muscle buffering capacity in this species.