Effects of modulatory agents on neurally-mediated responses of trout intestinal smooth musclein vitro

Mediators and mechanisms responsible for the inhibitory modulation of trout intestinal smooth muscle were examined using a series of putative mediators and substances known to modulate neurotransmission in mammalian systems. Frequency response relationships to transmural stimulation and concentration response relationships to 5-hydroxytryptamine, carbachol, and substance P were established on paired segments of rainbow trout intestinein vitro in the presence and absence of putative modulatory agents. Modulation of neurally-mediated contractions of trout intestine was achieved with dibutyryl cyclic AMP and forskolin, agents that increase intracellular levels of cyclic AMP. The effect appears to be at the level of the smooth muscle, since the adenylate cyclase activator, forskolin, inhibited muscarinic and serotoninergic contractions as well as transmurally stimulated contractions. Substance P-induced contractions were unaffected by forskolin. The endogenous agonists/neurotransmitters which would increase cyclic AMP levels in rainbow trout intestinal smooth muscle are as yet unknown. The effects do not appear to be modulated by vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), or agents that activate -adrenoceptors. Prostaglandin E₂ (PGE₂) and ₂-adrenergenic agonists are possible agents which will decrease contractility of the smooth muscle. They were only active in the proximal intestine and on transmurally stimulated contractions. The effects of both PGE₂ and ₂-agonists appear to be prejunctional, decreasing release of contractile neurotransmitters in the enteric nervous system.     

Changes in smooth muscle contractility of rainbow trout (Oncorhynchus mykiss Walbaum) intestine during acclimation to altered temperature

The effects of altered water temperature in vivo on in vitro smooth muscle contractility of rainbow trout intestine were investigated. Temperature has a significant effect on receptor-mediated intestinal smooth muscle contractility in the rainbow trout. The efficacy of 5-HT, carbachol, and transmural stimulation increased with temperatures above 10°C, with an optimal increase at 15°C. There was also a modest increase in the potency of 5-HT and carbachol within 2 days of establishing trout at 20°C. By day 8, most of these changes had either stabilized or were returning to control values, suggesting that acclimation changes in membranes and enzyme activities were taking effect. However, the contractile responses to carbachol and transmural stimulation were still increasing at this time. This may imply that the muscarinic receptors are more resistant to membrane acclimation changes and may take longer to adapt. Because these experiments were controlled for handling stress and seasonal changes that affect contractility, we have been able to demonstrate some early changes in smooth muscle contractility that occur during acclimation to altered temperature.     

Effects of calcium channel blockers on pharmacologically induced contractions of rainbow trout (Oncorhynchus mykiss) intestine

Calcium depletion/replacement studies were carried out to examine the role of calcium in contraction of trout intestinal smoot muscle in vitro. Three chemically distinct calcium channel blockers were used to determine whether voltage operated calcium channels (VOCs) were involved in calcium entry with either agonist or depolarization-induced contractions. Contractions induced by depolarizing intestinal smooth muscle with potassium were totally dependent on extracellular calcium, whereas receptor-mediated responses to 5-hydroxytryptamine (5-HT) and carbachol also relied on calcium derived from intracellular stores. The calcium channel blockers, verapamil, nitrendipine, and diltiazem, all shifted the calcium-response curve for potassium to the right, supporting the existence of VOCs in trout intestinal smooth muscle. The calcium-response curve for 5-HT was also shifted to the right, suggesting that 5-HT can induce calcium uptake into the smooth muscle via VOCs, in addition to mobilizing intracellular calcium. Verapamil also appeared to block 5-HT receptors directly. Carbachol-induced contractions were only reduced by diltiazem at low concentrations of calcium (0.1–1 mM), suggesting that diltiazem has some other mechanisms of action than the other calcium channel blockers. Activation of muscarinic receptors may induce calcium entry through channels other than the VOCs, in addition to mobilizing intracellular calcium.     

Withdrawal times of freshwater rainbow trout,Salmo gairdneri Richardson,after treatment with oxolinic acid,oxytetracycline and trimetoprim

Abstract. Oxolinic acid (OA), oxytetracyline (OTC) and trimetoprim (TMP) were investigated in whole gutted rainbow trout, Salmo gairdneri Richardson, and in their skin, muscle and blood, at 6, 12 and 18°C. Concentrations of drugs were measured by high-pressure liquid chromatography. It is recommended that withdrawal times for rainbow trout be based on whole gutted rainbow trout and not, as in earlier investigations, from residual concentrations in muscle. Any concentrations in rainbow trout feed were found to be stable during the pelleting process used and up to 81 days after pelleting. The absorption of the drugs in rainbow trout show different temperature dependencies. A quantitative comparison of our overall data from this work with individual results from earlier investigations was undertaken. Suggested withdrawal times for freshwater rainbow trout treated with these drugs are: for OTC, 90 days below 6°C, 70 days between 6 and 12°C and 60 days above 12°C; for TMP, 60 days below 8°C and 40 days above 8°C; for OA, 20 days at approximately 18°C.     

Effects of Oral Administration of Specific Antibodies to Rainbow Trout Oncorhynchus mykiss

A new product called oralized fish serum concentrate (OFSC) was evaluated for a possible effect against various bacterial pathogens in rainbow trout. The OFSC produced from immune trout sera was found to contain fully functional antibodies and complement component C3. The antibodies detected in the serum concentrate were specific to Vibrio anguillarum (O1 and O2) and Aeromonas salmonicida , which had been used for vaccination of the fish prior to serum collection. The functionality of the specific antibodies in OFSC was not reduced after 6 wk storage at -20 C, 5 C, and 20 C. The serum was mixed with commercial trout feed and used for feeding rainbow trout fry (first feed period). After oral delivery of OFSC to rainbow trout for 1 mo, samples of gut content and gut tissue contained functional antibodies. In gutted fish no functional antibodies were found. This suggests that antibodies from OFSC are unable to be transferred across the gut wall in a functional state. Oral administration of OFSC did not increase survival of rainbow trout in an immersion challenge with Vibrio anguillarum .     

Hypothalamic control of prolactin release in the rainbow trout,Salmo gairdneri: in vitro studies

Hypothalamic control of prolactin (PRL) release in immature rainbow troutSalmo gairdneri was investigated using anin vitro perifusion system of the rostral pars distalis. Hypothalamic extract of trout induced a dose-dependent stimulation of PRL release. A similar effect was observed when infusing the medium from a 24h static incubation of the hypothalamus. Extracts from different control tissues (muscle, liver, gut) did not changein vitro release, thus confirming the specificity of this stimulatory effect. Hypothalamic extract from adult male rat, known to contain PRL release inhibiting factors, stimulatedin vitro PRL secretion in rainbow trout. This suggests that PRL cells are predominantly influenced by PRL releasing factors. Measurement of TRH and serotonin content in trout hypothalamus indicated consistent physiological levels of these two factors. HPLC studies of hypothalamic extract showed that immunoreactive — TRH eluted at the same place as labelled TRH standard. Moreover, pizotifen, a serotonin antagonist, partially inhibited the stimulation observed with trout hypothalamic extract. These results suggest that, in immature rainbow trout, PRL release is under stimulatory hypothalamic control and that serotonin and probably TRH play a major role in this control.     

Cellular responses of leucocytes isolated from the gut of rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. Leucocytes were isolated from the gut of rainbow trout by collagcnase digestion. They were shown to be capable of producing a chemoattractant(s) for head kidney leucocytes and a macrophagc activating factor, following stimulation with calcium ionophore and T-cell mitogens, respectively. They also included low but significant numbers of phagocytes, as determined by phagocytosis of fluorescent latex microspheres and oxygen free radical production. However, migration of isolated gut cells in response to head kidney chemo-attractants was lacking. The significance of these results in relation to mucosal immunity is discussed.     

Control of gallbladder motility in the rainbow trout,Salmo gairdneri

Immunohistochemistry revealed nerves containing VIP-like and 5-HT-like material in both gallbladder wall and bile duct of the rainbow trout, while endocrine cells containing gastrin/CCK-like and substance P-like material were present in the mucosa of the bile duct and the duodenum. Fluorescence histochemistry showed adrenergic nerves close to the muscle layer of the gallbladder.Sulphated CCK8, caerulein and non-sulphated CCK8 (in this order of potency), 5-HT and acetylcholine were excitatory on isolated strip preparations, while VIP and adrenergic agonists were inhibitory. The adrenergic drugs were probably actingvia a beta-adrenergic receptor, while the effects of 5-HT and cholinergic drugs were antagonized by methysergide and atropine respectively.Electrical stimulation of the gallbladder nervesin situ failed to show any effect or under certain conditions induced a rebound effect.It is concluded that the motility control of the rainbow trout gallbladder may involve an inhibitory innervation by adrenergic and possibly VIP-releasing nerves, while 5-HT, acetylcholine and a CCK-like substance may be involved in the excitatory control.     

Stable isotopes and gut content show diet overlap among native and introduced piscivores in a large oligotrophic lake

Abstract – In past dietary studies kokanee Oncorhynchus nerka were prominent in the diet of Pend Oreille Lake's large piscivores: native bull trout Salvelinus confluentus, cutthroat trout O. clarki and northern pikeminnow Ptychocheilus oregonensis, and introduced lake trout S. namaycush and Kamloops rainbow trout O. mykiss gairdneri. However, kokanee have declined to 10–20% of their former abundance. We therefore initiated this study to understand current predation demands on kokanee and diet overlap among piscivores, using gut content samples and analysis of stable nitrogen (δ¹⁵N) and carbon (δ¹³C) isotopes from the lake's fish and invertebrate community. In gut content samples, kokanee were the main prey item of large [i.e., ≥400 mm total length (TL)] bull and lake trout; a conclusion that was affirmed by stable isotope analysis. Rainbow trout >500 mm TL consumed mostly kokanee, thus there was a high degree of diet overlap among large bull, lake and rainbow trout. Small (i.e., <400 mm TL) rainbow and cutthroat trout diets overlapped, and were composed mostly of littoral benthic invertebrates. However, gut content and stable isotope analysis did not accord for 400–500 mm TL rainbow trout, small lake trout, and large cutthroat trout. In these instances, a linear mixing model using stable isotope results predicted kokanee consumption for each species, but no kokanee were identified in rainbow or lake trout gut content. Gut content and stable isotope analysis of native northern pikeminnow indicated a diet of mostly littoral benthic invertebrates at smaller (100–150 mm TL) lengths, with kokanee becoming more prominent in the diet of individuals >300 mm TL. Percent of kokanee in the diet of northern pikeminnow has declined from a prior study; otherwise piscivore diets have apparently remained unchanged. In this study, judgments as to the feeding of some piscvores, based on gut content alone, would be tenuous because of small sample sizes, but stable isotope analysis provided an efficient means for confirming diets.     

Necrotic myositis of rainbow trout, Oncorhynchus mykiss (Walbaum): proteolytic characteristics of a crude extracellular preparation from Flavobacterium psychrophilum

A crude extracellular preparation (CEP) from a strain of Flavobacterium psychrophilum recovered from a case of necrotic myositis affecting rainbow trout was capable of causing severe muscle necrosis in rainbow trout following intramuscular injection. Cell wall-associated preparations, however, were unable to produce similar lesions in experimentally injected fish. The CEP degraded gelatin and type II collagen but not type I or type IV collagen. Furthermore, the CEP did not degrade 2-furanacryloyl- l -leucylglycyl- l -prolyl-alanine (FALGPA), chondroitin sulphates A, B or C, heparan sulphate, keratan sulphate, hyaluronic acid, elastin or rainbow trout erythrocytes. The addition of the protease inhibitors 1,10-phenanthroline, ethylenediamine-tetraacetic acid (EDTA) and EGTA to the CEP halted its ability to degrade gelatin in vitro and to produce muscle necrosis in rainbow trout in vivo . In vitro and in vivo activity was restored following the addition of 1 m m zinc chloride to the protease inhibitor-treated CEP, suggesting that this strain of F . psychrophilum secretes a protein complex with zinc metalloprotease-like activity. This protein complex, therefore, appears to be involved in the pathogenesis of necrotic myositis in rainbow trout.     

Alphavirus infections in salmonids--a review

The first alphavirus to be isolated from fish was recorded in 1995 with the isolation of salmon pancreas disease virus from Atlantic salmon, Salmo salar L., in Ireland. Subsequently, the closely related sleeping disease virus was isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), in France. More recently Norwegian salmonid alphavirus (SAV) has been isolated from marine phase production of Atlantic salmon and rainbow trout in Norway. These three viruses are closely related and are now considered to represent three subtypes of SAV, a new member of the genus Alphavirus within the family Togaviridae. SAVs are recognized as serious pathogens of farmed Atlantic salmon and rainbow trout in Europe. This paper aims to draw together both historical and current knowledge of the diseases caused by SAVs, the viruses, their diagnosis and control, and to discuss the differential diagnosis of similar pathologies seen in cardiomyopathy syndrome and heart and skeletal muscle inflammation of Atlantic salmon.     

The adhesion of Vibrio anguillarum to host tissues and its role in pathogenesis

Abstract. Vibrio anguillarum was shown to adhere to excised sections of rainbow trout gut in numbers which were a function of the concentration of bacterial cells and the time for which the tissue was exposed. Using a standardized method for assaying adhered bacteria the course of infection with vibriosis in rainbow trout was studied. It is suggested that the gut may be an important site of bacterial multiplication in slowly developing infections of mature fish. Bacteria adhere to gut sections of fish protected by vaccination substantially less than to sections from non-vaccinated fish.     

Metallothionein induction in cultured fibroblasts and liver of a marine flatfish,the turbot,Scophthalmus maximus

Intraperitoneal injection of turbot with Cd induced the synthesis of a low molecular weight hepatic Cd-binding protein and a 500bp mRNA, which hybridised to a plaice metallothionein (MT) cRNA probe. The Cd-binding protein displayed cross-reactivity in a competitive ELISA with antiserum raised against rainbow trout MT and had the characteristic amino acid composition, metal stoichiometry and spectral characteristics of a Class I MT. Only one isoform was apparent on ion exchange chromatography. Southern blot analysis of DNA cleaved with four restriction enzymes suggested that only a single MT gene is present in turbot.In an established turbot fibroblast cell line, Cd induced MT mRNA and MT levels in a dose and time-dependent manner. MT was also induced by Cu, Hg and Zn but not Pb exposure. Physiological concentrations of glucocorticoids and sex hormones did not induce MT synthesis, although at high concentrations a positive response to corticosterone, dexamethasone, hydrocortisone or progesterone was observedin vitro indicating the possible presence of a functional steroid regulatory element in the fish MT gene.Abbreviations used AMP adenosine monophosphate - CHSE chinook salmon embryo - DMSO dimethyl sulphoxide - HPLC high performance liquid chromatography - KB kenacid blue - MT metallothionein - Mr molecular weight (kDaltons) - NR neutral red - PEG polyethylene glycol - RTH rainbow trout hepatoma - SDS sodium dodecyl sulphate - SSC 0.15M NaCl, 0.015M sodium citrate - TF turbot fibroblast     

虹鳟PPARα基因克隆、序列分析及其组织表达分布

克隆了虹鳟(Oncorhynchus mykiss)过氧化物酶体增殖物激活受体(peroxisome proliferator activated receptor,PPAR)αmRNA全序列。对其序列分析发现,虹鳟PPARα与其他脊椎动物PPARα有较高的同源性,其中mRNA序列有66.4%~97.5%相同,氨基酸序列有69.2%~98.9%相同。DNA结合结构域和配体结合结构域从鱼类到人类高度保守,其中DNA结合结构域有88.0%~98.8%的氨基酸序列相同;配体结合结构域有74.6%~99.6%的氨基酸序列相同。系统发生树分析表明,虹鳟的PPARα基因与同属鲑科鱼类的大西洋鲑(Salmo salar)属于同一分支。实时定量RT-PCR方法分析虹鳟不同组织中的表达发现,PPARα基因在脂肪、肌肉、卵巢、肾脏和肠中表达量较高。     

Effects of dietary supplements on the availability of minerals in fish meal; preliminary observations

Preliminary studies were conducted to determine if several feed supplements with the potential to improve dietary mineral availabilities in fish meal had any measurable effect in fish feeds. In the first study with rainbow trout, 11 supplements were tested: citric acid; sodium citrate; potassium chloride; sodium chloride; histamine dihydrochloride; EDTA disodium salt; sodium bicarbonate; a mixture of amino acids; ascorbic acid; a mixture of inositol and choline; and cholecalciferol. Apparent availability of calcium, phosphorus, magnesium, sodium, iron, manganese and strontium in fish meal-based diets was determined using both yttrium oxide (Y₂O₃) and chromium oxide (Cr₂O₃) as inert dietary markers. Apparent availability was expressed as the fractional net absorption (%) of minerals from diets. After a 7-day acclimation period with test diets, fecal samples were collected for five consecutive days using passive collection systems. Apparent availability of calcium, phosphorus, magnesium, iron, manganese and strontium was increased by citric acid supplementation. Apparent availability of manganese also was increased by EDTA and sodium citrate. The other supplements had no measurable effect on the apparent availability of minerals in fish meal. In the second study, the effect of supplemental citric acid was further investigated using monogastric (rainbow trout) and agastric fish (goldfish). Fish were fed for 5 weeks (rainbow trout) or 3 weeks (goldfish) with fish meal-based diets containing either 0% (control), 2% or 5% citric acid on a dry basis. Feces were collected by settling and by stripping. Apparent availabilities of calcium and phosphorus were greatly affected by citric acid supplementation in rainbow trout but not in goldfish. Phosphorus levels in feces of fish fed a diet with 5% citric acid were approximately half of that of fish fed the control diet (0% citric acid) in the rainbow trout trial. This pattern was consistent during the 5-week feeding trial. A dietary supplement of citric acid as high as 5% did not reduce feed intake or appetite of rainbow trout. Conversely, this level of dietary acidification led to a marked reduction of feed intake in goldfish. Dietary supplementation of citric acid at 2% level did not reduce feed intake of goldfish; however, this level of dietary acidification had little effect on the apparent availability of major minerals in fish meal-based diet. Levels of non-fecal excretion of calcium and phosphorus, inorganic phosphorus in urine, and citric acid in feces were increased in rainbow trout fed 5% citric acid. The pH values of the feces and urine were decreased in rainbow trout fed citric acid. Plasma bicarbonate, plasma calcium and phosphorus, and blood pH of rainbow trout tended to increase by a 5% dietary supplementation of citric acid. The soluble inorganic phosphorus content increased in the diets and decreased in the feces of rainbow trout by supplementing the diet with 5% citric acid. Feces samples of rainbow trout collected by stripping provided similar availability values to data collected by settling for most elements except sodium, which had negative values in all dietary treatments.     

Distended, water-filled stomach in seawater farmed rainbow trout, Oncorhynchus mykiss (Walbaum), provoked experimentally by osmoregulatory stress

The present study evaluates if abdominal dis-tension caused by a water-filled stomach in seawater farmed rainbow trout, Oncorhynchus mykiss (Walbaum), and Atlantic salmon, Salmo salar L., could be provoked experimentally by live chilling in sea water (0.5 °C). Fifty rainbow trout and 50 salmon were visually classified, either as normal or suffering from the condition. Prior to chilling, no rainbow trout or salmon suffered from a water-filled stomach. During chilling, 25% of the rainbow trout and 2.5% of the salmon developed water-filled stomach. Affected rainbow trout had significantly higher blood plasma osmolality and significantly lower body weight than normal trout. The frequency of trout with a water-filled stomach increased significantly with increasing chilling time and increasing plasma osmolality. The regression coefficients revealed that the proportion of affected rainbow trout in-creased by 0.82% h⁻¹ of chilling and by 0.46% per unit increase in mosmol L^−1. The present study revealed that abdominal distension in seawater farmed rainbow trout was far more severe than in Atlantic salmon and could be provoked experimentally by osmoregulatory stress. This is consistent with the observation that water-filled stomach appear only occasionally in seawater farmed Atlantic salmon.     

A case history of whirling disease in a drainage system: Battle Creek drainage of the upper Sacramento River basin, California, USA

Abstract. Whirling disease was diagnosed in steelhead trout, Salmo gairdneri Richardson, during 1985 at Coleman National Fish Hatchery in the Battle Creek drainage of the upper Sacramento River basin, California, USA. Early confirmation of the aetiologic agent, Myxobolus cerebralis Hofer, 1903, was difficult. However, later investigation confirmed the identity of the agent and led to a management decision to destroy the 1985 brood year of steelhead trout at the hatchery. Sentinel fish and collections of feral trout were used to survey Battle Creek watershed to determine the source of infectivity. Feral rainbow trout, Salmo gairdneri Richardson, from South Fork Battle Creek were found to be infected with M. cerebralis and other infected trout were later found at two commercial rainbow trout hatcheries. No reinfection with M. cerebralis has been detected at the Coleman hatchery since ozonation was begun in early 1986.     

Urea and trimethylamine oxide in diets for seawater farmed rainbow trout: effect on fat belching,skin vesicle,winter ulcer and quality grading

Three experiments were conducted in order to evaluate the effects of urea and trimethylamine oxide (TMAO) supplementation in diets for seawater‐farmed rainbow trout Oncorhynchus mykiss. In all experiments, commercial extruded feed was used as basal control diet, whereas experimental diets were the basal diet supplemented with either 10 g kg^–1 TMAO, 10 g kg^–1 urea or 10 g kg^–1 TMAO and 10 g kg^–1 urea. The experiments were conducted both in research units and in a commercial fish farm. Quality was evaluated in three different commercial slaughterhouses, sorting the rainbow trout into superior, ordinary or processing grade. Significantly more TMAO was observed in muscle of rainbow trout given dietary supplementation of TMAO. However, the levels were low compared with levels in marine fish in general. Trimethylamine (TMA) was not found. No increase in muscle urea content was observed in trout fed urea in the diet. Significantly reduced fat belching, seen as reduced number of fat droplets surfacing in the net pen, was observed in groups given TMAO‐supplemented feeds. At least twice as many rainbow trout with skin vesicles were observed in the control group as in the group fed urea‐supplemented diet. In addition, skin vesicles developed into open sores in the winter only in rainbow trout fed control diet. In a large‐scale trial, quality grading of almost 200 000 market‐sized rainbow trout showed significantly more (88 vs. 83%) trout of superior grade in the group fed dietary urea compared with fish fed control diet. The main reason for downgrading from superior to ordinary and processing grade was skin lesions, supporting the observations made in the experimental units.     

Distribution of adipocyte-related cells in skeletal muscle of rainbow trout Oncorhynchus mykiss

Adiponectin is one of the adipokines secreted mainly from adipocytes in mammals. In rainbow trout, however, adiponectin is highly expressed in skeletal muscle. Although it has been suggested that fish skeletal muscle contains adipocytes, their endocrine function and distribution are poorly understood. Recently, an EST analysis of rainbow trout found that heart-type fatty acid binding protein (H-FABP), a member of the intracellular fatty acid binding protein family, encodes an adipose-specific gene. In this study, we produced anti-adiponectin and H-FABP antibodies and investigated the distribution of adipocytes and related cells in skeletal muscle of rainbow trout. The adiponectin signal was detected at around 75 kDa in muscle in Western blotting. Since the molecular mass of rainbow trout adiponectin is around 25 kDa, this 75 kDa band would be a trimer. For H-FABP, the signal band was detected at around 15 kDa. Immunohistochemistry of skeletal muscle sections indicated that adiponectin and H-FABP signals were present outside of muscle cells and throughout the muscle tissues, suggesting the existence of adipocyte-related cells in these regions. These results will contribute to our understanding of energy metabolism in fish skeletal muscle.     

Relationship between stream temperature, thermal refugia and rainbow trout Oncorhynchus mykiss abundance in arid-land streams in the northwestern United States

Abstract – Warm stream temperatures may effectively limit the distribution and abundance of Pacific salmon Oncorhynchus spp. in streams. The role of cold thermal refugia created by upwelling groundwater in mediating this effect has been hypothesized but not quantitatively described. Between June 21 and September 15, 1994, rainbow trout O. mykiss abundance within 12 northeast Oregon (USA) stream reaches was inversely correlated with mean ambient maximum stream temperatures ( r =−0.7, P <0.05). Some rainbow trout used thermal refugia (1–10 m² surface area) that were on average 3–8°C colder than ambient stream temperatures. Within the warmest reaches, high ambient stream temperatures (>22°C) persisted from mid-June through August, and on average 10–40% of rainbow trout were observed within thermal refugia during periods of midday maximum stream temperatures. Frequency of cold-water patches within reaches was not significantly associated with rainbow trout density after accounting for the influence of ambient stream temperature ( P =0.06; extra sum of squares F -test). Given prolonged high ambient stream temperatures in some reaches, the thermal refugia available in the streams we examined may be too small and too infrequent to sustain high densities of rainbow trout. However, these refugia could allow some rainbow trout to persist, although at low densities, in warm stream reaches.