Freshwater Trout Research Articles

Impact of glutaraldehyde versus glutaraldehyde-formaldehyde fixative on cell organization in fish corneal epithelium

The purpose of this study was to examine the impact of a low osmolality glutaraldehyde fixative and a high osmolality glutaraldehyde-formaldehyde fixative on the structural organization of a tissue that could be exposed to low and high osmolality environments. The corneas of freshwater trout were prepared for transmission and scanning electron microscopy using either a fixative of 2% glutaraldehyde in 60 mM cacodylate buffer (pH 7.8, 260 mOsm/l) or a fixative prepared by adding 2.5% glutaraldehyde to a solution of 1% formaldehyde and buffering the solution with 0.1 M cacodylate (pH 7.6, 850 mOsm/l; Karnovsky-type fixative). The corneal epithelial cell layer thickness was greater after glutaraldehyde compared to glutaraldehyde-formaldehyde fixation (67 vs 55 μm), as was the thickness of the superficial cells (5.1 vs 3.4 μm) and basal cells (43 vs 38 μm). The intermediate (wing) cells of the epithelium were, however, less thick after glutaraldehyde fixation (15 vs 18 μm). The width of the squamous, intermediate and basal cells was greater following glutaraldehyde fixation with the effect being greatest in the superficial layers and insignificant at the level of the basal cells. The results show that chemical fixatives with extremes of osmolality cannot only produce different cell sizes in a tissue but also determine the overall organization of the cells in a positional-dependent fashion.

Influence of spoilage and processing temperature on the quality of marine fish protein sources for salmonids

Press cake meals were prepared from previously frozen herring immediately following thawing and after storage for 8 or 12 days at 2-5°C. Each of the raw sources of herring was subjected to two processing temperatures, 75°C and 100°C, during meal preparation. Also, protein hydrolysates were prepared using ocean perch when fresh or after storage at 2-5°C for 4 or 8 days. Subsequently, each of the three hydrolysates was dried at 85°C or 93°C. In two separate experiments, each of the herring press cake meals and dried perch protein hydrolysates was blended with a reference diet in a 30:70 ratio (test protein source: reference diet). All diets contained 5 g kg−1 chromic oxide as an indigestible marker. The reference diet and all test diets were provided to satiation to chinook salmon in salt water and rainbow trout in fresh water, with digestibility of organic matter, protein and energy measured by difference. Digestibility of protein was also measured by the pH-stat and dilute pepsin solubility in vitro techniques. The results indicated that variation in processing temperature to a maximum of 100°C had little effect on digestibility of marine fish protein sources. By contrast, raw material storage for 8 days or more at 2-5°C prior to processing was found to reduce organic matter digestibility and sometimes nitrogen digestibility in salmonids. In vitro measures of digestibility were of little help in predicting the nutritive value of the test protein sources. Cadaverine level in herring press cake meal was shown to be a good indicator of spoilage in the raw material.

Parasite Infections in Danish Trout Farms

Samples from 5 Danish freshwater trout farms rearing rainbow trout (Oncorhynchus mykiss) were examined for parasite infections from October 1993 until November 1994 and recorded parasites are listed. In addition, results from an examination of a mariculture net cage system are presented as well. A total of 10 metazoan and 10 protozoan parasites were recorded. The metazoans included Gyrodactylus derjavini, Gyrodactylus salaris, Eubothrium crassum, Triaenophorus nodulosus, Proteocephalus sp., Diplostomum spathaceum, Tylodelphus clavata and Argulus foliaceus from the freshwater farms. The protozoans Hexamita salmonis, Ichthyobodo necator, Ichthyophthirius multifiliis, Apiosoma sp., Epistylis sp., Trichodina nigra, T. mutabilis, T. fultoni, Trichodinella epizootica, and an Ichthyophonus like intestinal parasite were also detected in the freshwater trout farms. Based on lectin binding studies, few fish were found positive for the myxosporean parasite PKX although no clinical cases were reported. In the mariculture system, Lepeophtheirus salmonis and Caligus elongatus were found.

Influence of improved feed quality and food conversion ratios on phosphorus loadings from cage culture of rainbow trout, Oncorhynchus rnykiss (Walbaum), in freshwater lakes

The expansion of freshwater cage culture in Scotland during the 1980s led to concern with respect to the impact of nutrient, particularly phosphorus (P), discharges on water quality in lakes. The primary route by which P enters the aquatic environment from cage farms is through the feed administered to the fish. In recent years, there have been considerable technological advances in feed manufacturing and improved feeding practices at farm level. This paper investigates how P inputs to freshwater lakes from cage farms may be reduced through changes in diets and improved food conversion ratios. The results demonstrate that there has been a significant improvement in the P content of freshwater trout diets in recent years and that it is possible to reduce waste P loadings from cage sites through the use of better-quality diets and improved feed management. In Scotland, production levels in freshwater cages are commonly set by the regulatory authority assuming a soluble waste P loading of 10 kg P tonne−1 of fish produced. This study suggests that actual soluble waste P loadings are likely to be far lower than this and that, as a result, increases in water column total P levels may be overestimated.

IMMUNOLOCALIZATION OF H+-ATPase IN THE GILL EPITHELIA OF RAINBOW TROUT

The localization of proton pumps (H+-ATPase) in gill epithelia of rainbow trout [Oncorhynchus mykiss (Walbaum)] was elucidated by immunofluorescence microscopy, using rabbit polyclonal antibodies against the 70 kDa subunit of H+-ATPase purified from clathrin-coated vesicles of bovine brain. In the gill epithelia of freshwater trout, the immunostaining was uniformly distributed along the lamellae and generally concentrated in apical regions. It is concluded, therefore, that H+-ATPase is located in the apex of both chloride cells and epithelial cells of freshwater fish. Hypercapnic treatment resulted in a non-polarized and restrictive distribution of H+-ATPase in the chloride cell. No fluorescent staining was observed in the gill epithelium of seawater-adapted rainbow trout, except in some unidentified anucleate surface material. The presence of the 70 kDa subunit in fish gill epithelia was confirmed by Western blot. These results support the proposed role of a proton pump in sodium uptake in freshwater fish and demonstrate that the H+-ATPase in fish gills is of the vacuolar type, antigenically similar to the H+-ATPase in mammalian brain and kidney.

AMMONIA EXCRETION IN FRESHWATER RAINBOW TROUT (ONCORHYNCHUS MYKISS) AND THE IMPORTANCE OF GILL BOUNDARY LAYER ACIDIFICATION: LACK OF EVIDENCE FOR Na+/NH4+ EXCHANGE

Net ammonia fluxes (JAmm) were measured in adult freshwater rainbow trout in vivo under a variety of conditions designed to inhibit unidirectional sodium uptake (JinNa; low external [NaCl], 10(-4) mol l-1 amiloride), alter transbranchial PNH3 and NH4+ gradients [24 h continuous (NH4)2SO4 infusion, or exposure to 1 mmol l-1 external total ammonia at pH 8] and prevent gill boundary layer acidification (5 mmol l-1 Hepes buffer). Inhibition of JinNa with amiloride or low external [NaCl] under normal conditions reduced JAmm by about 20 %, but did not prevent the net excretion of ammonia during exposure to high concentrations of external ammonia. Increasing the buffer capacity of the ventilatory water with Hepes buffer (pH 8) reduced JAmm by 36 % and abolished the effect of amiloride on ammonia excretion. No evidence could be found to support a directly coupled apical Na+/NH4+ exchange. We suggest that any dependence of ammonia excretion on sodium uptake is caused by alteration of transbranchial PNH3 gradients within the gill microenvironment secondary to changes in net H+ excretion. Under normal conditions (pH 8, low external ammonia) gill boundary layer acidification facilitates over one-third of the total ammonia excretion. During exposure to high concentrations of external ammonia in poorly buffered water, estimates of transbranchial PNH3 gradients from measurements of bulk water pH and total ammonia concentration (TAmm) may be grossly in error because of boundary layer acidification. Prevention of boundary layer acidification with Hepes buffer during exposure to high cocncentrations of external ammonia revealed that the local transbranchial PNH3 gradient at the gill may in fact be positive (blood to water), negating the need for an active NH4+ transport mechanism. In freshwater trout, NH3 diffusion may account for all ammonia excretion under all experimental conditions used in the present study.

Effects of seawater‐acclimatization and release sites on survival of hatchery–reared brown trout Salmo trutta

To test whether seawater–acclimatization of hatchery–reared anadromous and freshwater resident brown trout before release increased the survival of adults, smolts were retained 0, 2, 4 and 8 weeks in sea water before release. Total recapture rate increased for smolts retained 4 and 8 weeks in sea water before release relative to the controls. This trend was more pronounced for Freshwater resident than for anadromous stocks, Offspring of anadromous fish stayed longer at sea than offspring of freshwater resident fish. Recapture rates in fresh water were higher for brown trout released in the river than in the fjord in the R, Drammen area, but not in the R. Imsa. In both cases, most fish were recaptured in the sea. Moving into the R. Imsa (relative to other rivers) appeared higher for fish released at the mouth of the river (93%) than in the fjord (47%). Judged from the recapture rates, sea survival appeared to be the same whether released in the fjord or at the mouth of the river.

Induced breeding and larval rearing of the endangered Australian freshwater fish trout cod, Maccullochella macquariensis (Cuvier) (Percichthyidae): B. A. Ingram & M. A. Rimmer, Aquaculture & Fisheries Management, 24(1), 1993, pp 7–17

Induced breeding and larval rearing of the endangered Australian freshwater fish trout cod, Maccullochella macquariensis (Cuvier) (Percichthyidae): B. A. Ingram & M. A. Rimmer, Aquaculture & Fisheries Management, 24(1), 1993, pp 7–17

Distribution, growth and movement of River Usk brown trout (Salmo trutta)

The River Usk catchment in South Wales supports mainly freshwater resident brown trout, with few anadromous fish. Electric fishing and netting revealed that age‐class distribution differed between main river and tributary habitats, the latter environment acting as a nursery area for young fish. Few fry were found at main river sites. Age‐class distribution also differed between tributary systems, and possible reasons are discussed. Trapping experiments indicated that trout move to main river habitat at 2+ yr. Lengths at age (back‐calculated from scale reading) were similar for main river and tributary resident fish at 1 and 2 year, but main river fish were larger at 3 and 4 yr. This habitat shift and size difference is discussed with reference to current angling regulations.

Triiodothyronine is necessary for the action of growth hormone in acclimation to seawater of brown (Salmo trutta) and rainbow trout (Oncorhynchus mykiss)

Brown (BT) and rainbow trout (RT) in freshwater (FW) were treated with ovine growth hormone (GH), GH + iopanoic acid (IOP), and GH + IOP plus triiodothyronine (T3) for RT only. After 1 week of treatment, trout were transferred to 30 o/oo SW and treatment continued. In FW, GH treatment increased significantly plasma T3 level (BT) and T3/T4 ratio (BT and RT) by stimulating T4 to T3 deiodination. In the GH + IOP group, the plasma T3 levels and T3/T4 ratio fell significantly as T4 to T3 deiodination was inhibited. In GH + IOP + T3-treated RT, plasma T3 and T3/T4 ratios increased significantly relative to other groups. No mortality occurred and plasma osmolarity (PO) was not altered by any treatment in FW. After transfer to SW, all IOP + GH trout died within 2 (BT) or 3 days (RT). All GH-treated or control BT survived to the end of the experiment (6 days). RT survival rates tended to be improved in GH and GH + IOP + T3 groups relative to controls. Correlatively on day 1 the PO increase was significantly higher in IOP + GH groups (BT and RT) than in the other groups and significantly lower in GH and GH + IOP + T3 treated RT than in controls from days 1 to 6. These data confirm the requirement of T3 and deiodination of T4 to T3 for the development of hypoosmoregulatory mechanisms in SW as previously shown (Lebel and Leloup 1992). Furthermore, the suppression of the hypoosmoregulatory effect of GH, when conversion of T4 to T3 was inhibited by IOP and the reversal when T3 was added to IOP + GH treatment suggests that GH osmoregulatory action in SW acts via the simulation of T4-5' monodeiodination which increases T3 production.

H+ -ATPase Activity in Crude Homogenates of Fish Gill Tissue: Inhibitor Sensitivity and Environmental and Hormonal Regulation

ABSTRACT N-ethymaleimide-sensitive ATPase activity was measured in crude homogenates of gill tissue from rainbow trout using a coupled-enzyme ATPase assay in the presence of EGTA, ouabain and azide. This NEM-sensitive ATPase activity, determined to be about 1.5 μmolmg−1 protein h−1 at 15°C for freshwater trout, is also inhibited by other H+-ATPase blockers such as DCCD, DES, PCMBS and bafilomycin. It is concluded, therefore, that the NEM-sensitive ATPase activity was generated by a proton-translocating ATPase. Since this NEM-sensitive ATPase was also sensitive to the plasma membrane ATPase inhibitor vanadate, we conclude that the H+-ATPase in fish gill is of the plasma membrane type. The major role of the H+-ATPase in the gill epithelium is to facilitate Na+ uptake from fresh water. Sodium concentration in the external medium was the primary regulator of the H+-ATPase in fish gills, with low sodium levels being associated with high H+-ATPase activity. High external calcium concentration had a marked stimulatory effect on H+-ATPase activity in fish gills when the sodium level was low. Environmental hypercapnia induced a 70% increase in the H+-ATPase activity in fish gills. H+-ATPase activity was also elevated in freshwater fish after chronic cortisol infusion.

Digestibility of various feedstuffs by post-juvenile chinook salmon ( Oncorhynchus tshawytscha) in sea water. 1. Validation of technique

This study was undertaken to assess whether the Guelph system for measuring digestibility in rainbow trout in fresh water was suitable for determining digestibility in chinook salmon held in sea water. Feces were collected from chinook salmon by the Guelph system, the stripping technique (once per week) and intestinal dissection. Validation of the Guelph procedure also involved assessment of: (1) the digestibility coefficients based on total fecal collection versus those based on the indirect chromic oxide indicator method, (2) the extent of nutrient leaching when using the Guelph system, and (3) the effect of level of dietary intake and extent of fish handling (due to stripping of feces) on estimates of nutrient digestibility by the Guelph method. No significant differences were found between the complete collection and indicator methods for measuring apparent nutrient digestibilities. Apparent digestibility values calculated using feces collected directly from the fish by both stripping and dissection were consistently lower than the complete collection and indicator values derived using the Guelph system. The stripping technique yielded the lowest digestibility coefficients. Nutrient leaching occurred when the feces had been exposed to sea water for more than 6 h. This was attributed to incomplete entry of feces into the collection column. Fish handling and concomintant reduction of food intake depressed digestibility coefficients for nutrients and energy and increased fish mortality. It is concluded that the Guelph system of fecal collection involving the use of Cr 2O 3 as the indigestible marker is a potentially reliable digestibility procedure for chinook salmon in sea water. However, care must be exercised to ensure rapid and complete removal of feces to the collection column to minimize nutrient leaching.

Proton Transport and Chloride Cells in the Gill of Rainbow Trout (Oncorhynchus mykiss)

Particulate preparations from freshwater rainbow trout (Oncorhynchus mykiss) gill homogenates contain an active magnesium ion activated ATPase that transports protons into the vesicles. Oligomycin at a concentration of 20 μg∙mL−1had little effect on the proton transport, which was completely inhibited by N-ethylmaleimide. This inhibition was partly counteracted by dithiothreitol. Proton transport in freshwater trout gill submitochondrial particles was completely inhibited by oligomycin. When freshwater trout were kept for 40 min in water equilibrated with air containing 5% carbon dioxide, their gill lamellar epithelium contained many cells densely covered with erect microvilli. After changing to water bubbled with room air, the cells rapidly lost their microvilli. Control fish that were killed immediately after removal from the storage tank also had microvilli, but less erect and fewer in number. Labelled latex microspheres and dextran were used as markers for external medium uptake into gill epithelial cells. The fish were pretreated by bubbling the water by air containing 5% carbon dioxide, and the uptake took place when the water was bubbled with room air.

The physiological responses of freshwater rainbow trout, Oncorhynchus mykiss, during acutely lethal copper exposure

Acutely lethal (24 h) exposure of adult rainbow trout (Oncorhynchus mykiss) to 4.9 μmol copper·l-1 in fresh water (pH 7.9, [Ca2+]≈0.8 mEq·l-1) caused a rapid decline of plasma Na+ and Cl- and arterial O2 tension, and initially a pronounced tachycardia. The internal hypoxia probably resulted from histopathologies observed in the gills of fish exposed to copper, such as cell swelling, thickening and curling of the lamellae, and haematomas. Copper cannot therefore be considered purely as an ionoregulatory toxicant during acutely lethal conditions. Mortality during exposure to copper could not simply be explained by the plasma ionic dilution, nor by the internal hypoxia, since arterial O2 content remained relatively unchanged. Secondary to the ionoregulatory and respiratory disturbances were a number of deleterious physiological responses which included a massive haemoconcentration (haematocrit values as high as 60%) and a doubling of the mean arterial blood pressure. The time-course of these changes suggest that cardiac failure was the final cause of death. In this respect copper exposure resembles low pH exposure in freshwater trout (Milligan and Wood 1982). Copper and H+ appear to be similar in both the primary site of their toxic action (the gills) and the secondary physiological consequences which result from acutely lethal exposures. Furthermore, the acute toxicity syndrome observed may be common to many metals which cause ionoregulatory and/or respiratory problems in freshwater fish.

Induced breeding and larval rearing of the endangered Australian freshwater fish trout cod, Maccullochella macquariensis (Cuvier) (Percichthyidae)

. The endangered Australian freshwater fish trout cod, Maccullochella macquariensis (Cuvier), was the subject of a captive breeding programme to produce fry for reintroduction into the wild. Trout cod broodfish were maintained in earthen ponds for up to 5 years and underwent gonadal maturation each spring but did not spawn in the ponds. Infestations of the protozoan parasite Chilodonella hexasticha caused the death of at least 21 broodfish. Mature fish, removed from ponds when water temperatures had reached or exceeded 16°C, had a higher proportion of atretic oocytes and fewer fish spawned successfully compared to fish removed at lower temperatures. Ovulation was induced by a single injection of 1000–3000 iu/kg HCG. Between 1188 and 11338 eggs ranging from 2·5 to 3·6 mm in diameter were stripped from individual fish. Hatching commenced on days 5–9 and continued for up to 10 days (at 15·5–23°C). Larvae commenced feeding on days 21–25. Trout cod larvae were grown out to fry (363.3–48.6mm total length [TL]) in fertilized fry rearing ponds then released. Between 1986 and 1989, 8420 trout cod fry were released into several sites in the upper Murray River and upper Murrumbidgee River, and reports indicate that released fish are surviving.

Grey heron, Ardea cinerea L., predation at cage fish farms in Argyll, western Scotland

. The aims of the present study were to determine the number and age structure of grey herons, Ardea cinerea L., visiting a large freshwater cage trout farm in a study area in Argyll, western Scotland and to quantify heron predation at the site. Heron abundance was assessed on sea loch shores, running and standing freshwater bodies and at the trout farm between September 1985 and August 1987. Herons visited the farm almost exclusively at night or during twilight periods and, as a consequence, were seen more often by farm staff in winter than in summer. Herons selected cages containing small trout (<300g) and removed them from cages by standing on the top nets and fishing through the mesh. Stock losses were of two kinds; trout were either eaten directly or were dropped by herons during manipulation, when characteristic wounds increased susceptibility to disease or made the fish unmarketable. Adult herons were more successful at feeding than first-years and although younger birds spent more time feeding their intake rates remained lower than those of adults in terms of their total time at the farm. Within a cage, smaller fish aggregated closer to the surface than larger ones and so were more vulnerable to heron predation. In many cases, a high proportion of the fish attacked by herons were blind and/or in poor condition. By recording only wounded fish, farm staff considerably underestimated their losses to herons. Nevertheless, attacks sometimes appeared serious to farmers. However, such losses were small compared with other forms of fish mortality and loss.

17 alpha,20 beta-dihydroxy-4-pregnen-3-one 20-sulphate is a potent odorant in precocious male Atlantic salmon (Salmo salar L.) parr which have been pre-exposed to the urine of ovulated females.

Electrophysiological recordings from the olfactory epithelium have shown that 17 alpha,20 beta-dihydroxy-4-pregnen-3-one 20-sulphate (17,20 beta-P-sulphate; a conjugate of the oocyte-maturation-inducing steroid in teleosts) is a potent odorant in precocious male Atlantic salmon (Salmo salar L.) parr. However, the olfactory epithelium of these fish only appeared to be responsive to the steroid after stimulation with the urine of ovulated female Atlantic salmon. Immature fish did not respond at any time. Stimulation with urine from immature and precocious male Atlantic salmon parr did not make the olfactory epithelium of precocious male salmon parr responsive to the steroid. 17,20 beta-P-sulphate was found in the urines of ovulated females, precocious male parr and mature male Atlantic salmon. The findings are discussed in relation to the possible role of 17,20 beta-P-sulphate in the physiology of Atlantic salmon.

Transbranchial ammonia gradients and acid-base responses to high external ammonia concentration in rainbow trout (Oncorhynchus mykiss) acclimated to different salinities.

Transbranchial ammonia gradients and blood acid-base status have been examined in rainbow trout acclimated to fresh water (FW), 33% sea water (33% SW) and sea water (SW) and exposed to 1.0 mmol l-1 total ammonia (TAmm) at pH 7.9 for 24 h. At all three salinities trout maintained large negative (inwardly directed) NH3 and NH4+ gradients throughout the exposure, presumably by active excretion of NH4+ to counteract the passive inward diffusion of ammonia. Analysis of blood non-respiratory acid-base status (delta H+m) revealed an acid load in FW trout and a base load in SW trout following 24 h of exposure. This indicates that active NH4+/H+ exchange predominates in FW whereas NH4+/Na+ is the principal exchange utilised in SW under these experimental conditions. The plasma TAmm load incurred during ammonia exposure increased with salinity. Compared to FW trout, plasma TAmm values were 34 and 73% higher in the 33% SW and SW trout, respectively, after 24 h. This cannot be explained by differences in the prevailing transbranchial PNH3 gradient because ambient PNH3 was substantially lower at the higher salinities (due to higher pK' and solubility values). We interpret the difference between FW and SW trout as an increased permeability to NH4+ in fish acclimated to the higher-salinity environments. Transbranchial diffusion of NH4+ is, therefore, probably more important as a route for ammonia excretion in SW than in FW trout, especially considering the favourable transepithelial potentials normally found in SW teleosts. In addition, increased NH4+ permeability implies that the toxicity of ammonia will be greater in seawater than in freshwater teleosts and should not simply be measured as a function of the unionised ammonia concentration when considering seawater-adapted species.

Calcium transport by isolated skin of rainbow trout.

The skin overlying the cleithrum bone of freshwater-acclimated rainbow trout contains numerous mitochondria-rich (MR) cells, as detected by DASPEI fluorescence. This tissue was mounted in vitro in an Ussing-style chamber with fresh water on the mucosal surface and saline supplemented with bovine serum albumin on the serosal surface. The preparation developed a high transepithelial resistance and a small transepithelial potential (Vt), positive on the serosal side. Radioisotopic flux measurements indicated that the preparation actively transported Ca2+ from the mucosal to the serosal surface, as assessed by the Ussing flux ratio criterion. Ca2+ transport was positively correlated with MR cell density. Cortisol pretreatment in vivo reduced MR cell density and increased Vt but did not significantly alter Ca2+ fluxes. Ca2+ transport was unaffected by adrenergic agonists (10(-5) mol l-1 adrenaline, clonidine, isoprenaline) or cyclic AMP stimulants (10(-3) mol l-1 dibutyryl cyclic adenosine monophosphate, db-cAMP, plus 10(-4) mol l-1 isobutylmethylxanthine, IBMX) applied to the serosal surface. The Ca2+ ionophore ionomycin (1 x 10(-6)-3.2 x 10(-6) mol l-1 on the mucosal surface) increased both unidirectional Ca2+ fluxes and caused Ca2+ to accumulate within the epithelium. Lanthanum (10(-4) mol l-1) did not inhibit unidirectional Ca2+ fluxes, but apparently displaced Ca2+ from binding sites on the mucosal surface. Unlike Ca2+, movements of Na+ and Cl- across the epithelium were passive, as assessed by the flux ratio criterion, and neither adrenaline nor db-cAMP plus IBMX had any effect on Na+ or Cl- fluxes or electrical properties. These results indicate that ion transport across the skin mediated by MR cells ('chloride cells') contributes to Ca2+ but not to NaCl balance in freshwater trout.

Mechanisms of Cl- uptake through brush border membranes isolated from the posterior intestine of the freshwater trout, Oncorhynchus mykiss, R.

This paper presents a study of the mechanisms of Cl- transport through the brush border membranes of the posterior part of the intestine in the freshwater trout, Oncorhynchus mykiss. The mechanisms for Cl- transport in the posterior intestine are distinct from those in the middle intestine; an inwardly directed pH gradient stimulates Cl- uptake by brush border membrane vesicles, indicating a Cl-/OH- exchange. A pH-regulated Cl- conductance is present, which is not activated at normal intracellular pH. Cl- uptake is stimulated by an outwardly directed HCO3- gradient revealing the presence of a Cl-/HCO3- exchange but, conversely, Cl- is not exchanged against SO4(2-). In addition, carbonic anhydrase activities have been detected in both the intracellular and extracellular leaflets of the brush border membranes which favour the establishment of a bicarbonate gradient. A model of Cl- transport mechanisms through the brush-border membranes of the posterior intestine of the freshwater trout is proposed.