Studies were undertaken to determine the cardiac and ventilatory responses of restrained and unrestrained rainbow trout (Salmo gairdneri) to changes in inspired oxygen and carbon dioxide tensions. The role of blood oxygen carrying capacity in the control of ventilation was investigated, as well as the location and innervation of oxygen receptors activated by hypoxia. Ventilation volume (Vg) was measured directly in restrained fish using a ventilation chamber that separated inspired from expired water, the latter being collected in a graduated cylinder. In receptor localization experiments a wooden tongue depressor held vertically in the buccal cavity in the median plane divided water flows to the gills on each side of the fish. Thus, one set of gills could be irrigated with hyperoxic water to maintain arterial oxygen tension, while hypoxic water was passed over the other set of gills. Blood samples were obtained from cannulae implanted in both the dorsal aorta and right common cardinal vein. Vg increased in hypercapnea (inspired CO₂ tension (PICO₂) 0.5-2.0 kPa) due to increased "stroke volume (frequency remained constant), with higher levels of Vg recorded at higher C0₂ tensions. In fish exposed to PICO₂ levels of 0.5 and 0.9 kPa, raising the inspired oxygen tension (PIO₂) to 60.4 kPa eliminated the ventilatory response to hypercapnea. Hyperoxia had little or no effect on ventilatory responses to (PICO₂) levels of 1.5 and 2.0 kPa. Ventilation volume was inversely related to blood oxygen content (CaO₂) in trout. CaO₂ decreased and Vg increased during hypercapnea (PICO₂ 0.8 kPa), hypoxia (PIO₂12.4 kPa) and anaemia (haematocrit reduced from 22.3% to 14.3%), while CaO₂ increased and Vg decreased during hyperoxic hypercapnea (PIO₂ 60.4 kPa, PICO₂ 0.8 kPa). Increased Vg during hypercapnea is attributed to hypoxaemia produced by Bohr and Root off-shifts which result from increased blood CO₂ tension and reduced blood pH. Oxygen uptake remained constant during all experimental trials, indicating that the manoeuvre of increasing Vg is effective in relieving adverse effects of hypoxaemia. The significance of elevated Vg as a short-term adaptation to hypoxaemia is discussed, Heart rate decreased and ventilation increased in unrestrained fish exposed to gradual hypoxia (PIO₂ decreased from 20 kPa to 4 kPa) at 7°C and 16°C. The initial heart rate of fish acclimated to 16°C was higher than that of the 7°C group, but at the lowest level of PIO₂, heart rates of both groups dropped to the same level. Thus, the cardiac chronotropic response to hypoxia in trout is temperature independent. Receptors causing hypoxic bradycardia are located in the dorsal region of the first gill arch. Hypoxic bradycardia was eliminated by removing the first gill arch, or by sectioning the branches of cranial nerves IX and X innervating the arch. Blood flow through the arch does not appear to be necessary for this response, since ligation of the arch at its ventral insertion on the body wall did not affect hypoxic bradycardia. The pseudobranch has no role in cardiac control since interrupting the flow of blood through, and deafferentation of, the pseudobranch had no effect on the cardiac response to hypoxia. The biological significance of hypoxic bradycardia, and ventilatory-circulatory interaction during hypoxia, are discussed. Ventilatory responses to hypercapnea and hypoxia were unchanged after bilateral section of the nerves to the first gill arch. Receptors in the first gill arch thus have no role in control of ventilation during either hypercapnea or hypoxia. Possible locations for receptors responsible for control of ventilation are discussed. / Science, Faculty of / Zoology, Department of / Graduate

This study was conducted to satisfy three objectives. The first was to provide a detailed description of the differences between the prey contents of rainbow trout (Salmo qairdneri) and kokanee (Oncorhynchus nerka) compared either to each other or to the prey contents of the natural environment. The second was to determine how elements of anatomy, physiology and behaviour interact to promote the acquisition of species specific diets by trout and kokanee. The third was to precisely identify the foraging strategies of the two predators by assessing how different anatomical and behavioural characteristics serve as interrelated adaptations that suit each species to effectively use a specific habitat-prey complex. Matched samples of trout and kokanee from Marion Lake exhibit only modest A dietary overlap (mean of C λ = .462, range.136 to .881). Although kokanee appear to track the environmental abundance of prey more closely than trout, both predators exhibit pronounced patterns of "density independent" acquisition of prey from the total complex of prey that is apparently available in the lake. To test hypotheses about the factors that control these dietary patterns, I conducted a series of studies concerning where trout and kokanee choose to forage, when they choose to forage, how they search for prey, how they attack prey, and how experience in encountering various prey alters the predator's foraging behaviour. Temporal segregation of trout and kokanee foraging activities is not well-developed under field conditions and appears unlikely to promote strong patterns of food-resource partitioning. By contrast, spatial segregation is well-developed and clearly plays a major role in promoting the acquisition of relatively large numbers of nearshore benthic prey ( eg. planorbid snails or odonates ) by trout and of relatively large numbers of offshore, water-column prey ( eg. chironomid pupae ) by kokanee. Close inspection of the details of predator and prey distributions indicates that many aspects of food-resource partitioning are not logical outcomes of spatial segregation. Differences in predator search-techniques do not determine the presence or absence of various prey types in predator diets, however, differences in predator search behaviours do bias them to obtain different quantities of particular classes of prey. Kokanee search procedures allow them to detect prey in both exposed and concealed locations while trout detect only exposed prey. While searching for benthic or lake-surface prey, kokanee maintain search positions which allow them to detect prey of smaller sizes than trout. This clearly favours the trend for kokanee to include greater quantities of small prey (eg. Hyalella sp.) in their diets than trout. Differences in search procedures do not explain why kokanee obtain a greater proportion of their diet than trout from small zooplankton (≤ 1 mm ) in late summer or why kokanee seldom exploit any of the relatively large ( > 4 mm ), armoured prey that are common in the diet of similar sized trout. Differences in both behavioural and morphological characteristics involved in the attack phase of foraging by trout and kokanee serve as the basis for explanations of a number of differences between the diets of free-ranging predators. These differences include: the greater utilization of aerial prey by trout, the inclusion of large numbers of copepods in the diet of kokanee but not of trout, the generally greater utilization of zooplankton by kokanee compared to trout, and the relative-scarcity of large ( > 4 mm ), armoured prey, in the diet of kokanee. A series of laboratory experiments was used to examine the extent to which short term experience might influence food-resource partitioning by trout and kokanee. These experiments offered convincing evidence that differential effects of experience will amplify the trends in resource partitioning already set in motion by differences in habitat selection, search procedures, and attack procedures. I argue that the morphological and behavioural traits that control food "selection" by trout and kokanee in Marion Lake are a consequence of the evolution of mutually exclusive foraging strategies. Trout are portrayed as D-strategists that concentrate on relatively large, dispersed prey for the bulk of their energy requirements. Adaptations which enable trout to differentially exploit large prey include: procedures for area-extensive search; a predisposition to attack relatively large, armoured-prey; large mouth-size; and persistent responses to opportunities to attack large prey. An inability to sustain high attack rates on small prey ( < 1 mm ) at high density ( 35 per liter ) and a tendency to ignore or reject such prey suggest that trout are not well-adapted to exploit relatively small, morphologically-uniform,prey. Kokanee are portrayed as C-strategists which concentrate on relatively small, contagiously-distributed prey for the bulk of their energy requirements. Adaptations which enable kokanee to differentially exploit small prey include: procedures for area-intensive search; a predisposition to attack relatively small, morphologically-uniform prey; small mouth-size; well-developed gill-rakers; and an ability to sustain high attack rates on small planktonic prey. Low ingestion success with a variety of large benthic prey and a tendency to ignore or reject such prey under laboratory conditions where they serve as the sole source of food are evidence that kokanee are not well adapted to exploit large, armoured-prey. Adaptations associated with the search, capture, manipulation and ingestion phases of the foraging cycle appear in each instance to be evolutionary responses to specific features of a given habitat-prey complex. C-selected or D-selected foraging strategies appear to be mutually exclusive evolutionary avenues down which trout and kokanee have been directed by the fundamental nature of a given habitat-prey complex. / Science, Faculty of / Zoology, Department of / Graduate

The effects of steady-state, aerobic swimming exercise upon blood volume and flow distribution in the rainbow trout {Salmo gairdneri) were examined. Isotopic Rubidium-86, and radiolabelled microspheres were injected into trout forced to swim against a current at 80% of their critical velocity (U[sub=crit]) in a Brett-type water tunnel respirometer. The results gathered from experiments using these radioactive tracers within the circulatory system of the trout indicated that blood flow during exercise was redistributed to favour working muscles, at the expense of diminished blood flow to those organs and tissues in the systemic circulation which could tolerate periods of transient hypoxia. Active hyperaemia in the skeletal muscle and vasoconstriction of the coeliacomesenteric artery, via adrenergic receptor mechanisms are proposed as the main sites of the control for blood volume and flow redistribution in the systemic circulation in trout during exercise. The gills of these fish must be able to maintain adequate gas transfer in order to keep pace with the increased metabolic demands of the working muscles during exercise. An isolated, saline-perfused trout head preparation and a spontaneously ventilating, blood-perfused whole trout preparation were developed in order to study how increases in the pulsatility of input and increases in the cardiac output through these gills; cardiovascular alterations known to occur during exercise in vivo in these fish, affect fluid flow distribution through, and within the branchial vasculature, and gas exchange across the gills. Data from these preparations indicated that pulsatility of flow increased venolymphatic fluid drainage from within gill tissues, as well increasing the fluid flux/reflux across the branchial microvasculature. However, these changes in fluid distribution associated with increased pulse pressure did not significantly change the rate of gas transfer across the gills. Although gill vascular resistances to flow were very sensitive to alterations in pulse pressure and flow rate, only perfusion flow rate through the gills could cause significant changes in the rate of mass transfer of gases across the gills. The gills of trout therefore were found to be perfusion and not diffusion limited for gas transfer, under conditions which simulated those found at rest and during exercise, in vivo. It also was shown that, given oxygen uptake and cardiac output data from the literature, combined with those for blood flow redistribution during exercise from the present study, the working muscles, which were operational during steady-state, aerobic swimming exercise in rainbow trout, could account for nearly all the measured increase in the total oxygen uptake at this level of exercise. The circulatory system of the rainbow trout, both branchial and systemic, was shown to be quite efficient in its ability to take up and distribute oxygen to the tissues during prolonged, aerobic swimming exercise. The numerous cardiorespiratory adjustments noted during exercise account for this animal's ability to maintain swimming activity in the face of increased oxygen demands put upon the circulation by enforced activity. / Science, Faculty of / Zoology, Department of / Graduate

Memoria para optar al Título Profesional de ingeniero Agrónomo. Mención: Producción Animal / El color de la carne de salmónidos es uno de los parámetros de calidad más importante, pero su medición es compleja. En el presente estudio se evaluaron los sistemas de medición visual (cartilla Roche™ y regla Salmofan) e instrumental (fotocolorímetro triestímulo Minolta Chroma Meter) del color de la carne en salmónidos. Se estudió la precisión (repetibilidad y reproducibilidad) de ambos métodos, la relación entre estos y la relación entre la medición de color con el contenido de pigmento y lípidos. Además, se comparó la variabilidad entre dos fotocolorímetros con distinta área de medición. La precisión de ambos métodos se evaluó en tres plantas de proceso ubicadas en Chiloé, midiendo filetes de trucha arcoíris (Oncorhynchus mykiss) y salmón del Atlántico (Salmo salar) en forma repetida y aleatoria. Las mediciones se hicieron con la luz normal de la planta y luz D65 en cabina de iluminación controlada. El efecto del área de medición de los fotocolorímetros se evaluó sobre filetes de salmón del Atlántico en el centro Colaco de la empresa EWOS S.A. La relación entre contenido de pigmento, de grasa y color se midió en filetes de salmón Coho (Oncorhynchus kisutch) con ambos sistemas en el laboratorio de la empresa CETECSAL S.A. Además se analizó un set de datos de un proyecto FONDEF previo, en el cual se midió color, astaxantina y lípidos en “steaks” de salmón Coho. La repetibilidad del instrumento fue significativamente mayor a la cartilla Roche™ en la medición de trucha arcoíris y salmón del Atlántico. Las mediciones de salmón del Atlántico con regla SalmoFan no presentaron diferencias significativas con el instrumento. La reproducibilidad de la medición visual no presentó diferencias significativas al cambiar operarios ni al variar el tipo de luz. Las correlaciones entre ambos sistemas de medición de color, y de estos con el contenido de astaxantina y lípidos fueron de baja a mediana magnitud fluctuando entre 0 a 0,62, 0,07 a 0,57 y 0,01 a 0,63 respectivamente. Se concluye que la medición de color es un mal predictor del contenido de astaxantina, independiente del método utilizado. El fotocolorímetro de 50 mm de área de medición, para a*, b* y C* presentó una variabilidad (C.V. a* = 6%, b* = 7% y C* = 6%) significativamente menor al de 8 mm (C.V. a* = 20%, b* = 16% y C* = 16%). Para comparar resultados de medición de color, debe tomarse en cuenta el área de medición del fotocolorímetro utilizado, pues estos no son equivalentes. / Visual color with Roche™ card and Roche Salmofan™ ruler was compared with instrumental color (Minolta Chroma Meter™) measured in salmonid fillets. Precision (as repeatability and reproducibility) of methods, relationship between visual and instrumental measurements, and relationship between color, astaxanthin and lipid content were studied. In addition, variability between two colorimeters with different measurement area was compared. Repeatability of instrumental method was significantly higher to Roche ™ card in rainbow trout and Atlantic salmon samples. Measurements of Atlantic salmon fillet using Salmofan™ ruler did not show significant differences with the instrumental color measurements. There was not significant differences in reproducibility of visual measurement in rainbow trout and Atlantic salmon when changing personnel neither when varying illumination source. The relationship between both color systems, and between color systems, astaxanthin and fat content in Coho salmon ranged from low to medium magnitude (0-0.62, 0.07- 0.57 and 0.01-0.63 respectively). It was concluded that fillet color is a bad predictor of astaxanthin content, irrespective of the method used. Colorimeter with 50mm of measurement area, for a*, b* and C* showed a significant smaller variability compared with the 8mm area (C.V. a*=6, b*= and C*=7 v/s C.V. a*=16, b*=16 and C*=20). Measurement colorimeter area must be taken into account in order to compare results of color measurement .

Memoria para optar al título profesional de: Ingeniero Agrónomo, Mención Producción Animal. / El objetivo de este ensayo fue evaluar la recuperación del ácido eicosapentaenoico (EPA), el ácido docosahexaenoico (DHA) y la respuesta en comportamiento productivo, ante el reemplazo de aceite vegetal por aceite de pescado en la dieta de término de trucha arcoíris (Oncorhynchus mykiss) en condiciones comerciales. Este estudio corresponde a la segunda fase de un ensayo; en la primera fase los peces distribuidos en dos tratamientos, fueron alimentados durante un período de 10 semanas con 2 dietas que contenían diferentes proporciones de aceites vegetales (Dieta 1: 70% aceite de soya + 30% de aceite de pescado, dando origen al tratamiento 1; y Dieta 2: 57% aceite de soya + 13% de aceite de linaza + 30% de aceite de pescado, dando origen al tratamiento 2). Se adiciona un tercer grupo alimentado siempre con aceite de pescado el cual da origen al tratamiento control. En este ensayo, las truchas con un peso promedio de 2100g fueron alimentados con una dieta basada en aceite de pescado por un periodo de 12 semanas. Se analizaron los cambios en el perfil de ácidos grasos en el filete entero y en el aceite extraído, además del efecto en el comportamiento productivo. Se encontró que el contenido de EPA en el filete aumentó en los tratamientos 1 y 2 (37 y 24% respectivamente), con diferencias significativas (P-value<0,05) respecto al valor inicial sólo en tratamiento 1; sin embargo, ambos valores fueron inferiores al control, con diferencias significativas (0,59 tratamiento 1; 0,52 tratamiento 2 vs 0,73 Control). El contenido de DHA se mantuvo, sin diferencias significativas en el tiempo, ni respecto al control al término del ensayo (0,45 tratamiento 1; 0,47 tratamiento 2 vs 0,55 Control). La relación ω-3/ω-6 aumentó en el tiempo (57% tratamiento 1 y 31% tratamiento2) con diferencias significativas (P-value<0,05), siendo mayor la recuperación en las truchas alimentadas sin aceite de lino en el ensayo anterior. Los valores finales de la relación ω- 3/ω-6 fueron inferiores en un 13% al control, con diferencias significativas (P-value<0,05) (0,58 tratamiento 1; 0,59 tratamiento 2 vs 0,67 Control). Se puede concluir que la sustitución de diferentes proporciones de aceites vegetales por aceite de pescado durante 12 semanas previo a la cosecha es insuficiente para recuperar el perfil de ácidos grasos en el filete en truchas a peso de cosecha, identificándose un efecto residual de la alimentación con aceites vegetales de la etapa anterior sobre la resistencia al corte, el color instrumental y la ganancia de peso. / The objective of this study was to assess the recovery of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and the response in productive performance, with the replacement of vegetable oil for fish oil in finishing diet of rainbow trout (Oncorhynchus mykiss), under production conditions. This study is the second part of a trial, in the first phase, fishes were distributed in two treatments were fed over a period of 10 weeks with 2 diets containing different proportions of vegetable oils (Diet 1: 70% soybean oil + 30% fish oil, giving rise to treatment 1, and Diet 2: 57% soybean oil + 13% linseed oil + 30% fish oil, giving rise to treatment 2). It adds a third group always fed with fish oil which gives rise to the control treatment. In this trial, the trout with an average weight of 2100g were fed a diet based on fish oil for a period of 12 weeks. Changes in fatty acid profile in the whole steak and the oil extracted, and the effects on productive performance were analyzed. EPA content in the fillet increased in treatments 1 and 2 (37 and 24% respectively), with significant differences (P-value<0.05) from control only in treatment 1, but both values were lower than control, with significant differences (0.59 treatment 1; 0.52 treatment 2 vs. 0.73 control). The DHA content was maintained, with no significant differences in time, or concerning the control at the end (0.45 treatment 1; 0,47 treatment 2 vs. 0.55 control). The ω-3/ω-6 ratio increased over time (57% treatment 1 and 31% treatment 2) with significant differences (P-value <0.05), where the trouts fed the linseed oil in the previous diet, showed the higher value; nevertheless the final ω-3/ω-6 ratio were lower than control in a 13%, with significant differences (P-value <0.05) (0.58 treatment 1, 0.59 treatment 2 vs. 0.67 control). It can be concluded that the substitution of different proportions of vegetable oils for fish oil during 12 weeks before harvest is insufficient to recover the fatty acid profile in trout fillet, identifying a residual effect of the previous diet with vegetable oils over texture, instrumental color parameters and weight gain.

Kaliumkanäle ermöglichen es, den geladenen Kaliumionen selektiv die hydrophobe Lipidphase von Zellmembranen zu passieren. Sie operieren in ihren Funktionen als Antagonisten von Natrium- und Kalziumkanälen, um die elektrische Erregbarkeit von Zellen zu kontrollieren. Im ZNS der Vertebraten existiert eine Vielzahl verschiedener Kaliumkanäle, welche die zeitliche Struktur von Aktionspotentialen festlegen. Im Rahmen der vorliegenden Dissertation wurden bisher unbekannte Kanäle aus den Familien der spannungsgesteuerten Kv1-/Kv3- und der kalziumsensitiven SK-/BK-Kaliumkanäle aus dem ZNS der Regenbogenforelle kloniert. Diese wurden durch heterologe Expression in Insekten- und Säugerzellen einer biophysikalischen und pharmakologischen Charakterisierung zugeführt. Gegen fünf der bekannten Kanäle wurden spezifische Antikörper hergestellt. Mit Hilfe immunhistochemischen und PCR-Techniken konnte die gewebetypische und entwicklungsspezifische Expression der Kaliumkanäle untersucht werden. Mit den in dieser Arbeit präsentierten Daten konnte die Grundlage für eine differentielle Betrachtung der funktionellen Bedeutung und des Expressionsmusters der entsprechenden Kaliumkanäle im nativen Kontext gelegt werden.

Snack food development studies were conducted to iii utilize trout (Oncorhynchus mykiss) frames, a by-product of the filleting operation, using extrusion and conventional technology. Twin screw extrusion studies were conducted to study the effect of fish mince, non-fat dry milk, process temperature, and moisture content on the physicochemical properties of the extruded snack food. Response surfaces were plotted to understand the effects of the independent variables on dependent variables such as bulk density, expansion ratio, shear strength, and water absorption index. Quadratic models expressed the relationship between the dependent and independent variables. Based on the extrusion studies, conditions suitable for further development of a ready-to-eat snack food were obtained. Conventional technology was also studied in the development of a fish cracker called keropok. A well expanded, tasty snack food was obtained using the minced fish and tapioca starch. Physicochemical characteristics of the developed snack were determined. Taste panel ratings for texture and taste of the cracker indicated a good potential for acceptance of this product for production and sale by local fish processors. Further studies may be undertaken to develop a continuous process to prepare the crackers on a larger scale.

<p>Teleosts take up metals by two major pathways: gills and/or gut. Past research is heavily focused on branchial uptake despite evidence that the gastro-intestinal tract (GIT) is the dominant route in some natural environments. To address this information gap, my thesis characterizes uptake mechanisms of Cd and Ca along the GIT of <em>Oncorhynchus mykiss</em>. Toxic effects of Cd, protective effects of Ca against Cd uptake, and Cd distribution within fish after ingestion are also explored.</p> <p>Four-week dietary Cd exposure affected growth and Ca regulation, while causing toxicity at the subcellular level. Elevated Ca in diets protected against accumulation and altered subcellular handling of Cd. Pre-exposure to different diets changed unidirectional uptake and binding rates of Cd and Ca, although they remained highly correlated. Binding rates of Cd to mucus can predict absorption rates of Cd<strong>.</strong></p> <p>A variety of <em>in vitro </em>and <em>in vivo </em>experiments on four distinct GIT segments (stomach, anterior-, mid-, and posterior- intestine) were undertaken to determine specific mechanisms of Cd and Ca uptake in freshwater trout. Cd transport was unaffected by solvent drag, but was stretch- and temperature- sensitive. Strong evidence for a common pathway for Ca and Cd transport was obtained. Cd also appeared to be taken up in part by zinc (ZIP-like) transporters and the divalent metal transporter DMT1. Ca uptake along the GIT appeared to be carrier-mediated, time- and concentration- dependent, but was not affected by solvent drag, or by Na concentration. Mucosal Cd did not inhibit Ca uptake. Ca, but not Cd, uptake rates were much lower in sea water-acclimated trout. Seawater and freshwater fish accumulated similar whole body Cd concentrations when fed contaminated diets for three weeks, but the majority of Cd in seawater trout remained in the posterior intestine tissue, while freshwater trout had higher internal burdens.</p> <p>This thesis advances the knowledge of metal uptake along the GIT of trout.</p> / Doctor of Science (PhD)

<p> The aim of this research was to determine how freshwater fish respond to dietary lead (Pb) in their environment with a respect to both physiological and toxicological endpoints. Previous acute exposure studies have shown that waterborne Pb accumulates in the internal tissues, resulting in hematological alterations and disruptions in Na+, cr and Ca2+ ion-regulation, with waterborne Ca2+ being protective against Pb tissue burden. In comparison with other metals such as silver, zinc, cadmium and copper, very little is known about Pb toxicity via the trophic food chain. </p> <p> Using rainbow trout as the model freshwater fish species, it was discovered that dietary Pb accumulated mainly in the gastrointestinal tract, with the intestine and the stomach being suggested as the primary tissues involved in Pb toxicity. Mild physiological disruptions in terms of plasma Ca2+, Mg2+ and Na+ levels were evident. On a per fish weight basis, the bone accumulated the greatest Pb burden (38%) followed by the anterior intestine (19%) and white muscle (12%). The order of Pb accumulation reflected the exposure pathway of dietary Pb in the internal tissues after 42 days; anterior intestine> stomach> mid intestine> posterior intestine> bone> posterior kidney> anterior kidney> liver> spleen> gill> carcass> brain> white muscle (JJ.g Pb/g tissue wet weight). Despite this accumulation of Pb into the internal tissues, there were no effects on growth and survival rates, indicating that growth is not a sensitive indicator of dietary Pb toxicity in fish. Elevated dietary Ca2+ levels had an overall effect in reducing dietary Pb burdens in the whole body and most of the internal tissues analysed. The red blood cells (RBC) accumulated 99% of the Pb in the blood, with less than 1% in the plasma. The enzyme 8-aminolevulinic acid dehydratase (ALAD), important in hemoglobin synthesis was significantly inhibited by the end of the experiment in fish exposed to high concentrations of dietary Pb. In contrast to waterborne Pb studies, ALAD was not a sensitive biomarker of dietary Pb toxicity. </p> <p> Overall, this study has shown the mild physiological and toxicological disturbances of dietary Ph exposure to juvenile rainbow trout. It encourages studies on transport mechanisms at the intestine and in the blood of fish exposed to dietary Pb. It is the hope that the work from this thesis will be used in risk assessments and environmental policies that mitigate the toxic effects of dietary Pb in the aquatic environment. </p> / Thesis / Master of Science (MSc)

In aquatic environments Cu is both a vital nutrient and an important toxicant. Consequently fish require Cu as a micronutrient and can obtain this metal from either water or their diet. Inadequate intake of Cu is associated with reduced growth and development, while decreased growth rates, mortality and reduced swimming capacity have been reported in fish when Cu accumulates in excess of cellular needs. Characterization of Cu uptake is therefore critical in understanding the dynamics that govern toxicity and the risks associated with exposure to an aquatic contaminant. While mechanisms of waterborne uptake and toxicity are well understood, far less is known about gastrointestinal Cu uptake in fish. In vivo and in vitro techniques were therefore used in this study to investigate dietary Cu uptake in freshwater rainbow trout. The mid and posterior regions of the intestine emerged as important sites for Cu absorption in trout, while the role of the stomach and anterior intestine in Cu absorption requires further investigation. The intestinal uptake route was kinetically characterized as a low affinity absorption pathway as compared to the branchial route. Cu uptake appeared to occur via a hypoxia-resistant, carrier-mediated, saturable process which could be fueled by Cu(II)^2+ at concentrations typically found in the fluid phase of chyme in the trout intestine. Experimental manipulation of mucosal NaCl levels stimulated Cu uptake, Na2SO4 had an identical effect, implicating Na rather than the anion. These responses were unrelated to solvent drag, osmotic pressure or changes in TEP. The presence of excess luminal Ag and L-histidine stimulated Cu and Na uptake indicating that a portion of Cu transport was mediated by a Na-Cu co-transport system. Partial inhibition of Cu and Na uptake by phenamil and hypercapnia stimulated Na and Cu transport suggest Cu entry could also occur via the apical Na channel. The Na-dependent mechanism thus either involves more than one component or a unique Na-Cu co-transport mechanism with these combined characteristics mediates part of Cu uptake. Cu uptake was sensitive to pH and competed by Fe and Zn implicating DMT1 in the transport of Cu in the trout intestine. These factors had no effect on Na uptake, leading to the identification of a Na-independent mechanism for Cu uptake in the trout intestine. While the Na dependent nature of Cu uptake and Ag stimulated Cu transport argue against a role for Ctrl in this process, Cu transport characteristics identified in this study compare well with a recently identified Cu transporter in Ctr1 deficient mouse embryonic cells, indicating the existence of a similar transport mechanism in the trout intestine. / Thesis / Master of Science (MSc)