I investigated the effect of turbidity on the foraging behaviour of juvenile Chinook salmon (Oncorhynchus tshawytscha) in the laboratory. Specifically, I examined a behavioural "tradeoff" between visual ability and "perceived" risk. I assessed visual ability by measuring the reaction distance of juvenile Chinook to planktonic Artemia prey. I found Chinook exhibited a log-linear decline in reaction distance with increasing turbidity. To determine the effects of turbidity and microhabitat on foraging rate, I conducted separate experiments for surface (Drosophila), planktonic (Artemia), and benthic (Tubifex) prey across a range of turbidity levels (0, 25, 50, 100, 200, 400, 800 mg•L⁻¹). Foraging rates were reduced at higher turbidity conditions for all three prey. However, for surface and benthic prey, foraging rates were also low in clear water; highest rates were attained at intermediate turbidity levels (50-200 mg•L⁻¹). The degree to which intermediate turbidities were associated with higher foraging rates was size-dependent. Smaller individuals (150-57 mm FL) exhibited relatively higher foraging rates in clear conditions than did larger individuals. However, planktonic foraging rates by juveniles were consistently high in clear water, regardless of fish size. In experiments manipulating light level independent of turbidity, I allowed salmon to forage under conditions which were either turbid, or clear but with light intensity correspondingly reduced. Foraging rates were similar between the two treatments for planktonic prey, but differed for benthic and surface prey. Generally, foraging rates exhibited by juvenile Chinook salmon could not be explained on the basis of visual ability alone. I suggest that young salmon also exhibited foraging behaviour consistent with their perception of risk to predation. In arena experiments, juvenile Chinook distributed themselves randomly in turbid conditions; in clear conditions they associated with the bottom. When bird and fish predator models were introduced the fish altered their spatial distribution, occupying deeper regions regardless of turbidity. However, their response in turbid conditions was less marked and lasted for a shorter time. Turbidity apparently mitigated the perceived risk of predation in juvenile Chinook. I developed a conceptual tradeoff model that predicted the general effect of turbidity on foraging behaviour. Assuming differences in either prey quality or perceived risk of predation in three microhabitats (surface, water column, bottom), the model resolved the apparent dissimilarities between planktonic and other foraging behaviours. Perceived risk of Chinook to predation was significantly different between surface and water column microhabitats. When exposed to a non-visual "fixed-risk" stimulus (sound), salmon apparently perceived less risk as turbidity increased. I conclude that in turbid waters juvenile salmon exhibit foraging behaviour in a manner consistent with a tradeoff between their visual ability and perceived risk. / Science, Faculty of / Zoology, Department of / Graduate

Feed accounts for 40% to 60% of the operating costs of Pacific salmon farms. Presently, commercial diets for Pacific salmon in seawater are formulated according to the nutrient requirements of juvenile chinook salmon and digestibility information derived from studies on rainbow trout in fresh water. Information on feedstuff digestibility by Pacific salmon is completely lacking, regardless of life history stage. Hence, this thesis was undertaken to determine the apparent digestibility of organic matter, crude protein and energy in conventional and novel feedstuffs using post-juvenile chinook salmon (Oncorhynchus tshawytscha) in seawater. The validity of determining apparent nutrient digestibility using the "Guelph system" of fecal collection and chromic oxide (Cr₂O₃) as the indigestible indicator in the diet was assessed in experiment I. In this regard, fecal samples were collected either from a settling column affixed to each novelly designed digestibility tank at 6 and 18 hour intervals or directly from the terminal section of the intestinal tract by stripping or intestinal dissection. Digestibility coefficients were noted to be significantly increased (P<0.001) when the feces remained in the water for 18 hours instead of 6 hours, owing to nutrient leaching. The collection of feces directly from the fish resulted in lower digestibility coefficients than those found when feces were obtained from the "Guelph system". This was attributed to a flaw in the design of the digestibility tank drain system, whereby the feces did not settle quickly into the collection column. The problem was rectified before conducting the subsequent experiments described below. In three additional experiments on chinook salmon in seawater, the overall goal was to ascertain the organic matter, crude protein and energy digestibility coefficients and the digestible energy values for commercial sources of fish meal (herring meal, anchovy meal, menhaden meal, Norwegian low temperature fish meal), poultry by-product meal (two suppliers), feather meal, blood meal, dried whey, canola meal, soybean meal, soybean protein isolate, extruded wheat and wheat middlings. Also, three novel sources of rapeseed protein products (two types of glucosinolate-free canola meal and rapeseed protein concentrate) were evaluated in this regard. The initial minimum size of the chinook in these studies varied between 10.3 g and 40.5 g. The digestibility tank complex consisted of 27 open-circuit 150 L fiberglass digestibility tanks, each supplied with aerated, 8.0°C to 12.5°C filtered seawater at a rate of 6 L/min. The fish were fed by hand twice daily to satiation either a reference diet or a test diet (70% reference : 30% test ingredient). In some instances involving plant protein products, the test ingredient was included at two dietary levels (15% and 30%). Within each test, each diet treatment was assigned to three groups of fish using a completely random or randomized complete block design. Chromic oxide (0.5%) was included in all diets as the indigestible marker. At the end of each experiment, fish were sacrificed and their feces were removed either by stripping or intestinal dissection for comparisons of methodology. In general, the fish meals had higher available energy content for chinook salmon than the other animal and plant protein sources assessed. The importance of screening feedstuffs for potential nutritive value by digestibility measurements was particularly evident from an examination of the digestibility coefficients obtained for menhaden meal versus the other fish meal sources, the two sources of poultry by-product meal, wheat products, canola products and blood meal in this study. The assessment of soybean products, regardless of dietary inclusion level (15% or 30%), could not be ascertained in chinook salmon because of poor diet acceptance. Canola protein sources appear highly promising as partial or complete (in the case of rapeseed protein concentrate) replacements of fish meal, based on digestibility assessment. The use of the "Guelph system" for fecal collection resulted in organic matter digestibility coefficients similar to those obtained by intestinal dissection. Thus, nutrient leaching must have been minimal with the "Guelph system" and it is concluded that this is a satisfactory procedure for digestibility assessment using chinook salmon in seawater. / Land and Food Systems, Faculty of / Graduate

The Sacramento River watershed provides important rearing habitat for key aquatic species, including juvenile Chinook salmon (Oncorhynchus tshawytscha). Salmon rearing in the watershed may inhabit the mainstem river channel or a corresponding floodplain, the Yolo Bypass, before migrating to the ocean. Studies of juvenile salmon have indicated that floodplain rearing may be beneficial in terms of growth and survival, likely related to different trophic pathways of the river and floodplain. However, fish also encounter many anthropogenic stressors in these habitats, such as pesticides, which have well-documented use and environmental presence in the region. Rearing individuals are potentially exposed to pesticides via trophic transfer, which may vary based on utilized food webs due to hydrophobic pesticide fate and transport. To examine the food web structure of each system and the potential for pesticide exposure through dietary routes, a two-year field study was completed. First, to characterize dietary contributions, a three-tiered approach incorporating stable isotopes (δ13C, δ15N and δ34S), essential fatty acids and gut content analyses was employed. Subsequently, pesticides were extracted from prey items and salmon and analyzed to determine contaminant residues. Stable isotope analyses indicated that critically important components of juvenile Chinook diet were amphipoda and adult diptera in 2019 and 2020, respectively. Amphipoda groups had higher concentrations of the fatty acid docosahexaenoic acid (DHA), an important component for fish development, than juvenile diptera or oligochaeta. Diptera (larvae and adults) were frequently found in juvenile Chinook stomachs from both areas and years. Throughout the pesticide examination, organochlorines including the DDT group (p,p’-DDT, p,p’-DDD and p,p’-DDE) were prevalent in all examined biota. There were a significantly greater number of pyrethroid and phenylpyrazole detections and concentrations in zooplankton as compared to macroinvertebrates (Poisson regression, p < 0.05) across regions and years. Additionally, significantly higher concentrations of organochlorines were exhibited in floodplain rearing fish as compared to the Sacramento River (ANOVA, p < 0.05). These findings suggest that juvenile Chinook feeding primarily on zooplankton may be exposed to a greater range of pesticides than those exhibiting benthic feeding, but these pelagic prey were not demonstrated as a major dietary item during the two-year study. Additionally, the previously inferred benefits of floodplain rearing may come at a cost of increased organochlorine exposure. This research has allowed for a robust assessment of potential trophic transfer of pesticides to juvenile salmon, which may help inform future floodplain restoration efforts.

Since 1995, California State Fish Hatcheries (Feather River, Nimbus, and Mokelumne) and Coleman National Fish Hatchery have raised approximately 29 million 4 fall run Central Valley Chinook salmon (Oncorhynchus tshawytscha) per season for stock enhancement. From April through June, fish are acclimated in net-pens prior to release at one of three sites: the Carquinez Strait at Conoco Phillips (CP), the mouth of the Napa River at Mare Island (MI), and the San Joaquin River at Jersey Point (JP). Striped Bass, Marone saxatilis, are known to congregate at the release location to feed on the hatchery fish as they enter the Delta and Bay, and are suspected to be reducing numbers of Chinook recruitment. Dual-Frequency Identification Sonar (DIDSON) was used to capture video-like images to enumerate and estimate sizes of potential predators in the area. Stomach analysis was used to obtain consumption rate data and a simple model was used to estimate predator impacts on the hatchery fish. Data was collected in 2011 and 2012. In 2011 the striped bass population at CP was significantly larger than MI (p=0.009) and JP (p=0.038) and in 2011 , and MI (p=0.046) in 2012. Predators were significantly smaller (range 11.8-61.7 em, mean 34.6 em in 2011 ; 21-67 em, 42.9 in 2012) atJP (p<0.001). Average size predator at MI was 47.3 em (range 31-59 em) in 2011 and 50.9 em (range 33-73 em) in 20 12; and at CP was 48.3 em (range 16-77 em) in 2011 and 52.7 em (range 31-78 em) in 2012. On average an estimated 2.2% of hatchery fi sh are consumed each year by striped bass and predator impacts are greatest at CP (p<0.001). Changing the release site often could improve salmon survival by decreasing predator attraction to the site and reducing immediate predator-prey encounters.

This dissertation argues that the significance and extent of American animal “acclimatization”—the nineteenth-century term for the purposeful introduction of non-native wild animals—has been drastically underestimated in previous historiography. Far from a negligible “fad” that only briefly interested a small number of hunters and wildlife enthusiasts, American acclimatization was in fact a large-scale and enduring exercise in bioengineering that introduced dozens of new species to the nation over the course of more than a century. At first led by private individuals and organizations, American acclimatizers introduced several new birds and fish into the country from the mid-nineteenth century, including modern-day mainstays like the English sparrow, ring-necked pheasant, and German carp. While private organizations devoted to animal acclimatization mostly dissipated by the late nineteenth century, the federal government’s biologist-bureaucrats made the acclimatization of new animals a central component of vast efforts to supply America’s hunters and fishers deep into the twentieth century, a persistence that has been heretofore overlooked. In composing the first dedicated study of American animal acclimatization, I visited a dozen different archives and have brought hundreds of previously unexamined sources to bear. These revealed the enduring popularity of animal acclimatization and its persistence as a wildlife rejuvenation tool. These sources also laid bare the ideological motivations for animal acclimatization. Far from salving a nostalgic yearning for the fauna of Europe, Euro-Americans often saw animal acclimatization projects as progressive techniques of environmental management instead. Animal acclimatization projects, moreover, were intertwined with the Euro-American colonization of the American West. Settler-colonial ideology, that fusion of Euro-American racial supremacy with grandiose notions of national identity and expansion, runs through the rhetoric of many acclimatizers. More concretely, the United States Fish Commission effected the violent dispossession and subordination of the Winnemem Wintu People on California’s McCloud River in order to set up the nation’s first chinook salmon hatchery. The USFC used the hatchery to artificially spawn tens of millions of salmon to replenish American waters as well as establish chinook salmon in American and international watersheds where the fish had never existed before. Finally, I argue that the story of American acclimatization—what I call the American “acclimatization exchange”—offers important nuance and modification to the two most famous paradigms in environmental history: the conservation movement and Alfred Crosby’s “Columbian Exchange.” Massive parallel efforts in animal acclimatization indicate that the conservation era featured far more interventionist environmental management than usually appreciated. The early adoption of “fish culture” in 1860s American also suggests that the conservation era’s periodization should be significantly backdated. Furthermore, the sheer popularity and endurance of foreign species acclimatization in the nineteenth and twentieth centuries, plus the fact that Americans often obtained and exchanged species from Asia, India, and the broader Pacific World, temporally and geographically expands on Crosby’s notion of an Atlantic World “Columbian Exchange” in the wake of initial European discovery and colonization.

This study examined the distribution and behavior of adult spring chinook salmon (Oncorhynchus tshawytscha) related to patterns of stream temperature and physical habitat at channel unit, reach, and basin-wide spatial scales in both a wilderness stream and a disturbed stream in the John Day River basin in northeastern Oregon. Thermal remote sensing of holding and spawning reaches in the upper subbasins of the North Fork and Middle Fork John Day River provided spatially continuous maps of stream temperature. Multiscale associations between salmon and cool-water areas were assessed by overlaying thermal imagery with fish locations mapped during distributional surveys. Chinook salmon were distributed non-uniformly throughout each study area, indicating that salmon selected certain reaches within each subbasin. The coldest reaches available to salmon within the Middle Fork study areas were low gradient, unconstrained reaches where the cooling influence of groundwater flow was the most apparent. In the Middle Fork, the stream currently managed for grazing and timber harvest, water temperature differences were typically 1-2��C within riffle-pool sequences and 3-4��C among reaches. The reach level association between salmon distribution and stream temperature patterns at channel unit and reach level spatial scales was strongest in the warmest study reach, the Middle Fork, and weakest in the coldest study reach, the North Fork. Pools were the preferred habitat for adult spring chinook in both subbasins; however, riffles were used more in the North Fork, the coldest subbasin. This study identified the problems and also the benefits associated with stream temperature patchiness, or discontinuity, both in currently disturbed and in recovering riverine ecosystems. Connectivity among system components in aquatic ecosystems is generally considered necessary for maintaining long-term ecological health. However, it is heterogeneity in the landscape/hydrogeologic template that creates refuge patches in disturbed stream ecosystems, such as those in the John Day River basin. Our observations of thermal refugia occurring at multiple spatial scales, particularly in the Middle Fork John Day River, indicate that, although discontinuity may be an ecological warning sign, refuge patches in streams should also be viewed as expressions of restoration potential because they are functioning remnants of a once continuous, intact hydrologic system. / Graduation date: 1997

Early life stages of many marine and diadromous fish species use sandy beach surf zones, which occur along >50% of the world's marine coastlines. This extensive habitat can provide juvenile fishes with an abundant supply of potential prey and the ability to hide from predators in its shallow turbid waters. Chinook salmon is an anadromous species that migrates to the ocean during their first (subyearlings) or second (yearlings) year of life. The majority of subyearlings reside in estuaries during their first summer season; however, a small number of juveniles also use surf zones. Early marine residence is considered a critical period for Chinook salmon due to high mortality rates; however the role of surf zones in Chinook salmon life history is unclear. Therefore, I determined the distribution of juvenile Chinook salmon on beaches of the eastern North Pacific, compared the migration and growth patterns observed in surf zones and estuaries, identified the factors that accounted for variation in juvenile surf zone catch, explored the factors that influence growth rate variation in surf zones and estuaries, and modeled how growth rates in these coastal habitats may vary in the near future with predicted changes in climate. The majority (94%) of juveniles were caught in surf zones adjacent to estuaries with trough areas, which are beach sections where sand moved by currents and waves produce a trench-like shape. Surf zone fish were collected in significantly lower numbers than estuarine juveniles but entered brackish/ocean waters at similar sizes. Juveniles in surf zones consumed similar organisms (gammarid amphipods, crustacean larvae and insects) as in estuaries. Furthermore, stomach fullness indices (average = 2% of body weight) and growth rates (average = 0.4 mm day�����) were similar in surf zones and estuaries. At one surf zone, juvenile catch was positively correlated to short-term specific growth rates (14 days prior to capture). A bioenergetics modeling approach indicated that given current conditions, consumption rates accounted for more of the variation in growth than prey energetic content and temperature. Climate models predict future increases in fresh water temperature (1.5 to 5.8��C), sea surface temperature (1.2��C) and wave height (0.75 m) that could influence estuarine and surf zone use. Therefore, I developed a local mixing model based on these predictions to estimate future surf zone and estuarine water temperatures in two of the watersheds studied. Based on these temperature projections and the bioenergetics model, I predicted how juvenile specific growth rates would vary in both habitats. I determined that increases in water temperature in both habitats would reduce specific growth rates by 9 to 40% in surf zones and estuaries if diet composition and consumption rates remain similar to present conditions. To compensate for the decline in growth, juveniles may increase their consumption rates or consume more energetically rich prey, if available. If they are not able to compensate, their size at the end of the season may be reduced, which could reduce their overall survival. These results confirm that a small number of suyearling Chinook salmon use sandy beach surf zones, mostly adjacent to estuary mouths, where they experience growth conditions comparable to estuaries. My findings indicate that, in certain situations, juvenile Chinook salmon surf zone use can be influenced by surf zone growth conditions, while variation in growth rates are themselves most strongly influenced by variation in consumption rates in surf zones and estuaries. Predicted changes in coastal western North American climate will likely modify juvenile growth conditions in the next 50 years, and potentially reduce overall survival. Additional insights into the potential impacts of climate change on juvenile salmon will require estimates of changes in the composition, energetic quality and abundance of prey communities inhabiting coastal environments. / Graduation date: 2013

The mechanisms of mortality during critical life stages of fish are not well-understood and, for many species, it is not clear if the mechanisms are similar for naturally and artificially propagated individuals. For Chinook salmon (Oncorhynchus tshawytscha), natural fish potentially face negative interactions, such as competition, and survival disadvantages, such as smaller size, that may limit survival when in association with hatchery fish. To better understand the mechanisms of mortality for hatchery and natural Chinook salmon during the critical early marine residence stage, I: (1) developed a model to discriminate between hatchery and natural juveniles using otolith structure; (2) directly compared migratory patterns of hatchery and natural juveniles; and (3) determined if there was evidence for selective mortality during early marine residence. I followed two cohorts through space and time by collecting juveniles from May-September in the Columbia River estuary and off the coast in September of 2010 and 2011. I compared attributes of those juveniles when they firstentered marine waters with those of survivors after their first summer at sea. I used a combination of genetic stock identification, otolith chemistry and structure, and physical tags to determine stock of origin, size at and timing of freshwater emigration, marine growth, and production type (hatchery or natural). I focused on the subyearling life history of a federally managed genetic stock group (upper Columbia River summer and fall Chinook salmon, UCR Su/F) because: 1) it is an abundant stock group; 2) subyearlings may be more vulnerable to size-selective mortality than yearlings; and 3) it is currently impossible to assess impacts of hatchery production due to low rates of marking the hatchery fish within this stock group. The classification model included two metrics, the presence or absence of a previously unreported transfer check associated with hatchery rearing and variability in otolith increment width, and predicted production type with a 92% jack-knifed accuracy. Overall, timing of marine entry was similar for hatchery and natural UCR Su/F juveniles, which entered marine waters from May-September with a peak in July and August in both years. Estuarine residence times were brief: 80% of the individuals captured in the estuary had resided in saline waters for < 3 days and mean estuarine residence was significantly greater (7 ± 1.3 d) in 2010 than 2011 (1 ± 0.3 d). The only clear difference was that natural individuals captured in the estuary in 2011 migrated to saline waters earlier (July 13th ± 4 d) than hatchery conspecifics (August 10th ± 6 d). However, the timing of marine entry was similar (July 27th ± 1 d) between hatchery and natural fish collected later in the ocean. This observation could be due to differential survival related to the timing of marine entry. Alternatively, estuarine collections may not have adequately represented the emigrating population due to rapid emigration. I documented clear spatial overlap between production types during early marine residence but no difference in median size at marine entry (100 ± 3.5 mm), size at capture (152 ± 4.0 mm), or marine growth (0.94 ± 0.1 %b l d-1). There were also no significant differences in size at marine entry between estuary and ocean collections, which indicates that size-selective mortality had not occurred. Based on both external tags and the otolith classification model, the mean percentage of natural fish in ocean collections was 17% (± 4.8) greater than in the Columbia River estuary; this finding may indicate that estuarine collections are biased to hatchery fish or, more likely, that natural fish survived at higher rates than hatchery fish. Increased survival of natural fish may be related to greater selection pressure during freshwater rearing and prior experience with predators. This study provides the first direct stock-specific comparison of juvenile migratory behavior in natural and hatchery juvenile Columbia River Chinook salmon during early marine residence. Further research is needed to determine if natural fish consistently survive better than hatchery conspecifics and, if so, determine the specific traits and behaviors that afford a survival advantage. / Graduation date: 2013

Wild populations of fish are faced with a multitude of stressors, which may include human interaction, toxins, and disease. Benzo(a)pyrene (BaP), a known carcinogen and immunotoxin, has been reported in the stomach contents of juvenile chinook salmon, Oncorhynchus tshawytscha, in urban waterways. We investigated the impact of chronic dietary exposure of environmentally relevant levels of BaP on the immune system and cytochrome P4501A1 (CYP1A1) expression in juvenile chinook salmon. Two experiments were carried out in which juvenile fish were fed food treated with ethanol (control diet), low or high concentrations of BaP, or cortisol. In the first experiment we measured mitogen-stimulated proliferation of splenic leukocytes using flow cytometry and a colorimetric assay using Alamar Blue[superscript TM] Susceptibility to a microsporidian parasite, Loma salmonae, was evaluated in the second experiment by quantification of xenomas in the gills. Hepatic CYP1A1 and plasma cortisol were measured in both experiments. No significant trends were found in leukocyte mitogen activation or plasma cortisol between treatments or days. However, western blot analysis of CYP1A1 concentration in liver revealed interesting patterns of induction: in cortisol fed groups CYP1A1 was <20% of control on all days, groups fed low levels of BaP were 250% of control values on days 8 and 21 then dropped below control values on day 29, and groups fed high levels of BaP had less CYP1A1 than controls on all days. Similar patterns of CYP1A1 levels were found in the second experiment, and diseased control groups showed about a 55% decrease in CYP1A1 concentration when compared with non-diseased control groups. Susceptibility to L. salmonae was significantly higher in groups receiving cortisol. Whereas there was no effect of the high BaP dose, the low BaP dose appeared to increase disease susceptibility. This study supports concerns of stress and toxin induced immune dysfunction in wild populations of fish. / Graduation date: 2004

In this thesis, we examined the effects of the exposures to anthropogenic pollutants on the fish, primarily juvenile chinook salmon, immune system using newly and recently developed immune assays. In addition, we developed a new assay for measuring immunocompetence of fish. In the first chapter, the Alamar Blue assay was developed to quantify the proliferation of chinook salmon (Oncorhynchus tshawytscha) leukocytes. Isolated splenic and pronephric leukocytes were stimulated with different concentration of mitogens (LPS, PWM, and ConA) for various incubation times. Optimum cell culture conditions (cell density, mitogen concentration, and incubation time) for the Alamar Blue assay were evaluated by comparison with flow cytometric analysis. The Alamar Blue dye was non-toxic for leukocytes, and the assay proved to be able to quantify the mitogenic responses using LPS, but PWM and ConA. In the second chapter, we determined the effects and mechanisms by which p,p'- DDE exposure might affect the immune system of chinook salmon (Oncorhynchus tshawytscha). Isolated salmon splenic and pronephric leucocytes were incubated with different concentrations of p,p'-DDE, and cell viability, induction of apoptosis, and mitogenic responses were measured by flow cytometry and Alamar Blue assay. p,p'- DDE significantly reduced cell viability and proliferation and increased apoptosis. The effect of p,p'-DDE on pronephric leukocytes was more severe than on splenic leukocytes, likely because pronephric leucocytes had a higher proportion of granulocytes, cells that appear more sensitive to p,p'-DDE. The effect of p,p'-DDE on leucocytes appeared to vary between developmental stages or season. The mitogenic response of leukocytes of chinook salmon exposed to p,p'-DDE in vivo exhibited a biphasic dose-response relationship. Only leukocytes isolated from salmon treated with 59 ppm p,p'-DDE had a significantly lower percentage of Ig+ blasting cells than controls. Our results support the theory that exposure to chemical contaminants could lead to an increase in disease susceptibility and mortality of fish due to immune suppression. In the third chapter, we evaluated the direct effects of in vitro exposures to tributyltin (TBT), widely used biocide, on the cell mediated immune system of chinook salmon (Oncorhynchus tshawytscha). Splenic and pronephric leukocytes isolated from juvenile chinook salmon were exposed for 6, 24, or 96 hr to a concentration range of 0.03 0.1 mg TBT 1����� in cell cultures. Effects of TBT on cell viability, induction of apoptosis, and mitogenic responses were measured by flow cytometry. Splenic and pronephric leukocytes in the presence of TBT experienced a concentration-dependent decrease in the viability in cell cultures following the induction of apoptosis. In addition, pronephric lymphocytes exhibited a greater sensitivity to TBT exposure than pronephric granulocytes. The functional ability of splenic B-cells to undergo blastogenesis upon LPS stimulation was also significantly inhibited in the presence of 0.05, 0.07, or 0.10 mg 1����� of TBT in the cell cultures. Flow cytometric assay with the fluorescent conjugated monoclonal antibody against salmon surface immunoglobulin was employed for the conclusive identification of B-cell in the chinook salmon leukocytes. Our findings suggest that adverse effects of TBT on the function or development of fish immune systems could lead to an increase in disease susceptibility and its subsequent ecological implications. / Graduation date: 2004