Thesis (M.S.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains vi, 93 p. : ill. Vita. Includes abstract. Includes bibliographical references.

This study investigated the use of non-specific immunostimulants to alleviate stress-mediated suppression of defence mechanisms and subsequent susceptibility to bacterial pathogens in rainbow trout (Oncorhynchus mykiss). One yeast (1-3),)1-6)-β-glucan and a bacterial peptidoglycan were selected as immunostimulants from a panel of test substances on the basis of enhanced intracellular superoxide generation by kidney macrophages stimulated in vitro. Kidney macrophage effector activity was not affected after 1, 2, 3 or 4 weeks of in-feed treatment with 0.05% or 5% of glucan or peptidoglycan. However, production of bactericidal superoxide by inflammatory peritoneal macrophages did increase significantly after four weeks of oral treatment with 0.05% peptidoglycan. Although a single confinement of fish (93% reduction of water volume for five minutes) caused a physiological stress response, as indicated by hyperglycaemia in plasma, kidney and inflammatory macrophage activities were only affected after six daily confinements. Phagocytosis, intracellular superoxide production and killing of Aeromonas salmonicida in vitro by kidney macrophages were significantly reduced. Conversely, production of extracellular superoxide, which may be associated with damage to self, was enhanced. Peritoneal macrophages displayed a similar but less marked respiratory burst response after repeated confinement. Some of the alterations in macrophage function caused by daily confinement were prevented by feeding 0.05% peptidoglycan four weeks before the first confinement. The increase in kidney macrophage extracellular superoxide production caused by repeated confinement was significantly alleviated by in-feed peptidoglycan. Similarly, the decrease in intracellular production by peritoneal macrophages caused by repeated confinement was prevented by in-feed treatment with peptidoglycan. Neither peptidoglycan nor repetitive confinement had any effect on complement lytic activity. These results indicate that dietary peptidoglycan was able to reduce, by regulating macrophage function, the impact of stress on certain bactericidal defences and potential damage to self. However, there was no significant difference in the persistence of viable A. salmonicida in the spleen or blood of infected fish in any of the experimental treatments.

Determining the physical and biological habitat variables that influence the abundance of juvenile salmonids in British Columbia streams will improve management practices. Habitat models are tools that provide insight into organisms’ habitat needs and provide a more efficient mechanism for estimating population abundance than direct measurement. Models have been developed for salmonids in other jurisdictions, but very few have included invertebrate drift (a primary food source for juvenile salmonids) as a predictive variable. This is because temporal and spatial variation of drift abundance are widely assumed to be so high that drift cannot be reliably estimated without unreasonable effort. This thesis investigates the temporal and spatial variability of invertebrate drift and the impact of its inclusion in habitat models for juvenile salmonid abundance in two chapters. The first objective of the first chapter was to evaluate the temporal variability of invertebrate drift by comparing the seasonal and day-to-day variation in drift abundance to spatial variation within and between sites. The second objective was to develop predictive models for invertebrate drift abundance. Aquatic, terrestrial and total invertebrate drift abundances varied primarily between sites and very little between days or months at the same site, indicating that a single day of sampling is sufficient to assess drift abundance for comparison among sites. The abundance of invertebrate drift was related to productivity- and flow-related habitat variables. The objectives of the second chapter were to develop predictive models for juvenile salmonid abundance in southwestern BC using physical and biological habitat variables, to determine whether habitat variables differ between the Coast and Interior regions of BC, to determine the contribution of invertebrate drift to the relative predictive ability of the models, and to determine cost:benefit ratios for the predictive models and their component variables. The final models for predicting abundance of all young-of-year salmonids combined, and rainbow trout (Oncorhynchus mykiss) and coho salmon (O. kisutch) individually, included variables related to stream structure and productivity, and the models for rainbow and coho showed regional differences. Invertebrate drift did not improve model fit. / Science, Faculty of / Zoology, Department of / Graduate

Although empirical evidence suggests that interactions occur between serotonin (5- HT) and the hypothalamic-pituitary-interrenal (HPI) axis teleost fish, the mechanisms of serotonergic regulation of the HPI axis require elucidation. The hypothesis that 5-HT regulates the stress response in rainbow trout (Oncorhynchus mykiss) by acting at multiple levels in the HPI axis through tissue-specific 5-HT receptor subtypes was examined. Messenger RNA for 5-HT1A, 5-HT2 and 5-HT4 receptor subtypes was expressed in all HPI axis tissues. Administration of 5-HT in vivo in cannulated trout caused significant increases in plasma cortisol and glucose concentrations. In vitro head kidney preparations revealed a stimulatory effect of 5- HT, acting through the 5-HT4 receptor, on cortisol production. Collectively, these data suggest that 5-HT plays a role in HPI axis activation in rainbow trout, and that at the head kidney level, these effects likely are mediated by the 5-HT4 receptor.

Rainbow trout {Oncorhynchus mykiss) and coho salmon (0. kisutch) produce males which mature at an abnormally early age. The objectives of this study were to critically describe the role of growth rate in early sexual maturation of male salmonids by monitoring growth rate and precocious sexual development in individually identified fish. Underyearling rainbow trout were individually identified using coded tags. These fish were reared at 9°C and 15°C and growth was monitored twice a month in both groups over a 9 month period starting January 24, 1989. The number of precocious males in the 9°C and 15°C groups were 11% and 18% respectively. Yearling coho salmon were similarly tagged, reared at ambient temperature, and monitored for growth over an 8 month period starting March 22nd, 1989. Ten percent of the coho salmon matured precociously. In both experiments there was a similar relationship between fast growth in early spring and precocious maturation. The growth rate of fish that matured declined in autumn relative to non-maturing individuals. Condition factor (length to weight relationship) was greater among those fish that matured precociously. Plasma growth hormone was monitored in the coho salmon. Most samples fell below the detection limits of the assay (1.5. to 3.0 ng/ml). A strategy to reduce the number of precocious males in cultured salmonids is suggested. / Land and Food Systems, Faculty of / Graduate

The fry of the very similar salmonids, coho salmon (Oncorhynchus kisutch) and steelhead trout (Salmo gairdnerii), live in very close association with each other during their first year of life in fresh water. The present study was designed to measure the effects of competition between these species. Populations of different densities of coho and steelhead fry in four identical artificial stream-channels were studied. Observations were made on survival, growth rates, and some aspects of behavior. Among the various groups of fry, survival was apparently dependent upon interspecific factors; the presence of another similar species had no observable effect. Low-density populations survived well, even when another species was present at a high-density. Survival of the fish at high-densities was always depressed, even when the companion species was present at low density. Steelhead fry demonstrated a faster initial growth rate than the coho, enabling them to exceed the coho in growth despite the latter’s earlier hatching and consequent initial size advantage. Growth rates were inversely related to density, both inter- and intraspecific effects being noticeable. The two species tend to be spatially segregated, coho occupying positions in the middle and upper layers of the streams, and steelhead remaining close to the bottom. This stratification was reflected in their feeding behavior and diet. Emigration activity occurred to a greater degree among the coho than the steelhead. Emigrants of both species were observed, to undergo substantial weight loss subsequent to their disappearance from the stream-channels. It was concluded that coho and steelhead fry live in close association with one another without experiencing extensive interspecific competition. This is because of segregation of the species by having differing habitats, feeding habits, growth and survival rates, and consequent population dynamics. / Science, Faculty of / Zoology, Department of / Graduate

Various aspects of rainbow trout (Salmo gairdneri) predation on migrating sockeye salmon (Oncorhynchus nerka) fry and rainbow trout eggs were studied during 1970-71 in specially-constructed artificial streams, and in the laboratory. Tests involving sockeye fry as prey were conducted near Babine Lake, British Columbia, while those using rainbow trout eggs were done near Abbotsford, British Columbia. Examination of the effects of physical factors such as water velocity, water turbidity, and light intensity on predation on migrating sockeye fry showed mortality to be inversely related to water velocity and water turbidity; inversely related to light intensity at low light levels but directly related at very low levels. Other stream tests showed mortality to be inversely related to the amount of exposure of predators to fry, before the beginning of a nightly fry migration. Exposing predators to abnormal light for varying periods of time, immediately prior to fry migration, reduced over-all mortality during fry migration. Mortality was not proportional to the length of exposure of predators to abnormal light. Fry experience with predators was shown to increase the ability of fry to escape or avoid predation on subsequent predator encounters. Additional experience served to further increase their ability to escape or avoid predators. Conclusions drawn from stream tests and a behavioral study are that experienced fry migrate in a manner rendering them less susceptible to predation and the migration pattern (compact and in mass) is influenced in part, or completely, by encounter and escape from predator-prey interactions experienced earlier. "Handling" or other fright-evoking stimuli appeared to have similar effects. Rainbow trout feeding on colored fish eggs indicated color preference patterns which are influenced by background coloration (color contrast between food and background), and light intensity. Preference was for colors showing the most contrast with the background at a particular light intensity. At low light levels, on a pale-blue background, preference was for lighter colors, while it was for darker colors at high light levels. Mortality differences increased proportionately with contrast between colors. Finally, trout displayed what appears to be a behavioral preference for red, and possibly blue, regardless of surrounding environmental conditions. In a food deprivation study rainbow trout displayed an S-shaped hunger response curve when fed on eggs. Indications were that rainbow trout will feed to gut capacity when given the opportunity. Finally, beyond an upper limit of food deprivation, the amount of food eaten by an individual remains fairly constant. / Science, Faculty of / Zoology, Department of / Graduate

Mitochondria were isolated from heart and mosaic muscle of rainbow trout (Salmo qairdneri R.). State 3 respiratory rates were determined at 5 and 15°C using pyruvate, malate, lactate, glutamate or acetyl—carnitine as substrate. The final three substrates were used to generate pH profiles. Pyruvate was oxidized at high rates in all cases, indicating good potential for aerobic carbohydrate metabolism. At 15°C, malate was an equally good substrate for heart mitochondria, while all substrates were oxidized at similar rates to pyruvate in muscle mitochondria. Maximal oxidation rates of heart mitochondria were greater than or equal to those of muscle. State 3 Q₁₀ for oxidation of most substrates in heart was approximately 2, except for malate which had a Q₁₀ of 3. Mitochondrial oxidation tended to be more sensitive to decreased temperature in muscle than in heart, particularly with respect to acetyl—carnitine and glutamate oxidation which in muscle had Q₁₀ values of 4 and 7, respectively. Based on RCR values at 5 and 15°C, there was no indication that membrane permeability to H⁺ ions was altered by a 10°C change in temperature in mitochondria from either tissue. At pH above 7.6 respiratory rates decreased with increasing pH. State 3 respiratory rate increased in heart mitochondria as pH decreased, below 7.6 while in muscle mitochondria, no such pH dependence was observed. RCR values were above 4 in all experiments except at high pH. Muscle mitochondria were the more sensitive to extreme pH with respect to RCR. Heart mitochondria had higher oxidative rates than those of muscle and were less sensitive to decreased temperature, in keeping with the greater oxidative demands of that tissue relative to mosaic muscle. Muscle mitochondria which typically face larger fluctuations in extramitochondrial pH in vivo than do those of heart, were less sensitive to pH in vitro. It was concluded that substrate utilization patterns and response to changes in temperature and extramitochondrial pH in the two mitochondrial populations was different and reflected both the intracellular environment of the mitochondria and the different needs of each tissue for aerobic energy supply. / Science, Faculty of / Zoology, Department of / Graduate

The response of rainbow trout (Salmo gairdneri) to lures was investigated in trolling experiments at Loon Lake, British Columbia. The "action" of a lure was found to be an important parameter in determining its efficiency: of four actions tested, the flatfish caught the greatest number of fish. Although the color of lure was not significant, red lures were more efficient than yellow, green, and blue, and more efficient than various color patterns of red and white. The presence of a dodger with lures did not affect their efficiency, but larger fish were caught. No size selection occurred with either colors or actions of lures. In laboratory feeding experiments using dyed trout eggs as food, red was selected first or second more often than yellow, green, or blue. The color of background against which the fish were fed, and individual differences among fish caused significant changes in the preference shown for various colors of food. Combining two colors also affected the selection intensity, depending upon the contrast between the two colors. Preferences for different colors of food were not influenced by the hunger level of the fish, measured in terms of the quantity of food in the fish's gut. In the course of the experiments it was incidentally observed: (1) that rainbow trout possess a striking ability to match the hue of their skin (mainly in the dorsal region) to that of the background in which they are kept; (Z) the color of background affects the activity level of rainbow trout: yellow produces the highest level of activity and green the lowest. / Science, Faculty of / Zoology, Department of / Graduate

The role of carbonic anhydrase in carbon dioxide excretion and acid-base regulation in the rainbow trout, Salmo gairdneri has been investigated. While a significant amount of carbonic anhydrase was found in the blood of the trout, calculations based on red cell hemolysates suggest that the probable circulating levels of carbonic anhydrase activity in blood may not be sufficient to account for the observed carbon dioxide excretion. An analysis of carbonic anhydrase activity in whole blood from the trout revealed that intact fish erythrocytes, unlike mammalian erythrocytes totally fail to facilitate the dehydration of extracellular bicarbonate. The possible mechanism of this phenomenon has been examined; however the salient point was that fish red blood cells do not appear capable and therefore by implication apparently not necessary for the excretion of carbon dioxide at the gills of trout. The observed excretion of carbon dioxide in the trout was found to be accounted for by the gills and their compliment of carbonic anhydrase. This finding was based on the following observations. (1) Depletion of circulating blood carbonic anhydrase levels during severe anemia was without effect on carbon dioxide excretion rates or blood acid-base status. (2) Introduction of the carbonic anhydrase inhibitor, diamox into anemic fish produced a severe acid-base disturbance associated with a fall in observed carbon dioxide excretion. (3) Isolated perfused gill preparations excrete carbon dioxide at rates comparable to those observed in vivo from free swimming fish. (4) Carbon dioxide excretion in isolated gill preparations is abolished by diamox. The excretion of carbon dioxide in fish occurs via the movement of plasma bicarbonate into the branchial epithelium, where it is subsequently dehydrated into molecular carbon dioxide and excreted. A model is proposed and supportive evidence presented to account for the coupling of ionic exchange occurring across the gill with carbon dioxide excretion. The proposed model distinguishes between control of plasma hydrogen ion activity and regulation of plasma total carbon dioxide concentration per se. The functional significance of this pattern of carbon dioxide excretion for aquatic animals is discussed along with the implications for air breathing fish. / Science, Faculty of / Zoology, Department of / Graduate