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

Juvenile rainbow trout (Oncorhynchus mykiss) were fed either 5 or 20 μg 2,2′,4,4′,5,5′‐hexachlorobiphenyl (245‐HxCB)/g diet (wet wt.) for 4, 8, or 12 weeks. Hepatic xenobiotic‐metabolizing enzyme activities and dietary 245‐HxCB accumulation in liver, muscle, and remaining carcass were determined. Liver‐to‐body weight ratios were not altered by either of the two 245‐HxCB concentrations. Relative growth rate increased with time but was not altered by 245‐HxCB concentration. Bioaccumulation of 245‐HxCB was dose and time dependent in all tissues without reaching apparent steady state. Hepatic arylhydrocarbon hydroxylase (AHH) activities increased with 245‐HxCB dose and with time. Ethoxyresorufin‐O‐deethylase (EROD) activities also increased in fish fed 20 μg 245‐HxCB/g diet. No 245‐HxCB‐induced changes in uridine diphosphoglucuronosyl transferase (UDP‐GT) or NADPH‐cytochrome‐c reductase (NCCR) activities were determined. High‐resolution GC‐MS analysis of the 245‐HxCB standard revealed trace (0.4‐0.5%) contamination by two mono‐ortho pentachlorobiphenyls (PnCBs): 2,3,3′,4,4′‐PnCB and 2,3,4,4′,5‐PnCB. Total liver accumulation of these contaminants was inversely related with corresponding EROD and AHH activities and estimated to contribute minimally to their induction. Results from this study suggested that long‐term dietary 245‐HxCB exposures induced cytochrome P4501A activities in rainbow trout liver.

The data analysis indicated that travel distance, user age, and user income level are important determinants of use of trout streams. These variables were incorporated into regression analysis to develop a use-prediction model. This use-prediction model was used to determine predicted use levels of sample streams. The predicted use levels combined with travel costs and expenditures on new fishing equipment were used to derive a statistical estimate of the demand schedules for the sample streams. These statistical demand curves were subjected to consumer surplus procedures to determine the values of the sample streams.

The Diamond Fork River, and it’s tributary Sixth Water Creek, has been highly altered in terms of shape, function, and ecologicaly due to large, trans-basin flows additions to the system for irrigation starting in the early 1900s. Flows were exceptionally large for 80 years, after which they were reduced in 2004. Larger than natural flows during the low flow season were then added to the river in an effort to improve ecosystem health and recreational fishing opportunities. Since the prescription additional flow during low flow seasons, the river channel has undergone further change, most noticeably in the form of narrowing. With the channel change that has occurred over the past decade, it has been suggested that the additional flows, which augment baseflows, are too large and that key habitat elements, particularly pools, are lacking throughout the river. We evaluated trout habitat throughout the altered reaches of the Diamond Fork River and Sixth Water Creek which are still subject to flow additions. We used a model to estimate the number of fish a reach can support at a given flow in order to evaluate which flow produces the highest quality trout habitat. We found that the current summer baseflows of 80 cfs for the lower Diamond Fork and 32 cfs for Sixth Water Creek are less than desirable and that flows less than 40 cfs for the lower Diamond Fork and flows between 20 cfs and 30 cfs for Sixth Water Creek would increase the quality of trout habitat. We also evaluated the size and number of pools throughout the system and identified that pools are generally lacking both in size and number relative to standard requirements for trout success. We documented that as the channel narrowed, the number of pools/deep water disappeared. The lack of sediment and the limited number of flows capable of moving sediment were identified as key causes of the channel simplification and loss of pool habitat. Crucial components of future pool formation and maintenance are i) active sediment availability and ii) frequent sediment mobilizing flows. If the baseflow regime were to be lowered, we predict that the channel would narrow, which would increase the effectiveness of floods, increase mobility of sediment, and overall increase pool habitat. This research, as part of a larger, interdisciplinary project, lays the foundation for proposing new flows for the Diamond Fork River that targets ecological goals identified by stakeholders as well as providing information for future habitat restoration projects.

No description available.

No description available.

Trout anglers comprise about 20% of all anglers in Virginia and expended > 1-million angler-days in pursuit of Virginia trout. Stocked trout account for approximately 80% of the trout angling effort in Virginia (O'Neill 2001) through a program managed by the Virginia Department of Game and Inland Fisheries (VDGIF). Each year, the VDGIF stocks > 1-million catchable-sized trout in about 180 waters in Virginia. Despite the popularity of stocked trout fishing in Virginia, several factors potentially threaten the future of the stocked trout program in Virginia. First, previous surveys indicate that stocked trout anglers are less satisfied with their fishing experiences in Virginia than other species specialists. Additionally, the sale of stocked trout fishing licenses declined 31% between 2006 and 2013. Revenue generated from the sale of trout licenses is a significant source of income that helps fund the stocked trout program. Given the popularity of stocked trout fishing in Virginia and the considerable effort of VDGIF in managing the stocked trout program, my research objectives were to 1) identify homogenous angler groups seeking stocked trout in Virginia, 2) compare motivations, satisfaction, and preferences of stocked trout anglers in Virginia, and 3) examine the extent to which prior satisfaction and constraints relate to future participation. To address objectives 1 and 2, I administered a mail survey of 5,400 licensed trout anglers in Virginia. Using cluster analysis, I identified four distinct groups of stocked trout anglers: traditional anglers, generalists, occasional specialists, and specialists. Traditional anglers constituted the largest group, comprised of those who fished with bait, harvested the stocked trout they caught, fished more frequently than other groups, had low centrality, and invested less money in trout fishing than did other groups. Generalists included those who took fishing-related vacations, fished with a variety of terminal tackle, harvested their catch, and demonstrated low invest little in fishing for stocked trout. Occasional specialists fished infrequently, had low centrality, invested little money in fishing, fished with lures and flies, and released the stocked trout they caught. Specialists had high centrality and investment, fished with flies, and almost always released the stocked trout they caught. Psychological and natural-setting attributes were stronger motives to fish for stocked trout than were fishery and social motives. Anglers rated catching fish as being more important than keeping fish to eat. Overall, anglers were satisfied only somewhat with stocked trout fishing in Virginia. However, anglers rated satisfaction higher with activity-general characteristics, such as the setting and relaxing, than they did activity-specific factors, such as number or size of fish caught. Results of stated preference choice models indicated that anglers preferred unannounced stockings rather than stockings announced in advance. Furthermore, anglers preferred stocking of streams rather than lakes, stocking taking place during the spring, and catching six 10-inch trout rather than fewer, larger trout. Compared to weekday stockings, anglers did not show a preference for weekend stockings, despite comments made at public meetings suggesting that many anglers preferred weekend stockings. Although four specialization levels exist for Virginia's stocked trout anglers, choice models suggest that preferences coalesced into only two groups: preferences of specialists differed from those of traditional anglers, generalists, and occasional specialists. When compared to the status quo fishing trip, traditionalists, generalists, and occasional specialists preferred a prior announced stocking, whereas specialists preferred a delayed announcement and catching fewer, but larger, trout. To address the third objective, I surveyed 1,100 lapsed trout anglers (individuals who had purchased licenses twice between September 1, 2011, and August 31, 2013, but did not purchase one between September 1, 2013, and August 31, 2014). I found that anglers who placed high importance on non-catch-related factors (e.g., being outdoors, enjoying a relaxing experience, experiencing a natural setting) were more likely to fish for stocked trout again in the future. Constraints did not mediate the effect of prior satisfaction on future participation. However, structural constraints were related positively to future participation, suggesting that individuals who had more time and family commitments were more likely to negotiate those constraints and resume participation in the future. / Ph. D.

<p> Knowledge into uptake rate and interactions of the metals via the gastrointestinal tract of freshwater fish is vital, in order to provide tools to protect and to sustain aquatic biota. An in vitro stomach and gut sac technique was utilized to investigate uptake rates of essential metals (copper, zinc and nickel) and non-essential metals (cadmium, lead and nickel) at luminal concentrations of 50fJ.M via the gastrointestinal tract Metals had no effect on the fluid transport rates via the gastrointestinal tract except for copper at the stomach. The stomach emerged as small but important site for metal absorption and interaction. Essential metals were absorbed at approximately the same rate as nonessential metals via the gastrointestinal tract. Copper, zinc, nickel, silver, and lead showed statistical correlation between rate of absorption and mucus binding via gastrointestinal tract, an important first finding for the development of a Biotic Ligand Model (BLM) for the gastrointestinal tract of trout. </p> <p> There was an antagonistic effect of calcium on cadmium uptake at the stomach but not at the intestine. Zinc and calcium exhibited synergistic interaction at the stomach but no interaction at the intestine. These results showed the possibility of the transporters DMTl to mediate copper and cadmium uptake via the intestine; hZip 2 to mediate copper and zinc uptake at the stomach; and Mzip 4 or ZTLl to mediate zinc uptake via the gastrointestinal tract of trout. These results can be used to develop a BLM for the gastrointestinal tract of fish. </p> / Thesis / Master of Science (MSc)

<p> Juvenile rainbow trout (Oncorhynchus myldss) were chronically exposed (90 days) in synthetically softened water ([Ca2+]=50, [Na+]=100 J.tequiv·L-1), to sublethal low pH (5.2) and a simulated global warming scenario (+20C), added to the natural summer thermal cycle ofinshore Lake Ontario. Two studies were conducted over the periods June September 1993 and 1994 in order to examine the effects ofthe sublethal stressors under conditions ofunlimited food ration (satiation feeding twice daily= 10% dry body weight·day-1), and a limited food ration (4% dry body weight·day-1) respectively. The addition of 2°C and sulphuric acid was designed to result in four treatment conditions: i) Control conditions; ambient water temperature and pH; ii) ambient water temperature and pH 5.2; iii) simulated global warming at ambient pH; iv) a combination of simulated global warming and pH 5.2. Year to year variation in temperature provided the trout in the satiation-fed study with an ambient ~peraturerange of 13-24°C, while those in the limited ration study experienced a range of 16.5-21°C. Consequently, the trout in the treatments with an additional 2°C experienced temperatures close to the upper incipient lethal level, particularly the trout in the satiation feeding study. Apparent specific dynamic action raised routine metabolic rates in all treatments to -75% and 55% M02(max) in the satiation and limited ration studies respectively, the difference of 20% indicating the influence of an unlimited feeding regime on metabolism. Trout in the satiation feeding study increased in wet body mass by 30-50 g, while trout in the limited ration study increased by only 3-4 g. Whole body proximate composition of the trout fed to satiation changed over time with large increases in lipid content, small increases in protein content, and compensating decreases in water content in all treatments. No such changes occurred in the limited ration trout, although whole body lipid and carbohydrate were highly variable. In both studies, the addition of20C resulted in decreased growth, with an accompanying depression in appetite in the satiation fed trout, especially at peak temperatures. Surprisingly, trout exposed to low pH alone exhibited improved growth in both treatments. Energy budgets indicated that the addition of 2°C reduced gross energy intake and increased fecal (and Ulllidentified) energy losses resulting in lowered conversion efficiencies, while in limited ration trout, energy expenditure was slightly higher. Trout exposed to low pH exhibited higher gross energy intake and gain, and more efficient energy conversion under unlimited food conditions while trout with limited rations expended the least metabolic energy and exhibited lower nitrogen energy losses. The surprising lack of ionoregulatory disturbance in these pH 5.2 exposed trout in both studies suggests tbat the availability of NaCl in the diet was compensating for branchial ion losses, and perhaps driving appetite in the satiation fed trout Where ration was limited, reductions in activity level may have contributed to energy conservation and consequently improved growth. A 22Na+ flux experiment conducted at the end of the limited ration exposure, in which the fish were exposed to a challenge concentration of H+ (pH 4.2), provided evidence for improved recovery of ionoregulatory balance in trout which had been chronically exposed to low pH. Overall, the combination ofincreased global temperatures and sublethal low pH results in increased physiological costs for juvenile rainbow trout, most noticeably when summer temperatures peak. Ration level is of integral importance when considering the degree of impact of such environmental conditions. </p> / Thesis / Master of Science (MSc)

Largely through the use ofin vitro preparations there is now a generally accepted theory for ion transport for the seawater (SW) gill. However, to date there is no generally accepted freshwater (FW) model for the mechanisms of NaCl transport in the teleost gill. By using an Ussing chamber approach with the opercular epithelia of Fundulus heteroclitus and Oreochromis niloticus, and the urinary bladder of Oncorhynchus mykiss, all acclimated to FW, we hoped to establish one as a possible model for the study of FW ion regulation. FW Fundulus opercular epithelia displayed a serosal negative transepithelial potential (Vₜ) of-43.9 mV, transepithelial conductance (Gₜ) of 1.94 mS·cm⁻², and active transport of Cl⁻ from the mucosal FW against a strong electrochemical gradient. Na⁺ movement was dominated by passive diffusion. The opercular epithelia of Fundulus adapted to 10% SW exhibited properties similar to SW Fundulus by actively extruding Cl⁻ while Na⁺ moved passively into the mucosal 10% SW. With FW bathing the mucosal surface, FW Oreochromis opercular epithelia displayed a serosal positive Vₜ of +8.0 mV, Gₜ of 1.78 mS·cm⁻², and active reabsorption of Na⁺, Cl⁻ and Ca²⁺ against large electrical and/or chemical gradients. The FW Oreochromis opercular epithelia is the only FW in vitro preparation to date that exhibits active absorption (albeit at small absolute rates) of both Na⁺ and Cl⁻. FW 0. mykiss urinary bladders mounted in vitro under symmetrical saline conditions exhibited a transepithelial conductance (Gₜ) of-9.15 mS·cm⁻² and electroneutral active absorption of Na⁺ and Cl⁻ from the mucosal urine side. The transport of Na⁺ and Cl⁻ was a partially coupled process whereby removal of Na⁺ from the mucosal saline decreased Cl⁻ absorption by a 56% and removal of Cl⁻ inhibited Na⁺ absorption by 69%. However, active net absorption of both ions persisted when the counter-ion was replaced with a non-permeant ion. Under more realistic conditions with artificial urine bathing the mucosal surface, Vₜ increased to a serosal positive ~+7.6 mV and Gₜ decreased to ~1.47 mS·cm⁻² Unidirectional influx rates of both Na⁺ and Cl⁻ were much lower, but active absorption of both ions still occurred. Replacement of Na⁺ in the mucosal artificial urine caused no change in unidirectional influx of Cl⁻ and vice versa. The mucosal addition of DIDS, amiloride or bumetanide (10⁻⁴M) all had no affect on absorption rates of Na⁺ and/or Cl⁻, under either artificial urine or symmetrical saline conditions. When the mucosal surface was bathed in artificial urine, removal of mucosal Cl⁻· significantly reduced the maximum transport rate (Jₘₐₓ of Na⁺ (6.1 ~2.1 μmol·cm⁻²·h⁻¹) but had no effect on affinity for Na⁺ (Kₘ~27 mM). Similarly, removal of mucosal Na⁺ significantly reduced the Jₘₐₓ for Cl⁻ uptake (11.4 ~2.4 μmol·cm⁻²·h⁻¹) but had no effect on Cl⁻ Kₘ(~37 mM). The anterior portion of the urinary bladder transported Na⁺ and Cl⁻ at a faster rate than the posterior portion under symmetrical saline conditions, but there was no difference in measured Na⁺/K⁺ -ATPase activities between the two portions. In conclusion, opercular epithelia of Fundulus and Oreochromis demonstrated transport properties believed to be typical of FW teleosts; either preparation may eventually prove to be a good working model for studying FW transport mechanisms. The urinary bladder of Oncorhynchus mounted in vitro did not behave characteristically as the teleost gill is thought to function, but this model may be useful in characterizing various other mechanisms of ionic transport. / Thesis / Master of Science (MSc)