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The central objective of this thesis is to better understand early life history dynamics of salmonids in large regulated rivers. I studied spawning, incubating, and age-0 life stages of rainbow trout in the Lee’s Ferry reach of the Colorado River below Glen Canyon Dam, AZ. My first objective was to evaluate the effects of hourly fluctuations in flow on nearshore habitat use and growth of age-0 trout. Catch rates in nearshore areas were at least 2- to 4-fold higher at the daily minimum flow compared to the daily maximum and indicated that most age-0 trout do not maintain their position within immediate shoreline areas during the day when flows are high. Otolith growth increased by 25% on Sundays in one year of study, because it was the only day of the week when flows did not fluctuate. My second objective was to evaluate the effects of flow fluctuations on survival from fertilization to a few months from emergence (early survival). Fluctuations were predicted to result in incubation mortality rates of 24% in 2003 and 50% in 2004, when flow was experimentally manipulated to reduce trout abundance, compared to 5% in 2006 and 11% in 2007 under normal operations. Early survival increased by over 6-fold in 2006 when egg deposition decreased by at least 10-fold. Because of this strong compensatory dynamic, flow-dependent incubation mortality in experimental years was likely not large enough to reduce the abundance of age-0 trout. My final objective was to determine how flow, fish size and density effects habitat use, growth, and survival of age-0 trout. Apparent survival rates from July to November were 0.18 (2004), 0.19 (2006), and 0.32 (2007). A stock synthesis model was developed to jointly estimate parameters describing early life history dynamics, and indicated that early survival was lower for cohorts fertilized during the first half of the spawning period and was negatively correlated with egg deposition, that movement of age-0 trout from low- to high-angle shorelines increased with fish size, and that survival varied by habitat type and over time in response to flow changes from Glen Canyon Dam.

Co-variation in the genetic architecture, and co-localization of quantitative trait loci (QTL), for embryonic developmental rate and age at maturation in male rainbow trout (Oncorhynchus mykiss) was examined. Two lots (September and December spawning) of 2 x 5 (male x female) diallel crosses were performed and the resulting progeny were raised to maturity. Higher incidences of precocious maturation were observed in the September lot fish compared to December lot fish, which was uncoupled to developmental rate. However, precocious maturation and body size was higher in early hatching December lot fish. A genome scan was also completed across 4 linkage groups previously reported to possess embryonic developmental rate QTL and/or age at maturation QTL. No evidence of co-localization of QTL for the two traits was detected. However, the presence of QTL for embryonic developmental rate on RT-8 and RT-24 and one for age at maturation on RT-17 supports previous results.

I examined the influence of cyclic feeding regime on fillet yield and quality and their genetic architecture in rainbow trout (Oncorhynchus mykiss). Two lots of half-sib families produced in September and December were exposed to periods of reduced ration followed by increased ration. Fish fed cyclically reduced rations weighed less and had smaller fillets and body proportions relative to controls. They also had less estimated muscle and visceral lipid but also more proportional visceral mass. Lots and families within lots differed significantly for the majority of traits. Genotypic analysis with 23 microsatellite markers spanning 19 linkage groups detected QTL for all traits over multiple linkage groups that were not necessarily consistent between feeding regime and parental lot. Linkage groups RT-9 and 29 had the largest QTL effects for fillet composition traits and overall body size across the greatest number of parents sampled.

Corn gluten meal (CGM) is a high protein (60% crude protein), highly digestible feed ingredient widely used in diets for salmonids, however its use has been related to reduction in muscle pigmentation possibly due to pigment interaction. Therefore, laboratory scale and in vivo trials were conducted to reduce pigment level in CGM and to assess its effect on fish muscle pigmentation, respectively. In the first chapter, a bench-scale study was carried out to investigate factors that affect bleaching of carotenoids in CGM, using white soy flake flour (WSFF) as a lipoxygenase (LOX) source. Plackett-Burman and Box-Behnken designs were used to screen and optimize factors, respectively. Furthermore, a 12-week growth trial was conducted in order to assess the effects of dietary regular and pigment bleached CGM on growth and muscle pigmentation of rainbow. In the second chapter, a 24-week growth trial was carried out in order to assess the effects of increasing levels of CGM on growth and muscle pigment deposition in rainbow trout. In the third chapter, a bench-scale (10 g) corn wet milling procedure was used to assess the bleaching of carotenoids from CGM during steeping. Studies from this thesis confirmed the negative effects of CGM on fillet pigmentation and highlighted the need for evaluation of muscle quality traits such as colour in response to inclusion of new feed ingredients. Furthermore, this thesis gives insight on how to reduce pigments from corn gluten meal using cost-effective and practical bleaching approaches.

A standard methodology was developed for performing avoidance-preference tests, using Rainbow Trout (Salmo gairdneri) as the test organism. Experiments were conducted in a hydraulic channel, 9.15 m long by 0.30 m wide, partly divided along its length, and at a flow depth of 0.30 m. The design combined steep and shallow gradient characteristics. The toxicants investigated included Cu(II), Cr(III) and Cr(VI). The lowest avoidance threshold values were established at 2.1 $ mu$g/l for Cu(II) and 0.0026 mg/l and 0.026 mg/l for Cr(III) and Cr(VI) respectively while avoidance reactions increased with levels of toxicant in the channel. Similar experiments were performed with rainbow trout which were pre-exposed at sublethal levels to the toxicant, in order to assess the influence of toxic pre-exposure to the subsequent fish avoidance response. The length of pre-exposure varied between 7-20 weeks. Avoidance threshold values were correlated with safe levels of toxicant exposure. / Pre-exposed fish exhibited decreasing avoidance reactions compared to non-exposed populations. Increased tolerance to the toxicant, was suggested by the increase in avoidance threshold values with pre-exposure levels. Fish exposed to test concentrations matching their pre-exposure levels, clearly preferred this same concentration over the adjacent lower or higher test concentration. / A two mechanism avoidance model was proposed independent of toxicant used or level of pre-exposure. The toxicant concentration where the second mechanism begins to dominate was referred to as avoidance breakpoint, and was correlated to a MATC level for the toxicant in question. Olfactory responses were proposed to be associated with fish avoidance responses below the avoidance breakpoint, while hypoxic stress along with osmo- and iono regulatory stress appeared to be responsible for driving fish avoidance reactions beyond the avoidance breakpoint. / A clearance period of 7 days was sufficient to allow fish to recover normal avoidance behaviour following pre-exposure to Cr(VI) below the avoidance breakpoint.

No description available.

Municipal wastewater treatment plants (MWWTPs) are point sources of environmental contamination that can cause adverse effects on fish species exposed to their effluents (MWWEs). Contaminants, such as endocrine disrupting compounds (EDCs), commonly found in MWWEs have been shown to induce many adverse reproductive effects. In the Grand River watershed of southern Ontario, three MWWTPs from the cities of Guelph, Waterloo and Kitchener have been identified to adversely alter molecular and cellular responses in fish, which have been previously associated with exposure to estrogenic EDCs. Currently these systems are undergoing process upgrades to address aging infrastructure as well as expanding capacities to serve rapidly increasing populations. Studies in this thesis were conducted to determine whether the Guelph and Waterloo MWWEs, as well as surrogate pilot plant effluents that modeled possible treatment process upgrades for MWWTPs were estrogenic in vivo in controlled laboratory exposures. Unfortunately, the Kitchener MWWE (conventional activated sludge and lagoon) was acutely toxic and could not be included in laboratory exposures. Rainbow trout (Oncorhynchus mykiss) were exposed to 20% to 90% effluent from both Guelph and Waterloo, for 2, 8 and 14 days and fish plasma was analyzed for vitellogenin (VTG) a biomarker for estrogenic exposure. These exposures indicated that neither the Guelph nor the Waterloo MWWE were estrogenic in vivo under the exposure conditions in this study. In contrast the total estrogenic equivalence measured using the yeast estrogen screen (YES) of the Guelph, Waterloo and Kitchener MWWEs increased respectively, inversely related to the degree of treatment. Three pilot plant effluents (Burlington MWWTP) including conventional activated sludge (CAS), nitrifying with CAS (CAS-N) and biological nutrient removal with CAS (CAS-BNR) were also tested in rainbow trout in vivo for plasma VTG induction. Only the CAS-BNR effluent caused weak (2-3.5 fold) induction of VTG relative to controls. Although the estrogenicity (YES) of the effluent was variable the VTG induction corresponded with the slightly higher 17??-estradiol equivalents measured in these pilot plant effluents. An effect-directed analysis (EDA) for estrogenic substances was carried out on the Waterloo and Kitchener MWWEs and revealed that the estrogenicity (YES) of the Waterloo MWWE was associated with estrone, while the estrogenicity of the Kitchener MWWE was associated with several chemicals (estrone > 17??-estradiol > bisphenol A ~ testosterone).

Factors that can influence the transmission of aquatic viruses in fish production facilities and natural environment are the immune defense of host species, the ability of viruses to infect host cells, and the environmental persistence of viruses. In this thesis, fish cell lines were used to study different aspects of these factors. Five viruses were used in this study: viral hemorrhagic septicemia virus (VHSV) from the Rhabdoviridae family; chum salmon reovirus (CSV) from the Reoviridae family; infectious pancreatic necrosis virus (IPNV) from the Birnaviridae family; and grouper iridovirus (GIV) and frog virus-3 (FV3) from the Iridoviridae family. The first factor affecting the transmission of fish viruses examined in this thesis is the immune defense of host species. In this work, infections of marine VHSV-IVa and freshwater VHSV-IVb were studied in two rainbow trout cell lines, RTgill-W1 from the gill epithelium, and RTS11 from spleen macrophages. RTgill-W1 produced infectious progeny of both VHSV-IVa and -IVb. However, VHSV-IVa was more infectious than IVb toward RTgill-W1: IVa caused cytopathic effects (CPE) at a lower viral titre, elicited CPE earlier, and yielded higher titres. By contrast, no CPE and no increase in viral titre were observed in RTS11 cultures infected with either genotype. Yet in RTS11 all six VHSV genes were expressed and antiviral genes, Mx2 and Mx3, were up regulated by VHSV-IVb and -IVa. However, replication appeared to terminate at the translational stage as viral N protein, presumably the most abundant of the VHSV proteins, was not detected in either infected RTS11 cultures. In RTgill-W1, Mx2 and Mx3 were up regulated to similar levels by both viral genotypes, while VHSV-IVa induced higher levels of IFN1, IFN2 and LGP2A than VHSV-IVb. The second part of the thesis examined the ability of two Ranaviruses, GIV and FV3, to infect non-host fish cells. This is referred to as cellular tropism and is one of many host-virus interaction events required to established successful infection in new organisms. Grouper iridovirus (GIV), belonging to the Ranavirus genus of the Iridoviridae family, was demonstrated to differentially express viral genes and induce apoptosis in three non-host fish cell lines rainbow trout monocyte/macrophage (RTS11), Chinook salmon embryon (CHSE-214) and fathead minnow Epithelioma papulosum cyprinii (EPC). These cells were challenged with GIV and virus entry into all three cell lines was confirmed by the expression of viral immediate early genes. The expression of the late major capsid protein gene was detected in CHSE-214 and EPC, but not in RTS11, suggesting an earlier termination in the viral replication cycle in RTS11. Approximately 12 h after infection with GIV, cell death was prominent in all three non-host cell lines. Death was later confirmed to be apoptosis by the presence of chromosomal DNA fragmentation and phosphatidylserine externalization. To determine whether apoptosis was protein related or gene expression related, the three cell lines were infected with heat-inactivated GIV and UV-treated GIV (GIVUV). The heat inactivation abolished apoptosis in all three cell lines, but each cell line responded differently to GIVUV. Relative to GIV, GIVUV caused no apoptosis in CHSE-214, decreased apoptosis in RTS11, and increased apoptosis in EPC. These results suggest that early GIV gene expression was needed for apoptosis in CHSE-214 but impeded apoptosis in EPC. At the cellular level, only EPC was a permissive host as EPC was the only cell line of the three capable of producing a moderate increase in virus titre. The three non-host cell lines present a good system for potentially identifying different components of GIV-induced apoptotic pathways in future studies. Rainbow trout are not highly susceptible to frog virus 3 (FV3) induced diseases, and had been suggested to be a potential carrier for the virus. To determine which rainbow trout cell types are permissive for FV3 and act as a potential source for virus replication in vivo, the ability of rainbow trout cell lines from gonads (RTG-2), skin (RTHDF), liver (RTL-W1), gills (RTgill-W1), intestine (RTgut-GC) and spleen (RTS11), and primary leukocyte cultures from peripheral blood (PBL) and head kidney (HKL) to support FV3 infection was examined. RTG-2 supported a moderate level of FV3 replication while viral replication in RTL-W1 was minimal. The rest of the cell lines did not support viral replication but all succumbed to the infection and were killed by FV3. Lymphocyte-like cells from PBL and HKL were not killed by FV3 while macrophage-like cells were. Most of the cell lines died by an apoptosis-independent mechanism, presumably necrosis, while the monocyte/macrophage cell line, RTS11, died by an apoptosis-dependent mechanism. In addition, neoplastic macrophage-like human U937 cell line, and T lymphocyte-like PEER cell line were also infected with FV3 to compare their response to that of rainbow trout immune cells. U937 cells were killed by FV3 in an apoptosis-dependent manner; however, PEER T cells did not die from FV3 infection, a result similar to the lymphocyte-like fraction of rainbow trout PBL and HKL. In summary, most rainbow trout cell lines do not support significant FV3 replication; furthermore, cells of the lymphocyte origin appeared refractory to FV3 induced cell death while those of macrophage origin underwent apoptosis as a response to FV3. The last factor affecting the transmission of aquatic viruses examined in this thesis is the persistence of viruses in the aquatic environment. Virus persistence is influenced by natural environmental factors such as temperature, pH, desiccation and salinity, but the often unexplored anthropogenic factors can play a role. Therefore, the focus of this section was on the effect of one particular anthropogenic substance, Corexit 9500, on the infectivity of aquatic viruses with different physical characteristics. The effect of Corexit 9500, a dispersant used to clean up oil spills, on invertebrates, lower vertebrates, birds and human health have been examined but there is a significant lack of study on the effect of this dispersant on aquatic viruses. In this study, the effect of Corexit 9500 on four aquatic viruses of different structural composition was examined. Corexit 9500 reduced the titre of the enveloped viral hemorrhagic septicemia virus (VHSV) at all concentrations (10% to 0.001%) examined. The titre of frog virus 3 (FV3), a virus with both enveloped and non-enveloped virions, was only reduced at the high Corexit 9500 concentrations (10% to 0.1%). Corexit 9500 was unable to reduce the titre of non-enveloped infectious pancreatic necrosis virus (IPNV), but enhanced the titre of chum salmon reovirus (CSV) by 2-4 logs. With the ability to inactivate enveloped viruses and possibly enhance some non-enveloped viruses, Corexit 9500 has the potential to alter the aquatic virosphere.

Understanding how animals detect and discriminate different qualities of light is a key component of the study of visual ecology. My research investigated the use of three electrophysiological methods to assess the neuronal mechanisms involved in spectral and polarization sensitivity in one species of salmonid, the rainbow trout (Onchorynchus mykiss). 1 examined the neuronal mechanisms underlying polarization sensitivity using electroretinograms (ERG) and optic nerve compound action potential (CAP) recordings. Chromatic adaptation and pharmacological techniques were used to reveal opponent interaction at the cone-horizontal cell level and to provide the first evidence of retinal processing of polarization sensitivity. To facilitate additional research more suited to the exploration of neural networks and signaling, I developed the protocols and techniques necessary to investigate the spectral sensitivity of rainbow trout using whole-cell patch clamp (WPC) electrophysiology, and produced the first assessment of the ultraviolet component of spectral sensitivity in a vertebrate using this technique.