Thesis (MA)--University of Stellenbosch, 2006. / Bibliography. Also available via the Internet.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv., 2001. / Härtill 4 uppsatser.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv., 2007. / Härtill 4 uppsatser.

Thesis (Ph. D.)--Deakin University, 2001. / Title from PDF title page (viewed on May 16, 2005). Includes bibliographical references (p. 203-224).

Thesis (Ph. D.)--West Virginia University, 2010. / Title from document title page. Document formatted into pages; contains xii, 492 p. : ill. (some col.), maps. Includes abstract. Includes bibliographical references.

Thesis (Ph. D.)--Ohio State University, 2005. / Title from first page of PDF file. Document formatted into pages; contains xvi, 105 p.; also includes graphics (some col.) Includes bibliographical references (p. 95-105). Available online via OhioLINK's ETD Center

Thesis (M.S.)--Oregon State University, 2008. / Printout. Includes bibliographical references (leaves 40-46). Also available on the World Wide Web.

Replacement of fish meal as the primary protein source in diets for farmed carnivorous fish is a major priority for sustainability of the aquaculture industry. Three plant-based protein sources (soybeans, field peas, and canola) were investigated to compare their effects on the health and performance of rainbow trout (Oncorhynchus mykiss) and to identify significant anti-nutritional factors (ANFs). Six separate 8-week studies were conducted, over a period of one year, to assess the effects of protein source and processing level (meal versus protein concentrate) at dietary inclusion rates of 0 to 300 g kg-1. Abundance of inflammatory and immune marker transcripts including proliferating cell nuclear antigen (PCNA), immunoglobulin M (IgM), interleukin-1 beta (IL-1β), interleukin-8 (IL-8), and interleukin-10 (IL-10) was evaluated in distal intestinal tissue by quantitative PCR (qPCR) analysis. Activity of the pro-apoptotic enzyme caspase-3 was also assayed in distal intestinal tissue. Transcript abundance was highly variable and no suitable genes for the internal normalization of qPCR data could be identified. As a result, transcript copy numbers were reported per 50 ng of total RNA. At 300 g kg-1 inclusion, soybean meal (SBM) increased abundance of IL-8 and IgM, pea meal (PM) increased abundance of IL-10, and canola protein concentrate (CPC) increased abundance of IL-8. Pea protein concentrate (PPC) reduced IL-8 abundance and caspase-3 activity, while increasing abundance of IL-10. Canola meal (CM) and soy protein concentrate (SPC) did not significantly affect the transcript abundance of any assayed gene. Pearson correlation coefficients were determined between gene transcript abundance, performance parameters, protein source, inclusion level, and ANF content. Specific growth rate (SGR) was negatively correlated with the abundance of IL-1β and IgM. Dietary inclusion of SBM was positively correlated with all assayed proinflammatory markers and negatively correlated with SGR. Inclusion of PM was positively correlated with both SGR and the abundance of IL-10. The inclusion of CM was negatively correlated with average daily feed intake (ADFI) and with the abundance of both IL-8 and PCNA. Inclusion of PPC correlated positively with SGR and negatively with the activity of caspase-3. Correlation between transcript abundance and dietary content of putative ANFs suggested negative correlations between glucosinolate content, proinflammatory cytokine expression, SGR, and ADFI; whereas, isoflavone content was positively correlated with proinflammatory markers and negatively correlated with SGR. In conclusion, although high SBM and CM inclusion levels have been associated with reduced growth performance in trout, only SBM was associated with increased abundance of inflammatory marker transcripts. These contrasting responses may be mediated by CM glucosinolates, which could negatively affect palatability without inducing a pro-inflammatory response. Dietary PM was very well tolerated and may have promoted anti-inflammatory activity. Further processing of protein meals to concentrates markedly reduced any observable negative impact on performance parameters and the abundance of inflammatory marker mRNA transcripts. Interestingly, both PM and PPC were positively correlated with SGR and may contain a beneficial anti-inflammatory component.

Ecotoxicity testing of chemicals for environmental risk assessment is an area where a high number of vertebrates are used across a variety of industrial sectors. The application of the 3Rs in toxicity testing using fish address both the ethical and societal concerns around this issue in addition to the increasing legislative requests for the incorporation of animal alternatives. This thesis aims to highlight the potential of 3D cell culture models to "bridge the gap" between in vitro and in vivo screening procedures for testing of chemicals with the potential to persist or bioaccumulate thereby improving the predictive power of screening procedures. This thesis examines two alternative methods for their potential use as an intestinal based toxicokinetic tool for environmental risk assessment, utilising an in vitro fish cell line replacement tool (RTgutGC). In addition, for the first time a new intestinal primary cell culture based model was developed to address both intestine region specific response (pyloric, anterior, mid and posterior) and size related adaptability to toxins. Paramagnetic oximetry was used to measure oxygen content within 3D structures (spheroids) in order to better understand the microenvironment of these culture models. Using histology, immunohistochemistry, transepithelial electrical resistance (TEER), transmission electron microscopy (TEM), metabolic, fluorescence and gene expression assays, the comparability of this system to native intestinal response was established. Following exposure to carefully chosen environmental contaminants (Benzo[a]pyrene and Copper), the RTgutGC cell line demonstrated comparable responses to existing literature in terms of uptake, metabolism, DNA damage and the presence an equivalent saturable level. Primary enterocytes cultured on transwell inserts remained viable for upto six weeks, with permeability and metabolic activity comparable to native tissue (both in vitro and ex vivo). Taken in combination, these features of enterocytes represent a profile more closely representative of the intestine then the widely used "gut sac" method. With the potential advantages of incorporating complexity at differing levels (connective tissue layer, intestinal bacteria biome), the intestinal models described offer the potential to screen highly persistent toxins which may require prolonged incubation, in addition to the exploration of complex experimental designs which minimise animal usage (uptake, depuration, uptake). As a consequence, the models developed within this thesis significantly enrich the emerging fish based in vitro testing strategies.

Pharmaceutical pollutants have been aim of many recent aquatic environment studies. Due to human activities surface waters are polluted with plenty of substances, including products of personal care, which are ranked among a number of pharmaceuticals. The main goal of this work is to assess the impact of environmentally relevant concentrations of the synthetic glucocorticoid dexamethasone on aquatic organisms through biomarkers of oxidative stress. The principle of this experiment was chronic in vivo exposure model organisms (rainbow trout) to doses of this drug and the subsequent monitoring activities of antioxidant enzyme system in the liver and gill tissues. Experimental results have demonstrated a response enzymatic protection mechanism on the increased production of reactive oxygen species induced by the drug at environmentally relevant concentrations especially in gill tissue. Our results illustrate a real risk that this xenobiotic represents to aquatic organisms.