The objective of this study was to determine if the presence of two ALS inhibiting herbicide residues in three Saskatchewan soils would result in an additive, synergistic, or antagonistic interaction. This was determined through field trials where herbicides were applied sequentially over the course of two years and through dose-response modelling. The herbicides examined in these experiments were imazamethabenz, flucarbazone-sodium, sulfosulfuron, and florasulam, each in combination with imazamox/imazethapyr. The phytotoxicity and persistence of the herbicides in soil was assessed using an Oriental mustard root inhibition bioassay. The determination of herbicide interaction was made through the comparison of the experimentally observed values to theoretically expected values derived from a mathematical equation.<p>The dose response curves created by placing incremental concentrations of these herbicides in soil were compared using the I50 parameter, which is the concentration resulting in a 50% reduction in root length. It appeared that soil organic matter followed by soil pH had the greatest effect in reducing herbicide residue phytotoxicity in the tested soils. Based on the bioassay analysis of sequentially applied ALS inhibiting herbicides, it is proposed that the phytotoxic effect of herbicide residues in soil result in additive injury effects rather than synergistic or antagonistic interactions.

The overall objective of the work described in this thesis was to examine the effects of both waterborne and dietary routes of exposure to fathead minnow (Pimephales promelas) when exposed to complex metal mining mixtures. This was conducted using a 21-day, multi-trophic, short-term fathead minnow (FHM) reproductive bioassay. The endpoints that were measured were used to assess the effects on multiple levels of biological organization (sub-organismal to population endpoints). The first phase of this research was conducted in situ using environmentally realistic concentrations of 3 separate metal mining effluents [20% surface water effluent (SWE), 30% mine water effluent (MWE), 45% process water effluent (PWE)] from Sudbury, Ontario, Canada. Metals were analyzed in several media (water, sediments) and tissues (biofilm, Chironomus dilutus, female fathead minnow carcass, ovaries, liver and gills). The incorporation of the biofilm (primary producers) into the bioassay also added another level of organization that was novel to this study. Significant increases in metal concentrations were observed in the water and biofilm tissues in all treatments [SWE, MWE, PWE], compared to reference. Cobalt and nickel increased significantly in C. dilutus tissues in SWE (1.4-fold and 1.5-fold respectively), and copper and selenium in PWE (5.2-fold and 3.3-fold respectively), however no significant increases occurred in MWE compared to reference. There were no significant increases in metal concentrations in female FHM tissues (carcass, liver, gonads, gills) in any of the treatments, suggesting that metal bioavailability was reduced. Cumulative number of eggs per female per day increased significantly (+127%) after exposure to SWE and decreased significantly (-33%) after exposure to PWE when compared to the reference fish. Mean total number of days to hatch was also reduced in PWE compared to reference. In order to gain a better understanding of the routes of exposure causing toxicity in FHM, the second phase of this research examined the effects of exposure through diet, through water or through both using a fully factorial food exposure design in a laboratory setting. In this experiment we pre-exposed C. dilutus to both 45% PWE and laboratory control water until they reached the 3rd-4th instar stage of development (approximately 21 days) where they were collected and frozen until the start of the FHM reproductive bioassay. We further examined the role of food quality on fish toxicity by assessing differences between multi trophic (where fish were fed both a live and frozen diet of C. dilutus) in the laboratory. This research was conducted at the Toxicology Centre in Saskatoon, Saskatchewan, Canada. The results showed that significant effects were observed when fish were fed a live diet versus a frozen diet. Condition factor and body weight increased, although inconsistent effects were observed for liver somatic index (LSI) in fathead minnows in both experiments when exposed to one or both routes of exposure. Cumulative total egg production and cumulative spawning events were both significantly affected by both waterborne and dietborne exposures with the greatest effects seen in the multi-trophic streams and particularly when fish were fed a live diet. This significance of this research has demonstrated the importance of including both routes of exposure when assessing effects of mine effluent. This research also shows that the artificial stream technology is a useful tool in isolating the effects of a particular point source input (metal mining mixtures) when a system is highly confounded. The results suggest that under environmentally relevant exposure conditions, trophic transfer and live diet may lead to greater reproductive effects and increased fish toxicity. This also suggests that trophic transfer is an important route of exposure that is virtually impossible to attain using typical laboratory bioassay techniques (food-borne study using artificial diets or waterborne exposures only).

The present study deals with the comparison and establishment of methods for the detection of antibodies against recombinant human growth hormone (rhGH). Therefore, different methods for the detection of hGH-Abs were evaluated and compared in order to establish a test system that can be used for the detection of neutralizing antibodies against hGH, which could be developed under rhGH treatment. This manuscript describes in detail the validation of a newly developed biological assay, the neutralizing hGH-antibody assay (NAb assay). Therefore, a cell line transfected with the growth hormone receptor, that proliferates in the presence of hGH, was used. This proliferation was quantified by an increase of the optical density (OD/ absorbance) after addition of a colorimetric reagent, whereas the presence of hGH-antibodies leads to an inhibition of cell proliferation. To validate the test system for the detection of hGH-antibodies, we tested serum samples of 4 patients suffering from neurosecretory dysfunction (NSD) and samples taken from 6 patients with growth hormone deficiency (GHD) which were treated with rhGH and were highly suspected for a-hGH antibodies. These samples were tested in two different immunological assays, capable to screen sera for anti-hGH immunreactivity in the case of hGH-insensitivity during GH treatment. Using the NAb assay the neutralizing activity of specific hGH-antibodies was proved in serum samples of NSD and GHD type 1A patients. In case of neutralizing hGH-antibody activity, a clinically based decision can be made whether rhGH therapy should be stopped or the rhGH dosis should be increased. By the use of our test system, we offer the measurement of anti-hGH-antibody activity to other laboratories in cases when secondary hGH-insensitivity is assumed or observed.

Of the small percentage of organisms chemically investigated over the years as potential sources of natural products, much less is known about those from the marine realm. Despite the lack of attention they have received in comparison to terrestrial organisms, marine life have recently been found to represent a valuable source for novel bioactive compounds. Cold water marine habitats are home to a plethora of organisms that have the ability to produce secondary metabolites that exhibit a great deal of diversity in both their chemical structures and biological activities. The chemical investigation of these unique and relatively unstudied ecosystems is necessary to gain insight into the dynamics between predators and prey, while also making a significant impact in the field of drug discovery. Our laboratory has focused on the chemical investigation of invertebrates from the waters of Antarctica in search of bioactive secondary metabolites that can be used for the treatment of human pathogens. This dissertation reports a small portion of the progress made in our laboratory towards the exploration of Antarctic marine invertebrates. The chemical investigation of the circumpolar colonial tunicate Synoicum adareanum and the orange, encrusting sponge Artemisina plumosa will be discussed in detail in the following chapters.

The Deepwater Horizon blowout in 2010 released an estimated 4.9 million barrels of crude oil into the Gulf of Mexico in the 83 days between the initial explosion and the capping of the well. Response included extensive use of Corexit© oil dispersant. Although South Florida was spared exposure by currents, this event highlights the need for effective bioassay organisms for coral reefs. Amphistegina spp. are benthic foraminifers that host diatom symbionts in a relationship similar to that of coral and their zooxanthellae. Amphistegina spp. occur abundantly in reef communities nearly worldwide, are easily collected and maintained in culture, and are a key component of the FoRAM Index, a indicator of water and sediment quality in coastal waters. The major goals of this project were to develop protocols to test the acute and chronic responses of A. gibbosa to potentially toxic organic chemicals. Initial objectives were to determine lethal concentrations and effects ranges, as defined by the US Environmental Protection Agency, of two components of the Corexit© dispersants. Preliminary experiments indicated that many specimens exposed to propylene glycol (v/v) at concentrations of 2% or higher appeared to be dead following 48-hour exposure, resulting in apparent LC50 of 3% and an initial effects range of 2-4%. When placed in filtered seawater, after 72-hours the observed LC50 was 6%. All parameters assessed, including sub-lethal chronic effects (differences in growth and visible responses after 40 days), revealed an effects range of 0.5% to 12%, above which there was 100% mortality. For 2-butoxyethanol, the apparent LC50 after 48-hour exposure was 0.2%; after 72-hour recovery the LC50 was 1%. In all experiments, a 72-hour recovery period was sufficient to determine acute effects. A key discovery was the observation of inactivity during exposure to toxic substances, followed by recovery when placed in filtered seawater. This observation indicates the potential for dormancy in adult foraminifers exposed to toxic substances that has not previously been reported.

Reclamation success is highly dependent upon final soil quality of stockpiled soils, such as those found Cold Lake Oil Sands Region. Stockpiled soils, however, are generally poor in quality. Soils were amended with, biochar (BC), humalite (HU), a sub-bituminous coal; and peat (PT), as well as 50:50 blends of biochar:humalite (BCH) and biochar:peat (BCP). These amendments were applied at rates of 0, 6.55, 13.1 and 26.2 g C kg-1 each. Biochar, PT, and BCP applied at the rate of 26.2 g C kg-1 were found to reduce N2O emissions by 34, 54, and 70%, respectively, relative to the control. Within the bioassay, BC and PT amendment resulted in a 38 and 40% increase in dry matter yield (DMY) respectively. Finally, amendment typically resulted in significant net decreases in Olsen P values, while nitrate and ammonium concentrations were high in PT amended soils. In general, PT had the best overall performance. / October 2015

No description available.

Soil quality guidelines are currently based on total trace metal loads. There is a need to define indices of bioavailability to allow reasonable predictions for plant metal uptake and toxicity in soils. Trace metal toxicities to plants often correlate best with free metal ion activities. The first objective was to develop a plant bioassay that is sensitive to trace metals at concentrations realistic for soils. The root elongation of lettuce Lactuca sativa 'Buttercrunch' was used as toxicological endpoint. This endpoint was sensitive and reproducible to environmentally relevant concentrations of Cd, Cu, Ni, Pb and Zn. The second objective was to test whether free metal ion activities are constant predictors of metal toxicities in synthetic solutions and in soil extracts that differ in their concentrations of cations and ligands. The root elongation assay was used to test this hypothesis. In synthetic solutions, the rhizotoxicity of Cd, Cu, Ni, Pb and Zn decreased with increasing Ca and H concentrations. This could not be explained with the effect of higher cationic concentrations on root growth or on solution speciation. It was concluded that Ca and H inhibited the rhizotoxicity of all metals tested. The rhizotoxicity of Cu and Cd was further examined in soil extracts. Both metals became less rhizotoxic at higher H and dissolved organic matter concentrations. The rhizotoxicity endpoints from the experiments in synthetic solution were used to develop parameters for a Biotic Ligand Model (BLM) for Cd, Cu, Ni, Pb and Zn. The BLM accounts for solution speciation and interprets cationic inhibition of rhizotoxicity as competition of metals with Ca and H for potential sites of rhizotoxicity. The BLM predicted metal rhizotoxicity better than the free metal ion activity in synthetic solutions and in soil extracts. Different models were tested against literature rhizotoxicity data for metals at different Ca and H concentrations. Predictions for metal rhizotoxicity given by BLM, Gouy-Chapman-Stern model and Freundlich equation model were compared with predictions based on free metal ion activities in solution. The BLM predicted rhizotoxicity most accurately. The BLM seems promising for predictions of metal toxicity and metal bioavailability in soils to support site-specific environmental risk assessments.

In this study, the ability of various chemicals to be biotransformed into mutagens by plant cells was investigated. Two thiocarbamate herbicides, diallate and triallate, the sulfonylurea herbicide chlorsulfuron, and the aniline derivative m-phenylenediamine were tested for their ability to revert Salmonella typhimurium (strains TA100 and TA98) in the presence and absence of Nicotiana tabacum (TX1) cell cultures in liquid suspension. Chlorsulfuron and m-phenylenediamine were also tested in the presence and absence of Brassica napus cv. 'Topas' cells. Diallate was found to be activated by TX1 cells into a mutagen that induces base-pair substitution mutations. In the presence of the TX1 plant cell line, chlorsulfuron significantly increased the number of mutations on the strain TA98 of Salmonella. Tobacco TX1 cells did not activate triallate into a mutagen. m-Phenylenediamine was activated into a mutagen by TX1 and Brassica cells as detected by Salmonella TA98. This aniline derivative, in the absence of plant cells and at concentrations higher than 20 $ mu$ Moles/plate, was also able to significantly increase the number of TA98 revertants as compared to the control plants. Finally, Brassica napus cells activated chlorsulfuron into a mutagen that induces frameshift mutations.

The degradation of wetlands and loss of their associated ecosystem services is widely recognised in South Africa, however, at present there is no standard method of biologically assessing wetland health in this country. Internationally, particularly in the U.S.A and Australia, wetland bioassessment techniques using macroinvertebrates are well established. A number of these wetland bioassessment protocols have been derived from local river biomonitoring techniques, as there is a belief that river and wetland ecology and macroinvertebrate assemblages at family level are similar. However, some authors consider wetland macroinvertebrate assemblages and ecological processes to differ greatly from those found in rivers, and believe that such techniques are not transferable. South Africa has a well established macroinvertebrate biomonitoring protocol for rivers called SASS5 (South African Scoring System Version 5). This study is a preliminary investigation into the extent to which the SASS5 scoring system is applicable to the assessment of nutrient enriched wetland water quality. Macroinvertebrates are particularly suitable as biomonitoring tools: they respond to a variety of stressors, have life cycles that allow for integrated responses to episodic pollution, and are relatively easy to identify to family level. When selecting wetlands for the development of a biomonitoring protocol, wetlands should all be of the same; classification (Le. palustrine), geomorphological and climate setting, hydrological regime and dominant vegetation class. Sampling was restricted to sedge-dominated palustrine wetlands in the midlands of KwaZulu-Natal, with similar hydro-geomorphological settings. Due to wetlands and rivers having different biotopes (e.g. no riffles present in wetlands), the SASS5 sampling protocol could not be used, thus a pilot investigation was undertaken to derive a suitable sampling technique for \ collecting a representative and diagnostic sample of aquatic macroinvertebrates from a wetland. This technique was developed based on published methods. Both sweep net and activity trap sampling were conducted, and each evaluated for their effectiveness at macroinvertebrate collection. Sweep net sampling was tested over a range of sweep intensities (2-6 sweeps), and activity traps were placed at four different depths: at the water surface, just below the surface, 0.10.15m below surface and on the substrate. A total of 32 taxa identified to family level were identified in the samples. Taxon diversity and composition did not differ in the activity traps placed at the four depth locations. Taxon diversity did not differ significantly between different sweep intensities; however there was a significant difference in taxon composition between the different sweep intensities and between activity trap and sweep net samples (p<0.05). Sixty-eight percent of taxa appeared more frequently in sweep net sampling compared to activity trap sampling. Six taxa were found exclusively in sweep net samples, and two taxa were recorded exclusively in activity traps. There was no trend in either method collecting more or missing any unique trophic group. In conclusion, activity traps are not required to supplement sweep net data, and a technique using a sweep net with a sweep intensity of five would be suitable to collect a representative sample of wetland macroinvertebrates. Using the derived technique, four reference and three wetlands impacted by dairy effluent were sampled. Six macroinvertebrate samples were collected from each of the seven wetland, together with data for selected physico-chemical variables, macrohabitat condition, biotope suitability and organism detectability. For each sample, the macroinvertebrates were identified and assigned a predetermined SASS5 tolerance score between 1 and 15, with higher scores indicating increased sensitivity to poor water quality. 11 A total of 39 taxa, identified to family level, were collected during sampling. SASS5 scores ranged from 15-82. Five of the wetlands had mean SASS5 scores of between 46 and 59. Five of the wetlands had an intra-wetland SASS5 score range greater than 30. ASPT values ranged from 3.3 to 5.5, and few high scoring (~8) taxa were collected. There was no significant difference in SASS5 scores between samples collected above, at and downstream of an effluent discharge point within the same impacted wetland. SASS5 scores for reference wetlands were also not significantly higher than those recorded for impacted wetlands. Comparison of ranked SASS5 scores and environmental data did suggest a relationship between the variables, but was not significant. Based on the SASS5 score water quality guidelines, all sampled wetlands were considered to have impacted water quality; however, this was not supported by the macrohabitat and physico-chemical results. Possible reasons for the low SASS5 scores include: the lack of 'stones in/out current' biotopes in wetlands, lower levels of dissolved oxygen present compared to rivers, and the limited detectability of organisms due to large amounts of substrate in the samples. A wetland adaptation of SASS5 would require the reassignment of modified scores to certain taxa based on their distribution in wetlands of varying water quality. The SASS5 score level of 100 and the ASPT value of 6 (as specified in the SASS5 score water quality guidelines) were found to be inappropriate for wetlands. It is suggested that, either the range of taxa tolerance scores be increased (1 to >15), or the score level of 100 be lowered. The ASPT value should also be reduced. Although SASS5 appears unsuitable for assessing wetlands, variations in taxon composition between sampled wetlands, identified through CA analysis, suggests that macroinvertebrates are responsive to changes in wetland condition, and thus have potential as indicators of wetland water quality. Nine taxa responsive to the presence of nitrogen have been identified as being potentially good indicators. iii Further research should focus on the testing of SASS5 throughout the year, in a range of wetland types, and in wetlands moderately to severely impacted by pollutants other than dairy effluent. It is recommended that a habitat or biotope index be developed and used in conjunction with any future wetland macroinvertebrate bioassessment protocols. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.