K. mikimotoi has been classified under many names and has been mis-assigned to different species and genera in the North Atlantic and Pacific because of its morphological similarities to other Gymnodinoid species. It is now known to be widely distributed, but there remain unresolved questions about whether K. mikimotoi was introduced into the North Sea from Japanese waters, or whether it has always inhabited this region and been erroneously classified as Gymnodinium spp. or has been a part of the hidden flora prior to be recognised in a bloom off the Norwegian coast in 1966. To address questions about geographical genetic variation within K. mikimotoi and broader issues about its biogeography it was deemed important to develop a suitable diagnostic molecular marker that could then be used to monitor the presence/absence of different K. mikimotoi ecotypes over long time scales in European waters. This study showed that the partial rDNA LSU (D1-D2) was too conserved to separate the different strains of K. mikimotoi, while, the ITS region was better able to discriminate between the different strains. However, the rbcL gene was the most informative gene and contained sufficient substitutions to separate the different strains of K. mikimotoi. Specific PCR-primers were designed to amplify a variable region of the rbcL gene able to distinguish differences between K. mikimotoi isolates from the different regions. The innovative high resolution melting temperature (HRM) technique based on specific primer set allowed rapid discrimination of K. mikimotoi from distinct geographic localities (= sequence variants) that differed by only a single nucleotide. Moreover, this study used archival environmental samples collected from the Celtic Sea shelf-break region. The high resolution melting temperature assay successfully detected the European K. mikimotoi isolate within the south-western English Channel in a 1963 sample, which is prior to thefirst report of a K. mikimotoi bloom in Norwegian waters in 1966 and in the south-western English Channel in 1975 and in western Japan in 1965. HRM observations were further validated using clone libraries and sequencing. In summary, this data provided more information about the genotypes present over the analysed timescales, revealing that K. mikimotoi sub-species 2 (European and New Zealand strains) was present in south-western English Channel and south-west Ireland for over 47 years, with sub-species 1 (the Japanese isolate) being absent from all examined samples. This finding supports the hypothesis that K. mikimotoi isolates within Europe are not of Japanese origin and suggests that they are native species to the region.

Class of 2010 Abstract / OBJECTIVES: To determine if children ≤5 years of age come in contact with a greater number of harmful exposures, including medications during holidays compared to non-holidays from January 1, 2000- December 31, 2009. METHODS: This is a descriptive, retrospective study of children ≤5 years of age that are exposed to harmful substances. The independent variable will be holidays versus non-holidays. The primary dependent variable will be the number of harmful exposures. There will be two secondary dependent variables. The first being the number of medication exposures and the second will be the number of non-medication exposures. RESULTS: The proportion of total exposures (med and non-med) significantly increased during the period of 2005-2009 compared to 2000-2004 (1.1%, p<0.001). There were significantly fewer exposures on Friday, Saturday, and Sunday compared to the rest of the week (p<0.001). When comparing the number of exposures on holidays versus non-holidays, there were significantly fewer overall exposures for holidays versus non-holidays (p=0.039). Labor Day had significantly higher exposures than control data (p=0.019) while Christmas and New Year's had significantly less exposures than control data (p<0.0001 and p=0.007 respectively). Labor Day was associated with the highest average number of exposures (30.8) and Christmas was associated with the lowest average number of exposures (21.2) when comparing holidays to each other. A statistically significant greater number of exposures were due to non- medication versus medications for both holiday and non-holiday data (<0.001). The total number of exposures was significantly greater for boys (52.8%) versus girls (46.2%, p<0.001). CONCLUSIONS: Pediatric exposures for the pre-defined holidays are less common when compared to corresponding non-holidays. Interestingly, the same was found for weekends in general where Friday, Saturday, and Sunday yield fewer pediatric exposures than Monday through Thursday. The majority of exposures are due to non-medication sources with all exposures more common in boys than girls.

Harmful algal blooms (HABs) in the Gulf of Mexico are primarily caused by dense aggregations of the dinoflagellate species, Karenia brevis. Karenia brevis produces a highly toxic neurotoxin, brevetoxin which has been shown to cause Neurotoxic Shellfish Poisoning (NSP) and respiratory distress in humans in addition to a wide range of negative impacts upon natural ecosystems. Karenia mikimotoi is a co-existing species present during K. brevis blooms. K. mikimotoi has caused major HAB events in other parts of the ocean, but has not been recognized as a major contributor to toxicity of blooms in the Gulf of Mexico. K. brevis and K. mikimotoi have both been associated with the presence of unidentified hemolytic toxins. Production of hemolysins has not previously been investigated for either species to date in the Gulf of Mexico. Presence of hemolysins may affect toxicity and the overall impact of HABs. Therefore, detection of hemolysins is imperative for accurate identification of potential harmful impacts of such blooms. The primary goal of this research is to define whether either species is capable of producing hemolytic activity independent of brevetoxin activity; and to identify if there is significant differentiation between a variety of clonal isolates regarding toxicity and growth rate when subjected to variable experimental conditions.

Harmful algal blooms (HABs) in the Gulf of Mexico are primarily caused by dense aggregations of the dinoflagellate species, Karenia brevis. Karenia brevis produces a highly toxic neurotoxin, brevetoxin which has been shown to cause Neurotoxic Shellfish Poisoning (NSP) and respiratory distress in humans in addition to a wide range of negative impacts upon natural ecosystems. Karenia mikimotoi is a co-existing species present during K. brevis blooms. K. mikimotoi has caused major HAB events in other parts of the ocean, but has not been recognized as a major contributor to toxicity of blooms in the Gulf of Mexico. K. brevis and K. mikimotoi have both been associated with the presence of unidentified hemolytic toxins. Production of hemolysins has not previously been investigated for either species to date in the Gulf of Mexico. Presence of hemolysins may affect toxicity and the overall impact of HABs. Therefore, detection of hemolysins is imperative for accurate identification of potential harmful impacts of such blooms. The primary goal of this research is to define whether either species is capable of producing hemolytic activity independent of brevetoxin activity; and to identify if there is significant differentiation between a variety of clonal isolates regarding toxicity and growth rate when subjected to variable experimental conditions.

Harmful algal blooms (HABs) are becoming more prevalent throughout the world’s aquatic systems. These blooms have been the subjects of numerous studies because they can cause human health issues and economic impact through fish kills, contaminated shellfish and decreased tourism. Much research has focused on the “bottom-up” aspect of these blooms; namely, the potential role of increased nutrient input into coastal waters from anthropogenic sources causing increased growth in harmful algal species. However, there are also potential “top-down” controls affecting the rate at which harmful algal species are consumed by grazers. The aim of this project was to determine protozoan grazer population fluctuations and their grazing impact on HAB species through field monitoring and laboratory grazing experiments. Protozoan grazers were chosen because their growth rates could potentially keep up with those of HAB species. Declines in grazer populations before the onset of a bloom could be indicative of a release of the HAB from a “top-down” grazing control. Field samples taken during bloom and non-bloom events helped elucidate any microplankton community changes. After establishing that there appear to be changes to the grazer population before and after a bloom, ingestion experiments including direct epifluorescence microscopy and DNA analyses were conducted to determine if it is possible that a chosen protozoan grazer can ingest a HAB species. Finally, experiments were conducted to determine whether the HAB species was a favorable food source for the grazer. Population growth experiments in which grazers are fed a HAB species, 50:50 mixture, or normal culture food source were used to determine the survival and growth rate of the grazer. Although certain ciliates and heterotrophic dinoflagellates were found to feed on HAB species in the lab and in natural bloom samples, the HAB species as a food source produces lower grazer growth rates than on control food. Protozoan grazers may be a more effective control during bloom initiation than after the bloom has been established. / text

Background: It is estimated that adolescent offenders account for 65% of contact child sexual abuse. A range of explanatory theories and models of intervention have been proposed. These are mainly based on cognitive theories with Cognitive Behavioural Therapy (CBT) and psycho-education remaining as the dominant models of treatment intervention. This research explored the issue of harmful sexual behaviour from the psychosocial perspective, drawing on attachment theory and social constructionism. Aim: The research explored attachment strategies among young people with harmful sexual behaviour. It examined young people’s understanding and explanations regarding their sexually abusive behaviour. This included an exploration of how the participants’ accounts and defensive strategies were shaped by wider family and cultural discourses regarding masculinity and sexual activity. In selected cases, the family intergenerational patterns of attachment and emotional regulation were analysed. Furthermore, the research explored the prevalence of harmful sexual behaviour in the study area. This included an exploration of statutory responses and services which were offered to people who engaged in harmful sexual behaviour. Methods: The research employed three interconnected research methods: a) a quantitative audit data of fifty cases which had been reported to the Children’s Services; b) qualitative and descriptive quantitative exploration of eight Transition to Adulthood Attachment Interviews and semi-structured interviews; c) qualitative exploration of the family context and trans-generational patterns of defensive strategies by conducting an Adult Attachment Interview in three cases. The analysis of the attachment and open-ended interviews utilised a combination of the structured ‘discourse analysis’ developed for the Adult Attachment Interview and Critical Discourse Analysis. Findings: The research found that there was no clear and consistent pathway of social care for young people who engaged in harmful sexual behaviour. The analysis of psychological defensive processes found that all of the participants presented with complex insecure attachment strategies alongside intrusions of unresolved traumas or losses. The exploration of intergenerational patterns of attachment strategies identified that all parents showed complex avoidant attachment strategies with intrusion of unresolved trauma and loss. The qualitative study identified a range of discourse themes and strategies in the accounts the young people offered in order to explain their actions and to maintain their sense of self. The dominant discourse themes shared by the young people were: a) life is hard and violence is normal; b) a sense of powerlessness; c) sexual entitlement, d) bravado and denial of vulnerability. Conclusion: Complex insecure attachment strategies with intrusion of trauma and loss play a significant role in the development of harmful sexual behaviour. In addition, the study suggests that the complex relationship between care, comfort and sexual behaviour should be a central focus of clinical interventions with this group of young people and should be investigated further in future research studies.

Harmful algae studies, in particular toxic dinoflagellates, and their response to global warming in the Red Sea are still limited. This study was aimed to be the first to characterize the identity, thermal responses and toxin production of four Prorocentrum strains isolated from the Central Red Sea, Saudi Arabia. Morphological and molecular phylogenetic analysis identified the strains as P. elegans, P. rhathymum and P. emarginatum. However, the identity of strain P. sp.6 is currently unresolved, albeit sharing close affinity with P. leve. Growth experiments showed that all species could grow at 24-32°C, but only P. sp.6 survived the 34°C treatment. The optimum temperatures (Topt) estimated from the Gaussian model corresponded to 27.17, 29.33, 26.87, and 27.64°C for P. sp.6, P. elegans, P. rhathymum and P. emarginatum, respectively. However, some discrepancy with the Topt derived from the growth performance were observed for P. elegans and P. emarginatum, as thermal responses differed from the typical Gaussian fit. The Prorocentrum species examined showed a sharp decrease after the optimum temperature resulting in very high activation energies for the fall slope, especially for P. elegans and P. emarginatum. The minimum critical temperature limit for growth was not detected within the range of temperatures examined. Subsequently, high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis revealed all species as non okadaic acid (OA, common toxin of the Prorocentrum genus) producers at any temperature treatment. However, other forms of toxin (i.e. fast acting toxins) not examined here could be produced. Therefore, further investigations are required. The results of this study provided significant contribution to our knowledge regarding the presence, thermal response and toxin production of four Prorocentrum species from the Central Red Sea, Saudi Arabia.

Stress can be defined as factors which reduce an individual’s survival and/or reproductive ability. Certain stressors strike harder against individuals the more harmful mutations they carry, thus increasing selection against harmful mutations. The aim of this project is to test if exposure to stress, during many generations, decreases the frequency of harmful mutations and lead to more adapted individuals, or if specific adaptations to the stressor override this effect and results in reduced adaptation in the original environment through trade-offs. To test these hypotheses, I use selection lines of Drosophila melanogaster, where the flies during the larval state either have been exposed to media with reduced nutritional value or a heat shock treatment over 22 generations. The results from this experiment show that the selection lines have adapted to their novel environments, since survival of the larvae had increased in the novel environment they had been exposed to for 22 generations. However, survival of selection lines were if anything decreased rather than elevated in the original environment. A plausible explanation to these results is that adaptations to stressors occur both through trade-offs and reduced frequency of generally harmful mutations, but that the effect of the former possibly is larger than the latter. / Stress kan definieras som faktorer som minskar en individs överlevnad och/eller reproduktiva förmåga. Vissa stressmiljöer slår relativt hårdare mot individer ju fler skadliga mutationer de bär, och ökar därför selektionstrycket mot skadliga mutationer. Detta projekt har som mål att testa om exponering mot sådana stressmiljöer, under flera generationer, minskar frekvensen av skadliga mutationer och leder till generellt bättre anpassade individer, eller om specifika anpassningar till stressmiljön överskuggar denna effekt och via trade-offs leder till individer som är sämre anpassade till ursprungsmiljön. För att testa dessa hypoteser använder jag mig här av selektionslinjer av Drosophila melanogaster, där flugorna under larvstadiet antingen utsatts för en näringsfattig miljö eller en värmechock under 22 generationer. Resultaten från detta experiment visar en tydlig evolutionär respons, i och med att larvöverlevnaden ökat för de selekterade linjerna i den stressmiljö de utsatts för efter 22 generationer. Test av överlevnad i ursprungsmiljön visar dock ingen signifikant skillnad mot kontrollinjerna, men om något att de selekterade linjerna klarade sig något sämre. Dessa resultat tyder om något på att anpassningar som skett till den nya miljön på bekostnad av anpassningar i ursprungsmiljön (via trade-offs) överskuggar ökad anpassning via en minskad frekvens av generellt skadliga mutationer.

Interest in the effect of fish farming practices on the marine environment has arisen because there is concern that the wastes that fish farms produce may be contributing to eutrophication in coastal areas and the problem of harmful algal blooms. The focus of this thesis is an examination of phytoplankton distribution and abundance in relation to tuna fish farms in Boston Bay and near-shore Spencer Gulf. This is the first study in South Australia to define the short-term biomass fluctuations of chlorophyll and in vivo fluorescence, identify phytoplankton species distribution and abundance, including two potentially toxic dinoflagellates, and describe patch distribution relative to tuna fish farms in Boston Bay and the near-shore waters of Spencer Gulf. An ecological interpretation of phytoplankton distribution and abundance is determined and shows that community composition was different in lower Spencer Gulf compared to Boston Bay and upper Spencer Gulf sites. Pico- and nanophytoplankton were often the most abundant organisms. Diatoms and gymnoids were most common. Season and currents predominantly influenced the distribution of phytoplankton in Boston Bay and Spencer Gulf. Individual species may be influenced by inputs from the fish farms. Chlorophyll levels were different between the Spencer Gulf and Boston Bay sites and no differences were recorded, using mean levels of chlorophyll, between tuna cages and controls. Chlorophyll levels were higher east of Boston Island in autumn of 1999. Chlorophyll levels appeared to show a slight increase between years. This may have been an anomalous natural variation and future research may investigate this in the long term. In addition, Principal Components Analysis (PCA) was used to investigate differences between treatments and the functional grouping model supported an ecological interpretation of the factors from the PCA. A total of 131 taxa of phytoplankton were identified in this study. The 14 dominant taxa were used in the PCA and of these, 9 were diatoms. Phytoplankton abundance was not different between tuna cages and controls. However, when examining individual species, Karenia mikimotoi was more prevalent at tuna cages, close to shore, east and west of Boston Island than at other sites. PCA showed how different species bloomed together and were seasonal. Karenia brevis and K. mikimotoi featured predominantly in the PCA with K. brevis the dominant organism during summer and autumn along with Gyrodinium spp. and smaller gymnoids. K. brevis blooms were most likely influenced by water temperatures and fixation of nitrogen from a Trichodesmium erythaeum bloom. K. mikimotoi bloomed bimodally and may be influenced by ammonia excreted from fish from the tuna farms but , on the other hand, may be limited by the high salinities of South Australian waters. Currents in the region distribute both organisms. The final aspect of this study assessed finer temporal and spatial sampling using directional transects around tuna cages and controls using in-vivo fluorescence and size fractionated chlorophyll. The chlorophyll a sampling showed little spatial variability within a site in the 1000 m2 that the sampling area covers but far greater temporal variability (days). In contrast, fluorescence `mapping' expands the window of variability both spatially (within a site) and temporally (along transects and between days). This has given a spatial definition, which is unavailable from a single point sample, and thereby leaves room for much greater interpretation. Small patches are evident from the fluorescence mapping where this is impossible to detect from the single point samples. Therefore, the fluorescence `mapping' and patch definition show that the trend is widespread (spatially) and quite persistent (temporally) around the fish farm area. Size fractionated chlorophyll samples provided further insight into phytoplankton dynamics in this study where diatoms were favored over dinoflagellates and were responsible for the larger fraction of chlorophyll found at the tuna cage one (TC1) site. We suggest that seasonal fluctuations, high nutrient input from the farm activities and turbulence may be responsible for the different chlorophyll/fluorescent structures found at TC1. Future research may look at the long-term regional impact on phytoplankton size structure, biomass and communities from fish farm activities. As a good part of this journey involved counting phytoplankton using the Utërmohl technique, a short paper, published in the Journal of Plankton Research, on reducing the settling time of this method, is presented in Appendix.

Over the past two decades, Lake Winnipeg has been experiencing increasingly rapid eutrophication, and large cyanobacterial blooms now form in the North Basin in most years in late summer or fall. Cyanobacteria are considered a relatively poor food source compared with other phytoplankton, but the impacts of these blooms upon the primary consumers in the lake have not previously been researched. A microscopic analysis of whole water samples found cyanobacteria to be scarcely present in summer 2012, with nitrogen-fixing and non-fixing cyanobacteria comprising 11.2% and 8.4% of the basin-wide biovolume, respectively, and all but absent in fall. Gut content analysis of chironomids found that cyanobacteria made up an almost negligible part of their diet. Stable isotope analysis revealed that nitrogen-fixing cyanobacteria reduced phytoplankton δ15N values, and that this same reduction could be traced through the zooplankton, but not down to the sediments or chironomids.