Prevalence and occurrence of zoonotic bacterial pathogens in surface waters determined by quantitative PCR

The prevalence and concentrations of Campylobacter jejuni, Salmonella spp. and enterohaemorrhagic E. coli (EHEC) were investigated in surface waters in Brisbane, Australia using quantitative PCR (qPCR) based methodologies. Water samples were collected from Brisbane City Botanic Gardens (CBG) Pond, and two urban tidal creeks (i.e., Oxley Creek and Blunder Creek). Of the 32 water samples collected, 8 (25%), 1 (3%), 9 (28%), 14 (44%), and 15 (47%) were positive for C. jejuni mapA, Salmonella invA, EHEC O157 LPS, EHEC VT1, and EHEC VT2 genes, respectively. The presence/absence of the potential pathogens did not correlate with either E. coli or enterococci concentrations as determined by binary logistic regression. In conclusion, the high prevalence, and concentrations of potential zoonotic pathogens along with the concentrations of one or more fecal indicators in surface water samples indicate a poor level of microbial quality of surface water, and could represent a significant health risk to users. The results from the current study would provide valuable information to the water quality managers in terms of minimizing the risk from pathogens in surface waters.

Improved detection and tracking of objects in surveillance video

Surveillance networks are typically monitored by a few people, viewing several monitors displaying the camera feeds. It is then very difficult for a human operator to effectively detect events as they happen. Recently, computer vision research has begun to address ways to automatically process some of this data, to assist human operators. Object tracking, event recognition, crowd analysis and human identification at a distance are being pursued as a means to aid human operators and improve the security of areas such as transport hubs. The task of object tracking is key to the effective use of more advanced technologies. To recognize an event people and objects must be tracked. Tracking also enhances the performance of tasks such as crowd analysis or human identification. Before an object can be tracked, it must be detected. Motion segmentation techniques, widely employed in tracking systems, produce a binary image in which objects can be located. However, these techniques are prone to errors caused by shadows and lighting changes. Detection routines often fail, either due to erroneous motion caused by noise and lighting effects, or due to the detection routines being unable to split occluded regions into their component objects. Particle filters can be used as a self contained tracking system, and make it unnecessary for the task of detection to be carried out separately except for an initial (often manual) detection to initialise the filter. Particle filters use one or more extracted features to evaluate the likelihood of an object existing at a given point each frame. Such systems however do not easily allow for multiple objects to be tracked robustly, and do not explicitly maintain the identity of tracked objects. This dissertation investigates improvements to the performance of object tracking algorithms through improved motion segmentation and the use of a particle filter. A novel hybrid motion segmentation / optical flow algorithm, capable of simultaneously extracting multiple layers of foreground and optical flow in surveillance video frames is proposed. The algorithm is shown to perform well in the presence of adverse lighting conditions, and the optical flow is capable of extracting a moving object. The proposed algorithm is integrated within a tracking system and evaluated using the ETISEO (Evaluation du Traitement et de lInterpretation de Sequences vidEO - Evaluation for video understanding) database, and significant improvement in detection and tracking performance is demonstrated when compared to a baseline system. A Scalable Condensation Filter (SCF), a particle filter designed to work within an existing tracking system, is also developed. The creation and deletion of modes and maintenance of identity is handled by the underlying tracking system; and the tracking system is able to benefit from the improved performance in uncertain conditions arising from occlusion and noise provided by a particle filter. The system is evaluated using the ETISEO database. The dissertation then investigates fusion schemes for multi-spectral tracking systems. Four fusion schemes for combining a thermal and visual colour modality are evaluated using the OTCBVS (Object Tracking and Classification in and Beyond the Visible Spectrum) database. It is shown that a middle fusion scheme yields the best results and demonstrates a significant improvement in performance when compared to a system using either mode individually. Findings from the thesis contribute to improve the performance of semi-automated video processing and therefore improve security in areas under surveillance.

Microbial risk from rainwater tanks in South East Queensland

Quantitative Microbial Risk Assessment (QMRA) analysis was used to quantify the risk of infection associated with the exposure to pathogens from potable and non-potable uses of roof-harvested rainwater in South East Queensland (SEQ). A total of 84 rainwater samples were analysed for the presence of faecal indicators (using culture based methods) and zoonotic bacterial and protozoan pathogens using binary and quantitative PCR (qPCR). The concentrations of Salmonella invA, and Giardia lamblia β-giradin genes ranged from 65-380 genomic units/1000 mL and 9-57 genomic units/1000 mL of water, respectively. After converting gene copies to cell/cyst number, the risk of infection from G. lamblia and Salmonella spp. associated with the use of rainwater for bi-weekly garden hosing was calculated to be below the threshold value of 1 extra infection per 10,000 persons per year. However, the estimated risk of infection from drinking the rainwater daily was 44-250 (for G. lamblia) and 85-520 (for Salmonella spp.) infections per 10,000 persons per year. Since this health risk seems higher than that expected from the reported incidences of gastroenteritis, the assumptions used to estimate these infection risks are critically discussed. Nevertheless, it would seem prudent to disinfect rainwater for potable use.

Reliability driven reconfiguration of rural power distribution systems

This paper presents a reliability-based reconfiguration methodology for power distribution systems. Probabilistic reliability models of the system components are considered and Monte Carlo method is used while evaluating the reliability of the distribution system. The reconfiguration is aimed at maximizing the reliability of the power supplied to the customers. A binary particle swarm optimization (BPSO) algorithm is used as a tool to determine the optimal configuration of the sectionalizing and tie switches in the system. The proposed methodology is applied on a modified IEEE 13-bus distribution system.

Digital kids, analogue students : a mixed methods study of students' engagement with a school-based Web 2.0 learning innovation

The inquiry documented in this thesis is located at the nexus of technological innovation and traditional schooling. As we enter the second decade of a new century, few would argue against the increasingly urgent need to integrate digital literacies with traditional academic knowledge. Yet, despite substantial investments from governments and businesses, the adoption and diffusion of contemporary digital tools in formal schooling remain sluggish. To date, research on technology adoption in schools tends to take a deficit perspective of schools and teachers, with the lack of resources and teacher ‘technophobia’ most commonly cited as barriers to digital uptake. Corresponding interventions that focus on increasing funding and upskilling teachers, however, have made little difference to adoption trends in the last decade. Empirical evidence that explicates the cultural and pedagogical complexities of innovation diffusion within long-established conventions of mainstream schooling, particularly from the standpoint of students, is wanting. To address this knowledge gap, this thesis inquires into how students evaluate and account for the constraints and affordances of contemporary digital tools when they engage with them as part of their conventional schooling. It documents the attempted integration of a student-led Web 2.0 learning initiative, known as the Student Media Centre (SMC), into the schooling practices of a long-established, high-performing independent senior boys’ school in urban Australia. The study employed an ‘explanatory’ two-phase research design (Creswell, 2003) that combined complementary quantitative and qualitative methods to achieve both breadth of measurement and richness of characterisation. In the initial quantitative phase, a self-reported questionnaire was administered to the senior school student population to determine adoption trends and predictors of SMC usage (N=481). Measurement constructs included individual learning dispositions (learning and performance goals, cognitive playfulness and personal innovativeness), as well as social and technological variables (peer support, perceived usefulness and ease of use). Incremental predictive models of SMC usage were conducted using Classification and Regression Tree (CART) modelling: (i) individual-level predictors, (ii) individual and social predictors, and (iii) individual, social and technological predictors. Peer support emerged as the best predictor of SMC usage. Other salient predictors include perceived ease of use and usefulness, cognitive playfulness and learning goals. On the whole, an overwhelming proportion of students reported low usage levels, low perceived usefulness and a lack of peer support for engaging with the digital learning initiative. The small minority of frequent users reported having high levels of peer support and robust learning goal orientations, rather than being predominantly driven by performance goals. These findings indicate that tensions around social validation, digital learning and academic performance pressures influence students’ engagement with the Web 2.0 learning initiative. The qualitative phase that followed provided insights into these tensions by shifting the analytics from individual attitudes and behaviours to shared social and cultural reasoning practices that explain students’ engagement with the innovation. Six indepth focus groups, comprising 60 students with different levels of SMC usage, were conducted, audio-recorded and transcribed. Textual data were analysed using Membership Categorisation Analysis. Students’ accounts converged around a key proposition. The Web 2.0 learning initiative was useful-in-principle but useless-in-practice. While students endorsed the usefulness of the SMC for enhancing multimodal engagement, extending peer-topeer networks and acquiring real-world skills, they also called attention to a number of constraints that obfuscated the realisation of these design affordances in practice. These constraints were cast in terms of three binary formulations of social and cultural imperatives at play within the school: (i) ‘cool/uncool’, (ii) ‘dominant staff/compliant student’, and (iii) ‘digital learning/academic performance’. The first formulation foregrounds the social stigma of the SMC among peers and its resultant lack of positive network benefits. The second relates to students’ perception of the school culture as authoritarian and punitive with adverse effects on the very student agency required to drive the innovation. The third points to academic performance pressures in a crowded curriculum with tight timelines. Taken together, findings from both phases of the study provide the following key insights. First, students endorsed the learning affordances of contemporary digital tools such as the SMC for enhancing their current schooling practices. For the majority of students, however, these learning affordances were overshadowed by the performative demands of schooling, both social and academic. The student participants saw engagement with the SMC in-school as distinct from, even oppositional to, the conventional social and academic performance indicators of schooling, namely (i) being ‘cool’ (or at least ‘not uncool’), (ii) sufficiently ‘compliant’, and (iii) achieving good academic grades. Their reasoned response therefore, was simply to resist engagement with the digital learning innovation. Second, a small minority of students seemed dispositionally inclined to negotiate the learning affordances and performance constraints of digital learning and traditional schooling more effectively than others. These students were able to engage more frequently and meaningfully with the SMC in school. Their ability to adapt and traverse seemingly incommensurate social and institutional identities and norms is theorised as cultural agility – a dispositional construct that comprises personal innovativeness, cognitive playfulness and learning goals orientation. The logic then is ‘both and’ rather than ‘either or’ for these individuals with a capacity to accommodate both learning and performance in school, whether in terms of digital engagement and academic excellence, or successful brokerage across multiple social identities and institutional affiliations within the school. In sum, this study takes us beyond the familiar terrain of deficit discourses that tend to blame institutional conservatism, lack of resourcing and teacher resistance for low uptake of digital technologies in schools. It does so by providing an empirical base for the development of a ‘third way’ of theorising technological and pedagogical innovation in schools, one which is more informed by students as critical stakeholders and thus more relevant to the lived culture within the school, and its complex relationship to students’ lives outside of school. It is in this relationship that we find an explanation for how these individuals can, at the one time, be digital kids and analogue students.

Campylobacter jejuni and campylobacter coli genotyping by high-resolution melting analysis of a flaA fragment

The highly variable flagellin-encoding flaA gene has long been used for genotyping Campylobacter jejuni and Campylobacter coli. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The objective of this study was to apply HRM analysis to flaA-based genotyping. The initial aim was to identify a suitable flaA fragment. It was found that the PCR primers commonly used to amplify the flaA short variable repeat (SVR) yielded a mixed PCR product unsuitable for HRM analysis. However, a PCR primer set composed of the upstream primer used to amplify the fragment used for flaA restriction fragment length polymorphism (RFLP) analysis and the downstream primer used for flaA SVR amplification generated a very pure PCR product, and this primer set was used for the remainder of the study. Eighty-seven C. jejuni and 15 C. coli isolates were analyzed by flaA HRM and also partial flaA sequencing. There were 47 flaA sequence variants, and all were resolved by HRM analysis. The isolates used had previously also been genotyped using single-nucleotide polymorphisms (SNPs), binary markers, CRISPR HRM, and flaA RFLP. flaAHRManalysis provided resolving power multiplicative to the SNPs, binary markers, and CRISPR HRM and largely concordant with the flaA RFLP. It was concluded that HRM analysis is a promising approach to genotyping based on highly variable genes.

Prevalence of Staphylococcus aureus strains in an Australian cohort, 1989-2003 : evidence for the low prevalence of the toxic shock toxin and Panton-Valentine leukocidin genes

The purpose of this paper is to determine the prevalence of the toxic shock toxin gene (tst) and to enumerate the circulating strains of methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in Australian isolates collected over two decades. The aim was to subtype these strains using the binary genes pvl, cna, sdrE, pUB110 and pT181. Isolates were assayed using real-time polymerase chain reaction (PCR) for mecA, nuc, 16 S rRNA, eight single-nucleotide polymorphisms (SNPs) and for five binary genes. Two realtime PCR assays were developed for tst. The 90 MRSA isolates belonged to CC239 (39 in 1989, 38 in 1996 and ten in 2003), CC1 (two in 2003) and CC22 (one in 2003). The majority of the 210 MSSA isolates belonged to CC1 (26), CC5 (24) and CC78 (23). Only 18 isolates were tst-positive and only 15 were pvl-positive. Nine MSSA isolates belonged to five binary types of ST93, including two pvlpositive types. The proportion of tst-positive and pvl-positive isolates was low and no significant increase was demonstrated. Dominant MSSA clonal complexes were similar to those seen elsewhere, with the exception of CC78. CC239 MRSA (AUS-2/3) was the predominant MRSA but decreased significantly in prevalence, while CC22 (EMRSA-15) and CC1 (WA-1) emerged. Genetically diverse ST93 MSSA predated the emergence of ST93- MRSA (the Queensland clone).

Queer

The word “queer” is a slippery one; its etymology is uncertain, and academic and popular usage attributes conflicting meanings to the word. By the mid-nineteenth century, “queer” was used as a pejorative term for a (male) homosexual. This negative connotation continues when it becomes a term for homophobic abuse. In recent years, “queer” has taken on additional uses: as an all encompassing term for culturally marginalised sexualities – gay, lesbian, trans, bi, and intersex (“GLBTI”) – and as a theoretical strategy which deconstructs binary oppositions that govern identity formation. Tracing its history, the Oxford English Dictionary notes that the earliest references to “queer” may have appeared in the sixteenth century. These early examples of queer carried negative connotations such as “vulgar,” “bad,” “worthless,” “strange,” or “odd” and such associations continued until the mid-twentieth century. The early nineteenth century, and perhaps earlier, employed “queer” as a verb, meaning to “to put out of order,” “to spoil”, “to interfere with”. The adjectival form also began to emerge during this time to refer to a person’s condition as being “not normal,” “out of sorts” or to cause a person “to feel queer” meaning “to disconcert, perturb, unsettle.” According to Eve Sedgwick (1993), “the word ‘queer’ itself means across – it comes from the Indo-European root – twerkw, which also yields the German quer (traverse), Latin torquere (to twist), English athwart . . . it is relational and strange.” Despite the gaps in the lineage and changes in usage, meaning and grammatical form, “queer” as a political and theoretical strategy has benefited from its diverse origins. It refuses to settle comfortably into a single classification, preferring instead to traverse several categories that would otherwise attempt to stabilise notions of chromosomal sex, gender and sexuality.

Nasal carriage of Staphylococcus aureus, including community-associated methicillin-resistant strains, in Queensland adults

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections are emerging in southeast Queensland, Australia, but the incidence of carriage of CA-MRSA strains is unknown. The aim of this study was to assess the nasal carriage rate of S. aureus, including CA-MRSA strains, in the general adult population of southeast Queensland. 396 patients presenting to general practices in two Brisbane suburbs and 303 volunteers randomly selected from the electoral rolls in the same suburbs completed a medical questionnaire and had nasal swabs performed for S. aureus. All isolates of S. aureus underwent antibiotic susceptibility testing and single-nucleotide polymorphism (SNP) and binary typing, including determination of Panton–Valentine leukocidin (PVL). The nasal carriage rate of methicillin-susceptible S. aureus (MSSA) was 202/699 (28%), a rate similar to that found in other community-based nasal carriage studies. According to multivariate analysis, nasal carriage of S. aureus was associated with male sex, young adult age group and Caucasian ethnicity. Only two study isolates (one MSSA and one CA-MRSA) carried PVL. The nasal carriage rate of MRSA was low, at 5/699 (0.7%), and only two study participants (0.3%) had CA-MRSA strains. CA-MRSA is an emerging cause of infection in southeast Queensland, but as yet the incidence of carriage of CA-MRSA in the general community is low.

Investigation of highly regulated promoters for the production of recombinant proteins in plants

Plants have been identified as promising expression systems for the commercial production of recombinant proteins. Plant-based protein production or “biofarming” offers a number of advantages over traditional expression systems in terms of scale of production, the capacity for post-translation processing, providing a product free of contaminants and cost effectiveness. A number of pharmaceutically important and commercially valuable proteins, such as antibodies, biopharmaceuticals and industrial enzymes are currently being produced in plant expression systems. However, several challenges still remain to improve recombinant protein yield with no ill effect on the host plant. The ability for transgenic plants to produce foreign proteins at commercially viable levels can be directly related to the level and cell specificity of the selected promoter driving the transgene. The accumulation of recombinant proteins may be controlled by a tissue-specific, developmentally-regulated or chemically-inducible promoter such that expression of recombinant proteins can be spatially- or temporally- controlled. The strict control of gene expression is particularly useful for proteins that are considered toxic and whose expression is likely to have a detrimental effect on plant growth. To date, the most commonly used promoter in plant biotechnology is the cauliflower mosaic virus (CaMV) 35S promoter which is used to drive strong, constitutive transgene expression in most organs of transgenic plants. Of particular interest to researchers in the Centre for Tropical Crops and Biocommodities at QUT are tissue-specific promoters for the accumulation of foreign proteins in the roots, seeds and fruit of various plant species, including tobacco, banana and sugarcane. Therefore this Masters project aimed to isolate and characterise root- and seed-specific promoters for the control of genes encoding recombinant proteins in plant-based expression systems. Additionally, the effects of matching cognate terminators with their respective gene promoters were assessed. The Arabidopsis root promoters ARSK1 and EIR1 were selected from the literature based on their reported limited root expression profiles. Both promoters were analysed using the PlantCARE database to identify putative motifs or cis-acting elements that may be associated with this activity. A number of motifs were identified in the ARSK1 promoter region including, WUN (wound-inducible), MBS (MYB binding site), Skn-1, and a RY core element (seed-specific) and in the EIR1 promoter region including, Skn-1 (seed-specific), Box-W1 (fungal elicitor), Aux-RR core (auxin response) and ABRE (ABA response). However, no previously reported root-specific cis-acting elements were observed in either promoter region. To confirm root specificity, both promoters, and truncated versions, were fused to the GUS reporter gene and the expression cassette introduced into Arabidopsis via Agrobacterium-mediated transformation. Despite the reported tissue-specific nature of these promoters, both upstream regulatory regions directed constitutive GUS expression in all transgenic plants. Further, similar levels of GUS expression from the ARSK1 promoter were directed by the control CaMV 35S promoter. The truncated version of the EIR1 promoter (1.2 Kb) showed some differences in the level of GUS expression compared to the 2.2 Kb promoter. Therefore, this suggests an enhancer element is contained in the 2.2 Kb upstream region that increases transgene expression. The Arabidopsis seed-specific genes ATS1 and ATS3 were selected from the literature based on their seed-specific expression profiles and gene expression confirmed in this study as seed-specific by RT-PCR analysis. The selected promoter regions were analysed using the PlantCARE database in order to identify any putative cis elements. The seed-specific motifs GCN4 and Skn-1 were identified in both promoter regions that are associated with elevated expression levels in the endosperm. Additionaly, the seed-specific RY element and the ABRE were located in the ATS1 promoter. Both promoters were fused to the GUS reporter gene and used to transform Arabidopsis plants. GUS expression from the putative promoters was consitutive in all transgenic Arabidopsis tissue tested. Importantly, the positive control FAE1 seed-specific promoter also directed constitutive GUS expression throughout transgenic Arabidopsis plants. The constitutive nature seen in all of the promoters used in this study was not anticipated. While variations in promoter activity can be caused by a number of influencing factors, the variation in promoter activity observed here would imply a major contributing factor common to all plant expression cassettes tested. All promoter constructs generated in this study were based on the binary vector pCAMBIA2300. This vector contains the plant selection gene (NPTII) under the transcriptional control of the duplicated CaMV 35S promoter. This CaMV 35S promoter contains two enhancer domains that confer strong, constitutive expression of the selection gene and is located immediately upstream of the promoter-GUS fusion. During the course of this project, Yoo et al. (2005) reported that transgene expression is significantly affected when the expression cassette is located on the same T-DNA as the 35S enhancer. It was concluded, the trans-acting effects of the enhancer activate and control transgene expression causing irregular expression patterns. This phenomenon seems the most plausible reason for the constitutive expression profiles observed with the root- and seed-specific promoters assessed in this study. The expression from some promoters can be influenced by their cognate terminator sequences. Therefore, the Arabidopsis ARSK1, EIR1, ATS1 and ATS3 terminator sequences were isolated and incorporated into expression cassettes containing the GUS reporter gene under the control of their cognate promoters. Again, unrestricted GUS activity was displayed throughout transgenic plants transformed with these reporter gene fusions. As previously discussed constitutive GUS expression was most likely due to the trans-acting effect of the upstream CaMV 35S promoter in the selection cassette located on the same T-DNA. The results obtained in this study make it impossible to assess the influence matching terminators with their cognate promoters have on transgene expression profiles. The obvious future direction of research continuing from this study would be to transform pBIN-based promoter-GUS fusions (ie. constructs containing no CaMV 35S promoter driving the plant selection gene) into Arabidopsis in order to determine the true tissue specificity of these promoters and evaluate the effects of their cognate 3’ terminator sequences. Further, promoter truncations based around the cis-elements identified here may assist in determining whether these motifs are in fact involved in the overall activity of the promoter.

Resistance : re-imagining innovation in higher education teaching and learning

The call to innovate is ubiquitous across the Australian educational policy context. The claims of innovative practices and environments that occur frequently in university mission statements, strategic plans and marketing literature suggest that this exhortation to innovate appears to have been taken up enthusiastically by the university sector. Throughout the history of universities, a range of reported deficiencies of higher education have worked to produce a notion of crisis. At present, it would seem that innovation is positioned as the solution to the notion of crisis. This thesis is an inquiry into how the insistence on innovation works to both enable and constrain teaching and learning practices in Australian universities. Alongside the interplay between innovation and crisis is the link between resistance and innovation, a link which remains largely unproblematized in the scholarly literature. This thesis works to locate and unsettle understandings of a relationship between innovation and Australian higher education. The aim of this inquiry is to generate new understandings of what counts as innovation within this context and how innovation is enacted. The thesis draws on a number of postmodernist theorists, whose works have informed firstly the research method, and then the analysis and findings. Firstly, there is an assumption that power is capillary and works through discourse to enact power relations which shape certain truths (Foucault, 1990). Secondly, this research scrutinised language practices which frame the capacity for individuals to act, alongside the language practices which encourage an individual to adopt certain attitudes and actions as one’s own (Foucault, 1988). Thirdly, innovation talk is read in this thesis as an example of needs talk, that is, as a medium through which what is considered domestic, political or economic is made and contested (Fraser, 1989). Fourthly, relationships between and within discourses were identified and analysed beyond cause and effect descriptions, and more productively considered to be in a constant state of becoming (Deleuze, 1987). Finally, the use of ironic research methods assisted in producing alternate configurations of innovation talk which are useful and new (Rorty, 1989). The theoretical assumptions which underpin this thesis inform a document analysis methodology, used to examine how certain texts work to shape the ways in which innovation is constructed. The data consisted of three Federal higher education funding policies selected on the rationale that these documents, as opposed to state or locally based policy and legislation, represent the only shared policy context for all Australian universities. The analysis first provided a modernist reading of the three documents, and this was followed by postmodernist readings of these same policy documents. The modernist reading worked to locate and describe the current truths about innovation. The historical context in which the policy was produced as well as the textual features of the document itself were important to this reading. In the first modernist reading, the binaries involved in producing proper and improper notions of innovation were described and analysed. In the process of the modernist analysis and the subsequent location of binary organisation, a number of conceptual collisions were identified, and these sites of struggle were revisited, through the application of a postmodernist reading. By applying the theories of Rorty (1989) and Fraser (1989) it became possible to not treat these sites as contradictory and requiring resolution, but rather as spaces in which binary tensions are necessary and productive. This postmodernist reading constructed new spaces for refusing and resisting dominant discourses of innovation which value only certain kinds of teaching and learning practices. By exploring a number of ironic language practices found within the policies, this thesis proposes an alternative way of thinking about what counts as innovation and how it happens. The new readings of innovation made possible through the work of this thesis were in response to a suite of enduring, inter-related questions – what counts as innovation?, who or what supports innovation?, how does innovation occur?, and who are the innovators?. The truths presented in response to these questions were treated as the language practices which constitute a dominant discourse of innovation talk. The collisions that occur within these truths were the contested sites which were of most interest for the analysis. The thesis concludes by presenting a theoretical blueprint which works to shift the boundaries of what counts as innovation and how it happens in a manner which is productive, inclusive and powerful. This blueprint forms the foundation upon which a number of recommendations are made for both my own professional practice and broader contexts. In keeping with the conceptual tone of this study, these recommendations are a suite of new questions which focus attention on the boundaries of innovation talk as an attempt to re-configure what is valued about teaching and learning at university.

Gender

The OED informs us that “gender” has at its root the Latin genus, meaning “race, kind,” and emerges as early as the fifth century as a term for differentiating between types of (especially) people and words. In the following 1500 years, gender appears in linguistic and biological contexts to distinguish types of words and bodies from one another, as when words in Indo-European languages were identified as masculine, feminine, or neuter, and humans were identified as male or female. It is telling that gender has historically (whether overtly or covertly) been a tool of negotiation between our understandings of bodies, and meanings derived from and attributed to them. Within the field of children’s literature studies, as in other disciplines, gender in and of itself is rarely the object of critique. Rather, specific constructions of gender structure understandings of subjectivity; allow or disallow certain behaviors or experiences on the basis of biological sex; and dictate a specific vision of social relations and organization. Critical approaches to gender in children’s literature have included linguistic analysis (Turner-Bowker; Sunderland); analysis of visual representations (Bradford; Moebius); cultural images of females (Grauerholz and Pescosolido); consideration of gender and genre (Christian-Smith; Stephens); ideological (Nodelman and Reimer); psychoanalytic (Coats); discourse analysis (Stephens); and masculinity studies (Nodelman) among others. In the adjacent fields of education and literacy studies, gender has been a sustained point of investigation, often deriving from perceived gendering of pedagogical practices (Lehr) or of reading preferences and competencies, and in recent years, perceptions of boys as “reluctant readers” (Moss). The ideology of patriarchy has primarily come under critical scrutiny 2 because it has been used to locate characters and readers within the specific binary logic of gender relations that historically subordinated the feminine to the masculine. Just as feminism might be broadly defined as resistance to existing power structures, a gendered reading might be broadly defined as a “resistant reading” in that it most often reveals or contests that which a text assumes to be the norm.

Toward robust image detection of crown-of-thorns starfish for autonomous population monitoring

Robust texture recognition in underwater image sequences for marine pest population control such as Crown-Of-Thorns Starfish (COTS) is a relatively unexplored area of research. Typically, humans count COTS by laboriously processing individual images taken during surveys. Being able to autonomously collect and process images of reef habitat and segment out the various marine biota holds the promise of allowing researchers to gain a greater understanding of the marine ecosystem and evaluate the impact of different environmental variables. This research applies and extends the use of Local Binary Patterns (LBP) as a method for texture-based identification of COTS from survey images. The performance and accuracy of the algorithms are evaluated on a image data set taken on the Great Barrier Reef.

Development of rapid and highly resolving combinatorial genotyping schemes for Campylobacter jejuni and Campylobacter coli

Campylobacter jejuni followed by Campylobacter coli contribute substantially to the economic and public health burden attributed to food-borne infections in Australia. Genotypic characterisation of isolates has provided new insights into the epidemiology and pathogenesis of C. jejuni and C. coli. However, currently available methods are not conducive to large scale epidemiological investigations that are necessary to elucidate the global epidemiology of these common food-borne pathogens. This research aims to develop high resolution C. jejuni and C. coli genotyping schemes that are convenient for high throughput applications. Real-time PCR and High Resolution Melt (HRM) analysis are fundamental to the genotyping schemes developed in this study and enable rapid, cost effective, interrogation of a range of different polymorphic sites within the Campylobacter genome. While the sources and routes of transmission of campylobacters are unclear, handling and consumption of poultry meat is frequently associated with human campylobacteriosis in Australia. Therefore, chicken derived C. jejuni and C. coli isolates were used to develop and verify the methods described in this study. The first aim of this study describes the application of MLST-SNP (Multi Locus Sequence Typing Single Nucleotide Polymorphisms) + binary typing to 87 chicken C. jejuni isolates using real-time PCR analysis. These typing schemes were developed previously by our research group using isolates from campylobacteriosis patients. This present study showed that SNP + binary typing alone or in combination are effective at detecting epidemiological linkage between chicken derived Campylobacter isolates and enable data comparisons with other MLST based investigations. SNP + binary types obtained from chicken isolates in this study were compared with a previously SNP + binary and MLST typed set of human isolates. Common genotypes between the two collections of isolates were identified and ST-524 represented a clone that could be worth monitoring in the chicken meat industry. In contrast, ST-48, mainly associated with bovine hosts, was abundant in the human isolates. This genotype was, however, absent in the chicken isolates, indicating the role of non-poultry sources in causing human Campylobacter infections. This demonstrates the potential application of SNP + binary typing for epidemiological investigations and source tracing. While MLST SNPs and binary genes comprise the more stable backbone of the Campylobacter genome and are indicative of long term epidemiological linkage of the isolates, the development of a High Resolution Melt (HRM) based curve analysis method to interrogate the hypervariable Campylobacter flagellin encoding gene (flaA) is described in Aim 2 of this study. The flaA gene product appears to be an important pathogenicity determinant of campylobacters and is therefore a popular target for genotyping, especially for short term epidemiological studies such as outbreak investigations. HRM curve analysis based flaA interrogation is a single-step closed-tube method that provides portable data that can be easily shared and accessed. Critical to the development of flaA HRM was the use of flaA specific primers that did not amplify the flaB gene. HRM curve analysis flaA interrogation was successful at discriminating the 47 sequence variants identified within the 87 C. jejuni and 15 C. coli isolates and correlated to the epidemiological background of the isolates. In the combinatorial format, the resolving power of flaA was additive to that of SNP + binary typing and CRISPR (Clustered regularly spaced short Palindromic repeats) HRM and fits the PHRANA (Progressive hierarchical resolving assays using nucleic acids) approach for genotyping. The use of statistical methods to analyse the HRM data enhanced sophistication of the method. Therefore, flaA HRM is a rapid and cost effective alternative to gel- or sequence-based flaA typing schemes. Aim 3 of this study describes the development of a novel bioinformatics driven method to interrogate Campylobacter MLST gene fragments using HRM, and is called ‘SNP Nucleated Minim MLST’ or ‘Minim typing’. The method involves HRM interrogation of MLST fragments that encompass highly informative “Nucleating SNPS” to ensure high resolution. Selection of fragments potentially suited to HRM analysis was conducted in silico using i) “Minimum SNPs” and ii) the new ’HRMtype’ software packages. Species specific sets of six “Nucleating SNPs” and six HRM fragments were identified for both C. jejuni and C. coli to ensure high typeability and resolution relevant to the MLST database. ‘Minim typing’ was tested empirically by typing 15 C. jejuni and five C. coli isolates. The association of clonal complexes (CC) to each isolate by ‘Minim typing’ and SNP + binary typing were used to compare the two MLST interrogation schemes. The CCs linked with each C. jejuni isolate were consistent for both methods. Thus, ‘Minim typing’ is an efficient and cost effective method to interrogate MLST genes. However, it is not expected to be independent, or meet the resolution of, sequence based MLST gene interrogation. ‘Minim typing’ in combination with flaA HRM is envisaged to comprise a highly resolving combinatorial typing scheme developed around the HRM platform and is amenable to automation and multiplexing. The genotyping techniques described in this thesis involve the combinatorial interrogation of differentially evolving genetic markers on the unified real-time PCR and HRM platform. They provide high resolution and are simple, cost effective and ideally suited to rapid and high throughput genotyping for these common food-borne pathogens.

Implications of faecal indicator bacteria for the microbiological assessment of roof harvested rainwater quality in Southeast Queensland, Australia

The study aimed to evaluate the suitability of Escherichia coli, enterococci and C. perfringens to assess the microbiological quality of roof harvested rainwater, and to assess whether the concentrations of these faecal indicators can be used to predict the presence or absence of specific zoonotic bacterial or protozoan pathogens. From a total of 100 samples tested, respectively 58%, 83% and 46% of samples were found to be positive for E. coli, enterococci and C. perfringens spores, as determined by traditional culture based methods. Additionally, in the samples tested, 7%, 19%, 1%, 8%, 17%, and 15% were PCR positive for A. hydrophila lip, C. coli ceuE, C. jejuni mapA, L. pneumophila mip, Salmonella invA, and G. lamblia β-giardin genes. However, none of the samples was positive for E. coli O157 LPS, VT1, VT2 and C. parvum COWP genes. The presence or absence of these potential pathogens did not correlate with any of the faecal indicator bacterial concentrations as determined by a binary logistic regression model. The roof-harvested rainwater samples tested in this study appear to be of poor microbiological quality and no significant correlation was found between the concentration of faecal indicators and pathogenic microorganisms. The use of faecal indicator bacteria raises questions regarding their reliability in assessing the microbiological quality of water and particularly their poor correlation with pathogenic microorganisms. The presence of one or more zoonotic pathogens suggests that the microbiological analysis of water should be performed, and appropriate treatment measures should be undertaken especially in tanks where the water is used for drinking.