Development and validation of novel methods for microbial source tracking based on Escherichia coli as an indicator of water quality

Microbial pollution in water periodically affects human health in Australia, particularly in times of drought and flood. There is an increasing need for the control of waterborn microbial pathogens. Methods, allowing the determination of the origin of faecal contamination in water, are generally referred to as Microbial Source Tracking (MST). Various approaches have been evaluated as indicatorsof microbial pathogens in water samples, including detection of different microorganisms and various host-specific markers. However, until today there have been no universal MST methods that could reliably determine the source (human or animal) of faecal contamination. Therefore, the use of multiple approaches is frequently advised. MST is currently recognised as a research tool, rather than something to be included in routine practices. The main focus of this research was to develop novel and universally applicable methods to meet the demands for MST methods in routine testing of water samples. Escherichia coli was chosen initially as the object organism for our studies as, historically and globally, it is the standard indicator of microbial contamination in water. In this thesis, three approaches are described: single nucleotide polymorphism (SNP) genotyping, clustered regularly interspaced short palindromic repeats (CRISPR) screening using high resolution melt analysis (HRMA) methods and phage detection development based on CRISPR types. The advantage of the combination SNP genotyping and CRISPR genes has been discussed in this study. For the first time, a highly discriminatory single nucleotide polymorphism interrogation of E. coli population was applied to identify the host-specific cluster. Six human and one animal-specific SNP profile were revealed. SNP genotyping was successfully applied in the field investigations of the Coomera watershed, South-East Queensland, Australia. Four human profiles [11], [29], [32] and [45] and animal specific SNP profile [7] were detected in water. Two human-specific profiles [29] and [11] were found to be prevalent in the samples over a time period of years. The rainfall (24 and 72 hours), tide height and time, general land use (rural, suburban), seasons, distance from the river mouth and salinity show a lack of relashionship with the diversity of SNP profiles present in the Coomera watershed (p values > 0.05). Nevertheless, SNP genotyping method is able to identify and distinquish between human- and non-human specific E. coli isolates in water sources within one day. In some samples, only mixed profiles were detected. To further investigate host-specificity in these mixed profiles CRISPR screening protocol was developed, to be used on the set of E. coli, previously analysed for SNP profiles. CRISPR loci, which are the pattern of previous DNA coliphages attacks, were considered to be a promising tool for detecting host-specific markers in E. coli. Spacers in CRISPR loci could also reveal the dynamics of virulence in E. coli as well in other pathogens in water. Despite the fact that host-specificity was not observed in the set of E. coli analysed, CRISPR alleles were shown to be useful in detection of the geographical site of sources. HRMA allows determination of ‘different’ and ‘same’ CRISPR alleles and can be introduced in water monitoring as a cost-effective and rapid method. Overall, we show that the identified human specific SNP profiles [11], [29], [32] and [45] can be useful as marker genotypes globally for identification of human faecal contamination in water. Developed in the current study, the SNP typing approach can be used in water monitoring laboratories as an inexpensive, high-throughput and easy adapted protocol. The unique approach based on E. coli spacers for the search for unknown phage was developed to examine the host-specifity in phage sequences. Preliminary experiments on the recombinant plasmids showed the possibility of using this method for recovering phage sequences. Future studies will determine the host-specificity of DNA phage genotyping as soon as first reliable sequences can be acquired. No doubt, only implication of multiple approaches in MST will allow identification of the character of microbial contamination with higher confidence and readability.

Structural behaviour and design of cold-formed steel wall systems under fire conditions

In recent times, light gauge steel framed (LSF) structures, such as cold-formed steel wall systems, are increasingly used, but without a full understanding of their fire performance. Traditionally the fire resistance rating of these load-bearing LSF wall systems is based on approximate prescriptive methods developed based on limited fire tests. Very often they are limited to standard wall configurations used by the industry. Increased fire rating is provided simply by adding more plasterboards to these walls. This is not an acceptable situation as it not only inhibits innovation and structural and cost efficiencies but also casts doubt over the fire safety of these wall systems. Hence a detailed fire research study into the performance of LSF wall systems was undertaken using full scale fire tests and extensive numerical studies. A new composite wall panel developed at QUT was also considered in this study, where the insulation was used externally between the plasterboards on both sides of the steel wall frame instead of locating it in the cavity. Three full scale fire tests of LSF wall systems built using the new composite panel system were undertaken at a higher load ratio using a gas furnace designed to deliver heat in accordance with the standard time temperature curve in AS 1530.4 (SA, 2005). Fire tests included the measurements of load-deformation characteristics of LSF walls until failure as well as associated time-temperature measurements across the thickness and along the length of all the specimens. Tests of LSF walls under axial compression load have shown the improvement to their fire performance and fire resistance rating when the new composite panel was used. Hence this research recommends the use of the new composite panel system for cold-formed LSF walls. The numerical study was undertaken using a finite element program ABAQUS. The finite element analyses were conducted under both steady state and transient state conditions using the measured hot and cold flange temperature distributions from the fire tests. The elevated temperature reduction factors for mechanical properties were based on the equations proposed by Dolamune Kankanamge and Mahendran (2011). These finite element models were first validated by comparing their results with experimental test results from this study and Kolarkar (2010). The developed finite element models were able to predict the failure times within 5 minutes. The validated model was then used in a detailed numerical study into the strength of cold-formed thin-walled steel channels used in both the conventional and the new composite panel systems to increase the understanding of their behaviour under nonuniform elevated temperature conditions and to develop fire design rules. The measured time-temperature distributions obtained from the fire tests were used. Since the fire tests showed that the plasterboards provided sufficient lateral restraint until the failure of LSF wall panels, this assumption was also used in the analyses and was further validated by comparison with experimental results. Hence in this study of LSF wall studs, only the flexural buckling about the major axis and local buckling were considered. A new fire design method was proposed using AS/NZS 4600 (SA, 2005), NAS (AISI, 2007) and Eurocode 3 Part 1.3 (ECS, 2006). The importance of considering thermal bowing, magnified thermal bowing and neutral axis shift in the fire design was also investigated. A spread sheet based design tool was developed based on the above design codes to predict the failure load ratio versus time and temperature for varying LSF wall configurations including insulations. Idealised time-temperature profiles were developed based on the measured temperature values of the studs. This was used in a detailed numerical study to fully understand the structural behaviour of LSF wall panels. Appropriate equations were proposed to find the critical temperatures for different composite panels, varying in steel thickness, steel grade and screw spacing for any load ratio. Hence useful and simple design rules were proposed based on the current cold-formed steel structures and fire design standards, and their accuracy and advantages were discussed. The results were also used to validate the fire design rules developed based on AS/NZS 4600 (SA, 2005) and Eurocode Part 1.3 (ECS, 2006). This demonstrated the significant improvements to the design method when compared to the currently used prescriptive design methods for LSF wall systems under fire conditions. In summary, this research has developed comprehensive experimental and numerical thermal and structural performance data for both the conventional and the proposed new load bearing LSF wall systems under standard fire conditions. Finite element models were developed to predict the failure times of LSF walls accurately. Idealized hot flange temperature profiles were developed for non-insulated, cavity and externally insulated load bearing wall systems. Suitable fire design rules and spread sheet based design tools were developed based on the existing standards to predict the ultimate failure load, failure times and failure temperatures of LSF wall studs. Simplified equations were proposed to find the critical temperatures for varying wall panel configurations and load ratios. The results from this research are useful to both structural and fire engineers and researchers. Most importantly, this research has significantly improved the knowledge and understanding of cold-formed LSF loadbearing walls under standard fire conditions.

The relationship between chronic disease self-efficacy and nutritional status, functional ability and quality of life in older adults at risk of hospital readmission

Background and significance: Older adults with chronic diseases are at increasing risk of hospital admission and readmission. Approximately 75% of adults have at least one chronic condition, and the odds of developing a chronic condition increases with age. Chronic diseases consume about 70% of the total Australian health expenditure, and about 59% of hospital events for chronic conditions are potentially preventable. These figures have brought to light the importance of the management of chronic disease among the growing older population. Many studies have endeavoured to develop effective chronic disease management programs by applying social cognitive theory. However, limited studies have focused on chronic disease self-management in older adults at high risk of hospital readmission. Moreover, although the majority of studies have covered wide and valuable outcome measures, there is scant evidence on examining the fundamental health outcomes such as nutritional status, functional status and health-related quality of life. Aim: The aim of this research was to test social cognitive theory in relation to self-efficacy in managing chronic disease and three health outcomes, namely nutritional status, functional status, and health-related quality of life, in older adults at high risk of hospital readmission. Methods: A cross-sectional study design was employed for this research. Three studies were undertaken. Study One examined the nutritional status and validation of a nutritional screening tool; Study Two explored the relationships between participants. characteristics, self-efficacy beliefs, and health outcomes based on the study.s hypothesized model; Study Three tested a theoretical model based on social cognitive theory, which examines potential mechanisms of the mediation effects of social support and self-efficacy beliefs. One hundred and fifty-seven patients aged 65 years and older with a medical admission and at least one risk factor for readmission were recruited. Data were collected from medical records on demographics, medical history, and from self-report questionnaires. The nutrition data were collected by two registered nurses. For Study One, a contingency table and the kappa statistic was used to determine the validity of the Malnutrition Screening Tool. In Study Two, standard multiple regression, hierarchical multiple regression and logistic regression were undertaken to determine the significant influential predictors for the three health outcome measures. For Study Three, a structural equation modelling approach was taken to test the hypothesized self-efficacy model. Results: The findings of Study One suggested that a high prevalence of malnutrition continues to be a concern in older adults as the prevalence of malnutrition was 20.6% according to the Subjective Global Assessment. Additionally, the findings confirmed that the Malnutrition Screening Tool is a valid nutritional screening tool for hospitalized older adults at risk of readmission when compared to the Subjective Global Assessment with high sensitivity (94%), and specificity (89%) and substantial agreement between these two methods (k = .74, p < .001; 95% CI .62-.86). Analysis data for Study Two found that depressive symptoms and perceived social support were the two strongest influential factors for self-efficacy in managing chronic disease in a hierarchical multiple regression. Results of multivariable regression models suggested advancing age, depressive symptoms and less tangible support were three important predictors for malnutrition. In terms of functional status, a standard regression model found that social support was the strongest predictor for the Instrumental Activities of Daily Living, followed by self-efficacy in managing chronic disease. The results of standard multiple regression revealed that the number of hospital readmission risk factors adversely affected the physical component score, while depressive symptoms and self-efficacy beliefs were two significant predictors for the mental component score. In Study Three, the results of the structural equation modelling found that self-efficacy partially mediated the effect of health characteristics and depression on health-related quality of life. The health characteristics had strong direct effects on functional status and body mass index. The results also indicated that social support partially mediated the relationship between health characteristics and functional status. With regard to the joint effects of social support and self-efficacy, social support fully mediated the effect of health characteristics on self-efficacy, and self-efficacy partially mediated the effect of social support on functional status and health-related quality of life. The results also demonstrated that the models fitted the data well with relative high variance explained by the models, implying the hypothesized constructs under discussion were highly relevant, and hence the application for social cognitive theory in this context was supported. Conclusion: This thesis highlights the applicability of social cognitive theory on chronic disease self-management in older adults at risk of hospital readmission. Further studies are recommended to validate and continue to extend the development of social cognitive theory on chronic disease self-management in older adults to improve their nutritional and functional status, and health-related quality of life.

Validity and reliability of hallux valgus angle measured on digital photographs

STUDY DESIGN: Controlled laboratory study. OBJECTIVES: To investigate the reliability and concurrent validity of photographic measurements of hallux valgus angle compared to radiographs as the criterion standard. BACKGROUND: Clinical assessment of hallux valgus involves measuring alignment between the first toe and metatarsal on weight-bearing radiographs or visually grading the severity of deformity with categorical scales. Digital photographs offer a noninvasive method of measuring deformity on an exact scale; however, the validity of this technique has not previously been established. METHODS: Thirty-eight subjects (30 female, 8 male) were examined (76 feet, 54 with hallux valgus). Computer software was used to measure hallux valgus angle from digital records of bilateral weight-bearing dorsoplantar foot radiographs and photographs. One examiner measured 76 feet on 2 occasions 2 weeks apart, and a second examiner measured 40 feet on a single occasion. Reliability was investigated by intraclass correlation coefficients and validity by 95% limits of agreement. The Pearson correlation coefficient was also calculated. RESULTS: Intrarater and interrater reliability were very high (intraclass correlation coefficients greater than 0.96) and 95% limits of agreement between photographic and radiographic measurements were acceptable. Measurements from photographs and radiographs were also highly correlated (Pearson r = 0.96). CONCLUSIONS: Digital photographic measurements of hallux valgus angle are reliable and have acceptable validity compared to weight-bearing radiographs. This method provides a convenient and precise tool in assessment of hallux valgus, while avoiding the cost and radiation exposure associated with radiographs.

A new technique for optimal allocation and sizing of capacitors and setting of LTC

An iterative based strategy is proposed for finding the optimal rating and location of fixed and switched capacitors in distribution networks. The substation Load Tap Changer tap is also set during this procedure. A Modified Discrete Particle Swarm Optimization is employed in the proposed strategy. The objective function is composed of the distribution line loss cost and the capacitors investment cost. The line loss is calculated using estimation of the load duration curve to multiple levels. The constraints are the bus voltage and the feeder current which should be maintained within their standard range. For validation of the proposed method, two case studies are tested. The first case study is the semi-urban 37-bus distribution system which is connected at bus 2 of the Roy Billinton Test System which is located in the secondary side of a 33/11 kV distribution substation. The second case is a 33 kV distribution network based on the modification of the 18-bus IEEE distribution system. The results are compared with prior publications to illustrate the accuracy of the proposed strategy.

A critical regionalist approach to housing design in Vietnam : socio-environmental organisation of living spaces in pre- and post-reform houses

Vietnam has a unique culture which is revealed in the way that people have built and designed their traditional housing. Vietnamese dwellings reflect occupants’ activities in their everyday lives, while adapting to tropical climatic conditions impacted by seasoning monsoons. It is said that these characteristics of Vietnamese dwellings have remained unchanged until the economic reform in 1986, when Vietnam experienced an accelerated development based on the market-oriented economy. New housing types, including modern shop-houses, detached houses, and apartments, have been designed in many places, especially satisfying dwellers’ new lifestyles in Vietnamese cities. The contemporary housing, which has been mostly designed by architects, has reflected rules of spatial organisation so that occupants’ social activities are carried out. However, contemporary housing spaces seem unsustainable in relation to socio-cultural values because they has been influenced by globalism that advocates the use of homogeneous spatial patterns, modern technologies, materials and construction methods. This study investigates the rules of spaces in Vietnamese houses that were built before and after the reform to define the socio-cultural implications in Vietnamese housing design. Firstly, it describes occupants’ views of their current dwellings in terms of indoor comfort conditions and social activities in spaces. Then, it examines the use of spaces in pre-reform Vietnamese housing through occupants’ activities and material applications. Finally, it discusses the organisation of spaces in both pre- and post-reform housing to understand how Vietnamese housing has been designed for occupants to live, act, work, and conduct traditional activities. Understanding spatial organisation is a way to identify characteristics of the lived spaces of the occupants created from the conceived space, which is designed by designers. The characteristics of the housing spaces will inform the designers the way to design future Vietnamese housing in response to cultural contexts. The study applied an abductive approach for the investigation of housing spaces. It used a conceptual framework in relation to Henri Lefebvre’s (1991) theory to understand space as the main factor constituting the language of design, and the principles of semiotics to examine spatial structure in housing as a language used in the everyday life. The study involved a door-knocking survey to 350 households in four regional cities of Vietnam for interpretation of occupancy conditions and levels of occupants’ comfort. A statistical analysis was applied to interpret the survey data. The study also required a process of data selection and collection of fourteen cases of housing in three main climatic regions of the country for analysing spatial organisation and housing characteristics. The study found that there has been a shift in the relationship of spaces from the pre- to post-reform Vietnamese housing. It also indentified that the space for guest welcoming and family activity has been the central space of the Vietnamese housing. Based on the relationships of the central space with the others, theoretical models were proposed for three types of contemporary Vietnamese housing. The models will be significant in adapting to Vietnamese conditions to achieve socioenvironmental characteristics for housing design because it was developed from the occupants’ requirements for their social activities. Another contribution of the study is the use of methodological concepts to understand the language of living spaces. Further work will be needed to test future Vietnamese housing designs from the applications of the models.

Breastfeeding beliefs and practices of African women living in Brisbane and Perth, Australia

The purpose of this study was to explore the experience of breastfeeding among refugee women from Liberia, Sierra Leone, Burundi and the Democratic Republic of Congo living in two major capital cities in Australia. Participants were recruited from their relevant community associations and via a snowballing technique. Thirty-one women took part in either individual interviews or facilitated group discussions to explore their experiences of breastfeeding in their home country and in Australia. Thematic analysis revealed four main themes: cultural breastfeeding beliefs and practices; stigma and shame around breastfeeding in public; ambivalence towards breastfeeding and breastfeeding support. Women who originated from these four African countries highlighted a significant desire for breastfeeding and an understanding that it was the best method for feeding their infants. Their breastfeeding practices in Australia were a combination of practices maintained from their countries of origin and those adopted according to Australian cultural norms. They exemplified the complexity of breastfeeding behaviour and the relationship between infant feeding with economic status and the perceived social norms of the host country. The results illustrate the need for policy makers and health professionals to take into consideration the environmental, social and cultural contexts of the women who are purportedly targeted for the promotion of breastfeeding.

An assessment for cost effective maintenance and operation of the Callide Power Station electrical system, using reliability evaluation

The reliable operation of the electrical system at Callide Power Station is of extreme importance to the normal everyday running of the Station. This study applied the principles of reliability to do an analysis on the electrical system at Callide Power Station. It was found that the level of expected outage cost increased exponentially with a declining level of maintenance. Concluding that even in a harsh economic electricity market where CS Energy tries and push their plants to the limit, maintenance must not be neglected. A number of system configurations were found to increase the reliability of the system and reduce the expected outage costs. A number of other advantages were identified as a result of using reliability principles to do this study on the Callide electrical system configuration.

Design and modeling of collaboration architecture for security

Threats against computer networks evolve very fast and require more and more complex measures. We argue that teams respectively groups with a common purpose for intrusion detection and prevention improve the measures against rapid propagating attacks similar to the concept of teams solving complex tasks known from field of work sociology. Collaboration in this sense is not easy task especially for heterarchical environments. We propose CIMD (collaborative intrusion and malware detection) as a security overlay framework to enable cooperative intrusion detection approaches. Objectives and associated interests are used to create detection groups for exchange of security-related data. In this work, we contribute a tree-oriented data model for device representation in the scope of security. We introduce an algorithm for the formation of detection groups, show realization strategies for the system and conduct vulnerability analysis. We evaluate the benefit of CIMD by simulation and probabilistic analysis.

Modelling client puzzles and denial-of-service resistant protocols

Denial-of-service (DoS) attacks are a growing concern to networked services like the Internet. In recent years, major Internet e-commerce and government sites have been disabled due to various DoS attacks. A common form of DoS attack is a resource depletion attack, in which an attacker tries to overload the server's resources, such as memory or computational power, rendering the server unable to service honest clients. A promising way to deal with this problem is for a defending server to identify and segregate malicious traffic as earlier as possible. Client puzzles, also known as proofs of work, have been shown to be a promising tool to thwart DoS attacks in network protocols, particularly in authentication protocols. In this thesis, we design efficient client puzzles and propose a stronger security model to analyse client puzzles. We revisit a few key establishment protocols to analyse their DoS resilient properties and strengthen them using existing and novel techniques. Our contributions in the thesis are manifold. We propose an efficient client puzzle that enjoys its security in the standard model under new computational assumptions. Assuming the presence of powerful DoS attackers, we find a weakness in the most recent security model proposed to analyse client puzzles and this study leads us to introduce a better security model for analysing client puzzles. We demonstrate the utility of our new security definitions by including two hash based stronger client puzzles. We also show that using stronger client puzzles any protocol can be converted into a provably secure DoS resilient key exchange protocol. In other contributions, we analyse DoS resilient properties of network protocols such as Just Fast Keying (JFK) and Transport Layer Security (TLS). In the JFK protocol, we identify a new DoS attack by applying Meadows' cost based framework to analyse DoS resilient properties. We also prove that the original security claim of JFK does not hold. Then we combine an existing technique to reduce the server cost and prove that the new variant of JFK achieves perfect forward secrecy (the property not achieved by original JFK protocol) and secure under the original security assumptions of JFK. Finally, we introduce a novel cost shifting technique which reduces the computation cost of the server significantly and employ the technique in the most important network protocol, TLS, to analyse the security of the resultant protocol. We also observe that the cost shifting technique can be incorporated in any Diffine{Hellman based key exchange protocol to reduce the Diffie{Hellman exponential cost of a party by one multiplication and one addition.

Comparison of genomic DNA extraction techniques from whole blood samples : a time, cost and quality evaluation study

Genomic DNA obtained from patient whole blood samples is a key element for genomic research. Advantages and disadvantages, in terms of time-efficiency, cost-effectiveness and laboratory requirements, of procedures available to isolate nucleic acids need to be considered before choosing any particular method. These characteristics have not been fully evaluated for some laboratory techniques, such as the salting out method for DNA extraction, which has been excluded from comparison in different studies published to date. We compared three different protocols (a traditional salting out method, a modified salting out method and a commercially available kit method) to determine the most cost-effective and time-efficient method to extract DNA. We extracted genomic DNA from whole blood samples obtained from breast cancer patient volunteers and compared the results of the product obtained in terms of quantity (concentration of DNA extracted and DNA obtained per ml of blood used) and quality (260/280 ratio and polymerase chain reaction product amplification) of the obtained yield. On average, all three methods showed no statistically significant differences between the final result, but when we accounted for time and cost derived for each method, they showed very significant differences. The modified salting out method resulted in a seven- and twofold reduction in cost compared to the commercial kit and traditional salting out method, respectively and reduced time from 3 days to 1 hour compared to the traditional salting out method. This highlights a modified salting out method as a suitable choice to be used in laboratories and research centres, particularly when dealing with a large number of samples.

SettleMEN-2: Displaced twice? Impact of the Queensland floods on men from refugee backgrounds living in Brisbane and Toowoomba. Broadsheet No. 1: Exposure to and impact of the floods

Between 2008 and 2010, the SettleMEN study followed a group of 233 recently arrived men from refugee backgrounds living in urban and regional Southeast Queensland with the aim of documenting their health and settlement experiences. The study found that overall, these men bring important resources that may help them to cope better with the challenges of settlement: good levels of subjective health status, mental health and wellbeing; good family and social support; and good levels of engagement in tertiary/trade education in Australia. Over time, however, their levels of wellbeing decreased as they experienced barriers to social participation and inclusion within their host community, including: unemployment and difficulties securing good jobs (even for those with tertiary/trade qualifications obtained in Australia), financial stress, difficulties accessing housing, limited interactions with neighbours, and experiences of racism and discrimination. Importantly, although men living in the Toowoomba acknowledged some of the benefits of regional settlement, they faced greater barriers to participation in the labour market, reported lower job satisfaction, and were more likely to experience social exclusion overall. In 2012 method approach and a peer interviewer model, we were able to conduct a follow 141 (61%) of the original 233 SettleMEN participants to document the impact of the January 2011 Queensland floods on their health and settlement. This broadsheet focuses on participants’ degree of exposure to and impact of the floods, their perceptions of safety and security, and their vulnerability and adaptive capacity to extreme weather events.

"Talking with lips" : settlement, transnationalism and identity of Karen people from Burma living in Brisbane, Australia

This thesis explores, from an anthropological perspective, the settlement of Karen people from Burma now living in Brisbane. It critiques settlement constructs reified by public policy and settlement model-building using narratives of the lived experience of settlement. It gives voice to a typically voiceless group of people and challenges traditional conceptions of people with refugee backgrounds as passive and vulnerable, by bringing their experiences from the periphery to the centre. It explores transnationalism, identity work and Karen organisations to demonstrate how settlement can be done both to people through policy and by people through agency and self-determination.

Standard preanalytical coding for biospecimens: Review and implementation of the Sample PREanalytical Code (SPREC)

The first version of the Standard PREanalytical Code (SPREC) was developed in 2009 by the International Society for Biological and Environmental Repositories (ISBER) Biospecimen Science Working Group to facilitate documentation and communication of the most important preanalytical quality parameters of different types of biospecimens used for research. This same Working Group has now updated the SPREC to version 2.0, presented here, so that it contains more options to allow for recent technological developments. Existing elements have been fine tuned. An interface to the Biospecimen Reporting for Improved Study Quality (BRISQ) has been defined, and informatics solutions for SPREC implementation have been developed. A glossary with SPRECrelated definitions has also been added.