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.

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.

Applying the open innovation system concept to infrastructure projects

Purpose: The goal of this conceptual paper is to provide tools to help maximise the value delivered by infrastructure projects, by developing methods to increase adoption of innovative products during construction. Methods: The role of knowledge flows in determining innovation adoption rates is conceptually examined. A promising new approach is developed. Open innovation system theory is extended, by reviewing the role of three frameworks: (1) knowledge intermediaries, (2) absorptive capacity and (3) governance arrangements. Originality: We develop a novel open innovation system model to guide further research in the area of adoption of innovation on infrastructure projects. The open innovation system model currently lacks definition of core concepts, especially with regard to the impact of different degrees and types of openness. The three frameworks address this issue and add substance to the open innovation system model, addressing widespread criticism that it is underdeveloped. The novelty of our model is in the combination of the three frameworks to explore the system. These frameworks promise new insights into system dynamics and facilitate the development of new methods to optimise the diffusion of innovation. Practical Implications: The framework will help to reveal gaps in knowledge flows that impede the uptake of innovations. In the past, identifying these gaps has been difficult given the lack of nuance in existing theory. The knowledge maps proposed will enable informed policy advice to effectively harness the power of knowledge networks, increase innovation diffusion and improve the performance of infrastructure projects. The models developed in this paper will be used in planned empirical research into innovation on large scale infrastructure projects in the Australian built environment.

Evaluating sustainability of urban development and its contribution to a knowledge-base development

Purpose – The purpose of the paper is to discuss the components of urban sustainability as to their implications about knowledge-base economy and society Design/methodology/approach – An indexing model which can be used by the local government specifically in Australia is presented to generate sustainable urban development policies. The model consists of sustainable neighbourhood indicators and employs a spatial indexing method to measure the sustainability performance of the urban settings Originality/value – This methodology puts in evidence about the use of indexing methodology in the assessment of sustainable neighbourhood performance Practical implications – This model could be considered as a practical decision aid tool for local government planning agencies for the evaluation of development scenarios Keywords – knowledge-based urban development, sustainable urban development, sustainable transport, sustainability assessment Paper type – Academic Research Paper

Sustainable urban development : formulation of indicator-based residential sustainability assessment framework for local level

Purpose–The purpose of this paper is to formulate a conceptual framework for urban sustainability indicators selection. This framework will be used to develop an indicator-based evaluation method for assessing the sustainability levels of residential neighbourhood developments in Malaysia. Design/methodology/approach–We provide a brief overview of existing evaluation frameworks for sustainable development assessment. We then develop a conceptual Sustainable Residential Neighbourhood Assessment (SNA) framework utilising a four-pillar sustainability framework (environmental, social, economic and institutional) and a combination of domain-based and goal-based general frameworks. This merger offers the advantages of both individual frameworks, while also overcoming some of their weaknesses when used to develop the urban sustainability evaluation method for assessing residential neighbourhoods. Originality/value–This approach puts in evidence that many of the existing frameworks for evaluating urban sustainability do not extend their frameworks to include assessing housing sustainability at a local level. Practical implications–It is expected that the use of the indicator-based Sustainable Neighbourhood Assessment framework will present a potential mechanism for planners and developers to evaluate and monitor the sustainability performance of residential neighbourhood developments.

Towards a unified method for the knowledge based urban development framework

Purpose–The growing debate in the literature indicates that the initiative to implement Knowledge Based Urban Development (KBUD) approaches in urban development process is neither simple nor quick. Many research efforts has therefore, been put forward to the development of appropriate KBUD framework and KBUD practical approaches. But this has lead to a fragmented and incoherent methodological approach. This paper outlines and compares a few most popular KBUD frameworks selected from the literature. It aims to identify some key and common features in the effort to achieve a unified method of KBUD framework. Design/methodology/approach–This paper reviews, examines and identifies various popular KBUD frameworks discussed in the literature from urban planners’ viewpoint. It employs a content analysis technique i.e. a research tool used to determine the presence of certain words or concepts within texts or sets of texts. Originality/value–The paper reports on the key and common features of a few selected most popular KBUD frameworks. The synthesis of the results is based from a perspective of urban planners. The findings which encompass a new KBUD framework incorporating the key and common features will be valuable in setting a platform to achieve a unified method of KBUD. Practical implications –The discussion and results presented in this paper should be significant to researchers and practitioners and to any cities and countries that are aiming for KBUD. Keywords – Knowledge based urban development, Knowledge based urban development framework, Urban development and knowledge economy

Genotyping of enterococcus faecalis and enterococcus faecium isolates using a set of eight single nucleotide polymorphisms (SNPs)

A SNP genotyping method was developed for E. faecalis and E. faecium using the 'Minimum SNPs' program. SNP sets were interrogated using allele-specific real-time PCR. SNP-typing sub-divided clonal complexes 2 and 9 of E. faecalis and 17 of E. faecium, members of which cause the majority of nosocomial infections globally.

A comparative knowledge-based urban development analysis : Vancouver, Melbourne and Manchester vs. Boston

Purpose - The purpose of this paper is to introduce a knowledge-based urban development assessment framework, which has been constructed in order to evaluate and assist in the (re)formulation of local and regional policy frameworks and applications necessary in knowledge city transformations. Design/methodology/approach - The research reported in this paper follows a methodological approach that includes a thorough review of the literature, development of an assessment framework in order to inform policy-making by accurately evaluating knowledge-based development levels of cities, and application of this framework in a comparative study - Boston, Vancouver, Melbourne and Manchester. Originality/value - The paper, with its assessment framework, demonstrates an innovative way of examining the knowledge-based development capacity of cities by scrutinising their economic, socio-cultural, enviro-urban and institutional development mechanisms and capabilities. Practical implications - The paper introduces a framework developed to assess the knowledge-based development levels of cities; presents some of the generic indicators used to evaluate knowledge-based development performance of cities; demonstrates how a city can benchmark its development level against that of other cities, and; provides insights for achieving a more sustainable and knowledge-based development.

Sustainable urban infrastructure management : integration of urban computer modelling

Purpose - This paper seeks to examine the complex relationships between urban planning, infrastructure management, sustainable urban development, and to illustrate why there is an urgent need for local governments to develop a robust planning support system which integrates with advance urban computer modelling tools to facilitate better infrastructure management and improve knowledge sharing between the community, urban planners, engineers and decision makers. Design/methodology/approach - The methods used in this paper includes literature review and practical project case observations. Originality/value - This paper provides an insight of how the Brisbane's planning support system established by Brisbane City Council has significantly improved the effectiveness of urban planning, infrastructure management and community engagement through better knowledge management processes. Practical implications - This paper presents a practical framework for setting up a functional planning support system within local government. The integration of the Brisbane Urban Growth model, Virtual Brisbane and the Brisbane Economic Activity Monitoring (BEAM) database have proven initially successful to provide a dynamic platform to assist elected officials, planners and engineers to understand the limitations of the local environment, its urban systems and the planning implications on a city. With the Brisbane's planning support system, planners and decision makers are able to provide better planning outcomes, policy and infrastructure that adequately address the local needs and achieve sustainable spatial forms.

Evaluation of accessibility for knowledge-based cities

Purpose: The purpose of the paper is to develop a framework for evaluation of accessibility for knowledge based cities. ----- ----- Design/methodology/approach: This approach notifies common mistakes and problems in accessibility assessment for knowledge cities. ----- ----- Originality/value: Accessibility plays a key role in transport sustainability and recognizes the crucial links between transport and sustainable goals like air quality, environmental resource consumption & social equity. In knowledge cities, accessibility has significant effects on quality of life and social equity by improving the mobility of people and goods. Accessibility also influences patterns of growth and economic health by providing access to land. Accessibility is not only one of the components of knowledge cities but also affects other elements of knowledge cities directly or indirectly. ----- ----- Practical implications: The outcomes of the application will be helpful for developing particular methodologies for evaluating knowledge cities. On other words, this methodology attempts to develop an assessment procedure for examining accessibility of knowledge-based cities.

Preface [From The Third Knowledge City World Summit 2010 proceedings]

‘Knowledge’ is a resource, which relies on the past for a better future. In the 21st century, more than ever before, cities around the world depend on the knowledge of their citizens, their institutions and their firms and enterprises. The knowledge image, the human competence and the reputation of their public and private institutions and corporations profiles a city. It attracts investment, qualified labour and professionals, as well as students and researchers. And it creates local life spaces and professional milieus, which offer the quality of life to the citizens that are seeking to cope with the challenges of modern life in a competitive world. Integrating knowledge-based development in urban strategies and policies, beyond the provision of schools and locations for higher education, has become a new ambitious arena of city politics. Coming from theory to practice, and bringing together the manifold knowledge stakeholders in a city and preparing joint visions for the knowledge city is a new challenge for city managers, urban planners and leaders of the civic society . It requires visionary power, creativity, holistic thinking, the willingness to cooperate with all groups of the local civil society, and the capability to moderate communication processes to overcome conflicts and to develop joint action for a sustainable future.

Highly discriminatory single-nucleotide polymorphism interrogation of Escherichia coli by use of allele-specific real-time PCR and eBURST analysis

In total, 782 Escherichia coli strains originating from various host sources have been analyzed in this study by using a highly discriminatory single-nucleotide polymorphism (SNP) approach. A set of eight SNPs, with a discrimination value (Simpson's index of diversity [D]) of 0.96, was determined using the Minimum SNPs software, based on sequences of housekeeping genes from the E. coli multilocus sequence typing (MLST) database. Allele-specific real-time PCR was used to screen 114 E. coli isolates from various fecal sources in Southeast Queensland (SEQ). The combined analysis of both the MLST database and SEQ E. coli isolates using eight high-D SNPs resolved the isolates into 74 SNP profiles. The data obtained suggest that SNP typing is a promising approach for the discrimination of host-specific groups and allows for the identification of human-specific E. coli in environmental samples. However, a more diverse E. coli collection is required to determine animal- and environment-specific E. coli SNP profiles due to the abundance of human E. coli strains (56%) in the MLST database.

Investigation into the introduction of clonal strains of Pseudomonas aeruginosa to the cystic fibrosis population by consumption of raw salad vegetables

Most salad vegetables are eaten fresh by consumers. However, raw vegetables may pose a risk of transmitting opportunistic bacteria to immunocompromised people, including cystic fibrosis (CF) patients. In particular, CF patients are vulnerable to chronic Pseudomonas aeruginosa lung infections and this organism is the primary cause of morbidity and mortality in this group. Clonal variants of P. aeruginosa have been identified as emerging threats to people afflicted with CF; however it has not yet been proven from where these clones originate or how they are transmitted. Due to the organisms‟ aquatic environmental niche, it was hypothesised that vegetables may be a source of these clones. To test this hypothesis, lettuce, tomatoes, mushrooms and bean sprout packages (n = 150) were analysed from a green grocer, supermarket and farmers‟ market within the Brisbane region, availability permitting. The internal and external surfaces of the vegetables were separately analysed for the presence of clonal strains of P. aeruginosa using washings and homogenisation techniques, respectively. This separation was in an attempt to establish which surface was contaminated, so that recommendations could be made to decrease or eliminate P. aeruginosa from these foods prior to consumption. Soil and water samples (n = 17) from local farms were also analysed for the presence of P. aeruginosa. Presumptive identification of isolates recovered from these environmental samples was made based on growth on Cetrimide agar at 42°C, presence of the cytochrome-oxidase enzyme and inability to ferment lactose. P. aeruginosa duplex real-time polymerase chain reaction assay (PAduplex) was performed on all bacterial isolates presumptively identified as P. aeruginosa. Enterobacterial repetitive intergenic consensus strain typing PCR (ERIC-PCR) was subsequently performed on confirmed bacterial isolates. Although 72 P. aeruginosa were isolated, none of these proved to be clonal strains. The significance of these findings is that vegetables may pose a risk of transmitting sporadic strains of P. aeruginosa to people afflicted with CF and possibly, other immunocompromised people.

User-representative feature selection for keystroke dynamics

Continuous user authentication with keystroke dynamics uses characters sequences as features. Since users can type characters in any order, it is imperative to find character sequences (n-graphs) that are representative of user typing behavior. The contemporary feature selection approaches do not guarantee selecting frequently-typed features which may cause less accurate statistical user-representation. Furthermore, the selected features do not inherently reflect user typing behavior. We propose four statistical based feature selection techniques that mitigate limitations of existing approaches. The first technique selects the most frequently occurring features. The other three consider different user typing behaviors by selecting: n-graphs that are typed quickly; n-graphs that are typed with consistent time; and n-graphs that have large time variance among users. We use Gunetti’s keystroke dataset and k-means clustering algorithm for our experiments. The results show that among the proposed techniques, the most-frequent feature selection technique can effectively find user representative features. We further substantiate our results by comparing the most-frequent feature selection technique with three existing approaches (popular Italian words, common n-graphs, and least frequent ngraphs). We find that it performs better than the existing approaches after selecting a certain number of most-frequent n-graphs.

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.