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.

Corporate governance mechanisms and earnings management in transitional countries – evidence from Chinese listed firms

The objective of this thesis is to investigate whether the corporate governance practices adopted by Chinese listed firms are associated with the quality of earnings information. Based on a review of agency and institutional theory, this study develops hypotheses that predict the monitoring effectiveness of the board and the audit committee. Using a combination of univariate and multivariate analyses, the association between corporate governance mechanisms and earnings management are tested from 2004 to 2008. Through analysing the empirical results, a number of findings are summarised as below. First, board independence is weakened by the introduction of government officials as independent directors on the boards. Government officials acting as independent directors, claim that they meet the definition of independent director set by the regulation. However, they have some connection with the State, which is the controlling shareholder in listed SOEs affiliated companies. Consequently, the effect of the independent director’s expertise in constraining earnings management is mitigated as demonstrated by an insignificant association between board expertise and earnings management. An alternative explanation for the inefficiency of board independence may point to the pre-selection of independent directors by the powerful CEO. It is argued that a CEO can manipulate the board composition and choose the "desirable" independent directors to monitor themselves. Second, a number of internal mechanisms, such as board size, board activities, and the separation of the roles of the CEO and chair are found to be significantly associated with discretionary accruals. This result suggests that there are advantages in having a large and active board in the Chinese setting. This can offset the disadvantages associated with large boards, such as increased bureaucracy, and hence, increase the constraining effects of a large and resourceful board. Third, factor analysis identifies two factors: CEO power and board power. CEO power is the factor which consists of CEO duality and turnover, and board power is composed of board size and board activity. The results of CEO power show that if a Chinese listed company has CEO duality and turnover at the same time, it is more likely to have a high level of earnings management. The significant and negative relationship between board power and accruals indicate that large boards with frequent meetings can be associated with low level of earnings management. Overall, the factor analysis suggests that certain governance mechanisms complement each other to become more efficient monitors of opportunistic earnings management. A combination of board characteristics can increase the negative association with earnings management. Fourth, the insignificant results between audit committees and earnings management in Chinese listed firms suggests that the Chinese regulator should strengthen the audit committee functions. This thesis calls for listed firms to disclose more information on audit committee composition and activities, which can facilitate future research on the Chinese audit committee’s monitoring role. Fifth, the interactive results between State ownership and board characteristics show that dominant State ownership has a moderating effect on board monitoring power as the State totally controls 42% of the issued shares. The high percentage of State ownership makes it difficult for the non-controlling institutional shareholders to challenge the State’s dominant status. As a result, the association between non-controlling institutional ownership and earnings management is insignificant in most situations. Lastly, firms audited by the international Big4 have lower abnormal accruals than firms audited by domestic Chinese audit firms. In addition, the inverse U-shape relationship between audit tenure and earnings quality demonstrates the changing effects of audit quality after a certain period of appointment. Furthermore, this thesis finds that listing in Hong Kong Stock Exchanges can be an alternative governance mechanism to discipline Chinese firms to follow strict Hong Kong listing requirements. Management of Hong Kong listed companies are exposed to the scrutiny of international investors and Hong Kong regulators. This in turn reduces their chances of conducting self-interested earnings manipulation. This study is designed to fill the gap in governance literature in China that is related to earnings management. Previous research on corporate governance mechanisms and earnings management in China is not conclusive. The current research builds on previous literature and provides some meaningful implications for practitioners, regulators, academic, and international investors who have investment interests in a transitional country. The findings of this study contribute to corporate governance and earnings management literature in the context of the transitional economy of China. The use of alternative measures for earnings management yields similar results compared with the accruals models and produces additional findings.

Comparative study of coagulase negative Staphylococci (CoNS) from clinical isolates, skin and nasal sources

Staphylococci are important pathogenic bacteria responsible for a range of diseases in humans. The most frequently isolated microorganisms in a hospital microbiology laboratory are staphylococci. The general classification of staphylococci divides them into two major groups; Coagulase-positive staphylococci (e.g. Staphylococcus aureus) and Coagulase-negative staphylococci (e.g. Staphylococcus epidermidis). Coagulase-negative staphylococcal (CoNS) isolates include a variety of species and many different strains but are often dominated by the most important organism of this group, S. epidermidis. Currently, these organisms are regarded as important pathogenic organisms causing infections related to prosthetic materials and surgical wounds. A significant number of S. epidermidis isolates are also resistant to different antimicrobial agents. Virulence factors in CoNS are not very clearly established and not well documented. S. epidermidis is evolving as a resistant and powerful microbe related to nosocomial infections because it has different properties which independently, and in combination, make it a successful infectious agent, especially in the hospital environment. Such characteristics include biofilm formation, drug resistance and the evolution of genetic variables. The purpose of this project was to develop a novel SNP genotyping method to genotype S. epidermidis strains originating from hospital patients and healthy individuals. High-Resolution Melt Analysis was used to assign binary typing profiles to both clinical and commensal strains using a new bioinformatics approach. The presence of antibiotic resistance genes and biofilm coding genes were also interrogated in these isolates.

Development of a high-speed sensing and detection system for automatic removal of packages with leaky seals from a high-speed food processing and packaging line

Contamination of packaged foods due to micro-organisms entering through air leaks can cause serious public health issues and cost companies large amounts of money due to product recalls, consumer impact and subsequent loss of market share. The main source of contamination is leaks in packaging which allow air, moisture and microorganisms to enter the package. In the food processing and packaging industry worldwide, there is an increasing demand for cost effective state of the art inspection technologies that are capable of reliably detecting leaky seals and delivering products at six-sigma. The new technology will develop non-destructive testing technology using digital imaging and sensing combined with a differential vacuum technique to assess seal integrity of food packages on a high-speed production line. The cost of leaky packages in Australian food industries is estimated close to AUD $35 Million per year. Contamination of packaged foods due to micro-organisms entering through air leaks can cause serious public health issues and cost companies large sums of money due to product recalls, compensation claims and loss of market share. The main source of contamination is leaks in packaging which allow air, moisture and micro-organisms to enter the package. Flexible plastic packages are widely used, and are the least expensive form of retaining the quality of the product. These packets can be used to seal, and therefore maximise, the shelf life of both dry and moist products. The seals of food packages need to be airtight so that the food content is not contaminated due to contact with microorganisms that enter as a result of air leakage. Airtight seals also extend the shelf life of packaged foods, and manufacturers attempt to prevent food products with leaky seals being sold to consumers. There are many current NDT (non-destructive testing) methods of testing the seal of flexible packages best suited to random sampling, and for laboratory purposes. The three most commonly used methods are vacuum/pressure decay, bubble test, and helium leak detection. Although these methods can detect very fine leaks, they are limited by their high processing time and are not viable in a production line. Two nondestructive in-line packaging inspection machines are currently available and are discussed in the literature review. The detailed design and development of the High-Speed Sensing and Detection System (HSDS) is the fundamental requirement of this project and the future prototype and production unit. Successful laboratory testing was completed and a methodical design procedure was needed for a successful concept. The Mechanical tests confirmed the vacuum hypothesis and seal integrity with good consistent results. Electrically, the testing also provided solid results to enable the researcher to move the project forward with a certain amount of confidence. The laboratory design testing allowed the researcher to confirm theoretical assumptions before moving into the detailed design phase. Discussion on the development of the alternative concepts in both mechanical and electrical disciplines enables the researcher to make an informed decision. Each major mechanical and electrical component is detailed through the research and design process. The design procedure methodically works through the various major functions both from a mechanical and electrical perspective. It opens up alternative ideas for the major components that although are sometimes not practical in this application, show that the researcher has exhausted all engineering and functionality thoughts. Further concepts were then designed and developed for the entire HSDS unit based on previous practice and theory. In the future, it would be envisaged that both the Prototype and Production version of the HSDS would utilise standard industry available components, manufactured and distributed locally. Future research and testing of the prototype unit could result in a successful trial unit being incorporated in a working food processing production environment. Recommendations and future works are discussed, along with options in other food processing and packaging disciplines, and other areas in the non-food processing industry.

Molecular characterisation of environmental enterococci derived from water samples and assessment of associated public health hazards

Enterococci are versatile Gram-positive bacteria that can survive under extreme conditions. Most enterococci are non-virulent and found in the gastrointestinal tract of humans and animals. Other strains are opportunistic pathogens that contribute to a large number of nosocomial infections globally. Epidemiological studies demonstrated a direct relationship between the density of enterococci in surface waters and the risk of swimmer-associated gastroenteritis. The distribution of infectious enterococcal strains from the hospital environment or other sources to environmental water bodies through sewage discharge or other means, could increase the prevalence of these strains in the human population. Environmental water quality studies may benefit from focusing on a subset of Enterococcus spp. that are consistently associated with sources of faecal pollution such as domestic sewage, rather than testing for the entire genus. E. faecalis and E. faecium are potentially good focal species for such studies, as they have been consistently identified as the dominant Enterococcus spp. in human faeces and sewage. On the other hand enterococcal infections are predominantly caused by E. faecalis and E. faecium. The characterisation of E. faecalis and E. faecium is important in studying their population structures, particularly in environmental samples. In developing and implementing rapid, robust molecular genotyping techniques, it is possible to more accurately establish the relationship between human and environmental enterococci. Of particular importance, is to determine the distribution of high risk enterococcal clonal complexes, such as E. faecium clonal complex 17 and E. faecalis clonal complexes 2 and 9 in recreational waters. These clonal complexes are recognized as particularly pathogenic enterococcal genotypes that cause severe disease in humans globally. The Pimpama-Coomera watershed is located in South East Queensland, Australia and was investigated in this study mainly because it is used intensively for agriculture and recreational purposes and has a strong anthropogenic impact. The primary aim of this study was to develop novel, universally applicable, robust, rapid and cost effective genotyping methods which are likely to yield more definitive results for the routine monitoring of E. faecalis and E. faecium, particularly in environmental water sources. To fullfill this aim, new genotyping methods were developed based on the interrogation of highly informative single nucleotide polymorphisms (SNPs) located in housekeeping genes of both E. faecalis and E. faecium. SNP genotyping was successfully applied in field investigations of the Coomera watershed, South-East Queensland, Australia. E. faecalis and E. faecium isolates were grouped into 29 and 23 SNP profiles respectively. This study showed the high longitudinal diversity of E. faecalis and E. faecium over a period of two years, and both human-related and human-specific SNP profiles were identified. Furthermore, 4.25% of E. faecium strains isolated from water was found to correspond to the important clonal complex-17 (CC17). Strains that belong to CC17 cause the majority of hospital outbreaks and clinical infections globally. Of the six sampling sites of the Coomera River, Paradise Point had the highest number of human-related and human-specific E. faecalis and E. faecium SNP profiles. The secondary aim of this study was to determine the antibiotic-resistance profiles and virulence traits associated with environmental E. faecalis and E. faecium isolates compared to human pathogenic E. faecalis and E. faecium isolates. This was performed to predict the potential health risks associated with coming into contact with these strains in the Coomera watershed. In general, clinical isolates were found to be more resistant to all the antibiotics tested compared to water isolates and they harbored more virulence traits. Multi-drug resistance was more prevalent in clinical isolates (71.18% of E. faecalis and 70.3 % of E. faecium) compared to water isolates (only 5.66 % E. faecium). However, tetracycline, gentamicin, ciprofloxacin and ampicillin resistance was observed in water isolates. The virulence gene esp was the most prevalent virulence determinant observed in clinical isolates (67.79% of E. faecalis and 70.37 % of E. faecium), and this gene has been described as a human-specific marker used for microbial source tracking (MST). The presence of esp in water isolates (16.36% of E. faecalis and 19.14% of E. faecium) could be indicative of human faecal contamination in these waterways. Finally, in order to compare overall gene expression between environmental and clinical strains of E. faecalis, a comparative gene hybridization study was performed. The results of this investigation clearly demonstrated the up-regulation of genes associated with pathogenicity in E. faecalis isolated from water. The expression study was performed at physiological temperatures relative to ambient temperatures. The up-regulation of virulence genes demonstrates that environmental strains of E. faecalis can pose an increased health risk which can lead to serious disease, particularly if these strains belong to the virulent CC17 group. The genotyping techniques developed in this study not only provide a rapid, robust and highly discriminatory tool to characterize E. faecalis and E. faecium, but also enables the efficient identification of virulent enterococci that are distributed in environmental water sources.

An investigation on the accuracy of regional ocean models through field trials

Recent efforts in mission planning for underwater vehicles have utilised predictive models to aid in navigation, optimal path planning and drive opportunistic sampling. Although these models provide information at a unprecedented resolutions and have proven to increase accuracy and effectiveness in multiple campaigns, most are deterministic in nature. Thus, predictions cannot be incorporated into probabilistic planning frameworks, nor do they provide any metric on the variance or confidence of the output variables. In this paper, we provide an initial investigation into determining the confidence of ocean model predictions based on the results of multiple field deployments of two autonomous underwater vehicles. For multiple missions conducted over a two-month period in 2011, we compare actual vehicle executions to simulations of the same missions through the Regional Ocean Modeling System in an ocean region off the coast of southern California. This comparison provides a qualitative analysis of the current velocity predictions for areas within the selected deployment region. Ultimately, we present a spatial heat-map of the correlation between the ocean model predictions and the actual mission executions. Knowing where the model provides unreliable predictions can be incorporated into planners to increase the utility and application of the deterministic estimations.

Performance evaluation of onsite sewage treatment : Evaluation of current research

Efficient management of domestic wastewater is a primary requirement for human well being. Failure to adequately address issues of wastewater collection, treatment and disposal can lead to adverse public health and environmental impacts. The increasing spread of urbanisation has led to the conversion of previously rural land into urban developments and the more intensive development of semi urban areas. However the provision of reticulated sewerage facilities has not kept pace with this expansion in urbanisation. This has resulted in a growing dependency on onsite sewage treatment. Though considered only as a temporary measure in the past, these systems are now considered as the most cost effective option and have become a permanent feature in some urban areas. This report is the first of a series of reports to be produced and is the outcome of a research project initiated by the Brisbane City Council. The primary objective of the research undertaken was to relate the treatment performance of onsite sewage treatment systems with soil conditions at site, with the emphasis being on septic tanks. This report consists of a ‘state of the art’ review of research undertaken in the arena of onsite sewage treatment. The evaluation of research brings together significant work undertaken locally and overseas. It focuses mainly on septic tanks in keeping with the primary objectives of the project. This report has acted as the springboard for the later field investigations and analysis undertaken as part of the project. Septic tanks still continue to be used widely due to their simplicity and low cost. Generally the treatment performance of septic tanks can be highly variable due to numerous factors, but a properly designed, operated and maintained septic tank can produce effluent of satisfactory quality. The reduction of hydraulic surges from washing machines and dishwashers, regular removal of accumulated septage and the elimination of harmful chemicals are some of the practices that can improve system performance considerably. The relative advantages of multi chamber over single chamber septic tanks is an issue that needs to be resolved in view of the conflicting research outcomes. In recent years, aerobic wastewater treatment systems (AWTS) have been gaining in popularity. This can be mainly attributed to the desire to avoid subsurface effluent disposal, which is the main cause of septic tank failure. The use of aerobic processes for treatment of wastewater and the disinfection of effluent prior to disposal is capable of producing effluent of a quality suitable for surface disposal. However the field performance of these has been disappointing. A significant number of these systems do not perform to stipulated standards and quality can be highly variable. This is primarily due to houseowner neglect or ignorance of correct operational and maintenance procedures. The other problems include greater susceptibility to shock loadings and sludge bulking. As identified in literature a number of design features can also contribute to this wide variation in quality. The other treatment processes in common use are the various types of filter systems. These include intermittent and recirculating sand filters. These systems too have their inherent advantages and disadvantages. Furthermore as in the case of aerobic systems, their performance is very much dependent on individual houseowner operation and maintenance practices. In recent years the use of biofilters has attracted research interest and particularly the use of peat. High removal rates of various wastewater pollutants have been reported in research literature. Despite these satisfactory results, leachate from peat has been reported in various studies. This is an issue that needs further investigations and as such biofilters can still be considered to be in the experimental stage. The use of other filter media such as absorbent plastic and bark has also been reported in literature. The safe and hygienic disposal of treated effluent is a matter of concern in the case of onsite sewage treatment. Subsurface disposal is the most common and the only option in the case of septic tank treatment. Soil is an excellent treatment medium if suitable conditions are present. The processes of sorption, filtration and oxidation can remove the various wastewater pollutants. The subsurface characteristics of the disposal area are among the most important parameters governing process performance. Therefore it is important that the soil and topographic conditions are taken into consideration in the design of the soil absorption system. Seepage trenches and beds are the common systems in use. Seepage pits or chambers can be used where subsurface conditions warrant, whilst above grade mounds have been recommended for a variety of difficult site conditions. All these systems have their inherent advantages and disadvantages and the preferable soil absorption system should be selected based on site characteristics. The use of gravel as in-fill for beds and trenches is open to question. It does not contribute to effluent treatment and has been shown to reduce the effective infiltrative surface area. This is due to physical obstruction and the migration of fines entrained in the gravel, into the soil matrix. The surface application of effluent is coming into increasing use with the advent of aerobic treatment systems. This has the advantage that treatment is undertaken on the upper soil horizons, which is chemically and biologically the most effective in effluent renovation. Numerous research studies have demonstrated the feasibility of this practice. However the overriding criteria is the quality of the effluent. It has to be of exceptionally good quality in order to ensure that there are no resulting public health impacts due to aerosol drift. This essentially is the main issue of concern, due to the unreliability of the effluent quality from aerobic systems. Secondly, it has also been found that most householders do not take adequate care in the operation of spray irrigation systems or in the maintenance of the irrigation area. Under these circumstances surface disposal of effluent should be approached with caution and would require appropriate householder education and stringent compliance requirements. However despite all this, the efficiency with which the process is undertaken will ultimately rest with the individual householder and this is where most concern rests. Greywater too should require similar considerations. Surface irrigation of greywater is currently being permitted in a number of local authority jurisdictions in Queensland. Considering the fact that greywater constitutes the largest fraction of the total wastewater generated in a household, it could be considered to be a potential resource. Unfortunately in most circumstances the only pretreatment that is required to be undertaken prior to reuse is the removal of oil and grease. This is an issue of concern as greywater can considered to be a weak to medium sewage as it contains primary pollutants such as BOD material and nutrients and may also include microbial contamination. Therefore its use for surface irrigation can pose a potential health risk. This is further compounded by the fact that most householders are unaware of the potential adverse impacts of indiscriminate greywater reuse. As in the case of blackwater effluent reuse, there have been suggestions that greywater should also be subjected to stringent guidelines. Under these circumstances the surface application of any wastewater requires careful consideration. The other option available for the disposal effluent is the use of evaporation systems. The use of evapotranspiration systems has been covered in this report. Research has shown that these systems are susceptible to a number of factors and in particular to climatic conditions. As such their applicability is location specific. Also the design of systems based solely on evapotranspiration is questionable. In order to ensure more reliability, the systems should be designed to include soil absorption. The successful use of these systems for intermittent usage has been noted in literature. Taking into consideration the issues discussed above, subsurface disposal of effluent is the safest under most conditions. This is provided the facility has been designed to accommodate site conditions. The main problem associated with subsurface disposal is the formation of a clogging mat on the infiltrative surfaces. Due to the formation of the clogging mat, the capacity of the soil to handle effluent is no longer governed by the soil’s hydraulic conductivity as measured by the percolation test, but rather by the infiltration rate through the clogged zone. The characteristics of the clogging mat have been shown to be influenced by various soil and effluent characteristics. Secondly, the mechanisms of clogging mat formation have been found to be influenced by various physical, chemical and biological processes. Biological clogging is the most common process taking place and occurs due to bacterial growth or its by-products reducing the soil pore diameters. Biological clogging is generally associated with anaerobic conditions. The formation of the clogging mat provides significant benefits. It acts as an efficient filter for the removal of microorganisms. Also as the clogging mat increases the hydraulic impedance to flow, unsaturated flow conditions will occur below the mat. This permits greater contact between effluent and soil particles thereby enhancing the purification process. This is particularly important in the case of highly permeable soils. However the adverse impacts of the clogging mat formation cannot be ignored as they can lead to significant reduction in the infiltration rate. This in fact is the most common cause of soil absorption systems failure. As the formation of the clogging mat is inevitable, it is important to ensure that it does not impede effluent infiltration beyond tolerable limits. Various strategies have been investigated to either control clogging mat formation or to remediate its severity. Intermittent dosing of effluent is one such strategy that has attracted considerable attention. Research conclusions with regard to short duration time intervals are contradictory. It has been claimed that the intermittent rest periods would result in the aerobic decomposition of the clogging mat leading to a subsequent increase in the infiltration rate. Contrary to this, it has also been claimed that short duration rest periods are insufficient to completely decompose the clogging mat, and the intermediate by-products that form as a result of aerobic processes would in fact lead to even more severe clogging. It has been further recommended that the rest periods should be much longer and should be in the range of about six months. This entails the provision of a second and alternating seepage bed. The other concepts that have been investigated are the design of the bed to meet the equilibrium infiltration rate that would eventuate after clogging mat formation; improved geometry such as the use of seepage trenches instead of beds; serial instead of parallel effluent distribution and low pressure dosing of effluent. The use of physical measures such as oxidation with hydrogen peroxide and replacement of the infiltration surface have been shown to be only of short-term benefit. Another issue of importance is the degree of pretreatment that should be provided to the effluent prior to subsurface application and the influence exerted by pollutant loadings on the clogging mat formation. Laboratory studies have shown that the total mass loadings of BOD and suspended solids are important factors in the formation of the clogging mat. It has also been found that the nature of the suspended solids is also an important factor. The finer particles from extended aeration systems when compared to those from septic tanks will penetrate deeper into the soil and hence will ultimately cause a more dense clogging mat. However the importance of improved pretreatment in clogging mat formation may need to be qualified in view of other research studies. It has also shown that effluent quality may be a factor in the case of highly permeable soils but this may not be the case with fine structured soils. The ultimate test of onsite sewage treatment system efficiency rests with the final disposal of effluent. The implication of system failure as evidenced from the surface ponding of effluent or the seepage of contaminants into the groundwater can be very serious as it can lead to environmental and public health impacts. Significant microbial contamination of surface and groundwater has been attributed to septic tank effluent. There are a number of documented instances of septic tank related waterborne disease outbreaks affecting large numbers of people. In a recent incident, the local authority was found liable for an outbreak of viral hepatitis A and not the individual septic tank owners as no action had been taken to remedy septic tank failure. This illustrates the responsibility placed on local authorities in terms of ensuring the proper operation of onsite sewage treatment systems. Even a properly functioning soil absorption system is only capable of removing phosphorus and microorganisms. The nitrogen remaining after plant uptake will not be retained in the soil column, but will instead gradually seep into the groundwater as nitrate. Conditions for nitrogen removal by denitrification are not generally present in a soil absorption bed. Dilution by groundwater is the only treatment available for reducing the nitrogen concentration to specified levels. Therefore based on subsurface conditions, this essentially entails a maximum allowable concentration of septic tanks in a given area. Unfortunately nitrogen is not the only wastewater pollutant of concern. Relatively long survival times and travel distances have been noted for microorganisms originating from soil absorption systems. This is likely to happen if saturated conditions persist under the soil absorption bed or due to surface runoff of effluent as a result of system failure. Soils have a finite capacity for the removal of phosphorus. Once this capacity is exceeded, phosphorus too will seep into the groundwater. The relatively high mobility of phosphorus in sandy soils have been noted in the literature. These issues have serious implications in the design and siting of soil absorption systems. It is not only important to ensure that the system design is based on subsurface conditions but also the density of these systems in given areas is a critical issue. This essentially involves the adoption of a land capability approach to determine the limitations of an individual site for onsite sewage disposal. The most limiting factor at a particular site would determine the overall capability classification for that site which would also dictate the type of effluent disposal method to be adopted.

Maintenance optimisation of a multi-state series-parallel system considering economic dependence and state-dependent inspection intervals

This paper presents a maintenance optimisation method for a multi-state series-parallel system considering economic dependence and state-dependent inspection intervals. The objective function considered in the paper is the average revenue per unit time calculated based on the semi-regenerative theory and the universal generating function (UGF). A new algorithm using the stochastic ordering is also developed in this paper to reduce the search space of maintenance strategies and to enhance the efficiency of optimisation algorithms. A numerical simulation is presented in the study to evaluate the efficiency of the proposed maintenance strategy and optimisation algorithms. The simulation result reveals that maintenance strategies with opportunistic maintenance and state-dependent inspection intervals are more cost-effective when the influence of economic dependence and inspection cost is significant. The study further demonstrates that the optimisation algorithm proposed in this paper has higher computational efficiency than the commonly employed heuristic algorithms.

Hormone-dependent bacterial growth persistence and biofilm formation - A pilot study investigating human follicular fluid collected during IVF cycles

Human follicular fluid, considered sterile, is aspirated as part of an in vitro fertilization (IVF) cycle. However, it is easily contaminated by the trans-vaginal collection route and little information exists in its potential to support the growth of microorganisms. The objectives of this study were to determine whether human follicular fluid can support bacterial growth over time, whether the steroid hormones estradiol and progesterone (present at high levels within follicular fluid) contribute to the in vitro growth of bacterial species, and whether species isolated from follicular fluid form biofilms. We found that bacteria in follicular fluid could persist for at least 28 weeks in vitro and that the steroid hormones stimulated the growth of some bacterial species, specifically Lactobacillus spp., Bifidobacterium spp. Streptococcus spp. and E. coli. Several species, Lactobacillus spp., Propionibacterium spp., and Streptococcus spp., formed biofilms when incubated in native follicular fluids in vitro (18/24, 75%). We conclude that bacteria aspirated along with follicular fluid during IVF cycles demonstrate a persistent pattern of growth. This discovery is important since it can offer a new avenue for investigation in infertile couples.

Research report : computer surfaces and cross-infection

The computer is fast becoming part of the furniture in many hospital settings. Increasing reliance on the computer for documentation and dissemination of information in patient-care areas has increased the need to consider this equipment as a potential environmental reservoir for microorganisms. This paper reports on a small experimental study which investigated the potential role of computers in cross-infection. The results indicate that computer surfaces are similar to other environmental surfaces and carry the same risks for cross-infection.

The role of representative design in talent development : a comment on “Talent identification and promotion programmes of Olympic athletes”

A recent comment in the Journal of Sports Sciences (MacNamara & Collins, 2011) highlighted some major concerns with the current structure of talent identification and development (TID) programmes of Olympic athletes (e.g. Gulbin, 2008; Vaeyens, Gullich, Warr, & Philippaerts, 2009). In a cogent commentary, MacNamara and Collins (2011) provided a short review of the extant literature, which was both timely and insightful. Specifically, they criticised the ubiquitous one-dimensional ‘physically-biased’ attempts to produce world class performers, emphasising the need to consider a number of key environmental variables in a more multi-disciplinary perspective. They also lamented the wastage of talent, and alluded to the operational and opportunistic nature of current talent transfer programmes. A particularly compelling aspect of the comment was their allusion to high profile athletes who had ‘failed’ performance evaluation tests and then proceeded to succeed in that sport. This issue identifies a problem with current protocols for evaluating performance and is a line of research that is sorely needed in the area of talent development. To understand the nature of talent wastage that might be occurring in high performance programmes in sport, future empirical work should seek to follow the career paths of ‘successful’ and ‘unsuccessful’ products of TID programmes, in comparative analyses. Pertinent to the insights of MacNamara and Collins (2011), it remains clear that a number of questions have not received enough attention from sport scientists interested in talent development, including: (i) why is there so much wastage of talent in such programmes? And (ii), why are there so few reported examples of successful talent transfer programmes? These questions highlight critical areas for future investigation. The aim of this short correspondence is to discuss these and other issues researchers and practitioners might consider, and to propose how an ecological dynamics underpinning to such investigations may help the development of existing protocols...

Injuries leading to hospitalisation in the first year of life : analysis by trimester of age using coded data and textual description

Objective: To describe unintentional injuries to children aged less than one year, using coded and textual information, in three-month age bands to reflect their development over the year. Methods: Data from the Queensland Injury Surveillance Unit was used. The Unit collects demographic, clinical and circumstantial details about injured persons presenting to selected emergency departments across the State. Only injuries coded as unintentional in children admitted to hospital were included for this analysis. Results: After editing, 1,082 children remained for analysis, 24 with transport-related injuries. Falls were the most common injury, but becoming proportionately less over the year, whereas burns and scalds and foreign body injuries increased. The proportion of injuries due to contact with persons or objects varied little, but poisonings were relatively more common in the first and fourth three-month periods. Descriptions indicated that family members were somehow causally involved in 16% of injuries. Our findings are in qualitative agreement with comparable previous studies. Conclusion: The pattern of injuries varies over the first year of life and is clearly linked to the child's increasing mobility. Implications: Injury patterns in the first year of life should be reported over shorter intervals. Preventive measures for young children need to be designed with their rapidly changing developmental stage in mind, using a variety of strategies, one of which could be opportunistic developmentally specific education of parents. Injuries in young children are of abiding concern given their immediate health and emotional effects, and potential for long-term adverse sequelae. In Australia, in the financial year 2006/07, 2,869 children less than 12 months of age were admitted to hospital for an unintentional injury, a rate of 10.6 per 1,000, representing a considerable economic and social burden. Given that many of these injuries are preventable, this is particularly concerning. Most epidemiologic studies analyse data in five-year age bands, so children less than five years of age are examined as a group. This study includes only those children younger than one year of age to identify injury detail lost in analyses of the larger group, as we hypothesised that the injury pattern varied with the developmental stage of the child. The authors of several North American studies have commented that in dealing with injuries in pre-school children, broad age groupings are inadequate to do justice to the rapid developmental changes in infancy and early childhood, and have in consequence analysed injuries in shorter intervals. To our knowledge, no similar analysis of Australian infant injuries has been published to date. This paper describes injury in children less than 12 months of age using data from the Queensland Injury Surveillance Unit (QISU).

Genetic variability and antimicrobial resistance of Ureaplasma parvum in response to maternal erythromycin treatment : a study in pregnant sheep

Background: Ureaplasmas are the most frequently isolated microorganisms from the amniotic fluid (AF) of pregnant women and can cause chronic infections that are difficult to eradicate with standard macrolide treatment. We tested the effects of erythromycin treatment on phenotypic and genotypic markers of ureaplasmal antimicrobial resistance in sheep. Method: At 50 days of gestation (d, term=145d) 12 pregnant ewes received intra-amniotic injections of U. parvum serovar 3 (erythromycin-sensitive, 2x104 colony-forming-units). At 100d ewes received: erythromycin treatment (500 mg, q3h for 4 days, IM, n=6) or no treatment (n=6). Fetuses were delivered surgically (125d) and AF and chorioamnion were collected for: culture, minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) testing; 23S rRNA sequencing; and detection of macrolide-lincosamide-streptogramin resistance (MLSr) genes. Results: MICs of erythromycin, azithromycin and roxithromycin against AF isolates were low (range = 0.06 mg/L to 1.0 mg/L); however, chorioamnion isolates demonstrated increased resistance to roxithromycin (0.13 – 5.33 mg/L). 62.5% of chorioamnion ureaplasmas formed biofilms in vitro and mutations (125 nucleotides, 29.6%) were found in the 23S rRNA gene (domain V) of chorioamnion (but not AF) ureaplasmas. MLSr genes (ermB, msrC and msrD) were detected in 100% of chorioamnion isolates and only msrD was detected in AF isolates (40%). Conclusions: 23S rRNA mutations and MLSr genes occurred independently of erythromycin treatment, suggesting that the anatomical site of infection and microenvironment may exert selective pressures on ureaplasmas that cause genetic changes and alter antimicrobial sensitivity profiles. These results have serious implications for treatment of in utero infections.

Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization

Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA), a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic) and short term (acute) durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2x107 colony-forming-units (CFU) of U. parvum serovar 3 (Up) or media controls (C) were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n=8; C69, n=4); day 117 (7d Up, n=8; C7, n=2); and day 121 (3d Up, n=8; C3, n=2) of gestation (term=145-150d). At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i) snap frozen for subsequent ureaplasma culture, and (ii) fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up) or very few MBA variants (7d Up) were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion), during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

Water microbiology

Microbiology is the science devoted lo the study of organisms that are too small to be seen by the naked eye. These microorganisms are a large and diverse group of free-living forms that exist as single cells or cell clusters. Being free-living, microbial cells are distinct from the cells of animals and plants as the latter are not able to live alone in nature but only in characteristic groups. A single microbial cell, generally, is able to carry out its life processes of growth, respiration and reproduction independently of other cells, either of the same kind or of different kinds. There are five subdisciplines of microbiology: (a) the study of bacteria (bacteriology); (b) the study of viruses (virology); (c) the study of algae (phycology); (d) the study of fungi (mycology); and (e) the study of protozoa (protozoology). In the examination of the environment, all five areas of microbiology are studied. This becomes obvious when discussing the significance of each of these groups of organisms in relation to human health.