Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: Identification of genetic differences between Haemophilus parasuis isolates

Representational Difference Analysis (RDA) is an established technique used for isolation of specific genetic differences between or within bacterial species. This method was used to investigate the genetic basis of serovar-specificity and the relationship between serovar and virulence in Haemophilus parasuis. An RDA clone library of 96 isolates was constructed using H. parasuis strains H425(P) (serovar 12) and HS1967 (serovar 4). To screen such a large clone library to determine which clones are strain-specific would typically involved separately labelling each clone for use in Southern hybridisation against genomic DNA from each of the strains. In this study, a novel application of reverse Southern hybridisation was used to screen the RDA library: genomic DNA from each strain was labelled and used to probe the library to identify strain-specific clones. This novel approach represents a significant improvement in methodology that is rapid and efficient.

The process for implementation of a Quality Management System within a multi-functional cereal laboratory

The intent of this study was to design, document and implement a Quality Management System (QMS) into a laboratory that incorporated both research and development (R&D) and routine analytical activities. In addition, it was necessary for the QMS to be easily and efficiently maintained to: (a) provide documented evidence that would validate the system's compliance with a certifiable standard, (b) fit the purpose of the laboratory, (c) accommodate prevailing government policies and standards, and (d) promote positive outcomes for the laboratory through documentation and verification of the procedures and methodologies implemented. Initially, a matrix was developed that documented the standards' requirements and the necessary steps to be made to meet those requirements. The matrix provided a check mechanism on the progression of the system's development. In addition, it was later utilised in the Quality Manual as a reference tool for the location of full procedures documented elsewhere in the system. The necessary documentation to build and monitor the system consisted of a series of manuals along with forms that provided auditable evidence of the workings of the QMS. Quality Management (QM), in one form or another, has been in existence since the early 1900's. However, the question still remains: is it a good thing or just a bugbear? Many of the older style systems failed because they were designed by non-users, fiercely regulatory, restrictive and generally deemed to be an imposition. It is now considered important to foster a sense of ownership of the system by the people who use the system. The system's design must be tailored to best fit the purpose of the operations of the facility if maximum benefits to the organisation are to be gained.

Heterologous signals allow efficient targeting of a nuclear-encoded fusion protein to plastids and endoplasmic reticulum in diverse plant species

Approximately 30% of plant nuclear genes appear to encode proteins targeted to the plastids or endoplasmic reticulum (ER). The signals that direct proteins into these compartments are diverse in sequence, but, on the basis of a limited number of tests in heterologous systems, they appear to be functionally conserved across species. To further test the generality of this conclusion, we tested the ability of two plastid transit peptides and an ER signal peptide to target green fluorescent protein (GFP) in 12 crops, including three monocots (barley, sugarcane, wheat) and nine dicots (Arabidopsis, broccoli, cabbage, carrot, cauliflower, lettuce, radish, tobacco, turnip). In all species, transient assays following microprojectile bombardment or vacuum infiltration using Agrobacterium showed that the plastid transit peptides from tomato DCL (defective chloroplast and leaves) and tobacco RbcS [ribulose bisphosphate carboxylase (Rubisco) small subunit] genes were effective in targeting GFP to the leaf plastids. GFP engineered as a fusion to the N-terminal ER signal peptide from Arabidopsis basic chitinase and a C-terminal HDEL signal for protein retention in the ER was accumulated in the ER of all species. The results in tobacco were confirmed in stably transformed cells. These signal sequences should be useful to direct proteins to the plastid stroma or ER lumen in diverse plant species of biotechnological interest for the accumulation of particular recombinant proteins or for the modification of particular metabolic streams.

Implementation of an electronic nose for continuous odour monitoring in a poultry shed

Continuous odour monitoring technologies are necessary to understand the complex odour-generating mechanisms within poultry housing as well as to identify strategies to reduce the impact of odour emissions on local communities. To evaluate electronic nose (EN) technologies for continuously assessing odour concentration in poultry housing, a mobile laboratory containing an electronic nose and an associated sample delivery system was deployed to a commercial poultry farm and tested over a broiler production cycle. The results demonstrated that it was possible to develop a model to allow an electronic nose to provide a semi-continuous measurement of odour concentrations. The electronic nose was also able to demonstrate the influence of shed conditions on odour emissions.

EMS in the pastoral industries of western Queensland: From customisation to implementation

This paper outlines the customisation of Environmental Management Systems (EMS) for the pastoral industry of western Queensland, the recruitment and training of pastoral producers, and their development and implementation of EMS. EMS was simplified to a 7-step process and producers were recruited to trial this customised EMS. Producers from 40 properties received EMS training, either as groups or individually. Of these, 37 commenced Pastoral EMS development through a facilitated approach that allowed them to learn about EMS while developing an EMS for their property. EMS implementation has been more effective with producers who were trained in groups. At this stage, however, most producers do not see value in EMS as there are currently no strong drivers to warrant continued development and implementation. Key findings resulting from this work were that personal contact and assistance is vital to encourage producers to trial EMS, and that a staged approach to EMS implementation, commencing with a self-assessment, is recommended. EMS training is most successful in a group situation; however, an alternative method of delivery should be provided for those producers who, either by choice or isolation, have to work alone. A support network is also necessary to encourage and maintain progress with EMS development and implementation, particularly where no strong drivers exist.

Using bacteria in banana for efficient use of fertilizer

This project aims to develop sustainable banana production practices by improving efficiency of fertilizer use. We will investigate how we can reintroduce endophytic beneficial bacteria that can be established inside the banana host to provide lasting and durable benefits to growth. In the current research we have been able to isolate bacteria from inside banana and preliminary characterisation indicates isolates with a range of attributes for improved efficiency of fertilizer uptake. Experimentation will include evaluation in vitro and pot trials and field trials using bacteria most likely to increase plant access to nutrients as well to compare nutrient impacts from conventional verses slow release fertilizer.

Efficient organogenesis of an Australian passionfruit hybrid (Passiflora edulis x Passiflora edulis var. flavicarpa) suitable for gene delivery

An efficient regeneration protocol based on organogenesis from cotyledon explants and suitable for gene delivery has been developed for an Australian passionfruit hybrid. Multiple shoots were regenerated from 30-day-old cotyledon explants on Murashige and Skoog (MS) medium containing 6-benzylvaminopurine (BAP) and coconut water. Media pulsing experiments were conducted to investigate the effect on organogenesis of exposure time of the explants to MS containing 10 mu M BAP and 10% (v/v) coconut water, i.e. passionfruit regeneration medium (PRM). Continuous exposure of these explants to PRM maximised the number of shoots produced to 12.1 per explant. However, periods on hormone-free medium improved the appearance of the shoots and increased the number of explants with shoots from 75 to 84.6%. Further, shoots exposed for 7 days to half-strength MS supplemented with 10 mu M NAA (1-napthalene acetic acid) produced twice as many plantlets than those on half-strength MS alone. Transient GUS histochemical assays indicated delivery of the uidA gene via Agrobacterium tumefaciens.

Implementation costs of Agricultural Management Practices for Water Quality Improvement in the Great Barrier Reef Catchments. CSIRO: Water for Healthy Country National Research Flagship

Executive summary. In this report we analyse implementation costs and benefits for agricultural management practices, grouped into farming systems. In order to do so, we compare plot scale gross margins for the dominant agricultural production systems (sugarcane, grazing and banana cultivation) in the NRM regions Wet Tropics, Burdekin Dry Tropics and Mackay Whitsundays. Furthermore, where available, we present investment requirements for changing to improved farming systems. It must be noted that transaction costs are not captured within this project. For sugarcane, this economic analysis shows that there are expected benefits to sugarcane growers in the different regions through transitions to C and B class farming systems. Further transition to A-class farming systems can come at a cost, depending on the capital investment required and the length of the investment period. Obviously, the costs and benefits will vary for each individual grower and will depend on their starting point and individual property scenario therefore each circumstance needs to be carefully considered before making a change in management practice. In grazing, overall, reducing stocking rates comes at a cost (reduced benefits). However, when operating at low utilisation rates in wetter country, lowering stocking rates can potentially come at a benefit. With win-win potential, extension is preferred to assist farmer in changing management practices to improve their land condition. When reducing stocking rates comes at a cost, incentives may be applicable to support change among farmers. For banana cultivation, the results indicate that the transition to C and B class management practices is a worthwhile proposition from an economic perspective. For a change from B to A class farming systems however, it is not worthwhile from a financial perspective. This is largely due to the large capital investment associated with the change in irrigation system and negative impact in whole of farm gross margin. Overall, benefits will vary for each individual grower depending on their starting point and their individual property scenario. The results presented in this report are one possible set of figures to show the changes in profitability of a grower operating in different management classes. The results in this report are not prescriptive of every landholder. Landholders will have different costs and benefits from transitioning to improved practices, even if similar operations are practiced, hence it is recommended that landholders that are willing to change management undertake their own research and analysis into the expected costs and benefits for their own soil types and property circumstances.

NEESTIMATOR v2: re-implementation of software for the estimation of contemporary effective population size (N-e) from genetic data

NeEstimator v2 is a completely revised and updated implementation of software that produces estimates of contemporary effective population size, using several different methods and a single input file. NeEstimator v2 includes three single-sample estimators (updated versions of the linkage disequilibrium and heterozygote-excess methods, and a new method based on molecular coancestry), as well as the two-sample (moment-based temporal) method. New features include the following: (i) an improved method for accounting for missing data; (ii) options for screening out rare alleles; (iii) confidence intervals for all methods; (iv) the ability to analyse data sets with large numbers of genetic markers (10000 or more); (v) options for batch processing large numbers of different data sets, which will facilitate cross-method comparisons using simulated data; and (vi) correction for temporal estimates when individuals sampled are not removed from the population (Plan I sampling). The user is given considerable control over input data and composition, and format of output files. The freely available software has a new JAVA interface and runs under MacOS, Linux and Windows.

Identifying risk-efficient strategies using stochastic frontier analysis and simulation: An application to irrigated cropping in Australia

In irrigated cropping, as with any other industry, profit and risk are inter-dependent. An increase in profit would normally coincide with an increase in risk, and this means that risk can be traded for profit. It is desirable to manage a farm so that it achieves the maximum possible profit for the desired level of risk. This paper identifies risk-efficient cropping strategies that allocate land and water between crop enterprises for a case study of an irrigated farm in Southern Queensland, Australia. This is achieved by applying stochastic frontier analysis to the output of a simulation experiment. The simulation experiment involved changes to the levels of business risk by systematically varying the crop sowing rules in a bioeconomic model of the case study farm. This model utilises the multi-field capability of the process based Agricultural Production System Simulator (APSIM) and is parameterised using data collected from interviews with a collaborating farmer. We found sowing rules that increased the farm area sown to cotton caused the greatest increase in risk-efficiency. Increasing maize area also improved risk-efficiency but to a lesser extent than cotton. Sowing rules that increased the areas sown to wheat reduced the risk-efficiency of the farm business. Sowing rules were identified that had the potential to improve the expected farm profit by ca. $50,000 Annually, without significantly increasing risk. The concept of the shadow price of risk is discussed and an expression is derived from the estimated frontier equation that quantifies the trade-off between profit and risk.

Overcoming barriers to the successful implementation of a classical biological control strategy for the exotic invasive weed Mikania micrantha in the Asia-Pacific region

Mikania micrantha (Asteraceae) commonly known as mikania, is a major invasive alien plant (IAP) in the tropical humid agricultural and forest zones of the Asia-Pacific region. This fast-growing Neotropical vine is able to smother plants in agricultural ecosystems, agroforestry and natural habitats, reducing productivity and biodiversity. Fungal pathogens were first investigated for the classical biological control of this weed in 1996. This resulted in the selection and screening of the highly host-specific and damaging rust pathogen, Puccinia spegazzinii (Pucciniales). It was first released in India and China in 2005/6, although it is not believed to have established. Since then, it has been released successfully in Taiwan, Papua New Guinea (PNG), Fiji and most recently Vanuatu. The rust has established and is spreading rapidly after applying lessons learned from the first releases on the best rust pathotype and release strategy. In PNG, direct monitoring of vegetation change has demonstrated that the rust is having a significant impact on M. micrantha, with no unpredicted non-target impacts. Despite this, the authorities in many countries where mikania is a problem remain cautious about releasing the rust. In Western Samoa, introduction of the rust was not pursued because of a conflict of interest, and the perception that mikania suppresses even worse weeds. For some, ‘pathophobia’ is still a major obstacle. In Indonesia, where insects for weed CBC have been introduced, pathogens will currently not be considered. In other countries such as Bhutan and Myanmar, there are no baseline data on the presence and impact of IAPs and, with no history of CBC, no institutional framework for implementing this approach. Malaysia has a well-developed framework, but capacity needs to be built in the country. Overall, it remains critical to have champions at decision making levels. Hence, even with an effective ‘off-the-shelf’ agent available, implementation of mikania CBC still requires significant inputs tailored to the countries’ specific needs.

The need for speed: Timely prevention of the dispersal of noxious weeds in relief fodder using efficient sampling procedures

Invasive and noxious weeds are well known as a pervasive problem, imposing significant economic burdens on all areas of agriculture. Whilst there are multiple possible pathways of weed dispersal in this industry, of particular interest to this discussion is the unintended dispersal of weed seeds within fodder. During periods of drought or following natural disasters such as wild fire or flood, there arises the urgent need for 'relief' fodder to ensure survival and recovery of livestock. In emergency situations, relief fodder may be sourced from widely dispersed geographic regions, and some of these regions may be invaded by an extensive variety of weeds that are both exotic and detrimental to the intended destination for the fodder. Pasture hay is a common source of relief fodder and it typically consists of a mixture of grassy and broadleaf species that may include noxious weeds. When required urgently, pasture hay for relief fodder can be cut, baled, and transported over long distances in a short period of time, with little opportunity for prebaling inspection. It appears that, at the present time, there has been little effort towards rapid testing of bales, post-baling, for the presence of noxious weeds, as a measure to prevent dispersal of seeds. Published studies have relied on the analysis of relatively small numbers of bales, tested to destruction, in order to reveal seed species for identification and enumeration. The development of faster, more reliable, and non-destructive sampling methods is essential to increase the fodder industry's capacity to prevent the dispersal of noxious weeds to previously unaffected locales.

Understanding the barriers to the implementation of precision agriculture in the central region

There is an increasing requirement for more astute land resource management through efficiencies in agricultural inputs in a sugar cane production system. A precision agriculture (PA) approach can provide a pathway for a sustainable sugarcane production system. One of the impediments to the adoption of PA practices is access to paddock-scale mapping layers displaying variability in soil properties, crop growth and surface drainage. Variable rate application (VRA) of nutrients is an important component of PA. However, agronomic expertise within PA systems has fallen well behind significant advances in PA technologies. Generally, advisers in the sugar industry have a poor comprehension of the complex interaction of variables that contribute to within-paddock variations in crop growth. This is regarded as a significant impediment to the progression of PA in sugarcane and is one of the reasons for the poor adoption of VRA of nutrients in a PA approach to improved sugar cane production. This project therefore has established a number of key objectives which will contribute to the adoption of PA and the staged progression of VRA supported by relevant and practical agronomic expertise. These objectives include provision of base soils attribute mapping that can be determined using Veris 3100 Electrical Conductivity (EC) and digital elevation datasets using GPS mapping technology for a large sector of the central cane growing region using analysis of archived satellite imagery to determine the location and stability of yield patterns over time and in varying seasonal conditions on selected project study sites. They also include the stablishment of experiments to determine appropriate VRA nitrogen rates on various soil types subjected to extended anaerobic conditions, and the establishment of trials to determine nitrogen rates applicable to a declining yield potential associated with the aging of ratoons in the crop cycle. Preliminary analysis of archived yield estimation data indicates that yield patterns remain relatively stable overtime. Results also indicate the where there is considerable variability in EC values there is also significant variation in yield.

Strategic tillage in no-till farming systems in Australia’s northern grains-growing regions: I. Drivers and implementation

Development of no-tillage (NT) farming has revolutionized agricultural systems by allowing growers to manage greater areas of land with reduced energy, labour and machinery inputs to control erosion, improve soil health and reduce greenhouse gas emission. However, NT farming systems have resulted in a build-up of herbicide-resistant weeds, an increased incidence of soil- and stubble-borne diseases and enrichment of nutrients and carbon near the soil surface. Consequently, there is an increased interest in the use of an occasional tillage (termed strategic tillage, ST) to address such emerging constraints in otherwise-NT farming systems. Decisions around ST uses will depend upon the specific issues present on the individual field or farm, and profitability and effectiveness of available options for management. This paper explores some of the issues with the implementation of ST in NT farming systems. The impact of contrasting soil properties, the timing of the tillage and the prevailing climate exert a strong influence on the success of ST. Decisions around timing of tillage are very complex and depend on the interactions between soil water content and the purpose for which the ST is intended. The soil needs to be at the right water content before executing any tillage, while the objective of the ST will influence the frequency and type of tillage implement used. The use of ST in long-term NT systems will depend on factors associated with system costs and profitability, soil health and environmental impacts. For many farmers maintaining farm profitability is a priority, so economic considerations are likely to be a primary factor dictating adoption. However, impacts on soil health and environment, especially the risk of erosion and the loss of soil carbon, will also influence a grower’s choice to adopt ST, as will the impact on soil moisture reserves in rainfed cropping systems.