A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Characterisation and early detection of an offtype from micropropagated Lady Finger bananas

An offtype has been identified from micropropagated Lady Finger bananas (Musa spp., AAB group, Pome subgroup) that is characterised by its slow growth and poor bunch size. Bunch weights were approximately 25% those of normal Lady Finger plants and all of the fruit produced was unmarketable. This particular offtype is the most commonly encountered from micropropagated Lady Finger plants and, in 2 instances, blocks of 3000 and 1500 plants were entirely comprised of this single offtype. Detection of offtype plants was possible during establishment and growth of plants in the glasshouse by the presence of chlorotic streaks in the leaves. In more severe cases the streaks coalesced into chlorotic patches that developed thin, necrotic areas that eventually produced holes or splits in the leaves. Symptom expression was not ameliorated by the addition of fertiliser and even though symptoms were similar to severe Ca and B deficiency, both normal and offtype plants had similar levels of these elements in the leaves. The offtype plants were also slow growing in the glasshouse and produced significantly (P<0.05) smaller pseudostems and leaves than normal plants. Offtype plants could be readily detected after 4 weeks deflasking using the presence of chlorotic streaks in the leaves as the main selection criterion. Maximum discrimination was possible between weeks 5–7 and at the 6-leaf stage when all of the offtypes could be detected.

Development of hazard site surveillance programs for forest invasive species: A case study from Brisbane, Australia

Hazard site surveillance is a system for post-border detection of new pest incursions, targeting sites that are considered potentially at high risk of such introductions. Globalisation, increased volumes of containerised freight and competition for space at domestic ports means that goods are increasingly being first opened at premises some distance from the port of entry, thus dispersing risk away from the main inspection point. Hazard site surveillance acts as a backstop to border control to ensure that new incursions are detected sufficiently early to allow the full range of management options, including eradication and containment, to be considered. This is particularly important for some of the more cryptic forest pests whose presence in a forest often is not discovered until populations are already high and the pest is well established. General requirements for a hazard site surveillance program are discussed using a program developed in Brisbane, Australia, in 2006 as a case study. Some early results from the Brisbane program are presented. In total 67 species and 5757 individuals of wood-boring beetles have been trapped and identified during the program to date. Scolytines are the most abundant taxa, making up 83% of the catch. No new exotics have been trapped but 19 of the species and 60% of all specimens caught are exotics that are already established in Australia.

Hierarchical Bayesian modelling of early detection surveillance for plant pest invasions

Early detection surveillance programs aim to find invasions of exotic plant pests and diseases before they are too widespread to eradicate. However, the value of these programs can be difficult to justify when no positive detections are made. To demonstrate the value of pest absence information provided by these programs, we use a hierarchical Bayesian framework to model estimates of incursion extent with and without surveillance. A model for the latent invasion process provides the baseline against which surveillance data are assessed. Ecological knowledge and pest management criteria are introduced into the model using informative priors for invasion parameters. Observation models assimilate information from spatio-temporal presence/absence data to accommodate imperfect detection and generate posterior estimates of pest extent. When applied to an early detection program operating in Queensland, Australia, the framework demonstrates that this typical surveillance regime provides a modest reduction in the estimate that a surveyed district is infested. More importantly, the model suggests that early detection surveillance programs can provide a dramatic reduction in the putative area of incursion and therefore offer a substantial benefit to incursion management. By mapping spatial estimates of the point probability of infestation, the model identifies where future surveillance resources can be most effectively deployed.

Economic evaluation of precision controlled traffic farming in the Australian sugar industry: a case study of an early adopter

Controlled traffic has been identified as the most practical method of reducing compaction-related soil structural degradation in the Australian sugarcane industry. GPS auto-steer systems are required to maximize this potential. Unfortunately there is a perception that little economic gain will result from investing in this technology. Regardless, a number of growers have made the investment and are reaping substantial economic and lifestyle rewards. In this paper we assess the cost effectiveness of installing GPS guidance and using it to implement Precision Controlled Traffic Farming (PCTF) based on the experience of an early adopter. The Farm Economic Analysis Tool (FEAT) model was used with data provided by the grower to demonstrate the benefits of implementing PCTF. The results clearly show that a farming system based on PCTF and the minimum tillage improved farm gross margin by 11.8% and reduced fuel usage by 58%, compared to producers' traditional practice. PCTF and minimum tillage provide sugar producers with a tool to manage the price cost squeeze at a time of low sugar prices. These data provide producers with the evidence that investment in PCTF is economically prudent.

Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle

A competitive enzyme-linked immunosorbent assay (cELISA) based on a broadly conserved, species-specific, B-cell epitope within the C terminus of Babesia bigemina rhoptry-associated protein 1a was validated for international use. Receiver operating characteristic analysis revealed 16% inhibition as the threshold for a negative result, with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% but modestly decreased the sensitivity to 87.2%. By using 21% inhibition, the positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and the negative predictive values ranged from 97.0% (10% prevalence) to 48.2% (95% prevalence). The assay was able to detect serum antibody as early as 7 days after intravenous inoculation. The cELISA was distributed to five different laboratories along with a reference set of 100 defined bovine serum samples, including known positive, known negative, and field samples. The pairwise concordance among the five laboratories ranged from 100% to 97%, and all kappa values were above 0.8, indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

Managing mycotoxins in maize: Case studies

Mycotoxin contamination of Australian maize is neither common nor extensive, but has the capacity to seriously disrupt marketing. Low to moderate levels of aflatoxins and fumonisins can be widespread in some seasons, but zearalenone, nivalenol and deoxynivalenol are usually confined to small growing localities. Possible approaches to such situations were tested by an analysis of several case studies. It is concluded that communication and coordination across the industry, prediction and prevention of contamination, rapid detection and assessment of contamination, effective use of contaminated maize and breeding for resistance comprise a useful set of strategies for managing mycotoxins in maize.

Detection of nitrogen deficiency in wheat from spectral reflectance indices and basic crop eco-physiological concepts

We tested the capacity of several published multispectral indices to estimate the nitrogen nutrition of wheat canopies grown under different levels of water supply and plant density and derived a simple canopy reflectance index that is greatly independent of those factors. Planar domain geometry was used to account for mixed signals from the canopy and soil when the ground cover was low. A nitrogen stress index was developed, which adjusts shoot %N for plant biomass and area, thereby accounting for environmental conditions that affect growth, such as crop water status. The canopy chlorophyll content index (CCCi) and the modified spectral ratio planar index (mSRPi) could explain 68 and 69% of the observed variability in the nitrogen nutrition of the crop as early as Zadoks 33, irrespective of water status or ground cover. The CCCi was derived from the combination of 3 wavebands 670, 720 and 790 nm, and the mSRPi from 445, 705 and 750 nm, together with broader bands in the NIR and RED. The potential for their spatial application over large fields/paddocks is discussed.

Domestication for Conservation of an Endangered Species: The Case of the Wollemi Pine

A small population of tall slender conifers was discovered in 1994 in a deep rainforest canyon of the Wollemi National Park, New SouthWales, Australia. The living trees closely resembled fossils that were more than 65 million years old, and this ‘living fossil’ was recognised as a third extant genus in the Araucariaceae (Araucaria, Agathis and now Wollemia). The species was named the Wollemi pine (W. nobilis). Extensive searches uncovered very few populations, with the total number of adult trees being less than 100. Ex situ collections were quickly established in Sydney as part of the Wollemi Pine Recovery Plan. The majority of the ex situ population was later transferred to our custom-built facility in Queensland for commercial multiplication. Domestication has relied very heavily on the species’ amenability to vegetative propagation because seed collection from the natural populations is dangerous, expensive, and undesirable for conservation reasons. Early propagation success was poor, with only about 25% of cuttings producing roots. However, small increases in propagation success have a very large impact on a domestication program because plant production can be modelled on an exponential curve where each rooted cutting develops into a mother plant that, in turn, provides more rooted cuttings. An extensive research program elevated rooting percentages to greater than 80% and also provided in vitro methods for plant multiplication. These successes have enabled international release of the Wollemi pine as a new and attractive species for ornamental horticulture.

Specific detection of the Old World screwworm fly, Chrysomya bezziana, in bulk fly trap catches using real-time PCR.

The Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), is a myiasis-causing blowfly of major concern for both animals and humans. Surveillance traps are used in several countries for early detection of incursions and to monitor control strategies. Examination of surveillance trap catches is time-consuming and is complicated by the presence of morphologically similar flies that are difficult to differentiate from Ch. bezziana, especially when the condition of specimens is poor. A molecular-based method to confirm or refute the presence of Ch. bezziana in trap catches would greatly simplify monitoring programmes. A species-specific real-time polymerase chain reaction (PCR) assay was designed to target the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) of Ch. bezziana. The assay uses both species-specific primers and an OWS-specific Taqman MGB probe. Specificity was confirmed against morphologically similar and related Chrysomya and Cochliomyia species. An optimal extraction protocol was developed to process trap catches of up to 1000 flies and the assay is sensitive enough to detect one Ch. bezziana in a sample of 1000 non-target species. Blind testing of 29 trap catches from Australia and Malaysia detected Ch. bezziana with 100% accuracy. The probability of detecting OWS in a trap catch of 50 000 flies when the OWS population prevalence is low (one in 1000 flies) is 63.6% for one extraction. For three extractions (3000 flies), the probability of detection increases to 95.5%. The real-time PCR assay, used in conjunction with morphology, will greatly increase screening capabilities in surveillance areas where OWS prevalence is low.

A case study on the economics of overhead irrigation in the lower Burdekin

Converting from an existing irrigation system is often seen as high risk by the land owner. The significant financial investment and the long period over which the investment runs is also complicated by the uncertainty associated with long term input costs (such as energy), crop production, and the continually evolving natural resource management rules and policy. Irrigation plays a pivotal part in the Burdekin sugarcane farming system. At present the use of furrow irrigation is by far the most common form due to the ease of use, relatively low operating cost and well established infrastructure currently in place. The Mulgrave Area Farmer Integrated Action (MAFIA) grower group, located near Clare in the lower Burdekin region, identified the need to learn about sustainable farming systems with a focus on the environment, social and economic implications. In early 2007, Hesp Faming established a site to investigate the use of overhead irrigation as an alternative to furrow irrigation and its integration with new farming system practices, including Green Cane Trash Blanketing (GCTB). Although significant environmental and social benefits exist, the preliminary investment analysis indicates that the Overhead Low Pressure (OHLP) irrigation system is not adding financial value to the Hesp Farming business. A combination of high capital costs and other offsetting factors resulted in the benefits not being fully realised. A different outcome is achieved if Hesp Farming is able to realise value on the water saved, with both OHLP irrigation systems displaying a positive NPV. This case study provides a framework to further investigate the economics of OHLP irrigation in sugarcane and it is anticipated that with additional data a more definitive outcome will be developed in the future.

Detection of a putative hemolysin operon, hhdBA, of Haemophilus parasuis from pigs with Glasser disease

The aim of the current study was to investigate whether polymerase chain reaction amplification of 16S ribosomal (r)RNA and a putative hemolysin gene operon, hhdBA, can be used to monitor live pigs for the presence of Haemophilus parasuis and predict the virulence of the strains present. Nasal cavity swabs were taken from 30 live, healthy, 1- to 8-week-old pigs on a weekly cycle from a commercial Thai nursery pig herd. A total of 27 of these pigs (90%) tested positive for H. parasuis as early as week 1 of age. None of the H. parasuis-positive samples from healthy pigs was positive for the hhdBA genes. At the same pig nursery, swab samples from nasal cavity, tonsil, trachea, and lung, and exudate samples from pleural/peritoneal cavity were taken from 30 dead pigs displaying typical pathological lesions consistent with Glasser disease. Twenty-two of 140 samples (15.7%) taken from 30 diseased pigs yielded a positive result for H. parasuis. Samples from the exudate (27%) yielded the most positive results, followed by lung, tracheal swab, tonsil, and nasal swab, respectively. Out of 22 positive samples, 12 samples (54.5%) harbored hhdA and/or hhdB genes. Detection rates of hhdA were higher than hhdB. None of the H. parasuis-positive samples taken from nasal cavity of diseased pigs tested positive for hhdBA genes. More work is required to determine if the detection of hhdBA genes is useful for identifying the virulence potential of H. parasuis field isolates.

Reducing the risk of herbicide runoff in sugarcane farming through controlled traffic and early-banded application

The off-site transport of agricultural chemicals, such as herbicides, into freshwater and marine ecosystems is a world-wide concern. The adoption of farm management practices that minimise herbicide transport in rainfall-runoff is a priority for the Australian sugarcane industry, particularly in the coastal catchments draining into the World Heritage listed Great Barrier Reef (GBR) lagoon. In this study, residual herbicide runoff and infiltration were measured using a rainfall simulator in a replicated trial on a brown Chromosol with 90–100% cane trash blanket cover in the Mackay Whitsunday region, Queensland. Management treatments included conventional 1.5 m spaced sugarcane beds with a single row of sugarcane (CONV) and 2 m spaced, controlled traffic sugarcane beds with dual sugarcane rows (0.8 m apart) (2mCT). The aim was to simulate the first rainfall event after the application of the photosynthesis inhibiting (PSII) herbicides ametryn, atrazine, diuron and hexazinone, by broadcast (100% coverage, on bed and furrow) and banding (50–60% coverage, on bed only) methods. These events included heavy rainfall 1 day after herbicide application, considered a worst case scenario, or rainfall 21 days after application. The 2mCT rows had significantly (P < 0.05) less runoff (38%) and lower peak runoff rates (43%) than CONV rows for a rainfall average of 93 mm at 100 mm h−1 (1:20 yr Average Return Interval). Additionally, final infiltration rates were higher in 2mCT rows than CONV rows, with 72 and 52 mm h−1 respectively. This resulted in load reductions of 60, 55, 47, and 48% for ametryn, atrazine, diuron and hexazinone from 2mCT rows, respectively. Herbicide losses in runoff were also reduced by 32–42% when applications were banded rather than broadcast. When rainfall was experienced 1 day after application, a large percentage of herbicides were washed off the cane trash. However, by day 21, concentrations of herbicide residues on cane trash were lower and more resistant to washoff, resulting in lower losses in runoff. Consequently, ametryn and atrazine event mean concentrations in runoff were approximately 8 fold lower at day 21 compared with day 1, whilst diuron and hexazinone were only 1.6–1.9 fold lower, suggesting longer persistence of these chemicals. Runoff collected at the end of the paddock in natural rainfall events indicated consistent though smaller treatment differences to the rainfall simulation study. Overall, it was the combination of early application, banding and controlled traffic that was most effective in reducing herbicide losses in runoff. Crown copyright © 2012

Veneer Grade Analysis of Early to Mid-rotation Plantation Eucalyptus Species in Australia

Processing Australian hardwood plantations into rotary veneer can produce more acceptable marketable product recoveries compared to traditional processing techniques (e.g. sawmilling). Veneers resulting from processing trials from six commercially important Australian hardwood species were dominated by D-grade veneer. Defects such as encased knots, gum pockets, gum veins, surface roughness, splits, bark pockets, and decay impacted the final assigned grade. Four grading scenarios were adopted. The first included a change to the grade limitations for gum pockets and gum veins, while the second investigated the potential impact of effective pruning on grade recovery. Although both scenarios individually had a positive impact on achieving higher face grade veneer qualities, the third and fourth scenarios, which combined both, had a substantial impact, with relative veneer values increasing up to 18.2% using conservative calculations (scenario three) or up to 22.6% (scenario four) where some of the upgraded veneers were further upgraded to A-grade, which attracts superior value. The total change in veneer value was found to depend on the average billet diameter unless defects other than those relating to the scenarios (gum or knots) restricted the benefit of pruning and gum upgrading. This was the case for species prone to high levels of growth stress and related defects.

Novel vaccine acceptance in emerging infectious diseases: a case study of Hendra virus vaccine uptake in Australia

Background: The development of a horse vaccine against Hendra virus has been hailed as a good example of a One Health approach to the control of human disease. Although there is little doubt that this is true, it is clear from the underwhelming uptake of the vaccine by horse owners to date (approximately 10%) that realisation of a One Health approach requires more than just a scientific solution. As emerging infectious diseases may often be linked to the development and implementation of novel vaccines this presentation will discuss factors influencing their uptake; using Hendra virus in Australia as a case study. Methods: This presentation will draw on data collected from the Horse owners and Hendra virus: A Longitudinal cohort study To Evaluate Risk (HHALTER) study. The HHALTER study is a mixed methods research study comprising a two-year survey-based longitudinal cohort study and qualitative interview study with horse owners in Australia. The HHALTER study has investigated and tracked changes in a broad range of issues around early uptake of vaccination, horse owner uptake of other recommended disease risk mitigation strategies, and attitudes to government policy and disease response. Interviews provide further insights into attitudes towards risk and decision-making in relation to vaccine uptake. A combination of quantitative and qualitative data analysis will be reported. Results: Data collected from more than 1100 horse owners shortly after vaccine introduction indicated that vaccine uptake and intention to vaccinate was associated with a number of risk perception factors and financial cost factors. In addition, concerns about side effects and veterinarians refusing to treat unvaccinated horses were linked to uptake. Across the study period vaccine uptake in the study cohort increased to more than 50%, however, concerns around side effects, equine performance and breeding impacts, delays to full vaccine approvals, and attempts to mandate vaccination by horse associations and event organisers have all impacted acceptance. Conclusion: Despite being provided with a safe and effective vaccine for Hendra virus that can protect horses and break the transmission cycle of the virus to humans, Australian horse owners have been reluctant to commit to it. General issues pertinent to novel vaccines, combined with challenges in the implementation of the vaccine have led to issues of mistrust and misconception with some horse owners. Moreover, factors such as cost, booster dose schedules, complexities around perceived risk, and ulterior motives attributed to veterinarians have only served to polarise attitudes to vaccine acceptance.