Assessment of quality defects in macadamia kernels using NIR spectroscopy

Spectral data were collected of intact and ground kernels using 3 instruments (using Si-PbS, Si, and InGaAs detectors), operating over different areas of the spectrum (between 400 and 2500 nm) and employing transmittance, interactance, and reflectance sample presentation strategies. Kernels were assessed on the basis of oil and water content, and with respect to the defect categories of insect damage, rancidity, discoloration, mould growth, germination, and decomposition. Predictive model performance statistics for oil content models were acceptable on all instruments (R2 > 0.98; RMSECV < 2.5%, which is similar to reference analysis error), although that for the instrument employing reflectance optics was inferior to models developed for the instruments employing transmission optics. The spectral positions for calibration coefficients were consistent with absorbance due to the third overtones of CH2 stretching. Calibration models for moisture content in ground samples were acceptable on all instruments (R2 > 0.97; RMSECV < 0.2%), whereas calibration models for intact kernels were relatively poor. Calibration coefficients were more highly weighted around 1360, 740 and 840 nm, consistent with absorbance due to overtones of O-H stretching and combination. Intact kernels with brown centres or rancidity could be discriminated from each other and from sound kernels using principal component analysis. Part kernels affected by insect damage, discoloration, mould growth, germination, and decomposition could be discriminated from sound kernels. However, discrimination among these defect categories was not distinct and could not be validated on an independent set. It is concluded that there is good potential for a low cost Si photodiode array instrument to be employed to identify some quality defects of intact macadamia kernels and to quantify oil and moisture content of kernels in the process laboratory and for oil content in-line. Further work is required to examine the robustness of predictive models across different populations, including growing districts, cultivars and times of harvest.

Piggery pond sludge as a nitrogen source for crops. 1. Mineral N supply estimated from laboratory incubations and field application of stockpiled and wet sludge

Piggery pond sludge (PPS) was applied, as-collected (Wet PPS) and following stockpiling for 12 months (Stockpiled PPS), to a sandy Sodosol and clay Vertosol at sites on the Darling Downs of Queensland. Laboratory measures of N availability were carried out on unamended and PPS-amended soils to investigate their value in estimating supplementary N needs of crops in Australia's northern grains region. Cumulative net N mineralised from the long-term (30 weeks) leached aerobic incubation was described by a first-order single exponential model. The mineralisation rate constant (0.057/week) was not significantly different between Control and PPS treatments or across soil types, when the amounts of initial mineral N applied in PPS treatments were excluded. Potentially mineralisable N (No) was significantly increased by the application of Wet PPS, and increased with increasing rate of application. Application of Wet PPS significantly increased the total amount of inorganic N leached compared with the Control treatments. Mineral N applied in Wet PPS contributed as much to the total mineral N status of the soil as did that which mineralised over time from organic N. Rates of C02 evolution during 30 weeks of aerobic leached incubation indicated that the Stockpiled PPS was more stabilised (19-28% of applied organic C mineralised) than the WetPPS (35-58% of applied organic C mineralised), due to higher lignin content in the former. Net nitrate-N produced following 12 weeks of aerobic non-leached incubation was highly correlated with net nitrate-N leached during 12 weeks of aerobic incubation (R^2 = 0.96), although it was <60% of the latter in both sandy and clayey soils. Anaerobically mineralisable N determined by waterlogged incubation of laboratory PPS-amended soil samples increased with increasing application rate of Wet PPS. Anaerobically minemlisable N from field-moist soil was well correlated with net N mineralised during 30 weeks of aerobic leached incubation (R^2 =0.90 sandy soil; R^2=0.93 clay soil). In the clay soil, the amount of mineral N produced from all the laboratory incubations was significantly correlated with field-measured nitrate-N in the soil profile (0-1.5 m depth) after 9 months of weed-free fallow following PPS application. In contrast, only anaerobic mineralisable N was significantly correlated with field nitrate-N in the sandy soil. Anaerobic incubation would, therefore, be suitable as a rapid practical test to estimate potentially mineralisable N following applications of different PPS materials in the field.

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.

A universal test to determine the integrity of RNA, and its suitability for reverse transcription, in animal tissue laboratory specimens

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.

Detection of Chlamydiaceae DNA in veterinary specimens using a family-specific PCR

Aims: The aim of this work was to develop a rapid molecular test for the detection of the Chlamydiaceae family, irrespective of the species or animal host. Methods and Results: The method described herein is a polymerase chain reaction targeting the 16S rRNA gene of the Chlamydiaceae family, and the results demonstrate that the test reacts with five reference Chlamydiaceae but none of the 19 other bacterial species or five uninfected animal tissues tested. The results also indicate the enhanced sensitivity of this test when compared with conventional culture or serology techniques. This is demonstrated through parallel testing of six real clinical veterinary cases and confirmatory DNA sequence analysis. Conclusions, Significance and Impact of the Study: This test can be used by veterinary diagnostic laboratories for rapid detection of Chlamydiaceae in veterinary specimens, with no restriction of chlamydial species or animal host. The test does not differentiate chlamydial species, and if required, speciation must be carried out retrospectively using alternate methods. However, for the purpose of prescribing therapy for chlamydiosis, this test would be an invaluable laboratory tool.

Field and laboratory evaluations of fungicides for the control of brown spot (Corynespora cassiicola) and black spot (Asperisporium caricae) of papaya in far north Queensland, Australia

Several chemicals including strobilurins (pyraclostrobin and azoxystrobin), triazoles (difenoconazole and tebuconazole), dithiocarbamates (propineb, metiram, ziram and mancozeb) and the phthalimide chlorothalonil were evaluated in three field experiments in north Queensland, Australia, for the control of brown spot (caused by Corynespora cassiicola) and black spot (caused by Asperisporium caricae) of papaya. Chlorothalonil and pyraclostrobin were shown to be more effective than the industry standard, mancozeb, for the control of brown spot. In the black spot experiments, difenoconazole, pyraclostrobin and chlorothalonil used alone or in spray programs were as effective as, or better than, the industry standards, mancozeb and tebuconazole. Plants treated with pyraclostrobin and difenoconazole had more fruit unaffected by black spot (97% and 99% respectively) than plants treated with tebuconazole (51%), mancozeb (20%) and the untreated controls (1%). Laboratory tests also showed that A. caricae was more sensitive to difenoconazole (EC50 of 2ppm) than tebuconazole (EC50 of 14ppm). In 2007, off-label permits were obtained for chlorothalonil for control of brown spot and difenoconazole and chlorothalonil for the control of black spot of papaya.

Laboratory studies on Australian isolates of Metarhizium anisopliae as a biopesticide for the cattle tick Boophilus microplus

Thirty-one isolates of Metarhizium anisopliae were bioassayed against the cattle tick (Boophilus microplus). More than half of the isolates showed a high degree of virulence to ticks. Radial growth curves for growth between 20 °C and 40 °C were obtained for all isolates. This information together with information on virulence will be important for the selection of isolates suitable to kill ticks on the surface of cattle. A biopesticide for cattle ticks must kill ticks rapidly at temperatures within the upper end of most isolates' growth curves. It was also found that the time taken to achieve 100% tick mortality in vitro using a virulent isolate could be halved by applying conidia in a 10% oil emulsion. Scanning electron microscopy and light microscopy were used to investigate and compare the germination and penetration of conidia formulated in aqueous and oil formulations. It was found that conidia in both formulations were able to germinate and produce appressoria on the surface of ticks in less than 11 h. Marked weakness within 26 h, followed by extensive hyphal growth on the cuticle characterised the invasion of ticks by M. anisopliae.

Seed persistence in the field may be predicted by laboratory-controlled aging

Weed management is complicated by the presence of soil seed banks. The complexity of soil-seed interactions means that seed persistence in the field is often difficult to measure, let alone predict. Field trials, although accurate in their context, are time-consuming and expensive to conduct for individual species. Some ex situ techniques for estimating seed life expectancy have been proposed, but these fail to simulate the environmental complexity of the field. Also, it has been questioned whether techniques such as the controlled aging test (CAT) are useful indicators of field persistence. This study aimed to test the validity of the standard CAT (seed aging at 45 C and 60% relative humidity) in use at the Royal Botanic Gardens, Kew, U.K., for predicting field seed-persistence. Comparison of seed persistence and CAT data for 27 northwest European species suggested a significant positive correlation of 0.31. Subsequently, 13 species of emerging and common weeds of Queensland were assessed for their seed longevity using the CAT. The seed longevity data of these species in the CAT were linked with field seed-persistence data according to three broad seed-persistence categories: <1 yr, 1 to 3 yr, and >3 yr. We discuss the scope for using the CAT as a tool for rapid assignment of species to these categories. There is a need for further studies that compare predictions of seed persistence based on the CAT with seed persistence in the field for a larger range of species and environments.

Models for the field-based toxicity of copper and zinc salts to wheat in 11 Australian soils and comparison to laboratory-based models

Laboratory-based relationships that model the phytotoxicity of metals using soil properties have been developed. This paper presents the first field-based phytotoxicity relationships. Wheat(Triticum aestivum L) was grown at 11 Australian field sites at which soil was spiked with copper (Cu) and zinc (Zn) salts. Toxicity was measured as inhibition of plant growth at 8 weeks and grain yield at harvest. The added Cu and Zn EC10 values for both endpoints ranged from approximately 3 to 4760 mg/kg. There were no relationships between field-based 8-week biomass and grain yield toxicity values for either metal. Cu toxicity was best modelled using pH and organic carbon content while Zn toxicity was best modelled using pH and the cation exchange capacity. The best relationships estimated toxicity within a factor of two of measured values. Laboratory-based phytotoxicity relationships could not accurately predict field-based phytotoxicity responses.

Assessment of permeable covers for odour reduction in piggery effluent ponds. 1. Laboratory-scale trials

A variety of materials were trialed as supported permeable covers using a series of laboratory-scale anaerobic digesters. Efficacy of cover performance was assessed in terms of impact on odour and greenhouse gas emission rate, and the characteristics of anaerobic liquor. Data were collected over a 12-month period. Initially the covers reduced the rate of odour emission 40-100 times relative to uncovered digesters. After about three months, this decreased to about a threefold reduction in odour emission rate, which was maintained over the remainder of the trial. The covers did not alter methane emission rates. Carbon dioxide emission rates varied according to cover type. Performance of the covers was attributed to the physical characteristics of the cover materials and changes in liquor composition. The reductions in odour emission indicate that these covers offer a cost-effective method for odour control.

Successful management of colletotrichum crown and stolon rot in runner production in sub-tropical Australia

Crown, stolon, and petiole rots caused by Colletotrichum gloeosporioides (C.g.) were first identified in runner beds of the Queensland Approved Runner Scheme (QARS) in February 1989. The outbreaks occurred annually from 1990 to 1994. Minor losses in subsequent fruit crops occurred from 1990 to 1993, with 50% post-establishment losses occurring on fruit farms in southeast Queensland in 1994. The objective of this work was to provide a control strategy for the disease that would give stability to the QARS. Runner-bed trials in 1993-1994 showed that Octave® (462 g/kg prochloraz as the MnCl2 complex) was highly effective in reducing the incidence field symptoms and laboratory recovery of C.g. from symptomless petioles. A simple detached petiole laboratory test for measuring fungicide efficacy in runner bed trials and for laboratory screening of fungicides, is described. Scheme protocols were changed to require that only foundation plants from tissue culture were allowed onto QARS sites. These were to be symptomless and to have tested negative for the presence of C.g. The application of Octave® at fortnightly intervals in all QARS nurseries has reduced the level of visible symptoms and the laboratory recovery of C.g. from symptomless petioles to almost zero.

Laboratory information management software for genotyping workflows: Applications in high throughput crop genotyping

Background: With the advances in DNA sequencer-based technologies, it has become possible to automate several steps of the genotyping process leading to increased throughput. To efficiently handle the large amounts of genotypic data generated and help with quality control, there is a strong need for a software system that can help with the tracking of samples and capture and management of data at different steps of the process. Such systems, while serving to manage the workflow precisely, also encourage good laboratory practice by standardizing protocols, recording and annotating data from every step of the workflow Results: A laboratory information management system (LIMS) has been designed and implemented at the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) that meets the requirements of a moderately high throughput molecular genotyping facility. The application is designed as modules and is simple to learn and use. The application leads the user through each step of the process from starting an experiment to the storing of output data from the genotype detection step with auto-binning of alleles; thus ensuring that every DNA sample is handled in an identical manner and all the necessary data are captured. The application keeps track of DNA samples and generated data. Data entry into the system is through the use of forms for file uploads. The LIMS provides functions to trace back to the electrophoresis gel files or sample source for any genotypic data and for repeating experiments. The LIMS is being presently used for the capture of high throughput SSR (simple-sequence repeat) genotyping data from the legume (chickpea, groundnut and pigeonpea) and cereal (sorghum and millets) crops of importance in the semi-arid tropics. Conclusions: A laboratory information management system is available that has been found useful in the management of microsatellite genotype data in a moderately high throughput genotyping laboratory. The application with source code is freely available for academic users and can be downloaded from http://www.icrisat.org/bt-software-d-lims.htm

Development of a DNA based aging technique for use in fisheries assessments. Final report, Australian Fisheries Research and Development Corporation Project 2007/033.

The productivity of a fisheries resource can be quantified from estimates of recruitment, individual growth and natural and fisheries-related mortality, assuming the spatial extent of the resource has been quantified and there is minimal immigration or emigration. The sustainability of a fisheries resource is facilitated by management controls such as minimum and maximum size limits and total allowable catch. Minimum size limits are often set to allow individuals the opportunity to reproduce at least once before the chance of capture. Total allowable catches are a proportion of the population biomass, which is estimated based on known reproduction, recruitment, mortality and growth rates. In some fisheries, however, management actions are put in place without quantification of the resource through the stock assessment process. This occurs because species-specific information, for example individual growth, may not be available. In these circumstances, management actions need to be precautionary to protect against future resource collapse, but this often means that the resource is lightly exploited. Consequently, the productivity of the resource is not fully realised. Australia’s most valuable fisheries are invertebrate fisheries (Australian Department of Agriculture Fisheries and Forestry, 2008). For example, Australian fisheries (i.e. excluding aquaculture) production of crustaceans (largely prawns, rock lobster and crab) was 41,000 tonnes in 2006/7, worth $778 million. Production from mollusc (largely abalone, scallops, oysters and squid) fisheries was 39,000 tonnes, worth $502 million. Together, in 2006/7 crustacean and mollusc fisheries represented 58% of the total value of Australian wild fisheries production. Sustainable management of Australia’s invertebrate fisheries is frustrated by the lack of data on species-specific growth rates. This project investigated a new method to estimate age, and hence individual growth rates, in invertebrate fisheries species. The principle behind the new aging method was that telomeres (i.e. DNA end-caps of chromosomes) get shorter as an individual gets older. We studied commercial crustacean and molluscan species. A vertebrate fish species (silver perch, Bidyanus bidyanus) was used as a control to standardise our work against the literature. We found a clear relationship between telomere length and shell size for temperate abalone (Haliotis rubra). Further research is recommended before the method can be implemented to assist management of wildharvested abalone populations. Age needs to be substituted for shell size in the relationship and it needs to be studied for abalone from several regions. This project showed that telomere length declined with increasing age in Sydney rock oysters (Saccostrea glomerata) and was affected by regional variation. A relationship was not apparent between telomere length and age (or size as a surrogate for age) for crustacean species (school prawns, Metapenaeus macleayi; eastern rock lobster, Sagmariasus verreauxi; southern rock lobster, Jasus edwardsii; and spanner crabs, Ranina ranina). For school prawns, there was no difference between telomere length in males and females. Further research is recommended, however, as telomeric DNA from crustaceans was difficult to analyse using the terminal restriction fragment (TRF) assay. Telomere lengths of spanner crabs and lobsters were at the upper limit of resolution of the assay used and results were affected by degradation and possible contamination of telomeric DNA. It is possible that telomere length is an indicator of remaining lifespan in molluscan and crustacean individuals, as suggested for some vertebrate species (e.g. Monaghan, 2010). Among abalone of similar shell size and among lobster pueruli, there was evidence of individuals having significantly longer or shorter telomeres than the group average. At a population level, this may be a surrogate for estimates of future natural mortality, which may have usefulness in the management of those populations. The method used to assay telomere length (terminal restriction fragment assay) performed adequately for most species, but it was too expensive and time-consuming to be considered a useful tool for gathering information for fisheries management. Research on alternative methods is strongly recommended.

Best management practices for sustainable and safe playing surface of Australian Football League sports fields

In 2002, AFL Queensland and the Brisbane Lions Football Club approached the Department of Primary Industries and Fisheries (Queensland) for advice on improving their Premier League sports fields. They were concerned about player safety and dissatisfaction with playing surfaces, particularly uneven turf cover and variable under-foot conditions. They wanted to get the best from new investments in ground maintenance equipment and irrigation infrastructure. Their sports fields were representative of community-standard, multi-use venues throughout Australia; generally ‘natural’ soil fields, with low maintenance budgets, managed by volunteers. Improvements such as reconstruction, drainage, or regular re-turfing are generally not affordable. Our project aimed to: (a) Review current world practice and performance benchmarks; (b) Demonstrate best-practice management for community-standard fields; (c) Adapt relevant methods for surface performance testing; (d) Assess current soils, and investigate useful amendments; (e) Improve irrigation system performance; and (e) Build industry capacity and encourage patterns for ongoing learning. Most global sports field research focuses on elite, sand-based fields. We adjusted elite standards for surface performance (hardness, traction, soil moisture, evenness, sward cover/height) and maintenance programs, to suit community-standard fields with lesser input resources. In regularly auditing ground conditions across 12 AFLQ fields in SE QLD, we discovered surface hardness (measured by Clegg Hammer) was the No. 1 factor affecting player safety and surface performance. Other important indices were turf coverage and surface compaction (measured by penetrometer). AFLQ now runs regularly audits affiliated fields, and closes grounds with hardness readings greater than 190 Gmax. Aerating every two months was the primary mechanical practice improving surface condition and reducing hardness levels to < 110 Gmax on the renovated project fields. With irrigation installation, these fields now record surface conditions comparable to elite fields. These improvements encouraged many other sporting organisations to seek advice / assistance from the project team. AFLQ have since substantially invested in an expanded ground improvement program, to cater for this substantially increased demand. In auditing irrigation systems across project fields, we identified low maintenance (with < 65% of sprinklers operating optimally) as a major problem. Retrofitting better nozzles and adjusting sprinklers improved irrigation distribution uniformity to 75-80%. Research showed that reducing irrigation frequency to weekly, and preparedness to withhold irrigation longer after rain, reduced irrigation requirement by 30-50%, compared to industry benchmarks of 5-6 ML/ha/annum. Project team consultation with regulatory authorities enhanced irrigation efficiency under imposed regional water restrictions. Laboratory studies showed incorporated biosolids / composts, or topdressed crumb rubber, improved compaction resistance of soils. Field evaluations confirmed compost incorporation significantly reduced surface hardness of high wear areas in dry conditions, whilst crumb rubber assisted turf persistence into early winter. Neither amendment was a panacea for poor agronomic practices. Under the auspices of the project Trade Mark Sureplay®, we published > 80 articles, and held > 100 extension activities involving > 2,000 participants. Sureplay® has developed a multi-level curator training structure and resource materials, subject to commercial implementation. The partnerships with industry bodies (particularly AFLQ), frequent extension activities, and engagement with government/regulatory sectors have been very successful, and are encouraged for any future work. Specific aspects of sports field management for further research include: (a) Understanding of factors affecting turf wear resistance and recovery, to improve turf persistence under wear; (b) Simple tests for pinpointing areas of fields with high hardness risk; and (c) Evaluation of new irrigation infrastructure, ‘water-saving’ devices, and irrigation protocols, in improving water use and turf cover outcomes.

Preliminary investigation into the induction of reproduction in Ulva spp. in Southeast Queensland for mass cultivation purposes

Ulva is a common component of marine intertidal flora in Australia with many species frequently observed along the Queensland coastline. Three species of Ulva, U. lactuca, U. intestinalis and U. prolifera were found to naturally occur at the Bribie Island Research Centre (BIRC) in Southeast Queensland. Studies were undertaken to establish the most optimal conditions for growing Ulva in the BIRC laboratory. These tests were conducted in order to condition the algal material prior to the sporulation studies, offering more controlled material to assess treatment effects conclusively, and helping eliminate other potentially confounding environmental factors. Results showed that a stocking density of between 5-20 grams of Ulva per litre along with the addition of the soluble fertiliser Aquasol at a rate of 87 mg/L of seawater was ideal for achieving a desired doubling of growth per week. In the wild the formation of Ulva fragments occurs naturally in the ocean through wave and storm action. This breakage can trigger a survival response mechanism which stimulates the algae to form and release gametes. By chopping the tissue, this process could be artificially simulated in the laboratory and creating a simple and easy way to produce new individuals. Studies performed into inducing sporulation in Ulva through a combination of fragmentation and renewal of medium at BIRC showed that sporulation can be successfully induced in all three species of Ulva through these methods, however it was found to be to a degree that would not meet the demands of commercial production with on average a rate of only 33% achieved. While the current study did not find a method suitable for a commercial application the results presented here contribute to increasing our understanding about Ulva reproduction and set a platform for future work in to cultivating Ulva within Southeast Queensland.