The mycotoxins 4-deoxynivalenol, zearalenone and aflatoxin in weather-damaged wheat harvested 1983-1985 in south-east Queensland

A survey for various mycotoxins was carried out on samples of all wheat delivered to nine storage and marketing depots in south-eastern Queensland, selected as most likely to receive mycotoxin-contaminated grain. All wheat was surveyed during 1983, when the degree of weather damage was high. Samples of the poorest grade of wheat from these depots were also surveyed in 1984 and 1985. The surveys included all regions where head scab of wheat caused by Fusariurn graminearurn Schwabe Group 2 had been reported to occur at significant levels. 4-Deoxynivalenol was detected in nearly all pooled samples representing bulk wheat at concentrations ranging from traces of <0.01 up to 1.7 mg kg-1. The highest concentration of zearlenone detected in a pooled wheat sample was 0.04 mg kg-1. In a few samples representing individual wheat deliveries and with up to 2.8% by weight of pink grains, 4-deoxynivalenol concentrations ranged up to 11.7 mg kg-' and zearalenone up to 0.43 mg kg-l. Aflatoxins B,, B2, G1 and G2 were detected in only one pooled sample of wheat, at a total aflatoxin concentration of 0.003 mg kg-'. Ochratoxin A, sterigmatocystin and T-2 toxin were not detected. Higher concentrations of mycotoxins were found in the poorer grades of wheat.

Limitations of soil microbial biomass carbon as an indicator of soil pollution in the field

The size of the soil microbial biomass carbon (SMBC) has been proposed as a sensitive indicator for measuring the adverse effects of contaminants on the soil microbial community. In this study of Australian agricultural systems, we demonstrated that field variability of SMBC measured using the fumigation-extraction procedure limited its use as a robust ecotoxicological endpoint. The SMBC varied up to 4-fold across control samples collected from a single field site, due to small-scale spatial heterogeneity in the soil physicochemical environment. Power analysis revealed that large numbers of replicates (3-93) were required to identify 20% or 50% decreases in the size of the SMBC of contaminated soil samples relative to their uncontaminated control samples at the 0.05% level of statistical significance. We question the value of the routine measurement of SMBC as an ecotoxicological endpoint at the field scale, and suggest more robust and predictive microbiological indicators.

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.

The distribution and spread of citrus canker in Emerald, Australia

Citrus canker is a disease of citrus and closely related species, caused by the bacterium Xanthomonas citri subsp. citri. This disease, previously exotic to Australia, was detected on a single farm [infested premise-1, (IP1). IP is the terminology used in official biosecurity protocols to describe a locality at which an exotic plant pest has been confirmed or is presumed to exist. IP are numbered sequentially as they are detected] in Emerald, Queensland in July 2004. During the following 10 months the disease was subsequently detected on two other farms (IP2 and IP3) within the same area and studies indicated the disease first occurred on IP1 and spread to IP2 and IP3. The oldest, naturally infected plant tissue observed on any of these farms indicated the disease was present on IP1 for several months before detection and established on IP2 and IP3 during the second quarter (i.e. autumn) 2004. Transect studies on some IP1 blocks showed disease incidences ranged between 52 and 100% (trees infected). This contrasted to very low disease incidence, less than 4% of trees within a block, on IP2 and IP3. The mechanisms proposed for disease spread within blocks include weather-assisted dispersal of the bacterium (e.g. wind-driven rain) and movement of contaminated farm equipment, in particular by pivot irrigator towers via mechanical damage in combination with abundant water. Spread between blocks on IP2 was attributed to movement of contaminated farm equipment and/or people. Epidemiology results suggest: (i) successive surveillance rounds increase the likelihood of disease detection; (ii) surveillance sensitivity is affected by tree size; and (iii) individual destruction zones (for the purpose of eradication) could be determined using disease incidence and severity data rather than a predefined set area.

Application of a model to assess aflatoxin risk in peanuts.

When exposed to hot (22-35 degrees C) and dry climatic conditions in the field during the final 4-6 weeks of pod filling, peanuts (Arachis hypogaea L.) can accumulate highly carcinogenic and immuno-suppressing aflatoxins. Forecasting of the risk posed by these conditions can assist in minimizing pre-harvest contamination. A model was therefore developed as part of the Agricultural Production Systems Simulator (APSIM) peanut module, which calculated an aflatoxin risk index (ARI) using four temperature response functions when fractional available soil water was <0.20 and the crop was in the last 0.40 of the pod-filling phase. ARI explained 0.95 (P <= 0.05) of the variation in aflatoxin contamination, which varied from 0 to c. 800 mu g/kg in 17 large-scale sowings in tropical and four sowings in sub-tropical environments carried out in Australia between 13 November and 16 December 2007. ARI also explained 0.96 (P <= 0.01) of the variation in the proportion of aflatoxin-contaminated loads (>15 mu g/kg) of peanuts in the Kingaroy region of Australia during the period between the 1998/99 and 2007/08 seasons. Simulation of ARI using historical climatic data from 1890 to 2007 indicated a three-fold increase in its value since 1980 compared to the entire previous period. The increase was associated with increases in ambient temperature and decreases in rainfall. To facilitate routine monitoring of aflatoxin risk by growers in near real time, a web interface of the model was also developed. The ARI predicted using this interface for eight growers correlated significantly with the level of contamination in crops (r=095, P <= 0.01). These results suggest that ARI simulated by the model is a reliable indicator of aflatoxin contamination that can be used in aflatoxin research as well as a decision-support tool to monitor pre-harvest aflatoxin risk in peanuts.

A free-enzyme catalyst for the bioremediation of environmental atrazine contamination.

Herbicide contamination from agriculture is a major issue worldwide, and has been identified as a threat to freshwater and marine environments in the Great Barrier Reef World Heritage Area in Australia. The triazine herbicides are of particular concern because of potential adverse effects, both on photosynthetic organisms and upon vertebrate development. To date a number of bioremediation strategies have been proposed for triazine herbicides, but are unlikely to be implemented due to their reliance upon the release of genetically modified organisms. We propose an alternative strategy using a free-enzyme bioremediant, which is unconstrained by the issues surrounding the use of live organisms. Here we report an initial field trial with an enzyme-based product, demonstrating that the technology is technically capable of remediating water bodies contaminated with the most common triazine herbicide, atrazine.

Responses of tribolium castaneum to olfactory cues from cotton seeds, the fungi associated with cotton seeds, and cereals

We tested, in an olfactometer, whether or not Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) responds preferentially to the volatiles that emanate from the fungi associated with cotton [Gossypium hirsutum L. (Malvaceae)] seed over those that emanate from cereals, because cereals are usually portrayed as the primary resources of these beetles. Pairwise comparisons were conducted between cotton seed, wheat (Triticum aestivum L.), and sorghum [Sorghum bicolor (L.) Moench] (both Poaceae); volatiles were tested from intact seeds and from both water and ethanol extracts. The results demonstrate that T. castaneum is attracted more strongly to cotton seeds with its lint contaminated with fungi, than to the conventional resources of this species (i.e., wheat and sorghum). Further tests prove that it is the fungus on the lint that produces the active volatiles, because the beetles did not respond to sterilized cotton lint (i.e., without the fungi typically associated with it when cotton seed is stored). Tests with five fungal cultures (each representing an unidentified species that was isolated from the field-collected cotton lint) were variable across the cultures, with only one of them being significantly attractive to the beetles. The others were not attractive and one may even have repulsed the beetles. The results are consistent with the beetles having a strong ecological association with fungi and suggest it would be worth investigating the ecology of T. castaneum from this perspective. © 2012 The Netherlands Entomological Society.

An outbreak of Potato spindle tuber viroid in tomato is linked to imported seed

In 2011, an outbreak of the quarantine-regulated pathogen Potato spindle tuber viroid (PSTVd) occurred in a commercial glasshouse-grown tomato crop in Queensland, Australia. Phylogenetic studies showed that the genotype of this isolate grouped in a cluster of PSTVd genotypes from tomato and Physalis peruviana, and exhibited an interesting mutation (U257→A) that has previously been linked to lethal symptom expression in tomato. Transmission studies showed that the viroid could be mechanically transmitted from crushed fruit sap, but not from undamaged fruits. A low rate of asymptomatic infection was determined for plants in the affected glasshouse, demonstrating the efficacy of using symptoms to detect PSTVd infections in tomato. No PSTVd infections were detected in solanaceous weeds located outside of the infected glasshouse, excluding them from playing a role in the viroid epidemiology. Monitoring and subsequent testing of new tomato crops grown in the facility demonstrated successful eradication of the pathogen. A trace-back analysis linked the outbreak of PSTVd to an infected imported tomato seed-lot, indicating that PSTVd is transmitted internationally through contaminated seed

Estimating Resource Requirements to Staff a Response to a Medium to Large Outbreak of Foot and Mouth Disease in Australia

A recent report to the Australian Government identified concerns relating to Australia's capacity to respond to a medium to large outbreak of FMD. To assess the resources required, the AusSpread disease simulation model was used to develop a plausible outbreak scenario that included 62 infected premises in five different states at the time of detection, 28 days after the disease entered the first property in Victoria. Movements of infected animals and/or contaminated product/equipment led to smaller outbreaks in NSW, Queensland, South Australia and Tasmania. With unlimited staff resources, the outbreak was eradicated in 63 days with 54 infected premises and a 98% chance of eradication within 3 months. This unconstrained response was estimated to involve 2724 personnel. Unlimited personnel was considered unrealistic, and therefore, the course of the outbreak was modelled using three levels of staffing and the probability of achieving eradication within 3 or 6 months of introduction determined. Under the baseline staffing level, there was only a 16% probability that the outbreak would be eradicated within 3 months, and a 60% probability of eradication in 6 months. Deployment of an additional 60 personnel in the first 3 weeks of the response increased the likelihood of eradication in 3 months to 68%, and 100% in 6 months. Deployment of further personnel incrementally increased the likelihood of timely eradication and decreased the duration and size of the outbreak. Targeted use of vaccination in high-risk areas coupled with the baseline personnel resources increased the probability of eradication in 3 months to 74% and to 100% in 6 months. This required 25 vaccination teams commencing 12 days into the control program increasing to 50 vaccination teams 3 weeks later. Deploying an equal number of additional personnel to surveillance and infected premises operations was equally effective in reducing the outbreak size and duration.

Controlling Pythium in Ginger: Phase 2

The fungal disease Pythium Soft Rot is regarded by the ginger industry as their most serious disease threat. This project was developed to further investigate the factors contributing to the persistence and spread of Pythium Soft Rot on ginger farms and to identify measures for their control. The study demonstrated that the pathogen capable of causing Pythium Soft Rot in ginger was spread in contaminated ‘seed’, soil and water and that it can be managed through a combination of strategies that rely on early detection, reducing pathogen levels in soils, preventing water logging and restricting movement of contaminated ‘seed’, soil and water. However, in order to have an effective level of control, all strategies need to be integrated in an effective manner.

Indigofera linnaei-a problem legume across northern pastures

Indigofera linnaei (or Birdsville Indigo) is a native legume with widespread abundance in pastures across northern Australian, and occurs in all northern regions of Australia from the tropical Kimberleys and arid central Australia to subhumid coastal Queensland (Figure 1). I. linnaei in central Australia has been linked to canine fatalities due to the toxin indospicine. Indospicine, an analog of arginine, is an unusual non-protein amino acid found only in a number of Indigofera species including I. linnaei. Dogs are particularly sensitive to the heptatoxicity of indospicine, and while they do not themselves consume the plant, dogs have been poisoned indirectly through the consumption of indospicine-contaminated meat from horses and camels grazing in regions where I. linnaei is common (Hegarty and Pound 1988, FitzGerald et al 2011). I. linnaei is observed to occur in various forms from strongly prostrate in south-east Queensland to an erect shrub-like form growing to more than 50cm in height in some northern regions. It mostly occurs as a minor proportion of native pasture but denser stands develop under certain circumstances. The indospicine content of I. linnaei has not previously been reported outside of central Australia, and in this study we investigate the indospicine content of plant samples collected across various regions, including both prostrate and upright forms. All samples were collected in March-July, dried, milled and analysed by UPLC-MS/MS in an adaption of our method (Tan et al 2014). Indospicine was determined in all I. linnaei plant samples regardless of region or growth form (Table 1). Measured levels were in the range 159.5 to 658.8 mg/kg DM and indicate that this plant may pose a similar problem in all areas dependent on local seasonal abundance.

The Occurrence and Toxicity of Indospicine to Grazing Animals

Indospicine is a non-proteinogenic amino acid which occurs in Indigofera species with widespread prevalence in grazing pastures across tropical Africa, Asia, Australia, and the Americas. It accumulates in the tissues of grazing livestock after ingestion of Indigofera. It is a competitive inhibitor of arginase and causes both liver degeneration and abortion. Indospicine hepatoxicity occurs universally across animal species but the degree varies considerably between species, with dogs being particularly sensitive. The magnitude of canine sensitivity is such that ingestion of naturally indospicine-contaminated horse and camel meat has caused secondary poisoning of dogs, raising significant industry concern. Indospicine impacts on the health and production of grazing animals per se has been less widely documented. Livestock grazing Indigofera have a chronic and cumulative exposure to this toxin, with such exposure experimentally shown to induce both hepatotoxicity and embryo-lethal effects in cattle and sheep. In extensive pasture systems, where animals are not closely monitored, the resultant toxicosis may well occur after prolonged exposure but either be undetected, or even if detected not be attributable to a particular cause. Indospicine should be considered as a possible cause of animal poor performance, particularly reduced weight gain or reproductive losses, in pastures where Indigofera are prevalent.

In vitro degradation of hepatotoxic indospicine in indigofera spicata by camel foregut fluid

Indospicine toxicosis was reported in sheep, goats and cattle fed on Indigofera, a leguminous plant rich in indospicine. Recent death report on dogs as a result of dietary ingestion of indospicine contaminated camel meat has raised concern about the distribution of this toxin in camels fed on Indigofera. This in vitro study aimed at measuring the degradability of indospicine in Indigofera spicata by camel-foregut fluid and attempted at explaining indospicine accumulation in meat tissue. In the first experiment, in vitro dry matter digestibility and indospicine disappearance were evaluated by using foregut fluid from 15 feral camels. Foregut fluid was collected post mortem from a nearby abattoir. In the second experiment, a composite foregut fluid obtained from three feral camels was used to examine the time-dependent degradation of indospicine. Results indicated that 99 of the dietary indospicine was degraded after 48 h of incubation. The time-dependent degradation study showed rapid degradation (11 µg/h) during the first 18 h of incubation, followed by a much slower rate (2 µg/h) between 18-48 h. Results demonstrated the ability of the camel microbiota to degrade indospicine and suggest the presence of a by-pass mechanism that enables the toxin to escape degradation and reaches the intestine.