Effect of micro-propagation on the health status of strawberry planting material for commercial production of strawberry runners for Queensland

Plant tissue culture has been used for a number of years to produce micropropagated strawberry plants for planting into runner growing beds in the Stanthorpe (Queensland) and Bothwell (Tasmania) regions. This process has allowed the rapid release of new cultivars from the LAWS (Late Autumn, Winter, Spring) breeding program into the current runner production system. Micro-propagation in vitro allows plants to be produced during the autumn and winter months, when mother plants would normally be in a fruit production phase in the field in Queensland. The plants produced are of a high health status when they are planted. The subsequent arrival and build up of various diseases in the runner fields are closely monitored. Using tissue culture for the first generation reduces the time the plants spend in the field by twelve months, reducing disease incidence. To date, any disease outbreak has been successfully managed using early detection and rapid response methods.

ACIAR Improving soil health in support of sustainable development in the Pacific

The proposed project focuses on developing research-based indicators that growers and extensionists can use to assess soil health status (including key chemical, physical and biological variables), as well as extension approaches to communicate soil health.

Predicting survival of western rock lobsters Panulirus cygnus using discriminant analysis of hemolymph parameters taken immediately following simulated handling treatments

Instances of morbidity amongst rock lobsters (Panulirus cygnus) arriving at factories in Western Australia (WA) have been attributed to stress during post-harvest handling. This study used discriminant analysis to determine whether physiological correlates of stress following a period of simulated post-harvest handling had any validity as predictors of future rejection or morbidity of western rock lobsters. Groups of 230 western rock lobsters were stored for 6 h in five environments (submerged/flowing sea water, submerged/re-circulating sea water, humid air, flowing sea water spray, and re-circulated sea water spray). The experiment was conducted in late spring (ambient sea water 22°C), and repeated again in early autumn (ambient sea water 26°C). After 6 h treatment, each lobster was graded for acceptability for live export, numbered, and its hemolymph was sampled. The samples were analysed for a number of physiological and health status parameters. The lobsters were then stored for a week in tanks in the live lobster factory to record mortality. The mortality of lobsters in the factory was associated with earlier deviations in hemolymph parameters as they emerged from the storage treatments. Discriminant analysis (DA) of the hemolymph assays enabled the fate of 80-90% of the lobsters to be correctly categorised within each experiment. However, functions derived from one experiment were less accurate at predicting mortality when applied to the other experiments. One of the reasons for this was the higher mortality and the more severe patho-physiological changes observed in lobsters stored in humid air or sprays at the higher temperature. The analysis identified lactate accumulation during emersion and associated physiological and hemocyte-related effects as a major correlate of mortality. Reducing these deviations, for example by submerged transport, is expected to ensure high levels of survival. None of the indicators tested predicted mortality with total accuracy. The simplest and most accurate means of comparing emersed treatments was to count the mortality afterwards.

Effect of s-methylmethionine sulphonium chloride on oesophagogastric ulcers in pigs

Objective: To assess the value of s-methylmethionine sulphonium chloride (SMMSC) (200 mg/kg) on nutritional performance of pigs and as prevention or therapy for oesophagogastric ulcers. Design: Sixty pigs from a high health status herd with continuing oesophagogastric ulcer problems were endoscopically assessed for the presence or absence of oesophagogastric ulcers. Forty-eight pigs were then selected and allocated according to an initial oesophagogastric epithelial (ulcer score) classification to replicated treatment groups in a 2 × 2 factorial design. Weight gain and feed intake were measured over 49 d, after which pigs were killed and stomachs were collected, re-examined and scored for oesophagogastric ulceration. Results: There was no difference over the 49 d in weight gain, feed intake and backfat in pigs with and without SMMSC supplementation between pigs with or without fully developed oesophagogastric ulcers at the start of the study. In pigs with an initially low ulcer score, feeding SMMSC did not prevent further oesophagogastric ulcer development. No significant effect of SMMSC was apparent when final mean oesophagogastric ulcer scores were compared in pigs with existing high ulcer score. However, further analysis of the changes in individual pig oesophagogastric ulcer scores during the experiment showed that the observed reductions in scores of the high ulcer group was significantly different from all other groups. Conclusion: This study has indicated that supplementation of pig diets with SMMSC cannot be justified unless the slight ulcer score improvement observed could be translated to some commercial production advantage such as a reduction in pig mortalities due to oesophagogastric ulcers. This study has further confirmed the benefit of endoscopy as a tool to enable objective assessment of oesophageal gastric health.

Australian Bat Lyssavirus

In Chapter 1, the literature relating to rabies virus and the rabies like lyssaviruses is reviewed. In Chapter 2, data are presented from 1170 diagnostic submissions for ABLV testing by fluorescent antibody test (Centocor FAT). All 27 non-bat submissions were ABLV-negative. Of 1143 bat accessions 74 (16%) were ABLV-positive, including 69 of 974 (7.1%) flying foxes (Pteropus spp.), 5 of 7 (71.4%) Saccolaimus flaviventris (Yellow-bellied sheathtail bats), none of 151 other microchiropteran bats, and none of 11 unidentified bats. Statistical analysis of data from 868 wild Black, Grey-headed, Little Red and Spectacled flying foxes (Pteropus alecto, P. poliocephalus, P. scapulatus, and P. conspicillatus) indicated that three factors; species, health status and age were associated with significant (p< 0.001) differences in the proportion of ABLV-positive bats. Other factors including sex, whether the bat bit a person or animal, region, year, and season submitted, were not associated with ABLV. Case data for 74 ABLV-positive bats, including the circumstances in which they were found and clinical signs, is presented. In Chapter 3, the aetiological diagnosis was investigated for 100 consecutive flying fox submissions with neurological signs. ABLV (32%), spinal and head injuries (29%), and neuro-angiostrongylosis (18%) accounted for most neurological syndromes in flying foxes. No evidence of lead poisoning was found in unwell (n=16) or healthy flying foxes (n=50). No diagnosis was reached for 16 cases, all of which were negative for ABLV by TaqMan PCR. The molecular diversity of ABLV was examined in Chapter 4 by sequencing 36 bases of the leader sequence, the entire N gene, and start of the P gene of 28 isolates from pteropid bats and 3 isolates from Yellow-bellied sheathtail (YBST) bats. Phylogenetic analysis indicated all ABLV isolates clustered together as a discrete group within the Lyssavirus genera closely related to rabies virus and European bat lyssavirus-2 isolates. The ABLV lineage consisted of two variants; one (ybst-ABLV) consisted of isolates only from YBST bats, the other (pteropid-ABLV) was common to Black, Grey-headed and Little Red flying foxes. No associations were found between the sequences and either the geographical location or year found, or individual flying fox species. In Chapter 5, 15 inocula prepared from the brains or salivary glands of naturally-infected bats were evaluated by intracerebral (IC) and footpad (FP) inoculation of Quackenbush mice in order to select and characterize a highly virulent inoculum for further use in bats (Inoculum 5). In Chapter 6, nine Grey-headed flying foxes were inoculated with 105.2 to 105.5 MICED50 of Inoculum 5 divided into four sites, left footpad, pectoral muscle, temporal muscle and muzzle. Another bat was inoculated with half this dose divided into the footpad and pectoral muscle only. Seven of 10 bats developed clinical disease of 1 to 4 days duration between PI-days 10 and 19 and were shown to be ABL-positive by FAT, HAM immunoperoxidase staining, virus isolation in mice, and TaqMan PCR. Five of the seven bats displayed overt aggression, one died during a seizure, and one showed intractable agitation, pacing, tremors, and ataxia. Viral antigen was demonstrated throughout the central and peripheral nervous systems and in the epithelial cells of the submandibular salivary glands (n=4). All affected bats had mild to moderate non-suppurative meningoencephalitis and severe ganglioneuritis. No ABLV was detected in three bats that remained well until the end of the experiment on day 82. One survivor developed a strong but transient antibody response. In Chapter 7, the relative virulence of inocula prepared from the brains and salivary glands of experimentally infected flying foxes was evaluated in mice by IC and FP inoculation and TaqMan assay. The effects in mice were correlated to the TaqMan CT value and indicated a crude association between virulence and CT value that has potential application in the selection of inocula. In Chapter 8, 36 Black and Grey-headed flying foxes were vaccinated with one (day 0) or two (+ day 28) doses of Nobivac rabies vaccine and co-vaccinated with keyhole limpet haemocyanin (KLH). All bats responded to the Nobivac vaccine with a rabies-RFFIT titer > 0.5 IU/mL that is nominally indicative of protective immunity. Plasma from bats with rabies titres >2 IU/mL had cross-neutralising ABLV titres >1:154. A specifically developed ELISA detected a strong but transient response to KLH.

Pen parameters for improving the performance of sheep imported from Australia to Persian Gulf feedlots

Summer in the Persian Gulf region presents physiological challenges for Australian sheep that are part of the live export supply chain coming from the Australian winter. Many feedlots throughout the Gulf have very high numbers of animals during June to August in order to cater for the increased demand for religious festivals. From an animal welfare perspective it is important to understand the necessary requirements of feed and water trough allowances, and the amount of pen space required, to cope with exposure to these types of climatic conditions. This study addresses parameters that are pertinent to the wellbeing of animals arriving in the Persian Gulf all year round. Three experiments were conducted in a feedlot in the Persian Gulf between March 2010 and February 2012, totalling 44 replicate pens each with 60 or 100 sheep. The applied treatments covered animal densities, feed-bunk lengths and water trough lengths. Weights, carcass attributes and health status were the key recorded variables. Weight change results showed superior performance for animal densities of ≥1.2 m2/head during hot conditions (24-h average temperatures greater than 33 °C, or a diurnal range of around 29–37 °C). However the space allowance for animals can be decreased, with no demonstrated detrimental effect, to 0.6 m2/head under milder conditions. A feed-bunk length of ≥5 cm/head is needed, as 2 cm/head showed significantly poorer animal performance. When feeding at 90 ad libitum 10 cm/head was optimal, however under a maintenance feeding regime (1 kg/head/day) 5 cm/head was adequate. A minimum water trough allowance of 1 cm/head is required. However, this experiment was conducted during milder conditions, and it may well be expected that larger water trough lengths would be needed in hotter conditions. Carcass weights were determined mainly by weights at feedlot entry and subsequent weight gains, while dressing percentage was not significantly affected by any of the applied treatments. There was no demonstrated effect of any of the treatments on the number of animals that died, or were classified as unwell. However, across all the treatments, these animals lost significantly more weight than the healthy animals, so the above recommendations, which are aimed at maintaining weight, should also be applicable for good animal health and welfare. Therefore, best practice guidelines for managing Australian sheep in Persian Gulf feedlots in the hottest months (June–August) which present the greatest environmental and physical challenge is to allow feed-bunk length 5 cm/head on a maintenance-feeding program and 10 cm/head for 90 ad libitum feeding, and the space allowance per animal should be ≥1.2 m2/head. Water trough allocation should be at least 1 cm/head with provision for more in the summer when water intake potentially doubles.

Pen parameters for improving the performance of sheep imported from Australia to Persian Gulf feedlots

Summer in the Persian Gulf region presents physiological challenges for Australian sheep that are part of the live export supply chain coming from the Australian winter. Many feedlots throughout the Gulf have very high numbers of animals during June to August in order to cater for the increased demand for religious festivals. From an animal welfare perspective it is important to understand the necessary requirements of feed and water trough allowances, and the amount of pen space required, to cope with exposure to these types of climatic conditions. This study addresses parameters that are pertinent to the wellbeing of animals arriving in the Persian Gulf all year round. Three experiments were conducted in a feedlot in the Persian Gulf between March 2010 and February 2012, totalling 44 replicate pens each with 60 or 100 sheep. The applied treatments covered animal densities, feed-bunk lengths and water trough lengths. Weights, carcass attributes and health status were the key recorded variables. Weight change results showed superior performance for animal densities of ≥1.2 m2/head during hot conditions (24-h average temperatures greater than 33 °C, or a diurnal range of around 29–37 °C). However the space allowance for animals can be decreased, with no demonstrated detrimental effect, to 0.6 m2/head under milder conditions. A feed-bunk length of ≥5 cm/head is needed, as 2 cm/head showed significantly poorer animal performance. When feeding at 90% ad libitum 10 cm/head was optimal, however under a maintenance feeding regime (1 kg/head/day) 5 cm/head was adequate. A minimum water trough allowance of 1 cm/head is required. However, this experiment was conducted during milder conditions, and it may well be expected that larger water trough lengths would be needed in hotter conditions. Carcass weights were determined mainly by weights at feedlot entry and subsequent weight gains, while dressing percentage was not significantly affected by any of the applied treatments. There was no demonstrated effect of any of the treatments on the number of animals that died, or were classified as unwell. However, across all the treatments, these animals lost significantly more weight than the healthy animals, so the above recommendations, which are aimed at maintaining weight, should also be applicable for good animal health and welfare. Therefore, best practice guidelines for managing Australian sheep in Persian Gulf feedlots in the hottest months (June–August) which present the greatest environmental and physical challenge is to allow feed-bunk length 5 cm/head on a maintenance-feeding program and 10 cm/head for 90% ad libitum feeding, and the space allowance per animal should be ≥1.2 m2/head. Water trough allocation should be at least 1 cm/head with provision for more in the summer when water intake potentially doubles.

Automating Crop Damage Assessments with Unmanned Aerial Systems and Machine Learning

Agricultural pests are responsible for millions of dollars in crop losses and management costs every year. In order to implement optimal site-specific treatments and reduce control costs, new methods to accurately monitor and assess pest damage need to be investigated. In this paper we explore the combination of unmanned aerial vehicles (UAV), remote sensing and machine learning techniques as a promising methodology to address this challenge. The deployment of UAVs as a sensor platform is a rapidly growing field of study for biosecurity and precision agriculture applications. In this experiment, a data collection campaign is performed over a sorghum crop severely damaged by white grubs (Coleoptera: Scarabaeidae). The larvae of these scarab beetles feed on the roots of plants, which in turn impairs root exploration of the soil profile. In the field, crop health status could be classified according to three levels: bare soil where plants were decimated, transition zones of reduced plant density and healthy canopy areas. In this study, we describe the UAV platform deployed to collect high-resolution RGB imagery as well as the image processing pipeline implemented to create an orthoimage. An unsupervised machine learning approach is formulated in order to create a meaningful partition of the image into each of the crop levels. The aim of this approach is to simplify the image analysis step by minimizing user input requirements and avoiding the manual data labelling necessary in supervised learning approaches. The implemented algorithm is based on the K-means clustering algorithm. In order to control high-frequency components present in the feature space, a neighbourhood-oriented parameter is introduced by applying Gaussian convolution kernels prior to K-means clustering. The results show the algorithm delivers consistent decision boundaries that classify the field into three clusters, one for each crop health level as shown in Figure 1. The methodology presented in this paper represents a venue for further esearch towards automated crop damage assessments and biosecurity surveillance.