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.

Comparison of the indirect haemagglutination and gel diffusion test for serotyping Haemophilus parasuis

The aim of this study was to compare the use of indirect haemagglutination (IHA) and gel diffusion (GD) tests for serotyping Haemophilus parasuis by the Kielstein-Rapp-Gabrielson scheme. All 15 serovar reference strains, 72 Australian field isolates, nine Chinese field isolates, and seven isolates from seven experimentally infected pigs were evaluated with both tests. With the IHA test, 14 of the 15 reference strains were correctly serotyped – with serovar 10 failing to give a titre with serovar 10 antiserum. In the GD test, 13 reference strains were correctly serotyped – with antigen from serovars 7 and 8 failing to react with any antiserum. The IHA methodology serotyped a total of 45 of 81 field isolates while the GD methodology serotyped a total of 48 isolates. For 29 isolates, the GD and IHA methods gave discordant results. It was concluded that the IHA is a good additional test for the serotyping of H. parasuis by the KRG scheme if the GD methodology fails to provide a result or shows unusual cross-reactions.

Utilizing the assassin bug, Pristhesancus plagipennis (Hemiptera: Reduviidae), as a biological control agent within an integrated pest management programme for Helicoverpa spp. (Lepidoptera: Noctuidae) and Creontiades spp. (Hemiptera: Miridae) in cotton

Helicoverpa spp. and mirids, Creontiades spp., have been difficult to control biologically in cotton due to their unpredictable temporal abundance combined with a cropping environment often made hostile by frequent usage of broad spectrum insecticides. To address this problem, a range of new generation insecticides registered for use in cotton were tested for compatibility with the assassin bug, Pristhesancus plagipennis (Walker), a potential biological control agent for Helicoverpa spp. and Creontiades spp. Indoxacarb, pyriproxifen, buprofezin, spinosad and fipronil were found to be of low to moderate toxicity on P. plagipennis whilst emamectin benzoate, abamectin, diafenthiuron, imidacloprid and omethaote were moderate to highly toxic. Inundative releases of P. plagipennis integrated with insecticides identified as being of low toxicity were then tested and compared with treatments of P. plagipennis and the compatible insecticides used alone, conventionally sprayed usage practice and an untreated control during two field experiments in cotton. The biological control provided by P. plagipennis nymphs when combined with compatible insecticides provided significant (P<0.001) reductions in Helicoverpa and Creontiades spp. on cotton and provided equivalent yields to conventionally sprayed cotton with half of the synthetic insecticide input. Despite this, the utilization of P. plagipennis in cotton as part of an integrated pest management programme remains unlikely due to high inundative release costs relative to other control technologies such as insecticides and transgenic (Bt) cotton varieties.

Effect of irrigation amounts applied with subsurface drip irrigation on corn evapotranspiration, yield, water use efficiency, and dry matter production in a semiarid climate

Quantifying the local crop response to irrigation is important for establishing adequate irrigation management strategies. This study evaluated the effect of irrigation applied with subsurface drip irrigation on field corn (Zea mays L.) evapotranspiration (ETc), yield, water use efficiencies (WUE = yield/ETc, and IWUE = yield/irrigation), and dry matter production in the semiarid climate of west central Nebraska. Eight treatments were imposed with irrigation amounts ranging from 53 to 356 mm in 2005 and from 22 to 226 mm in 2006. A soil water balance approach (based on FAO-56) was used to estimate daily soil water and ETc. Treatments resulted in seasonal ETc of 580-663 mm and 466-656 mm in 2005 and 2006, respectively. Yields among treatments differed by as much as 22% in 2005 and 52% in 2006. In both seasons, irrigation significantly affected yields, which increased with irrigation up to a point where irrigation became excessive. Distinct relationships were obtained each season. Yields increased linearly with seasonal ETc (R 2 = 0.89) and ETc/ETp (R 2 = 0.87) (ETp = ETc with no water stress). The yield response factor (ky), which indicates the relative reduction in yield to relative reduction in ETc, averaged 1.58 over the two seasons. WUE increased non-linearly with seasonal ETc and with yield. WUE was more sensitive to irrigation during the drier 2006 season, compared with 2005. Both seasons, IWUE decreased sharply with irrigation. Irrigation significantly affected dry matter production and partitioning into the different plant components (grain, cob, and stover). On average, the grain accounted for the majority of the above-ground plant dry mass (≈59%), followed by the stover (≈33%) and the cob (≈8%). The dry mass of the plant and that of each plant component tended to increase with seasonal ETc. The good relationships obtained in the study between crop performance indicators and seasonal ETc demonstrate that accurate estimates of ETc on a daily and seasonal basis can be valuable for making tactical in-season irrigation management decisions and for strategic irrigation planning and management.

Baseline responses of Alphitobius diaperinus (Coleoptera : Tenebrionidae) to cyfluthrin and detection of strong resistance in field populations in eastern Australia

Resistance to cyfluthrin in broiler farm populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in eastern Australia was suspected to have contributed to recent control failures. In 2000-2001, beetles from 11 broiler farms were tested for resistance by comparing them to an insecticide-susceptible reference population by using topical application. Resistance was detected in almost all beetle populations (up to 22 times the susceptible at the LC50), especially in southeastern Queensland where more cyfluthrin applications had been made. Two from outside southeastern Queensland were found to be susceptible. Dose-mortality data generated from the reference population over a range of cyflutbrin concentrations showed that 0.0007% cyfluthrin at a LC99.9 level could be used as a convenient dose to discriminate between susceptible and resistant populations. Using this discriminating concentration, from 2001 to 2005, the susceptibilities of 18 field populations were determined. Of these, 11 did not exhibit complete mortality at the discriminating concentration (mortality range 2.8-97.7%), and in general, cyfluthrin resistance was directly related to the numbers of cyfluthrin applications. As in the full study, populations outside of southeastern Queensland were found to have lower levels of resistance or were susceptible. One population from an intensively farmed broiler area in southeastern Queensland exhibited low mortality despite having no known exposure to cyfluthrin. Comparisons of LC50 values of three broiler populations and a susceptible population, collected in 2000 and 2001 and recollected in 2004 and 2005 indicated that values from the three broiler populations had increased over this time for all populations. The continued use of cyfluthrin for control of A. diaperinus in eastern Australia is currently under consideration.

Diel variation in mangrove fish abundances and trophic guilds of northeastern Australian estuaries with a proposed trophodynamic model

Diel activity patterns of tropical fish assemblages in turbid, mangrove-dominated estuaries remain largely undocumented, leading to uncertainty about ecological processes in these systems. To capture active fishes by day and night, gill nets were set perpendicular to mangrove shorelines, in six northeastern Australian estuaries during 13 bimonthly trips. Fish were sampled with eight large mesh (102-151 mm) nets, set for 6 hrs (1500-2100), and checked hourly (1146 day, 635 dusk, 872 night checks). Four smaller mesh (19-51 mm) nets were also set for 1 hr before and after sunset (77 day, 78 night checks). Of 157 total species, 22 were netted exclusively before sunset and 47 exclusively after sunset. All of the top 26 species were present both day and night, but of these, 46% were primarily nocturnal (diel index > 0.65). An average of 77.2 fish hr−1 were netted by day vs 171.4 by night. Within the 400 km coastal region, assemblages differed between two northern wave-dominated (WD) estuaries and four southern tide-dominated ('I'D) estuaries. In all six estuaries Lates calcarifer (Bloch, 1790) dominated night assemblages. In 'I'D estuaries, night assemblages were also dominated by Thryssa hamiltoni Gray, 1835 and Eleutheronema tetradactylum (Shaw, 1804); while in WD estuaries Herklotsichthys castelnaui (Ogilby, 1897), Leiognathus equulus (Forsskål, 1775), and Megalops cyprinoids (Broussonet, 1782) were dominant at night. Nocturnal species included planktivores and carnivores, while daytime assemblages were dominated by detritivores (Mugillidae). Higher night catch rates are attributed to increased activity by mobile fishes moving from mangrove to adjacent habitats to forage, especially immediately post-sunset. Although day-night diets and forage resources have yet to be compared in mangrove systems, previously unrecognized trophic relationships involving variation in diel activity among important fishery species (Centropomidae, polynemidae, Carangidae) and their prey may be key ecological processes in these tropical mangrove estuaries. A proposed hypothesis explaining diel variation in mangrove fish assemblages of tropical estuaries is presented through a conceptual model.

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.

Defining and predicting the 'break of the season' for north-east Queensland grazing areas

For pasture growth in the semi-arid tropics of north-east Australia, where up to 80% of annual rainfall occurs between December and March, the timing and distribution of rainfall events is often more important than the total amount. In particular, the timing of the 'green break of the season' (GBOS) at the end of the dry season, when new pasture growth becomes available as forage and a live-weight gain is measured in cattle, affects several important management decisions that prevent overgrazing and pasture degradation. Currently, beef producers in the region use a GBOS rule based on rainfall (e. g. 40mm of rain over three days by 1 December) to define the event and make their management decisions. A survey of 16 beef producers in north-east Queensland shows three quarters of respondents use a rainfall amount that occurs in only half or less than half of all years at their location. In addition, only half the producers expect the GBOS to occur within two weeks of the median date calculated by the CSIRO plant growth days model GRIM. This result suggests that in the producer rules, either the rainfall quantity or the period of time over which the rain is expected, is unrealistic. Despite only 37% of beef producers indicating that they use a southern oscillation index (SOI) forecast in their decisions, cross validated LEPS (linear error in probability space) analyses showed both the average 3 month July-September SOI and the 2 month August-September SOI have significant forecast skill in predicting the probability of both the amount of wet season rainfall and the timing of the GBOS. The communication and implementation of a rigorous and realistic definition of the GBOS, and the likely impacts of anthropogenic climate change on the region are discussed in the context of the sustainable management of northern Australian rangelands.

Phylogenetic analysis of betanodavirus isolates from Australian finfish

In Australia, disease caused by betanodavirus has been reported in an increasing number of cultured finfish since the first report of mortalities in 1990. Partial coat protein gene sequences from the T2 or T4 regions of 8 betanodaviruses from barramundi Lates calcarifer, sleepy cod Oxyeleotris lineolata, striped trumpeter Latris lineata, barramundi cod Cromileptes altivelis, Australian bass Macquaria novemaculata and gold-spotted rockcod Epinephelus coioides from several Australian states were determined. Analysis of the 606 bp nucleotide sequences of the T2 region of 4 isolates demonstrated the close relationship with isolates from the red-spotted grouper nervous necrosis virus (RGNNV) genotype and the Cluster Ia subtype. Comparison of a smaller 289 bp sequence from the T4 region identified 2 distinct groupings of the Australian isolates within the RGNNV genotype. Isolates from barramundi from the Northern Territory, barramundi, sleepy cod, barramundi cod and gold-spotted rockcod from Queensland, and striped trumpeter from Tasmania shared a 96.2 to 99.7%, nucleotide identity with each other. These isolates were most similar to the RGNNV genotype Cluster Ia. Isolates from Australian bass from New South Wales and from barramundi from South Australia shared a 98.6% sequence identity with each other. However, these isolates only shared an 85.8 to 87.9%, identity with the other Australian isolates and representative RGNNV isolates. The closest nucleotide identity to sequences reported in the literature for the New South Wales and South Australian isolates was to an Australian barramundi isolate (Ba94Aus) from 1994. These 2 Australian isolates formed a new subtype within the RGNNV genotype, which is designated as Cluster Ic.

Queensland Coastal Wetland Resources: Sand Bay to Keppel Bay.

Protection of coastal wetland environments is an important prerequisite to effective and sustainable inshore fisheries management and conservation of habitats for use by future generations. Mangroves, saltmarshes, seagrasses and non vegetated habitats directly support local and regional inshore and offshore fisheries through the provision of food, shelter, breeding and nursery grounds. As such, these wetland environments have significant economic value as well as their intrinsic aesthetic and ecological values. This report summarises the results of the mapping undertaken in the Central Queensland Coast from Sand Bay to Keppel Bay (hereafter referred to as the Study Area). The study was undertaken in order to: 1. document and map the coastal wetland communities along the Queensland coastline from Sand Bay (20.93°S, 149.04°E) to Keppel Bay (23.65°S, 151.07°E); 2. document levels of existing disturbance to and protection of the wetlands; 3. examine existing recreational and commercial fisheries in the region; and 4. evaluate the conservation values of the areas investigated from the viewpoint of fisheries productivity and as habitat for important and/or threatened species. Dataset URL Link: Queensland Coastal Wetlands Resources Mapping data. [Dataset]

High-resolution melt-curve analysis of random-amplified-polymorphic-DNA markers, for the characterisation of pathogenic Leptospira.

A new test for pathogenic Leptospira isolates, based on RAPD-PCR and high-resolution melt (HRM) analysis (which measures the melting temperature of amplicons in real time, using a fluorescent DNA-binding dye), has recently been developed. A characteristic profile of the amplicons can be used to define serovars or detect genotypes. Ten serovars, of leptospires from the species Leptospira interrogans (serovars Australis, Robinsoni, Hardjo, Pomona, Zanoni, Copenhageni and Szwajizak), L. borgpetersenii (serovar Arborea), L. kirschneri (serovar Cynopteri) and L. weilii (serovar Celledoni), were typed against 13 previously published RAPD primers, using a real-time cycler (the Corbett Life Science RotorGene 6000) and the optimised reagents from a commercial kit (Quantace SensiMix). RAPD-HRM at specific temperatures generated defining amplicon melt profiles for each of the tested serovars. These profiles were evaluated as difference-curve graphs generated using the RotorGene software package, with a cut-off of at least 8 'U' (plus or minus). The results demonstrated that RAPD-HRM can be used to measure serovar diversity and establish identity, with a high degree of stability. The characterisation of Leptospira serotypes using a DNA-based methodology is now possible. As an objective and relatively inexpensive and rapid method of serovar identification, at least for cultured isolates, RAPD-HRM assays show convincing potentia.

Myelodysplasia as a cause of hindlimb ataxia in two beef calves.

Myelodysplasia is a general term referring to abnormal development of the spinal cord. Unless associated with vertebral malformations, it can be difficult to distinguish clinically from other causes of spinal cord disease. These case reports describe the clinical and pathological findings in two calves with a distinctive non-progressive pelvic limb ataxia. The syndrome was observed in two calves on a large, extensively managed beef cattle property near Richmond, north Queensland. Both calves had similar clinical signs, including hindlimb ataxia with swaying of the pelvis and a well-coordinated bilateral hopping-like action. The differential diagnoses are discussed. A focal or diffuse myelodysplasia should be suspected in calves that have exhibited a non-progressive hindlimb ataxia from birth.

Life history and host range of Alagoasa extrema (Coleoptera: Chrysomelidae: Alticinae), a potential biological control agent of Lantana spp. (Verbenaceae) in Australia

The life history and host range of the lantana beetle, Alagoasa extrema, a potential biocontrol agent for Lantana spp. were investigated in a quarantine unit at the Alan Fletcher Research Station, Brisbane, Australia. Adults feed on leaves and females lay batches of about 17 eggs on the soil surface around the stems of plants. The eggs take 16 days to hatch and newly emerged larvae move up the stem to feed on young leaves. Larvae feed for about 23 days and there are three instars. There is a prepupal non-feeding stage that lasts about 12 days and the pupal stage, which occurs in a cocoon in the soil, lasts 16 days. Teneral adults remain in the cocoon for 3 days to harden prior to emergence. Males live for about 151 days while females live for about 127 days. The pre-oviposition period is 19 days. In no-choice larval feeding trials, nine plant species, representing three families, supported development to adult. Three species, Aloysia triphylla, Citharexylum spinosum and Pandorea pandorana were able to support at least two successive generations. These results confirm those reported in South Africa and suggest that A. extrema is not sufficiently specific for release in Australia. Furthermore, it is not recommended for release in any other country which is considering biological control of lantana.

A multi-field bio-economic model of irrigated grain-cotton farming systems.

We present a participatory modelling framework that integrates information from interviews and discussions with farmers and consultants, with dynamic bio-economic models to answer complex questions on the allocation of limited resources at the farm business level. Interviews and discussions with farmers were used to: describe the farm business; identify relevant research questions; identify potential solutions; and discuss and learn from the whole-farm simulations. The simulations are done using a whole-farm, multi-field configuration of APSIM (APSFarm). APSFarm results were validated against farmers' experience. Once the model was accepted by the participating farmers as a fair representation of their farm business, the model was used to explore changes in the tactical or strategic management of the farm and results were then discussed to identify feasible options for improvement. Here we describe the modelling framework and present an example of the application of integrative whole farm system tools to answer relevant questions from an irrigated farm business case study near Dalby (151.27E - 27.17S), Queensland, Australia. Results indicated that even though cotton crops generates more farm income per hectare a more diversified rotation with less cotton would be relatively more profitable, with no increase in risk, as a more cotton dominated traditional rotation. Results are discussed in terms of the benefits and constraints from developing and applying more integrative approaches to represent farm businesses and their management in participatory research projects with the aim of designing more profitable and sustainable irrigated farming systems.

Characterization of north-eastern Australian environments using APSIM for increasing rainfed maize production

Recurring water stresses are a major risk factor for rainfed maize cropping across the highly diverse agro-ecological environments of Queensland (Qld) and northern New South Wales (NNSW). Enhanced understanding of such agro-ecological diversity is necessary to more consistently sample target production environments for testing and targeting release of improved germplasm, and to improve the efficiency of the maize pre-breeding and breeding programs of Qld and New South Wales. Here, we used the Agricultural Production Systems Simulator (APSIM) – a well validated maize crop model to characterize the key distinctive water stress patterns and risk to production across the main maize growing regions of Qld and NNSW located between 15.8° and 31.5°S, and 144.5° and 151.8°E. APSIM was configured to simulate daily water supply demand ratios (SDRs) around anthesis as an indicator of the degree of water stress, and the final grain yield. Simulations were performed using daily climatic records during the period between 1890 and 2010 for 32 sites-soils in the target production regions. The runs were made assuming adequate nitrogen supply for mid-season maize hybrid Pioneer 3153. Hierarchical complete linkage analyses of the simulated yield resulted in five major clusters showing distinct probability distribution of the expected yields and geographic patterns. The drought stress patterns and their frequencies using SDRs were quantified using multivariate statistical methods. The identified stress patterns included no stress, mid-season (flowering) stress, and three terminal stresses differing in terms of severity. The combined frequency of flowering and terminal stresses was highest (82.9%), mainly in sites-soils combinations in the west of Qld and NNSW. Yield variability across the different sites-soils was significantly related to the variability in frequencies of water stresses. Frequencies of water stresses within each yield cluster tended to be similar, but different across clusters. Sites-soils falling within each yield cluster therefore could be treated as distinct maize production environments for testing and targeting newly developed maize cultivars and hybrids for adaptation to water stress patterns most common to those environments.