The impacts and management of foxes Vulpes vulpes in Australia

1. The successful introduction of the red fox Vulpes vulpes into Australia in the 1870s has had dramatic and deleterious impacts on both native fauna and agricultural production. Historical accounts detail how the arrival of foxes in many areas coincided with the local demise of native fauna. Recent analyses suggest that native fauna can be successfully reintroduced to their former ranges only if foxes have been controlled, and several replicated removal experiments have confirmed that foxes are the major agents of extirpation of native fauna. Predation is the primary cause of losses, but competition and transmission of disease may be important for some species. 2. In agricultural landscapes, fox predation on lambs can cause losses of 1–30%; variation is due to flock size, health and management, as well as differences in the timing and duration of lambing and the density of foxes. 3. Fox control measures include trapping, shooting, den fumigation and exclusion fencing; baiting using the toxin 1080 is the most commonly employed method. Depending on the baiting strategy, habitat and area covered, baiting can reduce fox activity by 50–97%. We review patterns of baiting in a large sheep-grazing region in central New South Wales, and propose guidelines to increase landholder awareness of baiting strategies, to concentrate and coordinate bait use, and to maximize the cost-effectiveness of baiting programs. 4. The variable reduction in fox density within the baited area, together with the ability of the fox to recolonize rapidly, suggest that current baiting practices in eastern Australia are often ineffective, and that reforms are required. These might include increasing landholder awareness and involvement in group control programs, and the use of more efficient broadscale techniques, such as aerial baiting.

Reproductive Biology of Corymbia citriodora subsp. variegata and Effective Pollination across its Native Range.

The spotted gum species complex represents a group of four eucalypt hardwood taxa that have a native range that spans the east coast of Australia, with a morphological cline from Victoria to northern Queensland. Of this group, Corymbia citriodora subsp. variegata (CCV) is widespread in south-eastern Queensland and northern New South Wales. It is currently the most commonly harvested native hardwood in Queensland. However, little basic knowledge of the reproductive biology of the species is available to inform genetic improvement and resource management programmes. Here we take an integrative approach, using both field and molecular data, to identify ecological factors important to mating patterns in native populations of CCV. Field observation of pollinator visitation and flowering phenology of 20 trees showed that foraging behaviour of pollinator guilds varies depending on flowering phenology and canopy structure. A positive effect of tree mean flowering effort was found on insect visitation, while bat visitation was predicted by tree height and by the number of trees simultaneously bearing flowers. Moreover, introduced honeybees were observed frequently, performing 73% of detected flower visits. Conversely, nectar-feeding birds and mammals were observed sporadically with lorikeets and honeyeaters each contributing to 11% of visits. Fruit bats, represented solely by the grey-headed flying fox, performed less than 2% of visits. Genotyping at six microsatellite markers in 301 seeds from 17 families sampled from four of Queensland's native forests showed that CCV displays a mixed-mating system that is mostly outcrossing (tm = 0.899 ± 0.021). Preferential effective pollination from near-neighbours was detected by means of maximum-likelihood paternity analysis with up to 16% of reproduction events resulting from selfing. Forty to 48% of fertilising pollen was also carried from longer distance (>60 m). Marked differences in foraging behaviour and visitation frequency between observed pollinator guilds suggests that the observed dichotomy of effective pollen movement in spotted gums may be due to frequent visit from introduced honeybees favouring geitonogamy and sporadic visits from honeyeaters and fruit bats resulting in potential long-distance pollinations.

National screening for barley grain defects

Three defects on barley grain can impact on the price paid to grain growers. Black point (BP), kernel staining (KS) and pre-harvest sprouting (PHS) can result in malting barley being downgraded to feed. Resistance to these defects is the best option, and in this project hundreds of breeding lines grown over three years were screened for these traits. A number of lines exhibited resistance to each defect but very few had resistance to all defects. The results from the screening program have been provided to the Australian barley breeders through the Barley Breeding Australia (BBA) program.

DAQ00104 Sponge and dough bread quality of Australian wheat germplasm

The main grade of wheat targeted for the export sponge and dough (S&D) market is Australian prime hard (APH). By association, protein should be a key parameter relating to S&D quality, specifically loaf volume (LV). Surprisingly, the project revealed a low level of correlation between total protein content and LV. It appears that protein composition may be the key to understanding S&D quality, as the glutenin Glu D1 5+10 subunit contributed to the highest LVs. The current varieties KennedyA and SunzellA, together with several breeding lines, provided a consistently high quality over a number of seasons. These varieties performed as well as, if not better than, North American S&D varieties.

Simultaneous prediction of acidity parameters (pH and titratable acidity) in Kefir using near infrared reflectance spectroscopy.

Acidity in terms of pH and titratable acids influences the texture and flavour of fermented dairy products, such as Kefir. However, the methods for determining pH and titratable acidity (TA) are time consuming. Near infrared (NIR) spectroscopy is a non-destructive method, which simultaneously predicts multiple traits from a single scan and can be used to predict pH and TA. The best pH NIR calibration model was obtained with no spectral pre-treatment applied, whereas smoothing was found to be the best pre-treatment to develop the TA calibration model. Using cross-validation, the prediction results were found acceptable for both pH and TA. With external validation, similar results were found for pH and TA, and both models were found to be acceptable for screening purposes.

Evidence of bat origin for Menangle virus, a zoonotic paramyxovirus first isolated from diseased pigs

Menangle virus (MenPV) is a zoonotic paramyxovirus capable of causing disease in pigs and humans. It was first isolated in 1997 from stillborn piglets at a commercial piggery in New South Wales, Australia, where an outbreak of reproductive disease occurred. Neutralizing antibodies to MenPV were detected in various pteropid bat species in Australia and fruit bats were suspected to be the source of the virus responsible for the outbreak in pigs. However, previous attempts to isolate MenPV from various fruit bat species proved fruitless. Here, we report the isolation of MenPV from urine samples of the black flying fox, Pteropus alecto, using a combination of improved procedures and newly established bat cell lines. The nucleotide sequence of the bat isolate is 94% identical to the pig isolate. This finding provides strong evidence supporting the hypothesis that the MenPV outbreak in pigs originated from viruses in bats roosting near the piggery. © 2012 Printed in Great Britain.

Real-time PCR as a surveillance tool for the detection of Trichinella infection in muscle samples from wildlife

Trichinella nematodes are the causative agent of trichinellosis, a meat-borne zoonosis acquired by consuming undercooked, infected meat. Although most human infections are sourced from the domestic environment, the majority of Trichinella parasites circulate in the natural environment in carnivorous and scavenging wildlife. Surveillance using reliable and accurate diagnostic tools to detect Trichinella parasites in wildlife hosts is necessary to evaluate the prevalence and risk of transmission from wildlife to humans. Real-time PCR assays have previously been developed for the detection of European Trichinella species in commercial pork and wild fox muscle samples. We have expanded on the use of real-time PCR in Trichinella detection by developing an improved extraction method and SYBR green assay that detects all known Trichinella species in muscle samples from a greater variety of wildlife. We simulated low-level Trichinella infections in wild pig, fox, saltwater crocodile, wild cat and a native Australian marsupial using Trichinella pseudospiralis or Trichinella papuae ethanol-fixed larvae. Trichinella-specific primers targeted a conserved region of the small subunit of the ribosomal RNA and were tested for specificity against host and other parasite genomic DNAs. The analytical sensitivity of the assay was at least 100 fg using pure genomic T. pseudospiralis DNA serially diluted in water. The diagnostic sensitivity of the assay was evaluated by spiking log of each host muscle with T. pseudospiralis or T. papuae larvae at representative infections of 1.0, 0.5 and 0.1 larvae per gram, and shown to detect larvae at the lowest infection rate. A field sample evaluation on naturally infected muscle samples of wild pigs and Tasmanian devils showed complete agreement with the EU reference artificial digestion method (k-value = 1.00). Positive amplification of mouse tissue experimentally infected with T. spiralis indicated the assay could also be used on encapsulated species in situ. This real-time PCR assay offers an alternative highly specific and sensitive diagnostic method for use in Trichinella wildlife surveillance and could be adapted to wildlife hosts of any region. (C) 2012 Elsevier B.V. All rights reserved.

Correlation between NIRS generated and chemically measured feed quality data in barley (Hordeum vulgare), and potential use in QTL analysis identification

A study was performed to investigate the value of near infrared reflectance spectroscopy (NIRS) as an alternate method to analytical techniques for identifying QTL associated with feed quality traits. Milled samples from an F6-derived recombinant inbred Tallon/Scarlett population were incubated in the rumen of fistulated cattle, recovered, washed and dried to determine the in-situ dry matter digestibility (DMD). Both pre- and post-digestion samples were analysed using NIRS to quantify key quality components relating to acid detergent fibre, starch and protein. This phenotypic data was used to identify trait associated QTL and compare them to previously identified QTL. Though a number of genetic correlations were identified between the phenotypic data sets, the only correlation of most interest was between DMD and starch digested (r = -0.382). The significance of this genetic correlation was that the NIRS data set identified a putative QTL on chromosomes 7H (LOD = 3.3) associated with starch digested. A QTL for DMD occurred in the same region of chromosome 7H, with flanking markers fAG/CAT63 and bPb-0758. The significant correlation and identification of this putative QTL, highlights the potential of technologies like NIRS in QTL analysis.

Grain dormancy QTL identified in a doubled haploid barley population derived from two non-dormant parents

Grain dormancy provides protection against pre-harvest sprouting (PHS) in cereals. Composite interval mapping and association analyses were performed to identify quantitative trait loci (QTL) contributing grain dormancy in a doubled haploid (DH) barley population (ND24260 x Flagship) consisting of 321 lines genotyped with DArT markers. Harvest-ripe grain collected from three field experiments was germinated over a 7-day period to determine a weighted germination index for each line. DH lines displaying moderate to high levels of grain dormancy were identified; however, both parental lines were non-dormant and displayed rapid germination within the first two days of testing. Genetic analysis identified two QTL on chromosome 5H that were expressed consistently in each of the three environments. One QTL (donated by Flagship) was located close to the centromeric region of chromosome 5H (qSDFlag), accounting for up to 15% of the phenotypic variation. A second QTL with a larger effect (from ND24260) was detected on chromosome 5HL (qSDND), accounting for up to 35% of the phenotypic variation. qSDFlag and qSDND displayed an epistatic interaction and DH lines that had the highest levels of grain dormancy carried both genes. We demonstrate that qSDND in the ND24260 9 Flagship DH population is positioned proximal and independent to the well-characterised SD2 region that is associated with both high levels of dormancy and inferior malt quality. This indicates that it should be possible to develop cultivars that combine acceptable malting quality and adequate levels of grain dormancy for protection against PHS by utilizing these alternate QTL.

Estimating Hydrogen Cyanide in Forage Sorghum (Sorghum bicolor) by Near-Infrared Spectroscopy

Hydrogen cyanide (HCN) is a toxic chemical that can potentially cause mild to severe reactions in animals when grazing forage sorghum. Developing technologies to monitor the level of HCN in the growing crop would benefit graziers, so that they can move cattle into paddocks with acceptable levels of HCN. In this study, we developed near-infrared spectroscopy (MRS) calibrations to estimate HCN in forage sorghum and hay. The full spectral NIRS range (400-2498 nm) was used as well as specific spectral ranges within the full spectral range, i.e., visible (400-750 nm), shortwave (800-1100 nm) and near-infrared (NIR) (1100-2498 nm). Using the full spectrum approach and partial least-squares (PLS), the calibration produced a coefficient of determination (R-2) = 0.838 and standard error of cross-validation (SECV) = 0.040%, while the validation set had a R-2 = 0.824 with a low standard error of prediction (SEP = 0.047%). When using a multiple linear regression (MLR) approach, the best model (NIR spectra) produced a R-2 = 0.847 and standard error of calibration (SEC) = 0.050% and a R-2 = 0.829 and SEP = 0.057% for the validation set. The MLR models built from these spectral regions all used nine wavelengths. Two specific wavelengths 2034 and 2458 nm were of interest, with the former associated with C=O carbonyl stretch and the latter associated with C-N-C stretching. The most accurate PLS and MLR models produced a ratio of standard error of prediction to standard deviation of 3.4 and 3.0, respectively, suggesting that the calibrations could be used for screening breeding material. The results indicated that it should be feasible to develop calibrations using PLS or MLR models for a number of users, including breeding programs to screen for genotypes with low HCN, as well as graziers to monitor crop status to help with grazing efficiency.

QTL associated with barley (Hordeum vulgare) feed quality traits measured through in situ digestion

Barley (Hordeum vulgare) is a major feed source for the intensive livestock industry. Competitiveness against other cereal grains depends largely on the price per unit of expressed feed quality. The traits which contribute to feed quality in barley are largely quantitative in nature but little is known about their genetic control. A study to identify the quantitative trait loci (QTLs) associated with feed quality was performed using a F6-derived recombinant inbred barley population. Samples from each line were incubated in the rumen of fistulated cattle, recovered, washed and dried for determination of in situ dry matter digestibility. Additionally, both pre- and post-digestion samples were analysed to quantify the content of key quality components relating to acid detergent fibre, total starch and protein. The data was used to identify trait-associated QTLs. Genetic analysis identified significant QTLs on chromosomes 2H, 5H and 7H. Genetic markers linked to these QTL should provide an effective tool for the selection and improvement of feed barley in the future.

QTLs for water absorption and flour yield identified in the doubled haploid wheat population Lang/QT8766

Two key quality traits in milling wheat are flour yield (FY) and water absorption (WA). Ideally, breeders would prefer to use markers to select promising lines rather than time consuming rheology tests. In this study, we measured FY and WA on a wheat mapping population (Lang/QT8766) of 162 individuals grown in two replicated field experiments at three locations over 2 years. We also carried out near infrared reflectance spectroscopy (NIRS) predictions on the grain for these traits to see if NIRS phenotypic data could provide useful mapping results when compared to the reference phenotypic data. Several common QTLs were identified for FY and WA by both sets of data. The QTL on chromosome 4D was a consistently recurring QTL region for both traits. The QTL on chromosome 2A was positively linked to protein content which was supported by genetic correlation data. The results also indicated it was possible to obtain useful phenotypic data for mapping FY and WA using NIRS data. This would save time and costs as NIRS is quicker and cheaper than current rheology methods.

Barley genotype expressing "stay-green"-like characteristics maintains starch quality of the grain during water stress condition

The identification of "stay-green" in sorghum and its positive correlation with yield increases has encouraged attempts to incorporate "stay-green"-like traits into the genomes of other commercially important cereal crops. However, knowledge on the effects of "stay-green" expression on grain quality under extreme physiological stress is limited. This study examines impacts of "stay-green"-like expression on starch biosynthesis in barley (Hordeum vulgare L.) grain under mild, severe, and acute water stress conditions induced at anthesis. The proportions of long amylopectin branches and amylose branches in the grain of Flagship (a cultivar without "stay-green"-like characteristics) were higher at low water stress, suggesting that water stress affects starch biosynthesis in grain, probably due to early termination of grain fill. The changes in long branches can affect starch properties, such as the rates of enzymatic degradation, and hence its nutritional value. By contrast, grain from the "stay-green"-like cultivar (ND24260) did not show variation in starch molecular structure under the different water stress levels. The results indicate that the cultivar with "stay-green"-like traits has a greater potential to maintain starch biosynthesis and quality in grain during drought conditions, making the "stay-green"-like traits potentially useful in ensuring food security. (C) 2013 Elsevier Ltd. All rights reserved.

Does lethal control of top-predators release mesopredators? A re-evaluation of three Australian case studies

Top-predators can sometimes be important for structuring fauna assemblages in terrestrial ecosystems. Through a complex trophic cascade, the lethal control of top-predators has been predicted to elicit positive population responses from mesopredators that may in turn increase predation pressure on prey species of concern. In support of this hypothesis, many relevant research papers, opinion pieces and literature reviews identify three particular case studies as supporting evidence for top-predator control-induced release of mesopredators in Australia. However, many fundamental details essential for supporting this hypothesis are missing from these case studies, which were each designed to investigate alternative aims. Here, we re-evaluate the strength of evidence for top-predator control-induced mesopredator release from these three studies after comprehensive analyses of associated unpublished correlative and experimental data. Circumstantial evidence alluded to mesopredator releases of either the European Red Fox (Vulpes vulpes) or feral Cat (Felis catus) coinciding with Dingo (Canis lupus dingo) control in each case. Importantly, however, substantial limitations in predator population sampling techniques and/or experimental designs preclude strong assertions about the effect of lethal control on mesopredator populations from these studies. In all cases, multiple confounding factors and plausible alternative explanations for observed changes in predator populations exist. In accord with several critical reviews and a growing body of demonstrated experimental evidence on the subject, we conclude that there is an absence of reliable evidence for top-predator control-induced mesopredator release from these three case studies. Well-designed and executed studies are critical for investigating potential top-predator control-induced mesopredator release.

Isolation of multiple novel paramyxoviruses from pteropid bat urine

Bats have been found to harbor a number of new emerging viruses with zoonotic potential and there has been a great deal of interest in identifying novel bat pathogens to determine risk to human and animal health. Many groups have identified novel viruses in bats by detection of viral nucleic acid, however virus isolation is still a challenge and there are few reports of viral isolates from bats. In recent years, our group has developed optimized procedures for virus isolation from bat urine, including the use of primary bat cells. In previous reports we have described the isolation of Hendra virus, Menangle virus and Cedar virus, in Queensland, Australia. Here, we report the isolation of four additional novel bat paramyxoviruses from urine collected from beneath pteropid bat (flying fox) colonies in Queensland and New South Wales during 2009-2011.