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.

An exploratory study to assess the activity of the acarine growth inhibitor, fluazuron, against Sarcoptes scabei infestation in pigs

Background: The most common treatments for scabies in human and veterinary settings are topical 5% permethrin or systemic treatment with ivermectin. However, these treatments have very little activity against arthropod eggs, and therefore repeated treatment is frequently required. In-vitro, biochemical and molecular studies have demonstrated that human mites are becoming increasingly resistant to both acaricides. To identify alternate acaricides, we undertook a pilot study of the in vivo activity of the benzoylphenyl urea inhibitor of chitin synthesis, fluazuron, in pigs with sarcoptic mange. Findings: Pigs (n = 5) were infested with S. scabei var suis, and randomised to treatment at the start of peak infestation with fluazuron at a dose of 10 mg/kg/day per os for 7 days (n = 3) or no treatment (n = 2). Clinical scores, skin scrapings for mite counts and blood sampling for pharmacokinetic analysis were undertaken. Fluazuron was well absorbed in treated pigs with measureable blood levels up to 4 weeks post treatment. No adverse effects were observed. Modest acaricidal activity of the compound was observed, with a reduction in severity of skin lesions in treated pigs, as well as a reduction in number of scabies mite's early life stages. Conclusions: The moderate efficacy of fluazuron against scabies mites indicates a lead to the development of alternate treatments for scabies, such as combination therapies that maybe applicable for human use in the future.

Estimating mineralisation of organic nitrogen from biosolids and other organic wastes applied to soils in subtropical Australia

One major benefit of land application of biosolids is to supply nitrogen (N) for agricultural crops, and understanding mineralisation processes is the key for better N-management strategies. Field studies were conducted to investigate the process of mineralisation of three biosolids products (aerobic, anaerobic, and thermally dried biosolids) incorporated into four different soils at rates of 7-90 wet t/ha in subtropical Queensland. Two of these studies also examined mineralisation rates of commonly used organic amendments (composts, manures, and sugarcane mill muds). Organic N in all biosolids products mineralised very rapidly under ambient conditions in subtropical Queensland, with rates much faster than from other common amendments. Biosolids mineralisation rates ranged from 30 to 80% of applied N during periods ranging from 3.5 to 18 months after biosolids application; these rates were much higher than those suggested in the biosolids land application guidelines established by the NSW EPA (15% for anaerobic and 25% for aerobic biosolids). There was no consistently significant difference in mineralisation rate between aerobic and anaerobic biosolids in our studies. When applied at similar rates of N addition, other organic amendments supplied much less N to the soil mineral N and plant N pools during the crop season. A significant proportion of the applied biosolids total N (up to 60%) was unaccounted for at the end of the observation period. High rates of N addition in calculated Nitrogen Limited Biosolids Application Rates (850-1250 kg N/ha) resulted in excessive accumulation of mineral N in the soil profile, which increases the environmental risks due to leaching, runoff, or gaseous N losses. Moreover, the rapid mineralisation of the biosolids organic N in these subtropical environments suggests that biosolids should be applied at lower rates than in temperate areas, and that care must be taken with the timing to maximise plant uptake and minimise possible leaching, runoff, or denitrification losses of mineralised N.

Riverine flow and spawning requirements of Macquaria ambigua oriens: implications for conservation and management

Understanding the life-history attributes of aquatic species is integral to the development of environmental-flow strategies in regulated river systems. This is particularly important when species are under continual and increasing pressure from water-resource development. In this study, the water temperature and flow requirements for spawning of the Fitzroy River golden perch (Macquaria ambigua oriens) were investigated over 4 years at 22 sites in the Fitzroy River catchment. Eggs, larvae and young-of-year (YOY) M. ambigua oriens were sampled on a variety of flow events to determine the environmental requirements for spawning. Eggs and larvae of M. ambigua oriens were detected during natural flow events generally with a minimum of 1.5 m river rise and duration of 7 days. Spawning was associated with the peak and/or recession of the first or second post-winter flow event where water temperatures exceeded 248 degrees C. Our data suggests that it is important to protect a range of flows, not just flood flows, as previously documented for this species. The interaction of spawning flows with existing and future water-resource development should be considered to ensure maintenance of the population viability of M. ambigua oriens.

Life history of Anomalococcus indicus (Hemiptera: Lecanodiaspididae), a potential biological control agent for prickly acacia Vachellia nilotica ssp indica in Australia

Babul scale Anomalococcus indicus Ramakrishna Ayyar, a major pest of Vachellia nilotica (L.f.) P.J.H. Hurter & Mabb. on the Indian subcontinent, has been identified as a potential biocontrol agent for prickly acacia V. nilotica subsp. indica (Benth.) Kyal. & Boatwr. in Australia and was imported from southern India for detailed assessment. The life history of A. indicus under controlled glasshouse conditions was determined as a part of this assessment. Consistent with other scale species, A. indicus has a distinct sexual dimorphism which becomes apparent during the second instar. Females have three instars, developing into sexually mature nymphs after 52 days. The generation time from egg to egg was 89 days. Females are ovoviviparous, ovipositing mature eggs into a cavity underneath their body. An average of 802 +/- 114 offspring were produced per female. Reproductive output was closely associated with female size; larger females produced more than 1200 offspring. Crawlers emerged from beneath the female after an indeterminate period of inactivity. They have the only life stage at which A. indicus can disperse, though the majority settle close to their parent female forming aggregative distributions. In the absence of food, most crawlers died within three days. Males took 62 days to develop through five instars. Unlike females, males underwent complete metamorphosis. Adult males were small and winged, and lived for less than a day. Parthenogenesis was not observed in females excluded from males. The life history of A. indicus allows it to complement other biological control agents already established on prickly acacia in Australia.

Rhipicephalus (Boophilus) microplus tick in vitro feeding methods for functional (dsRNA) and vaccine candidate (antibody) screening

Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) ticks cause economic losses for cattle industries throughout tropical and subtropical regions of the world estimated at $US2.5 billion annually. Lack of access to efficacious long-lasting vaccination regimes and increases in tick acaricide resistance have led to the investigation of targets for the development of novel tick vaccines and treatments. In vitro tick feeding has been used for many tick species to study the effect of new acaricides on the transmission of tick-borne pathogens. Few studies have reported the use of in vitro feeding for functional genomic studies using RNA interference and/or the effect of specific anti-tick antibodies. In particular, in vitro feeding reports for the cattle tick are limited due to its relatively short hypostome. Previously published methods were further modified to broaden optimal tick sizes/weights, feeding sources including bovine and ovine serum, optimisation of commercially available blood anti-coagulant tubes, and IgG concentrations for effective antibody delivery. Ticks are fed overnight and monitored for ∼5–6 weeks to determine egg output and success of larval emergence using a humidified incubator. Lithium-heparin blood tubes provided the most reliable anti-coagulant for bovine blood feeding compared with commercial citrated (CPDA) and EDTA tubes. Although >30 mg semi-engorged ticks fed more reliably, ticks as small as 15 mg also fed to repletion to lay viable eggs. Ticks which gained less than ∼10 mg during in vitro feeding typically did not lay eggs. One mg/ml IgG from Bm86-vaccinated cattle produced a potent anti-tick effect in vitro (83% efficacy) similar to that observed in vivo. Alternatively, feeding of dsRNA targeting Bm86 did not demonstrate anti-tick effects (11% efficacy) compared with the potent effects of ubiquitin dsRNA. This study optimises R. microplus tick in vitro feeding methods which support the development of cattle tick vaccines and treatments.

Methods to determine resistance to anthelmintics when continuing larval development occurs

The in vivo faecal egg count reduction test (FECRT) is the most commonly used test to detect anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) of ruminants in pasture based systems. However, there are several variations on the method, some more appropriate than others in specific circumstances. While in some cases labour and time can be saved by just collecting post-drench faecal worm egg counts (FEC) of treatment groups with controls, or pre- and post-drench FEC of a treatment group with no controls, there are circumstances when pre- and post-drench FEC of an untreated control group as well as from the treatment groups are necessary. Computer simulation techniques were used to determine the most appropriate of several methods for calculating AR when there is continuing larval development during the testing period, as often occurs when anthelmintic treatments against genera of GIN with high biotic potential or high re-infection rates, such as Haemonchus contortus of sheep and Cooperia punctata of cattle, are less than 100% efficacious. Three field FECRT experimental designs were investigated: (I) post-drench FEC of treatment and controls groups, (II) pre- and post-drench FEC of a treatment group only and (III) pre- and post-drench FEC of treatment and control groups. To investigate the performance of methods of indicating AR for each of these designs, simulated animal FEC were generated from negative binominal distributions with subsequent sampling from the binomial distributions to account for drench effect, with varying parameters for worm burden, larval development and drench resistance. Calculations of percent reductions and confidence limits were based on those of the Standing Committee for Agriculture (SCA) guidelines. For the two field methods with pre-drench FEC, confidence limits were also determined from cumulative inverse Beta distributions of FEC, for eggs per gram (epg) and the number of eggs counted at detection levels of 50 and 25. Two rules for determining AR: (1) %reduction (%R) < 95% and lower confidence limit <90%; and (2) upper confidence limit <95%, were also assessed. For each combination of worm burden, larval development and drench resistance parameters, 1000 simulations were run to determine the number of times the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been declared. When continuing larval development occurs during the testing period of the FECRT, the simulations showed AR should be calculated from pre- and post-drench worm egg counts of an untreated control group as well as from the treatment group. If the widely used resistance rule 1 is used to assess resistance, rule 2 should also be applied, especially when %R is in the range 90 to 95% and resistance is suspected.

Freeze-drying of ovalbumin loaded mesoporous silica nanoparticle vaccine formulation increases antigen stability under ambient conditions

Amino functionalised mesoporous silica nanoparticles (AM-41) have been identified as a promising vaccine delivery material. The capacity of AM-41 to stabilise vaccine components at ambient temperature (23–27 °C) was determined by adsorbing the model antigen ovalbumin (OVA) to AM-41 particles (OVA-41). The OVA-41 was successfully freeze-dried using the excipients 5% trehalose and 1% PEG8000. Both the immunological activity of OVA and the nanoparticle structure were maintained following two months storage at ambient temperature. The results of immunisation studies in mice with reconstituted OVA-41 demonstrated the induction of humoral and cell-meditated immune responses. The capacity of AM-41 particles to facilitate ambient storage of vaccine components without loss of immunological potency will underpin the further development of this promising vaccine delivery platform.

Reproductive interference between honey bee species in artificial sympatry

Reproductive isolation between closely related species is often incomplete. The Western honey bee, Apis mellifera, and the Eastern hive bee, A. cerana have been allopatric for millions of years, but are nonetheless similar in morphology and behaviour. During the last century the two species were brought into contact anthropogenically, providing potential opportunities for interspecific matings. Hybrids between A. mellifera and A. cerana are inviable, so natural interspecific matings are of concern because they may reduce the viability of A. cerana and A. mellifera populations – two of the world's most important pollinators. We examined the mating behaviour of A. mellifera and A. cerana queens and drones from Caoba Basin, China and Cairns, Australia. Drone mating flight times overlap in both areas. Analysis of the spermathecal contents of queens with species-specific genetic markers indicated that in Caoba Basin, 14% of A. mellifera queens mated with at least one A. cerana male, but we detected no A. cerana queens that had mated with A. mellifera males. Similarly, in Cairns, no A. cerana queens carried A. mellifera sperm, but one third of A. mellifera queens had mated with at least one A. cerana male. No hybrid embryos were detected in eggs laid by interspecifically-mated A. mellifera queens in either location. However A. mellifera queens artificially inseminated with A. cerana sperm produced inviable hybrid eggs, or unfertilised drones. This suggests that reproductive interference will impact the viability of honey bee populations wherever A. cerana and A. mellifera are in contact. This article is protected by copyright. All rights reserved.

Strangers in a strange land: do life history traits differ for alien and native colonisers of novel environments?

Do alien invasive species exhibit life history characteristics that are similar to those of native species that have become pests in their continent of origin? We compared eucalypt specialists that have become pests in Australian plantations (natives) to those that have established overseas (aliens) using 13 life history traits and found that although traits that support rapid population build-up were shared, overall, aliens and native colonisers differed significantly. Distance from source (New Zealand vs. other) had no significant effect, but species that established more than 50 years ago exhibited different life history traits from those that established within the last 50 years, possibly because of more effective quarantine. Native and alien eucalypt insect invaders differed predominantly in traits that facilitate long-distance movement (pathway traits), compared to traits that facilitate establishment and spread. Aliens had longer adult flight seasons, were smaller and more closely host-associated (cryptic eggs and larvae), had lower incidence of diapause (i.e. were more seasonally plastic) and more generations per year than natives. Thus, studies of species invasive within their country of origin can shed light on alien invasions.

Rapid assessment of genotype-by-environment interactions and heritability for growth rate in aquaculture species using in vitro fertilisation and DNA tagging

Commercial environments may receive only a fraction of expected genetic gains for growth rate as predicted from the selection environment. This fraction is result of undesirable genotype-by-environment interactions (GxE) and measured by the genetic correlation (rg) of growth between environments. Rapid estimates of genetic correlation achieved in one generation are notoriously difficult to estimate with precision. A new design is proposed where genetic correlations can be estimated by utilising artificial mating from cryopreserved semen and unfertilised eggs stripped from a single female. We compare a traditional phenotype analysis of growth to a threshold model where only the largest fish are genotyped for sire identification. The threshold model was robust to differences in family mortality differing up to 30%. The design is unique as it negates potential re-ranking of families caused by an interaction between common maternal environmental effects and growing environment. The design is suitable for rapid assessment of GxE over one generation with a true 0.70 genetic correlation yielding standard errors as low as 0.07. Different design scenarios were tested for bias and accuracy with a range of heritability values, number of half-sib families created, number of progeny within each full-sib family, number of fish genotyped, number of fish stocked, differing family survival rates and at various simulated genetic correlation levels.

Life cycle and host range of Phycitasp. rejected for biological control of prickly acacia in Australia

Prickly acacia (Vachellia nilotica subsp. indica), a native of the Indian subcontinent, is a serious weed of the grazing areas of northern Australia and is a target for classical biological control. Native range surveys in India identified a leaf webber, Phycita sp. (Lepidoptera: Pyralidae) as a prospective biological control agent for prickly acacia. In this study, we report the life cycle and host-specificity test results Phycita sp. and highlight the contradictory results between the no-choice tests in India and Australia and the field host range in India. In no-choice tests in India and Australia, Phycita sp. completed development on two of 11 and 16 of 27 non-target test plant species, respectively. Although Phycita sp. fed and completed development on two non-target test plant species (Vachellia planifrons and V. leucophloea) in no-choice tests in India, there was no evidence of the insect on the two non-target test plant species in the field. Our contention is that oviposition behaviour could be the key mechanism in host selection of Phycita sp., resulting in its incidence only on prickly acacia in India. This is supported by paired oviposition choice tests involving three test plant species (Acacia baileyana, A. mearnsii and A. deanei) in quarantine in Australia, where eggs were laid only on prickly acacia. However, in paired oviposition choice trials, only few eggs were laid, making the results unreliable. Although oviposition choice tests suggest that prickly acacia is the most preferred and natural host, difficulties in conducting choice oviposition tests with fully grown trees under quarantine conditions in Australia and the logistic difficulties of conducting open-field tests with fully grown native Australian plants in India have led to rejection of Phycita sp. as a potential biological control agent for prickly acacia in Australia.

Zeaxanthin biofortification of sweet-corn and factors affecting zeaxanthin accumulation and colour change

Zeaxanthin, along with its isomer lutein, are the major carotenoids contributing to the characteristic colour of yellow sweet-corn. From a human health perspective, these two carotenoids are also specifically accumulated in the human macula, and are thought to protect the photoreceptor cells of the eye from blue light oxidative damage and to improve visual acuity. As humans cannot synthesise these compounds, they must be accumulated from dietary components containing zeaxanthin and lutein. In comparison to most dietary sources, yellow sweet-corn (Zea mays var. rugosa) is a particularly good source of zeaxanthin, although the concentration of zeaxanthin is still fairly low in comparison to what is considered a supplementary dose to improve macular pigment concentration (2 mg/person/day). In our present project, we have increased zeaxanthin concentration in sweet-corn kernels from 0.2 to 0.3 mg/100 g FW to greater than 2.0 mg/100 g FW at sweet-corn eating-stage, substantially reducing the amount of corn required to provide the same dosage of zeaxanthin. This was achieved by altering the carotenoid synthesis pathway to more than double total carotenoid synthesis and to redirect carotenoid synthesis towards the beta-arm of the pathway where zeaxanthin is synthesised. This resulted in a proportional increase of zeaxanthin from 22% to 70% of the total carotenoid present. As kernels increase in physiological maturity, carotenoid concentration also significantly increases, mainly due to increased synthesis but also due to a decline in moisture content of the kernels. When fully mature, dried kernels can reach zeaxanthin and carotene concentrations of 8.7 mg/100 g and 2.6 mg/100 g, respectively. Although kernels continue to increase in zeaxanthin when harvested past their normal harvest maturity stage, the texture of these 'over-mature' kernels is tough, making them less appealing for fresh consumption. Increase in zeaxanthin concentration and other orange carotenoids such as p-carotene also results in a decline in kernel hue angle of fresh sweet-corn from approximately 90 (yellow) to as low as 75 (orange-yellow). This enables high-zeaxanthin sweet-corn to be visually-distinguishable from standard yellow sweet-corn, which is predominantly pigmented by lutein.

Effect of drying, storage temperature and air exposure on astaxanthin stability from Haematococcus pluvialis

Astaxanthin is a powerful antioxidant with various health benefits such as prevention of age-related macular degeneration and improvement of the immune system, liver and heart function. To improve the post-harvesting stability of astaxanthin used in food, feed and nutraceutical industries, the biomass of the high astaxanthin producing alga Haematococcus pluvialis was dried by spray- or freeze-drying and under vacuum or air at − 20 °C to 37 °C for 20 weeks. Freeze-drying led to 41 higher astaxanthin recovery compared to commonly-used spray-drying. Low storage temperature (− 20 °C, 4 °C) and vacuum-packing also showed higher astaxanthin stability with as little as 12.3 ± 3.1 degradation during 20 weeks of storage. Cost-benefit analysis showed that freeze-drying followed by vacuum-packed storage at − 20 °C can generate AUD600 higher profit compared to spray-drying from 100 kg H. pluvialis powder. Therefore, freeze-drying can be suggested as a mild and more profitable method for ensuring longer shelf life of astaxanthin from H. pluvialis.

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.