Silica Vesicle Nanovaccine Formulations Stimulate Long-Term Immune Responses to the Bovine Viral Diarrhoea Virus E2 Protein

Bovine Viral Diarrhoea Virus (BVDV) is one of the most serious pathogen, which causes tremendous economic loss to the cattle industry worldwide, meriting the development of improved subunit vaccines. Structural glycoprotein E2 is reported to be a major immunogenic determinant of BVDV virion. We have developed a novel hollow silica vesicles (SV) based platform to administer BVDV-1 Escherichia coli-expressed optimised E2 (oE2) antigen as a nanovaccine formulation. The SV-140 vesicles (diameter 50 nm, wall thickness 6 nm, perforated by pores of entrance size 16 nm and total pore volume of 0.934 cm(3)g(-1)) have proven to be ideal candidates to load oE2 antigen and generate immune response. The current study for the first time demonstrates the ability of freeze-dried (FD) as well as non-FD oE2/SV140 nanovaccine formulation to induce long-term balanced antibody and cell mediated memory responses for at least 6 months with a shortened dosing regimen of two doses in small animal model. The in vivo ability of oE2 (100 mu g)/SV-140 (500 mu g) and FD oE2 (100 mu g)/SV-140 (500 mu g) to induce long-term immunity was compared to immunisation with oE2 (100 mu g) together with the conventional adjuvant Quil-A from the Quillaja saponira (10 mu g) in mice. The oE2/SV-140 as well as the FD oE2/SV-140 nanovaccine generated oE2-specific antibody and cell mediated responses for up to six months post the final second immunisation. Significantly, the cell-mediated responses were consistently high in mice immunised with oE2/SV-140 (1,500 SFU/million cells) at the six-month time point. Histopathology studies showed no morphological changes at the site of injection or in the different organs harvested from the mice immunised with 500 mu g SV-140 nanovaccine compared to the unimmunised control. The platform has the potential for developing single dose vaccines without the requirement of cold chain storage for veterinary and human applications.

Chemical containment and eradication of screw− worm incursions in Australia

Screwworms are obligate, invasive parasites of warm-blooded animals. The female flies lay batches of eggs at the edge of wounds or other lesions. These eggs hatch to larvae or screw-worms which feed on affected animals for 6-7 days, burrowing deeply into subcutaneous tissues and causing severe trauma to animals, production loss and potentially death. Susceptible sites include wounds resulting from management practices such as castration, de-horning and ear tagging and lesions caused by the activities of other parasites such as buffalo flies and ticks. The navels of the new born and the vulval region of their mothers following parturition are highly susceptible and body orifices such as nose and ears are also frequent targets for ovipositing screwworm flies. The Old World screw-worm, Chrysomya bezziana (OWS) is considered one of the most serious exotic insect pest threatening Australia's livestock industries and is endemic in a number of our closest neighbouring countries. New World screwworm (NWS), Cochliomyia hominivorax, endemic to South America, has also entered Australia on at least 2 occasions. Many tropical and subtropical areas of Australia are suitable for the establishment of OWS and the potential range is expected to increase with climate change. The Australian screwworm preparedness strategy indicates a program of containment with chemical treatments followed by eradication of OWS using sterile male release and parasiticides. However, there is no longer an operational OWS sterile insect screw-worm facility anywhere in the world and establishing a large scale production facility would most optimistically take at least 2 years. In the interim, containment would be almost totally dependent on the availability of effective chemical controls. A review of chemical formulations available for potential use against OWS in Australia found that currently only one chemical, ivermectin administered by subcutaneous injection (s.c.) is registered for use against OWS and that many of the chemicals previously shown to be effective against OWS were no longer registered for animal use in Australia.18 From this review a number of Australian-registered chemicals were recommended as a priority for testing against OWS. The Australian Pesticides and Veterinary Medicines Authority (APVMA) can issue an emergency use permit for use of pesticides if they are registered in Australia for other animal uses and shown to be effective against OWS. This project tested the therapeutic and prophylactic efficacy of chemicals with potential for use in the treatment and control of OWS.

Fertilising for yield and quality in sown grass pastures and forage crops

Sown pasture rundown and declining soil fertility for forage crops are too serious to ignore with losses in beef production of up to 50% across Queensland. The feasibility of using strategic applications of nitrogen (N) fertiliser to address these losses was assessed by analysing a series of scenarios using data drawn from published studies, local fertiliser trials and expert opinion. While N fertilser can dramatically increase productivity (growth, feed quality and beef production gains of over 200% in some scenarios), the estimated economic benefits, derived from paddock level enterprise budgets for a fattening operation, were much more modest. In the best-performing sown grass scenarios, average gross margins were doubled or tripled at the assumed fertiliser response rates, and internal rates of return of up to 11% were achieved. Using fertiliser on forage sorghum or oats was a much less attractive option and, under the paddock level analysis and assumptions used, forages struggled to be profitable even on fertile sites with no fertiliser input. The economics of nitrogen fertilising on grass pasture were sensitive to the assumed response rates in both pasture growth and liveweight gain. Consequently, targeted research is proposed to re-assess the responses used in this analysis, which are largely based on research 25-40 years ago when soils were generally more fertile and pastures less rundown.

Lesser auger beetle Heterobostrychus aequalis(Coleoptera: Bostrichidae) in Australia: absent or elusive?

The lesser auger beetle, Heterobostrychus aequalis (Waterhouse), is a serious pest of seasoned hardwood timber throughout the Oriental region and several areas beyond. Some early collection records of H. aequalis from Australia in the 1950s and 1960s indicated that the insect was present in northern Queensland, but no confirmed breeding population has been found in the past few decades suggesting either that it may have not established permanently or it is difficult to detect. The ambiguity about the breeding status of the pest in Australia has caused confusion for regulating authorities needing to respond to each new post-border detection. We examined records of H. aequalis in Australian insect collections and from intensive plant pest surveillance activities in Queensland and northern Australia over the past 48 years to resolve this confusion. Until very recently, available evidence suggested that H. aequalis was not established in Australia, despite multiple introductions and apparently suitable climate and hosts. Collection records of the pest are predominantly linked to intercepted items or are recorded as of unknown origin, and no established populations have been found during many years of targeted surveillance. However, a detection of H. aequalis in suburban Cairns, north Queensland, in late 2013 and two more in mid-2015 in the same general locality do not appear to be linked to any imported material, indicating that there is at least a tenuously established local population. Investigations are underway to confirm this, but the insect is not widely established in Australia and, if present, remains elusive. Our recommended response to any future detection of H. aequalis is to fumigate or destroy the infested material, conduct tracing enquiries and limit surveys to the immediate vicinity of the detection.

The host range of four new biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) from southern USA and their potential as biological control agents of Cylindropuntiaspp.(Cactaceae) in Australia: Part II.

Eight Cylindropuntia species have naturalised in Australia and pose serious economic, environmental and social impacts. Two biotypes of Dactylopius tomentosus have been used as bio-control agents to control different Cylindropuntia species. The host range of four additional biotypes of Dactylopius tomentosus from southern USA was investigated. Feeding and development were restricted to the genus Cylindropuntia. However, they showed differences in specificity within this genus and some biotypes discriminated between the provenances of C. rosea and C. tunicata. Efficacy trials were conducted to determine whether populations of each biotype could be sustained on the naturalised Cylindropuntia species and if these populations could retard the growth or kill these plants. The acanthocarpa biotype offers potential control of C. rosea (Lorne Station), while the cylindropuntia sp. biotype shows great potential to control C. rosea (Grawin). The cylindropuntia sp. biotype also had a high impact on C. kleiniae and C. imbricata, and a moderate impact on C. leptocaulis and C. prolifera. The acanthocarpa X echinocarpa biotype had its greatest impact on C. tunicata (Grawin), killing this plant in 18 weeks. A fourth biotype, leptocaulis, was damaging to some species, but was less effective than the other biotypes. Cylindropuntia spinosior is the only naturalised species in Australia where no effective biocontrol agent has been found.