Evaluation of the most effective measures of infectivity of a precocious line of Eimeria acervulina in poultry

Coccidiosis is an economically important parasitic disease of chickens that, in Australia, is caused by seven species of the genus Eimeria.1 The disease has traditionally been controlled by prophylactic drugs, but vaccination with attenuated lines of the parasites2–4 is rapidly gaining acceptance world wide. Live Eimeria vaccines are produced in batches which are not frozen and have a limited shelf life. The per cent infectivity of vaccine seed stocks and the vaccines produced from them must therefore be accurately monitored using standardised dose dependant assays to ensure that shelf life, quality control and vaccine release specifications are met. Infectivity for the chicken host cannot readily be determined by microscopic observation of oocysts or sporocyst hatching.5 Dose dependent parameters such as body weight gain, feed conversion ratio, visual lesion scores, mortality, oocysts production, clinical symptoms and microscopic lesion counts could be used as measures of infectivity.6–11 These parameters show significant dose dependant effects with field strains, but lines of vaccine parasites that have been selected for precocious development with associated reduced virulence and reproductive capability may not have the same effect.3,4 The aim of this trial was to determine which parameters provide the most effective measures of infective dose in birds inoculated with a precocious vaccine strain.

A method for mapping the distribution and density of rabbits and other vertebrate pests in Australia

The European wild rabbit has been considered Australia’s worst vertebrate pest and yet little effort appears to have gone into producing maps of rabbit distribution and density. Mapping the distribution and density of pests is an important step in effective management. A map is essential for estimating the extent of damage caused and for efficiently planning and monitoring the success of pest control operations. This paper describes the use of soil type and point data to prepare a map showing the distribution and density of rabbits in Australia. The potential for the method to be used for mapping other vertebrate pests is explored. The approach used to prepare the map is based on that used for rabbits in Queensland (Berman et al. 1998). An index of rabbit density was determined using the number of Spanish rabbit fleas released per square kilometre for each Soil Map Unit (Atlas of Australian Soils). Spanish rabbit fleas were released into active rabbit warrens at 1606 sites in the early 1990s as an additional vector for myxoma virus and the locations of the releases were recorded using a Global Positioning System (GPS). Releases were predominantly in arid areas but some fleas were released in south east Queensland and the New England Tablelands of New South Wales. The map produced appears to reflect well the distribution and density of rabbits, at least in the areas where Spanish fleas were released. Rabbit pellet counts conducted in 2007 at 54 sites across an area of south east South Australia, south eastern Queensland, and parts of New South Wales (New England Tablelands and south west) in soil Map Units where Spanish fleas were released, provided a preliminary means to ground truth the map. There was a good relationship between mean pellet count score and the index of abundance for soil Map Units. Rabbit pellet counts may allow extension of the map into other parts of Australia where there were no Spanish rabbit fleas released and where there may be no other consistent information on rabbit location and density. The recent Equine Influenza outbreak provided a further test of the value of this mapping method. The distribution and density of domestic horses were mapped to provide estimates of the number of horses in various regions. These estimates were close to the actual numbers of horses subsequently determined from vaccination records and registrations. The soil Map Units are not simply soil types they contain information on landuse and vegetation and the soil classification is relatively localised. These properties make this mapping method useful, not only for rabbits, but also for other species that are not so dependent on soil type for survival.

Characterization of the antibody response in birds following infection with wild-type and attenuated strains of Eimeria tenella and Eimeria necatrix.

Live vaccines containing attenuated parasite strains are increasingly used to control chicken coccidiosis. In this paper antibody responses elicited by infections with wild-type and attenuated strains of Eimeria tenella and E.necatrix were characterized by immunoblotting and ELISA with homologous and heterologous antisera. Few differences between antisera from birds infected with wild and attenuated strains of E. tenella were evident in immunoblots conducted with merozoite antigen preparations from both E. tenella strains, however the reactivity of sera raised in birds infected with the wild-type strain was noticeably more intense. In ELISAs conducted with merozoite antigen preparations, antisera from birds infected with the wild-type strains of E. tenella and E. necatrix consistently produced a significantly higher (P < 0.05) antibody response than antisera from birds infected with the attenuated strains. Likewise, avidity ELISAs conducted with the E. tenella strains demonstrated that antibodies in birds infected with the wild-type strain were of significantly higher avidity (P < 0.05) than antibodies in birds infected with the attenuated strain. The differences in the antibody responses are probably due to changes in the attenuated strain as a result of selection for precocious development and the less severe tissue damage and inflammation of the intestine resulting from infection with the attenuated strain.

Trends in therapeutic and prevention strategies for management of Bovine Mastitis: An overview

Mastitis is one of the most economically significant diseases for the dairy industry for backyard farmers in developing countries and high producing herds worldwide. Two of the major factors impeding reduction in the incidence of this disease is [a] the lack of availability of an effective vaccine capable of protecting against multiple etiological agents and [b] propensity of some of the etiological agents to develop persistent antibiotic resistance in biofilms. This is further complicated by the continuing revolving shift in the predominant etiological agents of mastitis, depending upon a multitude of factors such as variability in hygienic practices on farms, easy access leading to overuse of appropriate or inappropriate antibiotics at suboptimal concentrations, particularly in developing countries, and lack of compliance with the recommended treatment schedules. Regardless, Staphylococcus aureus and Streptococcus uberis followed by Escherichia coli, Streptococcus agalactiae has become the predominant etiological agents of bovine mastitis followed Streptococcus agalactiae, Streptococcus dysagalactiae, Klebsiella pneumonia and the newly emerging Mycoplasma bovis. Current approaches being pursued to reduce the negative economic impact of this disease are through early diagnosis of infection, immediate treatment with an antibiotic found to either inhibit or kill the pathogen(s) in vitro using planktonic cultures and the use of the currently marketed vaccines regardless of their demonstrated effectiveness. Given the limitations of breeding programs, including genetic selection to improve resistance against infectious diseases including mastitis, it is imperative to have the availability of an effective broad-spectrum, preferably cross-protective, vaccine capable of protecting against bovine mastitis for reduction in the incidence of bovine mastitis, as well as interrupting the potential cross-species transmission to humans. This overview highlights the major etiological agents, factors affecting susceptibility to mastitis, and the current status of antibiotic-based therapies and prototype vaccine candidates or commercially available vaccines against bovine mastitis as potential preventative strategies. © 2013 Tiwari JG, et al.

Agricultural intensification, priming for persistence and the emergence of Nipah virus: a lethal bat-borne zoonosis

Emerging zoonoses threaten global health, yet the processes by which they emerge are complex and poorly understood. Nipah virus (NiV) is an important threat owing to its broad host and geographical range, high case fatality, potential for human-to-human transmission and lack of effective prevention or therapies. Here, we investigate the origin of the first identified outbreak of NiV encephalitis in Malaysia and Singapore. We analyse data on livestock production from the index site (a commercial pig farm in Malaysia) prior to and during the outbreak, on Malaysian agricultural production, and from surveys of NiV's wildlife reservoir (flying foxes). Our analyses suggest that repeated introduction of NiV from wildlife changed infection dynamics in pigs. Initial viral introduction produced an explosive epizootic that drove itself to extinction but primed the population for enzootic persistence upon reintroduction of the virus. The resultant within-farm persistence permitted regional spread and increased the number of human infections. This study refutes an earlier hypothesis that anomalous El Nino Southern Oscillation-related climatic conditions drove emergence and suggests that priming for persistence drove the emergence of a novel zoonotic pathogen. Thus, we provide empirical evidence for a causative mechanism previously proposed as a precursor to widespread infection with H5N1 avian influenza and other emerging pathogens.

Efficacy of Rhipicephalus (Boophilus) microplus Bm86 against Hyalomma dromedarii and Amblyomma cajennense tick infestations in camels and cattle

The recombinant Bm86-based tick vaccines have shown their efficacy for the control of cattle ticks, Rhipicephalus (Boophilus) microplus and R. annulatus infestations. However, cattle ticks often co-exist with multi-host ticks such as Hyalomma and Amblyomma species, thus requiring the control of multiple tick infestations for cattle and other hosts. Vaccination trials using a R. microplus recombinant Bm86-based vaccine were conducted in cattle and camels against Hyalomma dromedarii and in cattle against Amblyomma cajennense immature and adult ticks. The results showed an 89% reduction in the number of H. dromedarii nymphs engorging on vaccinated cattle, and a further 32% reduction in the weight of the surviving adult ticks. In vaccinated camels, a reduction of 27% and 31% of tick engorgement and egg mass weight, respectively was shown, while egg hatching was reduced by 39%. However, cattle vaccination with Bm86 did not have an effect on A. cajennense tick infestations. These results showed that Bm86 vaccines are effective against R. microplus and other tick species but improved vaccines containing new antigens are required to control multiple tick infestations. (C) 2012 Elsevier Ltd. All rights reserved.

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.

Silica vesicles as nanocarriers and adjuvants for generating both antibody and T-cell mediated immune resposes to Bovine Viral Diarrhoea Virus E2 protein

Bovine Viral Diarrhoea Virus (BVDV) is widely distributed in cattle industries and causes significant economic losses worldwide annually. A limiting factor in the development of subunit vaccines for BVDV is the need to elicit both antibody and T-cell-mediated immunity as well as addressing the toxicity of adjuvants. In this study, we have prepared novel silica vesicles (SV) as the new generation antigen carriers and adjuvants. With small particle size of 50 nm, thin wall (similar to 6 nm), large cavity (similar to 40 nm) and large entrance size (5.9 nm for SV-100 and 16 nm for SV-140), the SV showed high loading capacity (similar to 250 mu g/mg) and controlled release of codon-optimised E2 (oE2) protein, a major immunogenic determinant of BVDV. The in vivo functionality of the system was validated in mice immunisation trials comparing oE2 plus Quil A (50 mu g of oE2 plus 10 mu g of Quil A, a conventional adjuvant) to the oE2/SV-140 (50 mu g of oE2 adsorbed to 250 mu g of SV-140) or oE2/SV-140 together with 10 mu g of Quil A. Compared to the oE2 plus Quil A, which generated BVDV specific antibody responses at a titre of 10(4), the oE2/SV-140 group induced a 10 times higher antibody response. In addition, the cell-mediated response, which is essential to recognise and eliminate the invading pathogens, was also found to be higher [1954-2628 spot forming units (SFU)/million cells] in mice immunised with oE2/SV-140 in comparison to oE2 plus Quil A (512-1369 SFU/million cells). Our study has demonstrated that SV can be used as the next-generation nanocarriers and adjuvants for enhanced veterinary vaccine delivery. (C) 2014 Elsevier Ltd. All rights reserved.

Novel vaccine acceptance in emerging infectious diseases: a case study of Hendra virus vaccine uptake in Australia

Background: The development of a horse vaccine against Hendra virus has been hailed as a good example of a One Health approach to the control of human disease. Although there is little doubt that this is true, it is clear from the underwhelming uptake of the vaccine by horse owners to date (approximately 10%) that realisation of a One Health approach requires more than just a scientific solution. As emerging infectious diseases may often be linked to the development and implementation of novel vaccines this presentation will discuss factors influencing their uptake; using Hendra virus in Australia as a case study. Methods: This presentation will draw on data collected from the Horse owners and Hendra virus: A Longitudinal cohort study To Evaluate Risk (HHALTER) study. The HHALTER study is a mixed methods research study comprising a two-year survey-based longitudinal cohort study and qualitative interview study with horse owners in Australia. The HHALTER study has investigated and tracked changes in a broad range of issues around early uptake of vaccination, horse owner uptake of other recommended disease risk mitigation strategies, and attitudes to government policy and disease response. Interviews provide further insights into attitudes towards risk and decision-making in relation to vaccine uptake. A combination of quantitative and qualitative data analysis will be reported. Results: Data collected from more than 1100 horse owners shortly after vaccine introduction indicated that vaccine uptake and intention to vaccinate was associated with a number of risk perception factors and financial cost factors. In addition, concerns about side effects and veterinarians refusing to treat unvaccinated horses were linked to uptake. Across the study period vaccine uptake in the study cohort increased to more than 50%, however, concerns around side effects, equine performance and breeding impacts, delays to full vaccine approvals, and attempts to mandate vaccination by horse associations and event organisers have all impacted acceptance. Conclusion: Despite being provided with a safe and effective vaccine for Hendra virus that can protect horses and break the transmission cycle of the virus to humans, Australian horse owners have been reluctant to commit to it. General issues pertinent to novel vaccines, combined with challenges in the implementation of the vaccine have led to issues of mistrust and misconception with some horse owners. Moreover, factors such as cost, booster dose schedules, complexities around perceived risk, and ulterior motives attributed to veterinarians have only served to polarise attitudes to vaccine acceptance.

A molecular survey of Eimeria in chickens across Australia

Coccidiosis is a costly enteric disease of chickens caused by protozoan parasites of the genus Eimeria. Disease diagnosis and management is complicated since there are multiple Eimeria species infecting chickens and mixed species infections are common. Current control measures are only partially effective and this, combined with concerns over vaccine efficacy and increasing drug resistance, demonstrates a need for improved coccidiosis diagnosis and control. Before improvements can be made, it is important to understand the species commonly infecting poultry flocks in both backyard and commercial enterprises. The aim of this project was to conduct a survey and assessment of poultry Eimeria across Australia using genetic markers, and create a collection of isolates for each Eimeria species. A total of 260 samples (faecal or caecal) was obtained, and survey results showed that Eimeria taxa were present in 98% of commercial and 81% of backyard flocks. The distribution of each Eimeria species was widespread across Australia, with representatives of all species being found in every state and territory, and the Eimeria species predominating in commercial flocks differed from those in backyard flocks. Three operational taxonomic units also occurred frequently in commercial flocks highlighting the need to understand the impact of these uncharacterised species on poultry production. As Eimeria infections were also frequent in backyard flocks, there is a potential for backyard flocks to act as reservoirs for disease, especially as the industry moves towards free range production systems. This Eimeria collection will be an important genetic resource which is the crucial first step in the development of more sophisticated diagnostic tools and the development of new live vaccines which ultimately will provide savings to the industry in terms of more efficient coccidiosis management.

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.

The multidimensional causal factors of ‘wet litter’ in chicken-meat production

The problem of ‘wet litter’, which occurs primarily in grow-out sheds for meat chickens (broilers), has been recognised for nearly a century. Nevertheless, it is an increasingly important problem in contemporary chicken-meat production as wet litter and associated conditions, especially footpad dermatitis, have developed into tangible welfare issues. This is only compounded by the market demand for chicken paws and compromised bird performance. This review considers the multidimensional causal factors of wet litter. While many causal factors can be listed it is evident that the critical ones could be described as micro-environmental factors and chief amongst them is proper management of drinking systems and adequate shed ventilation. Thus, this review focuses on these environmental factors and pays less attention to issues stemming from health and nutrition. Clearly, there are times when related avian health issues of coccidiosis and necrotic enteritis cannot be overlooked and the development of efficacious vaccines for the latter disease would be advantageous. Presently, the inclusion of phytate-degrading enzymes in meat chicken diets is routine and, therefore, the implication that exogenous phytases may contribute to wet litter is given consideration. Opinion is somewhat divided as how best to counter the problem of wet litter as some see education and extension as being more beneficial than furthering research efforts. However, it may prove instructive to assess the practice of whole grain feeding in relation to litter quality and the incidence of footpad dermatitis. Additional research could investigate the relationships between dietary concentrations of key minerals and the application of exogenous enzymes with litter quality.