Brian Cooke

Increased virulence of rabbit haemorrhagic disease virus associated with genetic resistance in wild Australian rabbits (Oryctolagus cuniculus)

The release of myxoma virus (MYXV) and Rabbit Haemorrhagic Disease Virus (RHDV) in Australia with the aim of controlling overabundant rabbits has provided a unique opportunity to study the initial spread and establishment of emerging pathogens, as well as their co-evolution with their mammalian hosts. In contrast to MYXV, which attenuated shortly after its introduction, rapid attenuation of RHDV has not been observed. By studying the change in virulence of recent field isolates at a single field site we show, for the first time, that RHDV virulence has increased through time, likely because of selection to overcome developing genetic resistance in Australian wild rabbits. High virulence also appears to be favoured as rabbit carcasses, rather than diseased animals, are the likely source of mechanical insect transmission. These findings not only help elucidate the co-evolutionary interaction between rabbits and RHDV, but reveal some of the key factors shaping virulence evolution.

Rabbit haemorrhagic disease: are Australian rabbits (Oryctolagus cuniculus) evolving resistance to infection with Czech CAPM 351 RHDV?

Rabbit haemorrhagic disease is a major tool for the management of introduced, wild rabbits in Australia. However, new evidence suggests that rabbits may be developing resistance to the disease. Rabbits sourced from wild populations in central and southeastern Australia, and domestic rabbits for comparison, were experimentally challenged with a low 60 ID50 oral dose of commercially available Czech CAPM 351 virus - the original strain released in Australia. Levels of resistance to infection were generally higher than for unselected domestic rabbits and also differed (0-73% infection rates) between wild populations. Resistance was lower in populations from cooler, wetter regions and also low in arid regions with the highest resistance seen within zones of moderate rainfall. These findings suggest the external influences of non-pathogenic calicivirus in cooler, wetter areas and poor recruitment in arid populations may influence the development rate of resistance in Australia.

Antibody status and survival of Australian wild rabbits challenged with rabbit haemorrhagic disease virus

In Australia, the epidemiology of rabbit haemorrhagic disease virus (RHDV) is complicated by non-pathogenic forms of calicivirus (bCV) co-circulating with RHDV and providing variable protection from RHDV. Currently no bCV virus-specific antibody tests exist; however, a series of four ELISAs used to detect antibodies to RHDV provided an indirect means to detect antibodies to bCV, enabling antibody categories of seronegative, maternal RHDV, RHDV or bCV to be determined. Rabbits (188) from four locations were challenged with RHDV and logistic regression models determined that, for rabbits <15 months old, survival was dependent on antibody titres alone and the relationship did not vary with age, capture site, gender, liveweight or reproductive status. All rabbits survived challenge after reaching 15 months of age, irrespective of their antibody titres. Where bCV antibodies were prevalent in young rabbits, the bCV category did not adequately summarise all information about rabbit survival that can be obtained from antibody titres. Within antibody categories, 95% of rabbits with RHDV, 33% with bCV, 40% with maternal RHDV and 22% with seronegative antibodies survived. The high survival rate of adults implies that natural outbreaks or controlled releases of RHDV will have little impact on adult breeding rabbits. Therefore, where RHDV and bCV are endemic, conventional rabbit-control programs targeting the immune breeding populations should provide the most predictable outcome for long-term maintenance of low rabbit populations.

A rapid survey method for estimating population density of European rabbits living in native vegetation

European rabbits are severe environmental pests in Australia but reporting of density-damage relationships has been hindered by a lack of simple methods to estimate the density of rabbit populations in native vegetation. A methodology for quantifying rabbit densities suitable for use in sparse populations of rabbits in conjunction with surveys of the condition of native vegetation is proposed. Dung pellets were counted in 11 629 regularly-spaced quadrats of 0.1 m2 in semiarid, coastal and cool-temperate areas of southern Australia. Mean pellet counts in latrines and the relationship between dung counts outside of latrines and the proportion of quadrats falling on latrines were quantified. This allowed density of dung pellets to be estimated by using a correction factor for latrines instead of attempting to count all pellets within quadrats that included parts of latrines. Rabbit density was calculated from pellet density based on mean pellet size, pellet breakdown rate and estimates of rabbits’ dry matter intake and digestive efficiency. Results were validated against estimates of rabbit density from long-term studies using a combination of spotlight transect counts and burrow entrance counts. The proposed methodology allows estimates of rabbit density in native vegetation to be obtained from just a few hours work and can be used in conjunction with surveys of the condition of native vegetation to quantify rabbit impacts. This methodology is seen as particularly useful in providing a tool to allow rabbit densities to be estimated and then compared with the thresholds, determined separately, at which damage occurs for given ecosystems.

Recovery of South Australian rabbit populations from the impact of rabbit haemorrhagic disease

Abstract Context. Recovery of Australian rabbit populations from the impact of rabbit haemorrhagic disease virus (RHDV) contrasts with more prolonged suppression of wild rabbits in Europe, and has been widely discussed in the scientific community, but not yet documented in formal scientific literature. The underlying causes of recovery remain unclear, but resistance to RHDV infection has been reported in laboratory studies of wild-caught rabbits. Aims. We document numerical changes in two South Australian wild rabbit populations that were initially suppressed by RHDV, and examine serological data to evaluate several alternative hypotheses for the cause of recovery. Methods. Rabbit numbers were assessed from spotlight transect counts and dung mass transects between 1991 and 2011, and age and RHDV antibody sero-prevalence were estimated from rabbits shot in late summer. Key results. Rabbit numbers were heavily suppressed by RHDV between 1995 and 2002, then increased 5- to 10-fold between 2003 and 2010. During the period of increase, annual RHDV infection rates remained stable or increased slightly, average age of rabbits remained stable and annual rainfall was below average. Conclusions. Rabbit populations recovered but neither avoidance of RHDV infection, gradual accumulation of long-lived RHD-immune rabbits, nor high pasture productivity were contributing factors. This leaves increased annual survival from RHDV infection as the most likely cause of recovery. Implications. Previously documented evidence of resistance to RHDV infection may be of little consequence to post-RHD recovery in rabbit numbers, unless the factors that influence the probability of infection also shape the course of infection and affect survival of infected rabbits.

Rabbit haemorrhagic disease: virus persistence and adaptation in Australia.

In Australia, the rabbit haemorrhagic disease virus (RHDV) has been used since 1996 to reduce numbers of introduced European rabbits (Oryctolagus cuniculus) which have a devastating impact on the native Australian environment. RHDV causes regular, short disease outbreaks, but little is known about how the virus persists and survives between epidemics. We examined the initial spread of RHDV to show that even upon its initial spread, the virus circulated continuously on a regional scale rather than persisting at a local population level and that Australian rabbit populations are highly interconnected by virus‐carrying flying vectors. Sequencing data obtained from a single rabbit population showed that the viruses that caused an epidemic each year seldom bore close genetic resemblance to those present in previous years. Together, these data suggest that RHDV survives in the Australian environment through its ability to spread amongst rabbit subpopulations. This is consistent with modelling results that indicated that in a large interconnected rabbit meta‐population, RHDV should maintain high virulence, cause short, strong disease outbreaks but show low persistence in any given subpopulation. This new epidemiological framework is important for understanding virus–host co‐evolution and future disease management options of pest species to secure Australias remaining natural biodiversity.

Establishing a serological surveillance protocol for rabbit hemorrhagic disease by combining mathematical models and field data: implication for rabbit conservation

Rabbit hemorrhagic disease (RHD) became endemic in wild rabbit (Oryctolagus cuniculus) populations in the Iberian Peninsula after its first arrival in 1988. This had significant implications for both the economy and environmental conservation because rabbits are one of the main game species in the Iberian Peninsula and a keystone species in the Mediterranean ecosystems. As a consequence, it is planned to include RHD surveillance in the Spanish Wildlife Disease Surveillance Strategy. Nevertheless, there is no practical methodology included in this program to help conservationists and gamekeepers understand the impact of disease on wild rabbit populations. Results from sera collected during the hunting season from 11 rabbit populations of Central and South Spain, which differed in their population abundance and trends, allowed us to use mathematical models to interpret the serological results gathered and determine the best strategy for finalizing a plan of RHD surveillance. Put simply, we focused our field surveys within the hunting season (October-January), and those times when the rabbit populations are at their highest (June or July). Field results showed that both rabbit abundance and population trend are closely related to the prevalence of RHD antibodies when rabbit abundance was at its annual low point (usually October-November). Rabbit population trends were positive only if antibody prevalence was high (>40%), and always negative if prevalence was low. Moreover, rabbit populations where abundance was low always showed low antibody prevalence. Since our models predicted a low variability in the prevalence obtained during the hunting season, it is suggested that future serological surveys should be carried out within this period to avoid problems related to the low sample size in low density rabbit populations.

Daily food intake of free-ranging wild rabbits in semiarid South Australia

Abstract Context. Although the daily food intake of wild rabbits is broadly known, precise field estimates have not been made. Moreover, regressions based on metabolic body size and food intake of grazing herbivores in general are too imprecise to provide close predictions. Using such values could result in substantial errors when estimating grazing equivalents to sheep or cattle and resultant economic losses, or estimating the numbers of rabbits per hectare based on rates of deposition of faeces. Aim. To re-analyse previously collected data on estimated food digestibility and food intake of rabbits, and provide a framework for better estimating economic and conservation losses attributable to wild rabbits. Methods. Food intake of wild free-ranging rabbits was calculated from past measurements of water turnover obtained from dilution of injected tritiated water and estimates of the water content and digestibility of the food eaten. Key results. During spring, male rabbits were estimated to eat 65.7 ± 12.5 g dry matter per corrected bodyweight (W–0.75) per day and lactating females ate 97.0 ± 19.4 g dry matter W–0.75 day–1. Similar results were obtained on repetition of trials at a second field site and from wild rabbits held in captivity. Conclusions. The estimates of food digestibility and intake obtained, although not precise, are an improvement on theoretical expectations alone and help put previously published data in better perspective. Implications. Improved estimates of food consumption provide more confidence in estimates of rabbit grazing pressure. Even moderate densities of rabbits (5 rabbits per ha) could remove about half the pasture produced in an average year in Australia’s arid-zone.

The future biological control of pest populations of European rabbits, Oryctolagus cuniculus

European rabbits are exotic pests in Australia, New Zealand, parts of South America and Europe, and on many islands. Their abundance, and the damage they cause, might be reduced by the release of naturally occurring or genetically modified organisms (GMOs) that act as biological control agents (BCAs). Some promising pathogens and parasites of European rabbits and other lagomorphs are discussed, with special reference to those absent from Australia as an example of the range of necessary considerations in any given case. The possibility of introducing these already-known BCAs into areas where rabbits are pests warrants further investigation. The most cost-effective method for finding potentially useful but as-yet undiscovered BCAs would be to maintain a global watch on new diseases and pathologies in domestic rabbits. The absence of wild European rabbits from climatically suitable parts of North and South America and southern Africa may indicate the presence there of useful BCAs, although other explanations for their absence are possible. Until the non-target risks of deploying disseminating GMOs to control rabbits have been satisfactorily minimised, efforts to introduce BCAs into exotic rabbit populations should focus on naturally occurring organisms. The development of safe disseminating GMOs remains an important long-term goal, with the possible use of homing endonuclease genes warranting further investigation.

Estimating density-dependent impacts of European rabbits on Australian tree and shrub populations

Introduced European rabbits, Oryctolagus cuniculus, can severely damage Australian native vegetation but the problem is difficult to quantify because simple methods to estimate rabbit impacts are lacking. Management decision-making is often uncertain because of unknown relationships between rabbit density and damage. We tested simple quantitative sampling methods using belt transects to detect differences in critical characteristics of perennial vegetation communities affected by rabbit browsing: damage to individual juvenile shrubs and trees, and loss of recruitment cohorts. Rabbit density and relative abundance of larger herbivores were estimated from dung pellet density. The prevalence of identifiable rabbit browse on juvenile plants increased with increasing rabbit density and was higher for plant species considered by previous authors to be highly palatable than for moderately palatable or unpalatable species. At densities of ≥0.5 rabbits ha–1, highly palatable plant species were severely damaged as juveniles and cohorts in 0.3–1.0 m height classes and 5–20 mm basal diameter classes were missing. Similar damage became apparent in moderately palatable species at 2 rabbits ha–1 but was rarely seen in unpalatable species. Within species, size cohort evenness was inversely related to the proportion of surviving juveniles with identifiable rabbit damage. The effect of rabbits on native vegetation condition can be recorded in a simple manner suitable for identifying density-damage relationships and changes in vegetation condition over time. It is particularly useful in setting target densities below which rabbits must be managed to maintain natural plant recruitment and ecosystem function in conservation reserves and pastoral grazing properties of southern Australia.