A population of European rabbits (Oryctolagus cuniculus) has been monitored since November 1996 through mark–recapture as part of a longitudinal epidemiological study into two Australian rabbit biocontrol agents, rabbit haemorrhagic disease (RHD) and myxomatosis. A female rabbit, first captured as a subadult in late November 1999, was recaptured 18 times before its final capture at the end of February 2007. The longevity of this rabbit, being from its calculated birth date to the date it was last captured, was 7.6 years. A review of the literature indicates this to be the longest lifespan recorded for a European rabbit in the wild.

Surveillance of wildlife virus impacts can be passive or active. Both approaches have their strengths and weaknesses, especially regarding cost and knowledge that can be gained. Monitoring of rabbit haemorrhagic disease virus (GI.1 and GI.2) in South Australia has utilised both strategies and their methods and gained insights are discussed. Active strategies to monitor the continuing impact of rabbit haemorrhagic disease virus 2 (GI.2) on susceptible lagomorphs in countries such as the USA, Mexico, South Africa, Spain, France and Portugal are encouraged to gain critical insights into the evolution, spread and impact of this virus. Furthermore, there are lessons here for the international monitoring of diseases in wildlife, particularly where there is a risk of them becoming zoonotic.

Lagovirus europaeus GI. 2, also commonly known as rabbit haemorrhagic disease virus 2, was first detected at two long-term monitoring sites for European rabbits, Oryctolagus cuniculus, in South Australia, in mid-2016. Numbers of rabbits in the following 12-18 months were reduced to approximately 20 per cent of average numbers in the preceding 10 years. The impact recorded at the two South Australian sites, if widespread in Australia and persistent for several years, is likely to be of enormous economic and environmental benefit.

1Global Ecology, College of Science and Engineering, Flinders University, Adelaide, South Australia, Australia 2School of Mathematical Sciences, University of Adelaide, Adelaide, South Australia, Australia 3Biosecurity South Australia, Department of Primary Industries and Regions, Adelaide, South Australia, Australia 4School of Biological Sciences, University of Adelaide, Adelaide, South Australia, Australia 5Institute for Applied Ecology, University of Canberra, Canberra, ACT, Australia

Veterinary RecordVolume 180, Issue 11 p. 280-280 Research RHDV2 overcoming RHDV immunity in wild rabbits (Oryctolagus cuniculus) in Australia D. Peacock B Ap Sc, DipNatRes, PhD, Corresponding Author D. Peacock B Ap Sc, DipNatRes, PhD David.Peacock@sa.gov.au Biosecurity SA, Adelaide, SA, AustraliaE-mail for correspondence: David.Peacock@sa.gov.auSearch for more papers by this authorJ. Kovaliski BSc, J. Kovaliski BSc Biosecurity SA, Adelaide, SA, AustraliaSearch for more papers by this authorR. Sinclair BSc, MSc, PhD, R. Sinclair BSc, MSc, PhD The University of Adelaide, Adelaide, SA, AustraliaSearch for more papers by this authorG. Mutze BSc, G. Mutze BSc Biosecurity SA, Adelaide, SA, AustraliaSearch for more papers by this authorA. Iannella BSc(Hons), A. Iannella BSc(Hons) The University of Adelaide, Adelaide, SA, AustraliaSearch for more papers by this authorL. Capucci BSc, L. Capucci BSc Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia, Brescia, ItalySearch for more papers by this author D. Peacock B Ap Sc, DipNatRes, PhD, Corresponding Author D. Peacock B Ap Sc, DipNatRes, PhD David.Peacock@sa.gov.au Biosecurity SA, Adelaide, SA, AustraliaE-mail for correspondence: David.Peacock@sa.gov.auSearch for more papers by this authorJ. Kovaliski BSc, J. Kovaliski BSc Biosecurity SA, Adelaide, SA, AustraliaSearch for more papers by this authorR. Sinclair BSc, MSc, PhD, R. Sinclair BSc, MSc, PhD The University of Adelaide, Adelaide, SA, AustraliaSearch for more papers by this authorG. Mutze BSc, G. Mutze BSc Biosecurity SA, Adelaide, SA, AustraliaSearch for more papers by this authorA. Iannella BSc(Hons), A. Iannella BSc(Hons) The University of Adelaide, Adelaide, SA, AustraliaSearch for more papers by this authorL. Capucci BSc, L. Capucci BSc Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia, Brescia, ItalySearch for more papers by this author First published: 18 March 2017 https://doi.org/10.1136/vr.104135Citations: 6 D. Peacock, J. Kovaliski, G. Mutze, are also at Invasive Animals Cooperative Research Centre, University of Canberra, Bruce, ACT, Australia Provenance: not commissioned; externally peer reviewed Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article.Citing Literature Volume180, Issue11March 2017Pages 280-280 RelatedInformation

Deciphering the genes involved in disease resistance is essential if we are to understand host-pathogen coevolutionary processes. The rabbit haemorrhagic disease virus (RHDV) was imported into Australia in 1995 as a biocontrol agent to manage one of the most successful and devastating invasive species, the European rabbit (Oryctolagus cuniculus). During the first outbreaks of the disease, RHDV caused mortality rates of up to 97%. Recently, however, increased genetic resistance to RHDV has been reported. Here, we have aimed to identify genomic differences between rabbits that survived a natural infection with RHDV and those that died in the field using a genomewide next-generation sequencing (NGS) approach. We detected 72 SNPs corresponding to 133 genes associated with survival of a RHD infection. Most of the identified genes have known functions in virus infections and replication, immune responses or apoptosis, or have previously been found to be regulated during RHD. Some of the genes identified in experimental studies, however, did not seem to play a role under natural selection regimes, highlighting the importance of field studies to complement the genomic background of wildlife diseases. Our study provides a set of candidate markers as a tool for the future scanning of wild rabbits for their resistance to RHDV. This is important both for wild rabbit populations in southern Europe where RHD is regarded as a serious problem decimating the prey of endangered predator species and for assessing the success of currently planned RHDV variant biocontrol releases in Australia.

The rabbit haemorrhagic disease virus (RHDV) was imported into Australia in 1995 as a biocontrol agent to manage one of the most successful and devastating invasive species, the European rabbit (Oryctolagus cuniculus cuniculus). During the first disease outbreaks, RHDV caused mortality rates of up to 97% and reduced Australian rabbit numbers to very low levels. However, recently increased genetic resistance to RHDV and strong population growth has been reported. Major histocompatibility complex (MHC) class I immune genes are important for immune responses against viruses, and a high MHC variability is thought to be crucial in adaptive processes under pathogen-driven selection. We asked whether strong population bottlenecks and presumed genetic drift would have led to low MHC variability in wild Australian rabbits, and if the retained MHC variability was enough to explain the increased resistance against RHD. Despite the past bottlenecks we found a relatively high number of MHC class I sequences distributed over 2–4 loci. We identified positive selection on putative antigen-binding sites of the MHC. We detected evidence for RHDV-driven selection as one MHC supertype was negatively associated with RHD survival, fitting expectations of frequency-dependent selection. Gene duplication and pathogen-driven selection are possible (and likely) mechanisms that maintained the adaptive potential of MHC genes in Australian rabbits. Our findings not only contribute to a better understanding of the evolution of invasive species, they are also important in the light of planned future rabbit biocontrol in Australia.

ABSTRACTThe effectiveness of invasive species control can be influenced by seasonal fluctuations in reproduction in response to environmental conditions. However, it is difficult to determine how demography and environmental conditions affect the efficacy of different control efforts from field trials alone. We incorporated an ontogenetic growth model into a hierarchical Bayesian mark‐recapture model to estimate age‐structured seasonal survival rates for European rabbits (Oryctolagus cuniculus) in Australia, based on a >15‐year data set. We integrated this demographic information into an individual‐based simulation model, which reproduces seasonal birth‐death processes, to test the effectiveness of pest‐management schemes that differed in intensity, specificity to age groups, and seasonal timing. Control measures that were simulated to affect only juveniles had a negligible effect on population size, whereas targeting subadults and adults led to considerable population declines when applied after the breeding season. Management that affected rabbits of all age groups caused significant population reductions. However, even repeated control efforts that caused 95% mortality each year only resulted in predictions of local population extirpation after an average of 119 calendar weeks in the absence of immigration. Our simulation study supports the use of pest rabbit control methods that account for demographic dynamics explicitly, and target those individuals with high reproductive potential. More broadly, we show that local and temporal population extirpation, or recovery, depends largely on the trade‐off between control intensity and frequency for species with recurrent population oscillations. © 2016 The Wildlife Society.

Mitochondria are critical for life, yet their underlying evolutionary biology is poorly understood. In particular, little is known about interaction between two levels of evolution: between individuals and within individuals (competition between cells, mitochondria or mitochondrial DNA molecules). Rapid evolution is suspected to occur frequently in mitochondrial DNA, whose maternal inheritance predisposes advantageous mutations to sweep rapidly though populations. Rapid evolution is also predicted in response to changed selection regimes after species invasion or removal of pathogens or competitors. Here, using empirical and simulated data from a model invasive bird species, we provide the first demonstration of rapid selection on the mitochondrial genome within individuals in the wild. Further, we show differences in mitochondrial DNA copy number associated with competing genetic variants, which may provide a mechanism for selection. We provide evidence for three rarely documented phenomena: selection associated with mitochondrial DNA abundance, selection on the mitochondrial control region, and contemporary selection during invasion.

Infectious diseases can exert a strong influence on the dynamics of host populations, but it remains unclear why such disease-mediated control only occurs under particular environmental conditions. We used 16 years of detailed field data on invasive European rabbits (Oryctolagus cuniculus) in Australia, linked to individual-based stochastic models and Bayesian approximations, to test whether (i) mortality associated with rabbit haemorrhagic disease (RHD) is driven primarily by seasonal matches/mismatches between demographic rates and epidemiological dynamics and (ii) delayed infection (arising from insusceptibility and maternal antibodies in juveniles) are important factors in determining disease severity and local population persistence of rabbits. We found that both the timing of reproduction and exposure to viruses drove recurrent seasonal epidemics of RHD. Protection conferred by insusceptibility and maternal antibodies controlled seasonal disease outbreaks by delaying infection; this could have also allowed escape from disease. The persistence of local populations was a stochastic outcome of recovery rates from both RHD and myxomatosis. If susceptibility to RHD is delayed, myxomatosis will have a pronounced effect on population extirpation when the two viruses coexist. This has important implications for wildlife management, because it is likely that such seasonal interplay and disease dynamics has a strong effect on long-term population viability for many species.

Although population genetic theory is largely based on the premise that loci under study are selectively neutral, it has been acknowledged that the study of DNA sequence data under the influence of selection can be useful. In some circumstances, these loci show increased population differentiation and gene diversity. Highly polymorphic loci may be especially useful when studying populations having low levels of diversity overall, such as is often the case with threatened or newly established invasive populations. Using common starlings Sturnus vulgaris sampled from invasive Australian populations, we investigated sequence data of the dopamine receptor D4 gene (DRD4), a locus suspected to be under selection for novelty-seeking behaviour in a range of taxa including humans and passerine birds. We hypothesised that such behaviour may be advantageous when species encounter novel environments, such as during invasion. In addition to analyses to detect the presence of selection, we also estimated population differentiation and gene diversity using DRD4 data and compared these estimates to those from microsatellite and mitochondrial DNA sequence data, using the same individuals. We found little evidence for selection on DRD4 in starlings. However, we did find elevated levels of within-population gene diversity when compared to microsatellites and mitochondrial DNA sequence, as well as a greater degree of population differentiation. We suggest that sequence data from putatively nonneutral loci are a useful addition to studies of invasive populations, where low genetic variability is expected.

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 Australia's remaining natural biodiversity.

The frequency and timing of rabbit haemorrhagic disease (RHD) epizootics and their impact on different age groups of rabbits were studied for 15 years in a recovering rabbit population in South Australia. We recorded the number and body size of rabbits dying during RHD epizootics, collected tissue for genetic analysis of rabbit haemorrhagic disease virus variants and compared the number of carcasses found to the number of susceptible rabbits present at the beginning of each epizootic. All RHD epizootics occurred between late winter and spring, but, progressively, epizootics started earlier and became more frequent and prolonged, fewer susceptible adult rabbits were present during epizootics, and the age of rabbits dying of RHD declined. Increased infection and virus shedding in juvenile rabbits offers the most plausible explanation for those epidemiological changes; the disease is now increasingly transmitted through populations of kittens, starting before young-of-the-year reach adult size and persisting late in the breeding season, so that most rabbits are challenged in their year of birth. These changes have increased juvenile mortality due to RHD but reduced total mortality across all age groups, because age-specific mortality rates are lower in young rabbits than in older rabbits. We hypothesise that this may be the proximate cause of recovery in rabbit populations across Australia and possibly elsewhere.

Rabbit Haemorrhagic Disease Virus (RHDV) was introduced into Australia in 1995 as a biological control agent against the wild European rabbit (Oryctolagus cuniculus). We evaluated its evolution over a 16‐year period (1995–2011) by examining 50 isolates collected throughout Australia, as well as the original inoculum strains. Phylogenetic analysis of capsid protein VP60 sequences of the Australian isolates, compared with those sampled globally, revealed that they form a monophyletic group with the inoculum strains (CAPM V‐351 and RHDV351INOC). Strikingly, despite more than 3000 rereleases of RHDV351INOC since 1995, only a single viral lineage has sustained its transmission in the long‐term, indicative of a major competitive advantage. In addition, we find evidence for widespread viral gene flow, in which multiple lineages entered individual geographic locations, resulting in a marked turnover of viral lineages with time, as well as a continual increase in viral genetic diversity. The rate of RHDV evolution recorded in Australia −4.0 (3.3–4.7) × 10−3 nucleotide substitutions per site per year – was higher than previously observed in RHDV, and evidence for adaptive evolution was obtained at two VP60 residues. Finally, more intensive study of a single rabbit population (Turretfield) in South Australia provided no evidence for viral persistence between outbreaks, with genetic diversity instead generated by continual strain importation.

Large-scale international monitoring studies are important to assess emission patterns and environmental distributions of organohalogenated contaminants (OHCs) on a worldwide scale. In this study, the presence of OHCs was investigated on three continents (Europe, North America and Australasia), using eggs of starlings (Sturnus vulgaris and Sturnus unicolor) to assess their suitability for large-scale monitoring studies. To the best of our knowledge, this is the first study using bird eggs of the same species as a biomonitor for OHCs on an intercontinental scale. We found significant differences in OHC concentrations of the eggs among sampling locations, except for hexachlorocyclohexanes (HCHs). Mean concentrations of sum polychlorinated biphenyls (PCBs) in eggs ranged from 78 ± 26 ng/g lipid weight (lw) in Australia to 2900 ± 1300 ng/g lw in the United States. The PCB profile was dominated by CB 153 and CB 138 in all locations, except for New Zealand, where the contribution of CB 95, CB 101 and CB 149 was also high. The highest mean sum polybrominated diphenyl ether (PBDE) concentrations were found in Canada (4400 ± 830 ng/g lw), while the lowest mean PBDE concentrations were measured in Spain (3.7 ± 0.1 ng/g lw). The PBDE profile in starling eggs was dominated by BDE 47 and BDE 99 in all countries, but in Belgium, the higher brominated PBDEs had a higher contribution compared to other countries. For the organochlorine pesticides (OCPs), dichlorodiphenyltrichloroethanes (DDTs) ranged from 110 ± 16 ng/g lw in France to 17,000 ± 3400 ng/g lw in New Zealand, while HCHs and hexachlorobenzene were generally in low concentrations in all sampling locations. Chlordanes were remarkably high in eggs from the United States (2500 ± 1300 ng/g lw). The OCP profile in all countries was largely dominated by p,p′-DDE. In general, the worldwide trends we observed in starling eggs were in accordance with the literature on human and environmental OHC data, which suggests that there is potential for using starling eggs as a biomonitoring tool on a large geographical scale.

The European rabbit, Oryctolagus cuniculus, is threatened within its native range, yet it is a highly successful colonizing pest species across its worldwide introduced range, causing large economic losses and widespread environmental degradation. To date, there has been no long-term empirical evidence documenting the relative roles of climatic, epidemiological and biological factors in limiting life-history determinants of rabbit range and abundance. Using 12 years of capturemarkrecapture data from their exotic range in Australia, we constructed candidate CormackJollySeber models to test the influence of environmental, competition and disease conditions on rabbit survival and recruitment. Our results show that: (i) population-level disease infection rate has the largest overall impact on rabbit survival, explaining 80% of variance in survival rates; (ii) environmental as well as epidemiological conditions constrain rabbit survival, especially for younger animals; (iii) temporal variation in rabbit kitten recruitment patterns are best described by a combination of climate, competition and disease settings (accounting for 68% of variance), while temperature alone has a strong negative influence on kitten recruitment; and (iv) recruitment responds positively to rabbit haemorrhagic disease, but negatively to myxomatosis the former, probably being mediated through a disease driven effect on intraspecific competition for food. A strengthened understanding of climate change impacts on rabbit range and abundance can be achieved by accounting explicitly for potential synergisms between disease dynamics and climate. In this analysis, we provide the first step towards such an attempt for this important mammal species. Integrated approaches of this kind are essential for future forecasts of rabbit range and abundance, offsetting the conservation threat faced by O. cuniculus in its native range, and achieving effective management in exotic habitats.

Rabbit haemorrhagic disease virus (RHDV) is a highly virulent lagovirus endemic in Europe and Australasian populations of the European rabbit, Oryctolagus cuniculus. It has also caused several unexplained disease outbreaks in domestic European rabbits in North America. Non‐pathogenic spread of RHDV leading to persistent infection which later reactivated has recently been proposed as the cause of overt disease and death of a pet rabbit in Canada, the first confirmed case of Rabbit haemorrhagic disease in that country. We suggest that there is little evidence to support non‐pathogenic spread of virulent RHDV, some evidence that is contradictory, and evidence to support a simpler alternative hypothesis. RHDV can be spread over long distances between sparse rabbit populations by fomites or flying insects. Although highly pathogenic, RHDV can be limited in its spread within rabbit populations, or its presence masked by closely related but non‐pathogenic lagoviruses which can provide protection against acute disease. In the absence of any evidence from clinical studies to support reactivation of persistent RHDV infection, the simpler explanation seems more likely to be correct.

Abstract Context. Worldwide, invasive fauna species present one of the most intractable problems for agriculture and natural systems. Our ability to improve control techniques to combat the global invasive species predicament is constrained within the bounds of both economic and ethical considerations. In south-eastern Australia, the common starling (Sturnus vulgaris) is an established invasive avian pest that is now making incursions into areas of Western Australia (WA) that are currently free of this species. The most cost-effective and widely implemented starling control tool is trapping with live-lure birds. In recent years, the use of live-lure birds has been questioned on both economic and ethical grounds, and consequently alternative lure methods need investigating. Aims. To evaluate the effectiveness of different trap and lure combinations for the capture of starlings in western South Australia (SA). Methods. Modified Australian Crow (MAC) traps, used traditionally in WA to trap starlings, and Myna traps, originally designed for trapping common mynas (Sturnus tristis), were set during the peaks in starling flocking activity (Austral summer, 2007) using three different lure types: (1) live lure (live starlings); (2) moving water; and (3) acoustic lures. A trapping grid consisting of a single Myna trap with live lure and three MAC traps, each with one type of lure (live, water or acoustic) was established at five sites on the Eyre Peninsula in SA and monitored twice daily for 28 days. Key results. Live lures were significantly more effective at attracting starlings into traps compared with both water and sound lures. We also trapped at an additional three sites and showed that Myna traps caught ∼1.5 times more starlings than MAC traps when both traps were fitted with live-lure birds. Conclusions. Neither moving water nor acoustic play-back lures proved suitable replacements for the use of live-lure birds to capture starlings. The efficacy of alternative lure types may depend on several factors and may include neophobic response(s) to novel signals and also the length of time that an invasive population has been established. Implications. We recommend that use of live lures is continued in ongoing starling control programs, and that MAC traps currently in use be modified to capitalise on known starling behaviour. Further research and development of traps that do not contain live lures will improve the welfare of invasive species control programs.

Mitochondrial DNA (mtDNA) can be a powerful genetic marker for tracing origins and history of invasive populations. Here, we use mtDNA to address questions relevant to the understanding of invasion pathways of common starlings (Sturnus vulgaris) into Western Australia (WA) and discuss the utility of this marker to provide information useful to invasive species management. Mitochondrial sequence data indicate two geographically restricted genetic groups within Australia. Evidence of dispersal from genetically distinct sources outside the sampled range of starlings in Australia suggests increased vigilance by management agencies may be required to prevent further incursions from widely separated localities. Overall, genetic diversity in Australia was lower than in samples from the native range. Within Australia, genetic diversity was lowest in the most recently colonized area in the west, indicating that demographic bottlenecks have occurred in this area. Evidence of restricted dispersal between localities on the edge of the range expansion (ERE) in WA and other Australian sampling localities suggests that localized control within the ERE may be effective in preventing further range expansion. Signatures of spatial and demographic expansion are present in mismatch analyses from sampling localities located at the ERE, but neutrality indices did not support this finding, suggesting that the former may be more sensitive to recent expansion. Additionally, mismatch analyses support the presence of admixture, which is likely to have occurred pre-introduction. We compare our findings with those from a microsatellite study of the same samples and discuss how the mtDNA analyses used here offer valuable and unique insights into the invasion history of introduced species.

Context. European rabbits are serious environmental and agricultural pests throughout their range in Australia. Rabbit haemorrhagic disease virus (RHDV) greatly reduced rabbit numbers in arid central Australia but had less impact in cooler, higher-rainfall areas. RHDV-like benign caliciviruses (bCVs) have been implicated in limiting the impact of RHDV in the higher-rainfall regions of Australia and also in Europe. Aims. Experimental releases of RHDV on bait were tested as a means of initiating disease outbreaks. Serological evidence of antibodies to bCVs was examined to determine whether they reduce mortality rates and/or spread of the released RHDV, and how that might influence the effectiveness of future RHDV releases for rabbit management. Methods. Four experimental releases were conducted in high-rainfall and coastal regions of southern Australia. Virus activity was implied from recapture rates and serological changes in marked rabbits, and genetic sequencing of virus recovered from dead rabbits. Changes in rabbit abundance were estimated from spotlight transect counts. Key results. Release of RHDV on bait produced disease outbreaks that challenged almost all animals within the general release area and spread up to 4 km beyond the release sites. Recapture rates were high in marked rabbits that possessed antibodies from previous exposure to RHDV and extremely low amongst rabbits that lacked any detectable antibodies. Rabbits carrying antibodies classified as being due to previous infection with bCVs had recapture rates that were dependent on circulating antibody titre and were ~55% of recapture rates in rabbits with clear antibodies to RHDV. Conclusions. This is the first quantified evidence that antibodies produced against bCVs provide significant protection against RHD outbreaks in field populations of rabbits. Implications. bCVs can greatly reduce the impact of RHDV on wild-rabbit populations in Australia and presumably elsewhere. RHDV can be effectively released on bait although further releases are likely to be of minor or inconsistent benefit for controlling rabbit numbers where bCVs are common.