The potential of genomics for restoring ecosystems and biodiversity.

Billions of hectares of natural ecosystems have been degraded through human actions. The global community has agreed on targets to halt and reverse these declines, and the restoration sector faces the important but arduous task of implementing programmes to meet these objectives. Existing and emerging genomics tools offer the potential to improve the odds of achieving these targets. These tools include population genomics that can improve seed sourcing, meta-omics that can improve assessment and monitoring of restoration outcomes, and genome editing that can generate novel genotypes for restoring challenging environments. We identify barriers to adopting these tools in a restoration context and emphasize that regulatory and ethical frameworks are required to guide their use.

Leveraging a natural murine meiotic drive to suppress invasive populations.

Invasive rodents are a major cause of environmental damage and biodiversity loss, particularly on islands. Unlike insects, genetic biocontrol strategies including population-suppressing gene drives with biased inheritance have not been developed in mice. Here, we demonstrate a gene drive strategy (tCRISPR) that leverages super-Mendelian transmission of the t haplotype to spread inactivating mutations in a haplosufficient female fertility gene (Prl). Using spatially explicit individual-based in silico modeling, we show that tCRISPR can eradicate island populations under a range of realistic field-based parameter values. We also engineer transgenic tCRISPR mice that, crucially, exhibit biased transmission of the modified t haplotype and Prl mutations at levels our modeling predicts would be sufficient for eradication. This is an example of a feasible gene drive system for invasive alien rodent population control.

Environment and shipping drive environmental DNA beta-diversity among commercial ports.

The spread of nonindigenous species by shipping is a large and growing global problem that harms coastal ecosystems and economies and may blur coastal biogeographical patterns. This study coupled eukaryotic environmental DNA (eDNA) metabarcoding with dissimilarity regression to test the hypothesis that ship-borne species spread homogenizes port communities. We first collected and metabarcoded water samples from ports in Europe, Asia, Australia and the Americas. We then calculated community dissimilarities between port pairs and tested for effects of environmental dissimilarity, biogeographical region and four alternative measures of ship-borne species transport risk. We predicted that higher shipping between ports would decrease community dissimilarity, that the effect of shipping would be small compared to that of environment dissimilarity and shared biogeography, and that more complex shipping risk metrics (which account for ballast water and stepping-stone spread) would perform better. Consistent with our hypotheses, community dissimilarities increased significantly with environmental dissimilarity and, to a lesser extent, decreased with ship-borne species transport risks, particularly if the ports had similar environments and stepping-stone risks were considered. Unexpectedly, we found no clear effect of shared biogeography, and that risk metrics incorporating estimates of ballast discharge did not offer more explanatory power than simpler traffic-based risks. Overall, we found that shipping homogenizes eukaryotic communities between ports in predictable ways, which could inform improvements in invasive species policy and management. We demonstrated the usefulness of eDNA metabarcoding and dissimilarity regression for disentangling the drivers of large-scale biodiversity patterns. We conclude by outlining logistical considerations and recommendations for future studies using this approach.

Gene drives for vertebrate pest control: Realistic spatial modelling of eradication probabilities and times for island mouse populations.

Invasive alien species continue to threaten global biodiversity. CRISPR-based gene drives, which can theoretically spread through populations despite imparting a fitness cost, could be used to suppress or eradicate pest populations. We develop an individual-based, spatially explicit, stochastic model to simulate the ability of CRISPR-based homing and X chromosome shredding drives to eradicate populations of invasive house mice (Mus muculus) from islands. Using the model, we explore the interactive effect of the efficiency of the drive constructs and the spatial ecology of the target population on the outcome of a gene-drive release. We also consider the impact of polyandrous mating and sperm competition, which could compromise the efficacy of some gene-drive strategies. Our results show that both drive strategies could be used to eradicate large populations of mice. Whereas parameters related to drive efficiency and demography strongly influence drive performance, we find that sperm competition following polyandrous mating is unlikely to impact the outcome of an eradication effort substantially. Assumptions regarding the spatial ecology of mice influenced the probability of and time required for eradication, with short-range dispersal capacities and limited mate-search areas producing 'chase' dynamics across the island characterized by cycles of local extinction and recolonization by mice. We also show that highly efficient drives are not always optimal, when dispersal and mate-search capabilities are low. Rapid local population suppression around the introduction sites can cause loss of the gene drive before it can spread to the entire island. We conclude that, although the design of efficient gene drives is undoubtedly critical, accurate data on the spatial ecology of target species are critical for predicting the result of a gene-drive release.

Rodent gene drives for conservation: opportunities and data needs.

Invasive rodents impact biodiversity, human health and food security worldwide. The biodiversity impacts are particularly significant on islands, which are the primary sites of vertebrate extinctions and where we are reaching the limits of current control technologies. Gene drives may represent an effective approach to this challenge, but knowledge gaps remain in a number of areas. This paper is focused on what is currently known about natural and developing synthetic gene drive systems in mice, some key areas where key knowledge gaps exist, findings in a variety of disciplines relevant to those gaps and a brief consideration of how engagement at the regulatory, stakeholder and community levels can accompany and contribute to this effort. Our primary species focus is the house mouse, Mus musculus, as a genetic model system that is also an important invasive pest. Our primary application focus is the development of gene drive systems intended to reduce reproduction and potentially eliminate invasive rodents from islands. Gene drive technologies in rodents have the potential to produce significant benefits for biodiversity conservation, human health and food security. A broad-based, multidisciplinary approach is necessary to assess this potential in a transparent, effective and responsible manner.

Bonamia in Ostrea angasi: Diagnostic performance, field prevalence and intensity.

Bonamia spp. parasites threaten flat oyster (Ostrea spp.) farming worldwide. Understanding test performance is important for designing surveillance and interpreting diagnostic results. Following a pilot survey which found low Bonamia sp. intensity in farmed Ostrea angasi, we tested further oysters (n = 100-150) from each of three farms for Bonamia sp. using heart smear, histology and qPCR. We used a Bayesian Latent Class Model to assess diagnostic sensitivity (DSe) and specificity (DSp) of these tests individually or in combination, and to assess prevalence. Histology was the best individual test (DSe 0.76, DSp 0.93) compared to quantitative polymerase chain reaction (qPCR) (DSe 0.69, DSp 0.93) and heart smear (DSe 0.61, DSp 0.60). Histology combined with qPCR and defining a positive from either test as an infected case maximized test performance (DSe 0.91, DSp 0.88). Prevalence was higher at two farms in a high-density oyster growing region than at a farm cultivating oysters at lower density. Parasite intensities were lower than in New Zealand and European studies, and this is probably contributed to differences in the performance of test when compared to other studies. Understanding diagnostic test performance in different populations can support the development of improved Bonamia surveillance programs.

High accuracy at low frequency: detailed behavioural classification from accelerometer data.

Accelerometers are a valuable tool for studying animal behaviour and physiology where direct observation is unfeasible. However, giving biological meaning to multivariate acceleration data is challenging. Here, we describe a method that reliably classifies a large number of behaviours using tri-axial accelerometer data collected at the low sampling frequency of 1 Hz, using the dingo (Canis dingo) as an example. We used out-of-sample validation to compare the predictive performance of four commonly used classification models (random forest, k-nearest neighbour, support vector machine, and naïve Bayes). We tested the importance of predictor variable selection and moving window size for the classification of each behaviour and overall model performance. Random forests produced the highest out-of-sample classification accuracy, with our best-performing model predicting 14 behaviours with a mean accuracy of 87%. We also investigated the relationship between overall dynamic body acceleration (ODBA) and the activity level of each behaviour, given the increasing use of ODBA in ecophysiology as a proxy for energy expenditure. ODBA values for our four 'high activity' behaviours were significantly greater than all other behaviours, with an overall positive trend between ODBA and intensity of movement. We show that a random forest model of relatively low complexity can mitigate some major challenges associated with establishing meaningful ecological conclusions from acceleration data. Our approach has broad applicability to free-ranging terrestrial quadrupeds of comparable size. Our use of a low sampling frequency shows potential for deploying accelerometers over extended time periods, enabling the capture of invaluable behavioural and physiological data across different ontogenies.

Dodging silver bullets: good CRISPR gene-drive design is critical for eradicating exotic vertebrates.

Self-replicating gene drives that can spread deleterious alleles through animal populations have been promoted as a much needed but controversial 'silver bullet' for controlling invasive alien species. Homing-based drives comprise an endonuclease and a guide RNA (gRNA) that are replicated during meiosis via homologous recombination. However, their efficacy for controlling wild populations is threatened by inherent polymorphic resistance and the creation of resistance alleles via non-homologous end-joining (NHEJ)-mediated DNA repair. We used stochastic individual-based models to identify realistic gene-drive strategies capable of eradicating vertebrate pest populations (mice, rats and rabbits) on islands. One popular strategy, a sex-reversing drive that converts heterozygous females into sterile males, failed to spread and required the ongoing deployment of gene-drive carriers to achieve eradication. Under alternative strategies, multiplexed gRNAs could overcome inherent polymorphic resistance and were required for eradication success even when the probability of NHEJ was low. Strategies causing homozygotic embryonic non-viability or homozygotic female sterility produced high probabilities of eradication and were robust to NHEJ-mediated deletion of the DNA sequence between multiplexed endonuclease recognition sites. The latter two strategies also purged the gene drive when eradication failed, therefore posing lower long-term risk should animals escape beyond target islands. Multiplexing gRNAs will be necessary if this technology is to be useful for insular extirpation attempts; however, precise knowledge of homing rates will be required to design low-risk gene drives with high probabilities of eradication success.

Environmental effects and individual body condition drive seasonal fecundity of rabbits: identifying acute and lagged processes.

The reproduction of many species is determined by seasonally-driven resource supply. But it is difficult to quantify whether the fecundity is sensitive to short- or long-term exposure to environmental conditions such as rainfall that drive resource supply. Using 25 years of data on individual fecundity of European female rabbits, Oryctolagus cuniculus, from semiarid Australia, we investigate the role of individual body condition, rainfall and temperature as drivers of seasonal and long-term and population-level changes in fecundity (breeding probability, ovulation rate, embryo survival). We built distributed lag models in a hierarchical Bayesian framework to account for both immediate and time-lagged effects of climate and other environmental drivers, and possible shifts in reproduction over consecutive seasons. We show that rainfall during summer, when rabbits typically breed only rarely, increased breeding probability immediately and with time lags of up to 10 weeks. However, an earlier onset of the yearly breeding period did not result in more overall reproductive output. Better body condition was associated with an earlier onset of breeding and higher embryo survival. Breeding probability in the main breeding season declined with increased breeding activity in the preceding season and only individuals in good body condition were able to breed late in the season. Higher temperatures reduce breeding success across seasons. We conclude that a better understanding of seasonal dynamics and plasticity (and their interplay) in reproduction will provide crucial insights into how lagomorphs are likely to respond and potentially adapt to the influence of future climate and other environmental change.

Empirical tests of harvest-induced body-size evolution along a geographic gradient in Australian macropods.

Life-history theory predicts the progressive dwarfing of animal populations that are subjected to chronic mortality stress, but the evolutionary impact of harvesting terrestrial herbivores has seldom been tested. In Australia, marsupials of the genus Macropus (kangaroos and wallabies) are subjected to size-selective commercial harvesting. Mathematical modelling suggests that harvest quotas (c. 10-20% of population estimates annually) could be driving body-size evolution in these species. We tested this hypothesis for three harvested macropod species with continental-scale distributions. To do so, we measured more than 2000 macropod skulls sourced from wildlife collections spanning the last 130 years. We analysed these data using spatial Bayesian models that controlled for the age and sex of specimens as well as environmental drivers and island effects. We found no evidence for the hypothesized decline in body size for any species; rather, models that fit trend terms supported minor body size increases over time. This apparently counterintuitive result is consistent with reduced mortality due to a depauperate predator guild and increased primary productivity of grassland vegetation following European settlement in Australia. Spatial patterns in macropod body size supported the heat dissipation limit and productivity hypotheses proposed to explain geographic body-size variation (i.e. skull size increased with decreasing summer maximum temperature and increasing rainfall, respectively). There is no empirical evidence that size-selective harvesting has driven the evolution of smaller body size in Australian macropods. Bayesian models are appropriate for investigating the long-term impact of human harvesting because they can impute missing data, fit nonlinear growth models and account for non-random spatial sampling inherent in wildlife collections.

An ecological regime shift resulting from disrupted predator-prey interactions in Holocene Australia.

The mass extinction events during human prehistory are striking examples of ecological regime shifts, the causes of which are still hotly debated. In Australia, human arrival approximately 50 thousand years ago was associated with the continental-scale extinction of numerous marsupial megafauna species and a permanent change in vegetation structure. An alternative stable state persisted until a second regime shift occurred during the late Holocene, when the largest two remaining marsupial carnivores, the thylacine and devil, disappeared from mainland Australia. These extinctions have been widely attributed to the human-assisted invasion of a competing predator, the dingo. In this unusual case, the simultaneous effects of human "intensification" (population growth and technological advances) and climate change (particularly increased ENSO variability) have been largely overlooked. We developed a dynamic model system capable of simulating the complex interactions between the main predators (humans, thylacines, devils, dingoes) and their marsupial prey (macropods), which we coupled with reconstructions of human population growth and climate change for late-Holocene Australia. Because the strength of important interspecific interactions cannot be estimated directly, we used detailed scenario testing and sensitivity analysis to identify robust model outcomes and investigate competing explanations for the Holocene regime shift. This approach identified human intensification as the most probable cause, while also demonstrating the potential importance of synergies with the effects of climate change. Our models indicate that the prehistoric impact of humans on Australian mammals was not limited to the late Pleistocene (i.e., the megafaunal extinctions) but extended into the late Holocene.

A population model for predicting the successful establishment of introduced bird species.

One of the strongest generalities in invasion biology is the positive relationship between probability of establishment and the numbers of individuals introduced. Nevertheless, a number of significant questions remain regarding: (1) the relative importance of different processes during introduction (e.g., demographic, environmental, and genetic stochasticity, and Allee effects); (2) the relative effects of propagule pressure (e.g., number of introductions, size of introductions, and lag between introductions); and (3) different life history characteristics of the species themselves. Here, we adopt an individual-based simulation modeling approach to explore a range of such details in the relationship between establishment success and numbers of individuals introduced. Our models are developed for typical exotic bird introductions, for which the relationship between probability of establishment and the numbers of individuals introduced has been particularly well documented. For both short-lived and long-lived species, probability of establishment decreased across multiple introductions (compared with a single introduction of the same total size), and this decrease was greater when inbreeding depression was included. Sensitivity analyses revealed four predictors that together accounted for >95% of model performance. Of these, R 0 (the average number of daughters produced per female over her lifetime) and propagule pressure were of primary importance, while random environmental effects and inbreeding depression exerted lesser influence. Initial founder size is undoubtedly going to be important for ensuring the persistence of introduced populations. However, we found the demographic traits, which influence how introduced individuals behave, to have the greatest effect on establishment success.

No need for disease: testing extinction hypotheses for the thylacine using multi-species metamodels.

Population viability analysis (PVA) is widely used to assess the extinction risk of threatened species and to evaluate different management strategies. However, conventional PVA neglects important biotic interactions and therefore can fail to identify important threatening processes. We designed a new PVA approach that includes species interactions explicitly by networking species models within a single 'metamodel'. We demonstrate the utility of PVA metamodels by employing them to reinterpret the extinction of the carnivorous, marsupial thylacine Thylacinus cynocephalus in Tasmania. In particular, we test the claim that well-documented impacts of European settlement cannot account for this extinction and that an unknown disease must have been an additional and necessary cause. We first constructed a classical, single-species PVA model for thylacines, which was then extended by incorporation within a dynamic predator-herbivore-vegetation metamodel that accounted for the influence of Europeans on the thylacine's prey base. Given obvious parameter uncertainties, we explored both modelling approaches with rigorous sensitivity analyses. Single-species PVA models were unable to recreate the thylacine's extinction unless a high human harvest, small starting population size or low maximum population growth rate was assumed, even if disease effects were included from 1906 to 1909. In contrast, we readily recreated the thylacine's demise using disease-free multi-species metamodels that simulated declines in native prey populations (particularly due to competition with introduced sheep). Dynamic, multi-species metamodels provide a simple, flexible framework for studying current species declines and historical extinctions caused by complex, interacting factors.

Quantum computing: a new paradigm for ecology.

A global technology arms race is underway to build evermore powerful and precise quantum computers. Quantum computers have the potential to tackle certain quantitative problems quicker than classical computers. The current focus of quantum computing is on pushing the boundaries of fundamental quantum information and commercial applications in industrial sectors, financial services, and other profit-led sectors, particularly where improvements in optimisation and sampling can improve increased economic return. We believe that ecologists could exploit the computational power of quantum computers because the statistical approaches commonly used in ecology already have proven pathways on quantum computers. Moreover, quantum computing could ultimately leapfrog our understanding of complex ecological systems, if the hardware, opportunity, and creativity of quantitative ecologists all align.

Measuring habitat quality for waterbirds: A review.

Quantifying habitat quality is dependent on measuring a site's relative contribution to population growth rate. This is challenging for studies of waterbirds, whose high mobility can decouple demographic rates from local habitat conditions and make sustained monitoring of individuals near-impossible. To overcome these challenges, biologists have used many direct and indirect proxies of waterbird habitat quality. However, consensus on what methods are most appropriate for a given scenario is lacking. We undertook a structured literature review of the methods used to quantify waterbird habitat quality, and provide a synthesis of the context-dependent strengths and limitations of those methods. Our search of the Web of Science and Scopus databases returned a sample of 666 studies, upon which our review was based. The reviewed studies assessed habitat quality by either measuring habitat attributes (e.g., food abundance, water quality, vegetation structure), or measuring attributes of the waterbirds themselves (e.g., demographic parameters, body condition, behavior, distribution). Measuring habitat attributes, although they are only indirectly related to demographic rates, has the advantage of being unaffected by waterbird behavioral stochasticity. Conversely, waterbird-derived measures (e.g., body condition, peck rates) may be more directly related to demographic rates than habitat variables, but may be subject to greater stochastic variation (e.g., behavioral change due to presence of conspecifics). Therefore, caution is needed to ensure that the measured variable does influence waterbird demographic rates. This assumption was usually based on ecological theory rather than empirical evidence. Our review highlighted that there is no single best, universally applicable method to quantify waterbird habitat quality. Individual project specifics (e.g., time frame, spatial scale, funding) will influence the choice of variables measured. Where possible, practitioners should measure variables most directly related to demographic rates. Generally, measuring multiple variables yields a better chance of accurately capturing the relationship between habitat characteristics and demographic rates.

Evolution of yolk protein genes in the Echinodermata.

Vitellogenin genes (vtg) encode large lipid transfer proteins (LLTPs) that are typically female-specific, functioning as precursors to major yolk proteins (MYPs). Within the phylum Echinodermata, however, the MYP of the Echinozoa (Echinoidea + Holothuroidea) is expressed by an unrelated transferrin-like gene that has a reproductive function in both sexes. We investigated egg proteins in the Asterozoa (Asteroidea + Ophiuroidea), a sister clade to the Echinozoa, showing that eggs of the asteroid Parvulastra exigua contain a vitellogenin protein (Vtg). vtg is expressed by P. exigua, a species with large eggs and nonfeeding larvae, and by the related asterinid Patiriella regularis which has small eggs and feeding larvae. In the Asteroidea, therefore, the reproductive function of vtg is conserved despite significant life history evolution. Like the echinozoan MYP gene, asteroid vtg is expressed in both sexes and may play a role in the development of both ovaries and testes. Phylogenetic analysis indicated that a putative Vtg from the sea urchin genome, a likely pseudogene, does not clade with asteroid Vtg. We propose the following sequence as a potential pathway for the evolution of YP genes in the Echinodermata: (1) the ancestral echinoderm produced YPs derived from Vtg, (2) bisexual vtg expression subsequently evolved in the echinoderm lineage, (3) the reproductive function of vtg was assumed by a transferrin-like gene in the ancestral echinozoan, and (4) redundant echinozoan vtg was released from stabilizing selection.

Novel passive detection approach reveals low breeding season survival and apparent lactation cost in a critically endangered cave bat.

Capture-mark-recapture/resight (CMR) methods are used for survival-rate studies, yet are often limited by small sample sizes. Advances in passive integrated transponder (PIT) technology have enabled passive detection or 'resight' of marked individuals using large antennas with greater read-ranges than previously possible. We used passively-detected resight data and CMR models to study survival rates of the southern bent-winged bat Miniopterus orianae bassanii, a critically endangered, cave-dwelling bat. Over three years, we used PIT-tagging to monitor 2966 individuals at the species' largest breeding aggregation, using daily detection data (> 1.6 million detections) to estimate seasonal survival probabilities, structured by age, sex and reproductive status, and parameterise population projection matrices. This has hitherto been impossible using traditional CMR methods due to disturbance risk and low recapture rates. Bats exhibited lowest apparent seasonal survival over summer and autumn, particularly for reproductive females in summer (when lactating) and juveniles in autumn (after weaning), and high survival in winter. Lowest survival rates coincided with severe drought in summer-autumn 2016, suggesting that dry conditions affect population viability. Under all likely demographic assumptions, population projection matrices suggested the population is in deterministic decline, requiring urgent action to reduce extinction risk. Passively-collected resight data can now be used in combination with CMR models to provide extensive, robust information for targeted wildlife population management.

Accelerometer informed time-energy budgets reveal the importance of temperature to the activity of a wild, arid zone canid.

BACKGROUND: Globally, arid regions are expanding and becoming hotter and drier with climate change. For medium and large bodied endotherms in the arid zone, the necessity to dissipate heat drives a range of adaptations, from behaviour to anatomy and physiology. Understanding how apex predators negotiate these landscapes and how they balance their energy is important as it may have broad impacts on ecosystem function. METHODS: We used tri-axial accelerometry (ACC) and GPS data collected from free-ranging dingoes in central Australia to investigate their activity-specific energetics, and activity patterns through time and space. We classified dingo activity into stationary, walking, and running behaviours, and estimated daily energy expenditure via activity-specific time-energy budgets developed using energy expenditure data derived from the literature. We tested whether dingoes behaviourally thermoregulate by modelling ODBA as a function of ambient temperature during the day and night. We used traditional distance measurements (GPS) as well as fine-scale activity (ODBA) data to assess their daily movement patterns. RESULTS: We retrieved ACC and GPS data from seven dingoes. Their mass-specific daily energy expenditure was significantly lower in summer (288 kJ kg- 1 day- 1) than winter (495 kJ kg- 1 day- 1; p = 0.03). Overall, dingoes were much less active during summer where 91% of their day was spent stationary in contrast to just 46% during winter. There was a sharp decrease in ODBA with increasing ambient temperature during the day (R2 = 0.59), whereas ODBA increased with increasing Ta at night (R2 = 0.39). Distance and ODBA were positively correlated (R = 0.65) and produced similar crepuscular patterns of activity. CONCLUSION: Our results indicate that ambient temperature may drive the behaviour of dingoes. Seasonal differences of daily energy expenditure in free-ranging eutherian mammals have been found in several species, though this was the first time it has been observed in a wild canid. We conclude that the negative relationship between dingo activity (ODBA) and ambient temperature during the day implies that high heat gain from solar radiation may be a factor limiting diurnal dingo activity in an arid environment.

Serological evidence of exposure to a coronavirus antigenically related to severe acute respiratory syndrome virus (SARS-CoV-1) in the Grey-headed flying fox (Pteropus poliocephalus).

Many infectious pathogens can be transmitted by highly mobile species, like bats that can act as reservoir hosts for viruses such as henipaviruses, lyssaviruses and coronaviruses. In this study, we investigated the seroepidemiology of protein antigens to Severe acute respiratory syndrome virus (SARS-CoV-1) and Middle eastern respiratory syndrome virus (MERS-CoV) in Grey-headed flying foxes (Pteropus poliocephalus) in Adelaide, Australia sampled between September 2015 and February 2018. A total of 301 serum samples were collected and evaluated using a multiplex Luminex binding assay, and median fluorescence intensity thresholds were determined using finite-mixture modelling. We found evidence of antibodies reactive to SARS-CoV-1 or a related antigen with 42.5% (CI: 34.3%-51.2%) seroprevalence but insufficient evidence of reactivity to MERS-CoV antigen. This study provides evidence that the Grey-headed flying foxes sampled in Adelaide have been exposed to a SARS-like coronavirus.