Olfactory misinformation provides refuge to palatable plants from mammalian browsing.

Mammalian herbivores browse palatable plants of ecological and economical value. Undesirable neighbours can reduce browsing to these plants by providing 'associational refuge', but they can also compete for resources. Here we recreated the informative odour emitted by undesirable plants. We then tested whether this odour could act as virtual neighbours, providing browsing refuge to palatable eucalyptus tree seedlings. We found that protection using this method was equivalent to protection provided by real plants. Palatable seedlings were 17-20 times more likely to be eaten by herbivores without virtual, or real, neighbours. Because many herbivores use plant odour to forage, virtual neighbours could provide a useful practical management approach to help protect valued plants.

Grow up, be persistent, and stay focused: keys for solving foraging problems by free-ranging possums.

Individuals within a species often vary in both their problem-solving approach and ability, affecting their capacity to access novel food resources. Testing problem-solving in free-ranging individuals is crucial for understanding the fundamental ecological implications of problem-solving capacity. To examine the factors affecting problem-solving in free-ranging animals, we presented three food-extraction tasks of increasing difficulty to urban common brushtail possums (Trichosurus vulpecula). We quantified two measures of problem-solving performance: trial outcome (success/failure) and time to solve and tested the influence of a range of potential drivers, including individual traits (personality, body weight, sex, and age), mechanistic behaviors that quantify problem-solving approach (work time, functional behavior time, behavioral diversity, and flexibility), and prior experience with the puzzles. We found that mechanistic behaviors were key drivers of performance. Individuals displaying greater persistence (higher work and functional behavior time) were more likely to solve a food-extraction task on their first attempt. Individuals also solved problems faster if they were more persistent and had lower behavioral flexibility. Personality indirectly affected time to solve one of the three problems by influencing time allocated to functional behaviors. Finally, adults solved the most difficult problem faster than juveniles. Overall, our study provides rare insight into the drivers underlying the problem-solving performance of wild animals. Such insight could be used to improve management strategies and conservation efforts, such as food or bait deployment, tailored to suit the innovative foraging abilities of target individuals in new and changing environments.

To eat, or not to eat: a phantom decoy affects information-gathering behavior by a free-ranging mammalian herbivore.

When foraging, making appropriate food choices is crucial to an animal's fitness. Classic foraging ecology theories assume animals choose food of greatest benefit based on their absolute value across multiple dimensions. Consequently, poorer options are considered irrelevant alternatives that should not influence decision-making among better options. But heuristic studies demonstrate that irrelevant alternatives (termed decoys) can influence the decisions of some animals, indicating they use a relative rather than absolute evaluation system. Our aim was to test whether a decoy influenced the decision-making process-that is, information-gathering and food choice-of a free-ranging mammalian herbivore. We tested swamp wallabies, Wallabia bicolor, comparing their behavior toward, and choice of, two available food options over time in the absence or presence of the decoy. We used a phantom decoy-unavailable option-and ran two trials in different locations and seasons. Binary preferences (decoy absent) for the two available food options differed between trials. Irrespective of this difference, across both trials the presence of the decoy resulted in animals more likely to overtly investigate available food options. But, the decoy only shifted food choice, weakly, in one trial. Our results indicate that the decoy influenced the information-gathering behavior during decision-making, providing the first evidence that decoys can affect decision-making process of free-ranging mammalian herbivores in an ecologically realistic context. It is premature to say these findings confirm the use of relative evaluation systems. Whether the foraging outcome is more strongly affected by other decoys, food dimensions, or ecological contexts, is yet to be determined.

The ethics of intervening in animal behaviour for conservation.

Conservation behaviour is a growing field that applies insights from the study of animal behaviour to address challenges in wildlife conservation and management. Conservation behaviour interventions often aim to manage specific behaviours of a species to solve conservation challenges. The field is often viewed as offering approaches that are less intrusive or harmful to animals than, for example, managing the impact of a problematic species by reducing its population size (frequently through lethal control). However, intervening in animal behaviour, even for conservation purposes, may still raise important ethical considerations. We discuss these issues and develop a framework and a decision support tool, to aid managers and researchers in evaluating the ethical considerations of conservation behaviour interventions against other options.

An invasive human commensal and a native marsupial maintain tick populations at the urban fringe.

Ticks (Acari: Ixodidae) are major disease vectors globally making it increasingly important to understand how altered vertebrate communities in urban areas shape tick population dynamics. In urban landscapes of Australia, little is known about which native and introduced small mammals maintain tick populations preventing host-targeted tick management and leading to human-wildlife conflict. Here, we determined (1) larval, nymphal, and adult tick burdens on host species and potential drivers, (2) the number of ticks supported by the different host populations, and (3) the proportion of medically significant tick species feeding on the different host species in Northern Sydney. We counted 3551 ticks on 241 mammals at 15 sites and found that long-nosed bandicoots (Perameles nasuta) hosted more ticks of all life stages than other small mammals but introduced black rats (Rattus rattus) were more abundant at most sites (33%-100%) and therefore important in supporting larval and nymphal ticks in our study areas. Black rats and bandicoots hosted a greater proportion of medically significant tick species including Ixodes holocyclus than other hosts. Our results show that an introduced human commensal contributes to maintaining urban tick populations and suggests ticks could be managed by controlling rat populations on urban fringes.

A new conceptual and quantitative approach to exploring and defining potential open-access olfactory information.

All organisms emit odour, providing 'open-access' olfactory information for any receiver with the right sensory apparatus. Characterizing open-access information emitted by groups of organisms, such as plant species, provides the means to answer significant questions about ecological interactions and their evolution. We present a new conceptual framework defining information reliability and a practical method to characterize and recover information from amongst olfactory noise. We quantified odour emissions from two tree species, one focal group and one outgroup, to demonstrate our approach using two new R statistical functions. We explore the consequences of relaxing or tightening criteria defining information and, from thousands of odour combinations, we identify and quantify those few likely to be informative. Our method uses core general principles characterizing information while incorporating knowledge of how receivers detect and discriminate odours. We can now map information in consistency-precision reliability space, explore the concept of information, and test information-noise boundaries, and between cues and signals.

Metatranscriptomic profiling reveals diverse tick-borne bacteria, protozoans and viruses in ticks and wildlife from Australia.

Tick-borne zoonoses are emerging globally due to changes in climate and land use. While the zoonotic threats associated with ticks are well studied elsewhere, in Australia, the diversity of potentially zoonotic agents carried by ticks and their significance to human and animal health is not sufficiently understood. To this end, we used untargeted metatranscriptomics to audit the prokaryotic, eukaryotic and viral biomes of questing ticks and wildlife blood samples from two urban and rural sites in New South Wales, Australia. Ixodes holocyclus and Haemaphysalis bancrofti were the main tick species collected, and blood samples from Rattus rattus, Rattus fuscipes, Perameles nasuta and Trichosurus vulpecula were also collected and screened for tick-borne microorganisms using metatranscriptomics followed by conventional targeted PCR to identify important microbial taxa to the species level. Our analyses identified 32 unique tick-borne taxa, including 10 novel putative species. Overall, a wide range of tick-borne microorganisms were found in questing ticks including haemoprotozoa such as Babesia, Theileria, Hepatozoon and Trypanosoma spp., bacteria such as Borrelia, Rickettsia, Ehrlichia, Neoehrlichia and Anaplasma spp., and numerous viral taxa including Reoviridiae (including two coltiviruses) and a novel Flaviviridae-like jingmenvirus. Of note, a novel hard tick-borne relapsing fever Borrelia sp. was identified in questing H. bancrofti ticks which is closely related to, but distinct from, cervid-associated Borrelia spp. found throughout Asia. Notably, all tick-borne microorganisms were phylogenetically unique compared to their relatives found outside Australia, and no foreign tick-borne human pathogens such as Borrelia burgdorferi s.l. or Babesia microti were found. This work adds to the growing literature demonstrating that Australian ticks harbour a unique and endemic microbial fauna, including potentially zoonotic agents which should be further studied to determine their relative risk to human and animal health.

The bacterial biome of ticks and their wildlife hosts at the urban-wildland interface.

Advances in sequencing technologies have revealed the complex and diverse microbial communities present in ticks (Ixodida). As obligate blood-feeding arthropods, ticks are responsible for a number of infectious diseases that can affect humans, livestock, domestic animals and wildlife. While cases of human tick-borne diseases continue to increase in the northern hemisphere, there has been relatively little recognition of zoonotic tick-borne pathogens in Australia. Over the past 5 years, studies using high-throughput sequencing technologies have shown that Australian ticks harbour unique and diverse bacterial communities. In the present study, free-ranging wildlife (n=203), representing ten mammal species, were sampled from urban and peri-urban areas in New South Wales (NSW), Queensland (QLD) and Western Australia (WA). Bacterial metabarcoding targeting the 16S rRNA locus was used to characterize the microbiomes of three sample types collected from wildlife: blood, ticks and tissue samples. Further sequence information was obtained for selected taxa of interest. Six tick species were identified from wildlife: Amblyomma triguttatum, Ixodes antechini, Ixodes australiensis, Ixodes holocyclus, Ixodes tasmani and Ixodes trichosuri. Bacterial 16S rRNA metabarcoding was performed on 536 samples and 65 controls, generating over 100 million sequences. Alpha diversity was significantly different between the three sample types, with tissue samples displaying the highest alpha diversity (P<0.001). Proteobacteria was the most abundant taxon identified across all sample types (37.3 %). Beta diversity analysis and ordination revealed little overlap between the three sample types (P<0.001). Taxa of interest included Anaplasmataceae, Bartonella, Borrelia, Coxiellaceae, Francisella, Midichloria, Mycoplasma and Rickettsia. Anaplasmataceae bacteria were detected in 17.7% (95/536) of samples and included Anaplasma, Ehrlichia and Neoehrlichia species. In samples from NSW, 'Ca. Neoehrlichia australis', 'Ca. Neoehrlichia arcana', Neoehrlichia sp. and Ehrlichia sp. were identified. A putative novel Ehrlichia sp. was identified from WA and Anaplasma platys was identified from QLD. Nine rodent tissue samples were positive for a novel Borrelia sp. that formed a phylogenetically distinct clade separate from the Lyme Borrelia and relapsing fever groups. This novel clade included recently identified rodent-associated Borrelia genotypes, which were described from Spain and North America. Bartonella was identified in 12.9% (69/536) of samples. Over half of these positive samples were obtained from black rats (Rattus rattus), and the dominant bacterial species identified were Bartonella coopersplainsensis and Bartonella queenslandensis. The results from the present study show the value of using unbiased high-throughput sequencing applied to samples collected from wildlife. In addition to understanding the sylvatic cycle of known vector-associated pathogens, surveillance work is important to ensure preparedness for potential zoonotic spillover events.

Behavioural drivers of survey bias: interactive effects of personality, the perceived risk and device properties.

Detecting small mammal species for wildlife research and management typically depends on animals deciding to engage with a device, for instance, by entering a trap. While some animals engage and are detected, others do not, and we often lack a mechanistic understanding of what drives these decisions. As trappability can be influenced by traits of personality, personality has high potential to similarly influence detection success for non-capture devices (chew-track cards, tracking tunnels, etc.). We present a conceptual model of the detection process where animal behaviours which are detected by different devices are grouped into tiers based on the degree of intimacy with a device (e.g., approach, interact, enter). Each tier is associated with an increase in the perceived danger of engaging with a device, and an increase in the potential for personality bias. To test this model, we first surveyed 36 populations of free-living black rats (Rattus rattus), a global pest species, to uniquely mark individuals (n = 128) and quantify personality traits. We then filmed rat behaviour at novel tracking tunnels with different risk-reward treatments. As predicted, detection biases were driven by personality, the bias increased with each tier and differed between the risk treatments. Our findings suggest that personality biases are not limited to live-capture traps but are widespread across devices which detect specific animal behaviours. In showing that biases can be predictable, we also show biases can be managed. We recommend that studies involving small mammal sampling report on steps taken to manage a personality-driven bias.

Misinformation tactics protect rare birds from problem predators.

Efficient decision-making integrates previous experience with new information. Tactical use of misinformation can alter choice in humans. Whether misinformation affects decision-making in other free-living species, including problem species, is unknown. Here, we show that sensory misinformation tactics can reduce the impacts of predators on vulnerable bird populations as effectively as lethal control. We repeatedly exposed invasive mammalian predators to unprofitable bird odors for 5 weeks before native shorebirds arrived for nesting and for 8 weeks thereafter. Chick production increased 1.7-fold at odor-treated sites over 25 to 35 days, with doubled or tripled odds of successful hatching, resulting in a 127% increase in modeled population size in 25 years. We demonstrate that decision-making processes that respond to changes in information reliability are vulnerable to tactical manipulation by misinformation. Altering perceptions of prey availability offers an innovative, nonlethal approach to managing problem predators and improving conservation outcomes for threatened species.

A mechanistic understanding of prebaiting to improve interaction with wildlife management devices.

BACKGROUND: Prebaiting is a technique involving early deployment of 'unarmed' devices (e.g. baits and traps) to increase efficacy of wildlife management. Although commonly used, the mechanisms by which prebaiting works are poorly understood. We propose three mechanisms by which prebaiting may increase device interaction probabilities; (1) overcoming neophobia towards novel devices, (2) a 'trickle in' effect increasing time for animals to encounter devices; and (3) social information transfer about rewards associated with devices. We conducted a survey of 100 articles to understand how prebaiting has been used. We then tested our proposed prebaiting mechanisms using a global pest (black rats, Rattus rattus) examining how uniquely marked free-living rats responded to a common yet novel monitoring technique (tracking tunnels). RESULTS: No studies in our dataset tested how prebaiting functioned. Most studies (61%) did not propose a mechanism for prebaiting, but overcoming neophobia was most commonly mentioned. We only found partial support for the overcoming neophobia hypothesis in our field test. We found the dominant mechanism operating in our system to be the 'trickle in' effect with the proportion of individuals visiting the device increasing over time. We found no support for social information transfer as a mechanism of prebaiting. CONCLUSION: Applying a mechanistic understanding of how prebaiting functions will improve the efficacy of management devices. Our results suggest that prebaiting allows time for more rats to encounter a device, hence surveys in our system would benefit from long prebaiting periods. © 2021 Society of Chemical Industry.

Testing transgenerational transfer of personality in managed wildlife populations: a house mouse control experiment.

Pest species control operations are most effective if every individual in a population is targeted. Yet, individual personality drives variation in animal responses to devices such as traps and baits. Failing to account for differences in behavior during control operations may drive a selective removal, resulting in residual animals with biased expressions of personality. If these biased traits are passed onto offspring, control operations would become increasingly problematic. To test if biased trait expressions in founding populations are passed on to offspring, we quantified personality traits in wild-caught house mice (Mus musculus) and created founder populations selected for biased (high, low) or intermediate expressions of activity. We released the behaviorally biased populations into outdoor yards to breed to the F1 generation and, 10 weeks later, removed the mice and quantified the personality traits of the offspring. Despite the strong personality bias in founder populations, we observed no transgenerational transfer of personality and detected no personality bias in the F1 generation. Our results provide reassuring evidence that a single intensive control operation that selects for survivors with a personality bias is unlikely to lead to a recovering population inherently more difficult to eradicate, at least for house mice.

Haemoprotozoan surveillance in peri-urban native and introduced wildlife from Australia.

Vector-borne haemoprotozoans comprise a diverse group of eukaryote single-celled organisms transmitted by haematophagous (blood-feeding) invertebrates. They can cause debilitating diseases that impact wildlife, livestock, companion animals and humans. Recent research has shown that Australian wildlife host a diverse range of haemoprotozoan species; however, to date this work has primarily been confined to a few host species or isolated populations in rural habitats. There has been little investigation into the presence of these blood parasites in wildlife inhabiting urban and peri-urban areas. In this study, blood and tissue samples and ticks were collected from wildlife in New South Wales and Western Australia. Extracted DNA samples were screened with pan-specific molecular assays to determine the presence of haemoprotozoans using amplicon metabarcoding and Sanger sequencing approaches. In addition, light microscopy was performed on blood films. Eight haemoprotozoans were identified in the present study, which included species of Babesia, Hepatozoon, Theileria and Trypanosoma. Blood samples were collected from 134 animals; 70 black rats (Rattus), 18 common brush-tailed possums (Trichosurus vulpecula), two bush rats (Rattus fuscipes), 22 chuditch (Dasyurus geoffroii), 20 long-nosed bandicoots (Perameles nasuta), one quenda (Isoodon fusciventer) and one swamp rat (Rattus lutreolus). Molecular screening of DNA extracted from blood samples identified 52.2% (95% CI: 43.8-60.5%) of individuals were positive for at least one haemoprotozoan species, with 19.4% (95% CI: 13.4-26.7%) positive for more than one species. The present study provides the first sequences of Theileria cf. peramelis from black rats and long-nosed bandicoots. Babesia lohae was identified from brush-tailed possums. Two Hepatozoon genotypes were identified from black rats and bush rats. Black rats showed the highest haemoprotozoan diversity, with five species identified. No known human pathogens that have been described in the northern hemisphere were identified in the present study, and future work is required to understand the zoonotic potential of these microbes in Australia. This work represents the first large-scale body of research using molecular tools to investigate haemoprotozoans in animals at the urban-wildland interface. Further research is needed to investigate potential consequences of infection in wildlife, particularly effects of pathogen spillover from invasive black rats to native wildlife.

Leveraging Motivations, Personality, and Sensory Cues for Vertebrate Pest Management.

Managing vertebrate pests is a global conservation challenge given their undesirable socio-ecological impacts. Pest management often focuses on the 'average' individual, neglecting individual-level behavioural variation ('personalities') and differences in life histories. These differences affect pest impacts and modify attraction to, or avoidance of, sensory cues. Strategies targeting the average individual may fail to mitigate damage by 'rogues' (individuals causing disproportionate impact) or to target 'recalcitrants' (individuals avoiding standard control measures). Effective management leverages animal behaviours that relate primarily to four core motivations: feeding, fleeing, fighting, and fornication. Management success could be greatly increased by identifying and exploiting individual variation in motivations. We provide explicit suggestions for cue-based tools to manipulate these four motivators, thereby improving pest management outcomes.

Invasive mammalian predators habituate to and generalize avian prey cues: a mechanism for conserving native prey.

Invasive mammalian predators can cause the decline and extinction of vulnerable native species. Many invasive mammalian predators are dietary generalists that hunt a variety of prey. These predators often rely upon olfaction when foraging, particularly at night. Little is understood about how prey odor cues are used to inform foraging decisions. Prey cues can vary spatially and temporally in their association with prey and can either reveal the location of prey or lead to unsuccessful foraging. Here we examine how two wild-caught invasive mammalian bird predator species (European hedgehogs Erinaceus europaeus and ferrets Mustela putorius furo) respond to unrewarded bird odors over successive exposures, first demonstrating that the odors are perceptually different using house mice (Mus musculus) as a biological olfactometer. We aim to test if introduced predators categorize odor cues of similar prey together, a tactic that could increase foraging efficiency. We exposed house mice to the odors using a standard habituation/dishabituation test in a laboratory setting, and wild-caught European hedgehogs and ferrets in an outdoor enclosure using a similar procedure. Mice discriminated among all bird odors presented, showing more interest in chicken odor than quail or gull odor. Both predator species showed a decline in interest toward unrewarded prey odor (i.e., habituation), but only ferrets generalized their response from one unrewarded bird odor to another bird odor. Hedgehog responses to unrewarded bird odors were highly variable between individuals. Taken together, our results reveal interspecific and intraspecific differences in response to prey odors, which we argue are a consequence of different diet breadth, life and evolutionary histories, and the conditions in each experiment. Generalization of prey odors may have enabled some species of invasive predators to efficiently hunt a range of intraguild prey species, for example, ground-nesting shorebirds. Olfactory manipulation of predators may be a useful conservation tool for threatened prey if it reduces the conspicuousness of vulnerable prey.

Invasive rabbits host immature Ixodes ticks at the urban-forest interface.

Introduced wildlife may be important alternative hosts for generalist ticks that cause health issues for humans and companion animals in urban areas, but to date are rarely considered as part of the tick-host community compared to native wildlife. In Australia, European rabbits, Oryctolagus cuniculus, are a widespread and abundant invasive species common to a range of human-modified ecosystems. To understand the potential role of rabbits in the life cycle of Australian ticks, we investigated the seasonal abundance of all tick life stages (larva, nymph, and adult) on rabbits collected from pest control programs in two urban forest remnants in Sydney, Australia. We also recorded whether larvae, nymphs, and adults were attached to the head, body, or limbs of rabbits to reveal patterns of tick attachment. Of the 2426 Ixodes ticks collected from 42 rabbits, larvae were by far the most abundant life stage (2360), peaking in abundance in autumn, while small numbers of nymphs (62) and adults (4) were present in winter and summer respectively. Larvae were found all over the body, whereas adults and nymphs were predominantly attached to the head, suggesting that the mature life stages use the host landscape differently, or that adults or nymphs may be groomed off the body. The most abundant tick species, as determined by morphology and DNA sequencing, was Ixodes holocyclus, a generalist tick responsible for significant human and companion animal health concerns in Australia. Our findings highlight the importance of understanding the role of introduced wildlife in tick dynamics particularly in novel ecosystems where non-native hosts may be more abundant than native hosts.

Molecular identification of the Trypanosoma (Herpetosoma) lewisi clade in black rats (Rattus rattus) from Australia.

Invasive rodent species are known hosts for a diverse range of infectious microorganisms and have long been associated with the spread of disease globally. The present study describes molecular evidence for the presence of a Trypanosoma sp. from black rats (Rattus rattus) in northern Sydney, Australia. Sequences of the 18S ribosomal RNA (rRNA) locus were obtained in two out of eleven (18%) blood samples with subsequent phylogenetic analysis confirming the identity within the Trypanosoma lewisi clade.

Bacterial community profiling highlights complex diversity and novel organisms in wildlife ticks.

Ticks Acari:Ixodida transmit a greater variety of pathogens than any other blood-feeding group of arthropods. While numerous microbes have been identified inhabiting Australian Ixodidae, some of which are related to globally important tick-borne pathogens, little is known about the bacterial communities within ticks collected from Australian wildlife. In this study, 1,019 ticks were identified on 221 hosts spanning 27 wildlife species. Next-generation sequencing was used to amplify the V1-2 hypervariable region of the bacterial 16S rRNA gene from 238 ticks; Amblyomma triguttatum (n = 6), Bothriocroton auruginans (n = 11), Bothriocroton concolor (n = 20), Haemaphysalis bancrofti (n = 10), Haemaphysalis bremneri (n = 4), Haemaphysalis humerosa (n = 13), Haemaphysalis longicornis (n = 4), Ixodes antechini (n = 29), Ixodes australiensis (n = 26), Ixodes fecialis (n = 13), Ixodes holocyclus (n = 37), Ixodes myrmecobii (n = 1), Ixodes ornithorhynchi (n = 10), Ixodes tasmani (n = 51) and Ixodes trichosuri (n = 3). After bioinformatic analyses, over 14 million assigned bacterial sequences revealed the presence of recently described bacteria 'Candidatus Borrelia tachyglossi', 'Candidatus Neoehrlichia australis', 'Candidatus Neoehrlichia arcana' and 'Candidatus Ehrlichia ornithorhynchi'. Furthermore, three novel Anaplasmataceae species were identified in the present study including; a Neoehrlichia sp. in I. australiensis and I. fecialis collected from quenda (Isoodon fusciventer) (Western Australia), an Anaplasma sp. from one B. concolor from echidna (Tachyglossus aculeatus) (New South Wales), and an Ehrlichia sp. from a single I. fecialis parasitising a quenda (WA). This study highlights the diversity of bacterial genera harboured within wildlife ticks, which may prove to be of medical and/or veterinary importance in the future.

Counting Ticks (Acari: Ixodida) on Hosts Is Complex: A Review and Comparison of Methods.

Locating and counting parasites on a host is a fundamental aspect of ecological research and basic veterinary and clinical practice. Variability in the biology and behavior of both hosts and parasites creates many methodological, logistical, and ethical considerations that must be made to collect this deceptively simple measurement. We identified methods that are used to count ticks (Acari: Ixodida, Leach 1815) on hosts by reviewing the methods sections of relevant published studies. Unfortunately, there is no best method agreed upon by scientists to collect ticks from hosts. In general, we suggest that studies focusing purely on counting ticks on hosts should use more sensitive methods to determine patterns of tick distribution on the surfaces of unconscious or deceased hosts in order to provide host body regions to target in future studies to maximize tick detection ability and limit the costs of research for researchers and the host animals involved. As ticks are counted on hosts for many different reasons, researchers must be goal oriented and chose methods that are appropriate for addressing their specific aims.