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Healthy adult horses can balance accumulation and dissipation of body heat to maintain their body temperature between 37.5 and 38.5 °C, when they are in their thermoneutral zone (5 to 25 °C). However, under some circumstances, such as following strenuous exercise under hot, or hot and humid conditions, the accumulation of body heat exceeds dissipation and horses can suffer from heat stress. Prolonged or severe heat stress can lead to anhidrosis, heat stroke, or brain damage in the horse. To ameliorate the negative effects of high heat load in the body, early detection of heat stress and immediate human intervention is required to reduce the horse’s elevated body temperature in a timely manner. Body temperature measurement and deviations from the normal range are used to detect heat stress. Rectal temperature is the most commonly used method to monitor body temperature in horses, but other body temperature monitoring technologies, percutaneous thermal sensing microchips or infrared thermometry, are currently being studied for routine monitoring of the body temperature of horses as a more practical alternative. When heat stress is detected, horses can be cooled down by cool water application, air movement over the horse (e.g., fans), or a combination of these. The early detection of heat stress and the use of the most effective cooling methods is important to improve the welfare of heat stressed horses.

Animal welfare is important for the humane treatment of animals under our care. Zoos and rescue centres manage various charismatic animals, such as big cats, with limited resources. It is therefore essential for caretakers to understand the needs of an individual big cat to ensure its welfare. However, these needs may differ due to a big cat's personality, which may be identified by its coping style in a stressful situation. In addition, stress is one of the major factors affecting animal welfare. There is limited evidence showing strong associations between personality and stress physiology in big cats. This review focuses on the integration of personality and stress physiology of captive big cats, to highlight possible improvements in their husbandry. Our review identifies key factors that may influence big cat responses to stressors. These influencing factors include: i) social interactions; ii) environment; iii) life history and evolutionary traits; iv) genetics; and v) health. The first two factors are relatively well covered in the literature; however, the final three are potentially very promising avenues for future research to better understand how we can improve big cat welfare.

Wildlife translocation is increasingly utilized as a conservation management action, to mitigate the immediate negative effects of habitat loss and fragmentation (e.g. from land clearing or bushfires). Previous research has shown that stress responses can help or hinder survival in translocated wildlife and determine the efficacy of translocation as a conservation action. Yet these translocated animals are only one side of the equation, with translocation also potentially impacting the animals in the recipient population. We measured physiological markers of stress (faecal cortisol metabolite concentrations and neutrophil-lymphocyte ratios) and assessed health condition in a wild koala population one year after a major translocation of bushfire-rescued koalas on Kangaroo Island. We expected to find a high population density at the site (>0.75 koalas per hectare) and that resident koalas would show signs of chronic stress and ill health as a result of territorial conflict over food trees and reproductive opportunities. In contrast, we found that only one-fifth of the population remaining at the site were translocated koalas. The overall population density was also much lower (0.21 koalas per hectare) than anticipated. With no evidence of mass mortality at the site, we suggest that the majority of translocated koalas dispersed away from the site. Our stress marker measurements did not differ between the wild koalas and a sample of captive (non-display) koalas at the nearby Kangaroo Island Wildlife Park and were generally low compared to other studies. Veterinary examinations found that most koalas were in good body condition with very few diagnostic indicators of systemic ill health. Overall, our results suggest that, if there is adequate landscape-scale habitat connectivity and opportunity for dispersal, translocated koalas are likely to disperse from the site of release, with limited impacts on recipient koala populations at translocation release sites.

Orphaned koala joeys constitute a substantial number of wildlife rescues. Mortality is highly prevalent in rehabilitating joeys, with little knowledge about the causes of mortality. The hypothalamic-pituitary-adrenal axis plays a vital role in mediating stress by producing glucocorticoids (e.g. cortisol), however, no studies have quantified glucocorticoids in koala joeys. Traditional cortisol enzyme immuno-assay (e.g. R4866) are limited in supply and are process intensive, whereas, modern enzyme immuno-assay (EIA) kits (e.g. Arbor Assay cortisol kit) are available world-wide and provide rapid results. Biological validation is unsuitable to be performed in recuperating joeys due to ethical considerations, hence, we compared the results from biologically validated R4866 assay with the commercially available Arbor Assay cortisol kit. Thirty-four faecal samples were collected, processed and analysed for faecal cortisol metabolites (FCM) using both, R4866 assay and Arbor Assay kit. The joeys presented a suite of clinical conditions which provided the natural variation in stress response for comparing the assay sensitivity and range. The results indicated that there were no significant differences between the FCM values measured by both the assays. Furthermore, the Bland-Altman plot indicated a very strong agreement between the FCM concentrations measured by the two assays. This study is only a step towards recommending the routine use of commercial kit in clinical settings with basic resources, for rapid quantification of stress in koala patients. It is crucial for future studies to perform laboratory validation procedures to confirm the efficacy of the commercial kit before practical use for FCM monitoring in koalas.

Reproductive wastage is a major economic burden in sheep production globally, especially within Australia as livestock production systems face increased pressure from climatic variability (e.g. prolonged droughts or flooding). Sheep are sensitive to acute changes in their environment such as heat stress, which if not adequately monitored will result in significant production losses such as reproductive failure, increased parasite and worm burden, morbidity and mortality risks. Through basic and applied research in the areas of stress and reproductive physiology our team has made significant advancements in the understanding of sheep behaviour and physiological responses to acute and chronic stressors. Using minimally invasive hormone monitoring technology in combination with field based assessment of sheep health and productivity traits, our team has delivered new knowledge on how sheep react to acute environmental stress and how it impacts on sheep reproduction. In this chapter, we evaluated the fertility rates and embryo quality of Merino ewes under AI breeding program. We discuss factors such as heat stress that can impact on ewe and offspring quality.

Glucocorticoids (GCs) are primarily involved in mobilising energy reserves to various physiological processes including reproduction. During situations of stress, GCs also help in coping with stress by diverting energy away from processes such as growth and reproduction. Hence, studying annual and seasonal changes in GCs of wild populations can help in understanding their role in stress management and reproduction. The quantification of GCs in wild animals involves capturing, handling and restraining, which could be stressful. Moreover, different species may exhibit differential sensitivity to different stressors. Hence, determining species-specific sensitivities and responses to different stressors may help in developing effective conservation measures. In this context, we studied the annual and seasonal variations in corticosterone metabolites of the Bombay night frog, Nyctibatrachus humayuni. In addition, the effects of handling, marking and short-term captivity (24 h) on corticosterone metabolite levels of N. humayuni were determined. Our results show that urinary corticosterone metabolites (UCM) varied significantly annually and between the sexes; in males, the levels were highest during the breeding season, whilst in females, the levels were highest just before the breeding season. Interestingly, UCM levels of both the sexes were not affected by tagging with visual implant elastomer (VIE), and by short-term captivity, suggesting that these manipulations were not stressful in terms of corticosterone responses.

In this study, we determined the effect(s) of early shearing on Australian Merino ewes (Ovis aries) and their lambs. To test this research question, we used a suite of field and laboratory methods including GPS collars, wool cortisol, and epigenetic change between ewes and lambs identified using Illumina NovaSeq RRBS. Once shorn ewes (n = 24) were kept on their full fleece throughout the entire gestation period, whereas twice (early) shorn ewes (n = 24) had their wool shorn pre-joining. Top-knot wool sample was taken from ewes during pre-joining, day 50 (mid-gestation), and day 90 (late gestation) for laboratory analysis. Ewes were pregnancy scanned at mid-gestation to determine whether they were early or late parturition (this confirmation is provided by the pregnancy scanner based on fetus size). Top-knot wool sample was also taken from the lambs at weaning for hormone and wool quality testing. Ear tissue was taken from ewes at day 50 (mid-gestation) and from lambs at lamb marking for DNA analysis. Results showed that twice or early shorn ewes grazed 10% higher and maintained stronger body condition than once shorn ewes. Wool cortisol levels were also significantly lower in the early shorn ewes between mid- and late gestation. Lambs bred from twice shorn ewes had on average better visual wool quality parameters in terms of micron, spin finesses, and curvature. For the DNA methylation results, when comparing a group of once sheared with twice sheared ewes, we have discovered one locus (Chr20:50404014) that was significantly differentially methylated [False Discovery Rate (FDR) = 0.005]. This locus is upstream of a protein-coding gene (ENSOARG00000002778.1), which shows similarities to the forkhead box C1 (FOXC1) mRNA using BLAST searches. To further our understanding of the potential interaction between pregnancy status and shearing frequency of the ewes, we performed further differential methylation analysis using a combination of shearing treatment and pregnancy scanning status. The comparisons (1) late pregnancy vs. early pregnancy for ewes with one shearing treatment and (2) late pregnancy vs. early pregnancy for sheep with two shearing treatments were carried out to identify associations between loci and pregnancy duration for sheep with either one or two shearing events. We discovered that 36 gene loci were significantly modulated either between different shearing treatments or late vs. early pregnancy status of ewes. This result suggests that maternal pregnancy and nutritional status during gestation influence DNA methylation. We further investigated DNA methylation in lambs and identified 16 annotated gene loci that showed epigenetic modulation as a result of being born from an early or late stage pregnancy. From the genomics data, we pointed out that ewes go through epigenetic modifications during gestation, and there is a degree of intra-individual variation in the reproductive performance of ewes, which could be due to combination of intrinsic (genetic and physiological) and extrinsic (management and climatic) factors. Collectively, this research provides novel dataset combining physiological, molecular epigenetics, and digital tracking indices that advances our understanding of how Merino ewes respond to shearing frequency, and this information could guide further research on Merino sheep breeding and welfare.

Koala (Phascolarctos cinereus) is an iconic folivorous marsupial native to the sclerophyll forests and woodlands of Australia. Due to the ever-changing habitat, this species is highly vulnerable to anthropogenic factors such as habitat loss and fragmentation, and this is reflected in the increasing number of injured and/or diseased koalas over the years. The majority of adult koalas admitted at wildlife hospitals are deceased, either due to natural causes, or have to be euthanized. Thus, orphaned koala joeys constitute a substantial number of wildlife rescues, and mortality is also prevalent in koala joeys being hand-reared/rehabilitated, with little knowledge about the causes of such high rates of mortality. Wildlife hospitals/rehabilitation centres are inherently stressful, and although the hypothalamic–pituitary–adrenal (HPA) axis plays a vital role in mediating the stress endocrine function (by producing glucocorticoids such as cortisol), there are no studies quantifying glucocorticoids in koala joeys. To contribute to this dearth of research, we sampled a total of seven individuals residing at Port Macquarie Koala Hospital and noted their clinical information. Faecal samples were collected from all seven koala joeys during routine cage cleaning. In total, 123 faecal samples were collected, processed and analysed for cortisol using enzyme-immunoassay (EIA). We used the iterative baseline approach to determine baseline and peak concentrations of FCM in koala joeys. Baseline concentrations ranged between 14.11 ng/g – 51.10 ng/g (healthy – sick), whereas, peak FCM concentrations ranged between 25.65 ng/g – 56.58 ng/g (healthy – sick). There was a significant difference (p < 0.05) between FCM concentrations of healthy and impaired individuals. Healthy individuals displayed relatively consistent FCM concentrations, whereas, diseased individuals displayed a significant increase in FCM concentrations over time. Our study provides the first record of baseline and peak FCM concentrations in rescued koala joeys with their associated clinical condition. Future studies can use the iterative baseline approach to determine FCM concentration in wild koala joeys that can serve as a baseline to compare glucocorticoid levels of rescued joeys.

Koalas (Phascolarctos cinereus) are one of Australia’s most charismatic native small marsupial species. Unfortunately, populations of koalas are rapidly declining throughout Australia as they continue to face increasing pressure from a changing ecosystem. All wildlife species to some degree will use their hypothalamic–pituitary–adrenal (HPA) axis in response to stress. Depending on the duration of activation, the stress response can lead to either acute or chronic side effects and is modulated through the neuroendocrine stress system with the release of catecholamines and glucocorticoids (e.g., cortisol). It is well known that rehabilitation sanctuaries are inherently stressful for all animals, in particular for rescued wild koalas, as it is an unfamiliar environment where the animals cannot predict or control what will happen to them. In this pilot study, we set out to quantify faecal and fur cortisol metabolites in wild rescued koalas undergoing wildlife rehabilitation. Absolute levels of acute and chronic stress were indexed non-invasively, with faecal samples taken to evaluate acute stress, and fur samples taken to evaluate chronic stress. Sampling occurred sporadically over four months (the start of September 2018 to the end of December 2018), and was performed on three rescued koalas (Maree, Tai, and Solstice) being held at the rehabilitation centre. Results of this study show that between the three koalas, the highest recorded faecal cortisol result was 241 ng/g, and the lowest recorded faecal cortisol result was 4 ng/g, whereas the highest recorded fur cortisol result was 1.75 ng/g, and the lowest recorded fur cortisol result was 0.10 ng/g. Statistically, there was a significant difference between all three koalas and their faecal cortisol responses, as well as their fur cortisol responses. Statistically for Maree and Solstice, there was a significant difference in their faecal cortisol response between days when a stressor was recorded, and days when a stressor was not recorded. However, statistically for Tai, this was not the case, as there was no significant difference in his faecal cortisol response between days when a stressor was recorded, and days when a stressor was not recorded. In summary, the hypothesis that faecal glucocorticoids and fur glucocorticoids between koalas will differ based on individual responses to stressors was true as a whole, but individually, this hypothesis was true for Maree and Solstice, but untrue for Tai. The use of biological samples such as faeces and fur to obtain readings of glucocorticoids is a method of measuring absolute levels of physiological stress that is still evolving for koalas, and there is no current glucocorticoid baseline with which to compare the results of this study; although, measuring faecal and fur glucocorticoids is the first step in understanding how koalas undergoing wildlife rehabilitation respond to stressors.

In anurans, vocalisation is used to communicate within and between the sexes during reproduction. Knowledge of vocal repertoire and its diversity is essential in understanding its significance in sexual selection. In this context, we studied the vocal behaviour of Nyctibatrachus humayuni and quantified the urinary metabolites of androgens and corticosterone to understand the associations among hormones, body condition index and vocalisation. Our results show that males of N. humayuni produce an amplitude-modulated call consisting of a train of pulses. Interestingly, a high degree of inter-individual variation was observed in the call parameters. The dominant frequency and pulse interval were negatively correlated with male body size, whereas pulse duration and pulse duty cycle were positively correlated with it. Dominant frequency was negatively correlated with androgens and corticosterone, whereas pulse interval was negatively correlated with corticosterone. Further, pulse duty cycle, pulse duration and pulses per call were positively associated with corticosterone. Surprisingly, none of the acoustic parameters was associated with male body temperature and body condition. A significant variation was evident in the vocal effort, dominant frequency and amplitude of advertisement calls produced under different social situations. Calls produced during territory establishment had lower dominant frequency but higher vocal effort, whereas those produced during courtship had higher amplitude and vocal effort than the calls produced in the absence of a competitor/female. Our results suggest that greater inter-male variability in the advertisement call of N. humayuni could be important in female choice as it may help females to assess the quality of a male. Further, androgens and corticosterone could be important in modulating crucial parameters of the advertisement call.

Herpetological research involving amphibians is increasingly using mark and recapture methods, employing various techniques such as toe-clipping and visible implant alphanumeric tags. Visual Implant Elastomer (VIE) is a new method available for herpetological surveys, involving a coloured dye inserted into the epidermal skin surface of frogs. Previously, there has been only one study which demonstrated that the VIE method does not generate a significant physiological stress response (using a faecal glucocorticoid method) in a captive amphibian species. Physiological stress can also be quantified non-invasively using urinary corticosterone metabolite (UCM) enzyme-immunoassay in amphibians. In this study, we tested the physiological stress response of a common amphibian species, wild caught Asian toads (Duttaphrynus melanostictus), by comparing UCM responses to capture handling, sham control or VIE marking method. Adult males (n = 38) were captured and sampled for baseline UCM (t = 0 h) then marked either using the VIE or sham (saline control), or only handled during capture. Subsequently, urine samples were collected at t = 2, 12 and 24 h for toads within each group. UCM levels were quantified using an enzyme immunoassay (EIA) to determine differences among treatment groups and over time following capture. Toads generated acute stress responses to all three groups, showing a change in UCMs between baseline samples, 12 h, and 24 h samples. The mean UCM levels were not significantly different between the VIE method and the control groups (capture handling or sham operated). These results indicate that VIE method of tagging is no more stressful than routine handling of amphibians, hence in this context, the method does not have any additional welfare implications. Future research should explore the limitations of VIE tagging for long-term mark recapture studies, however, our current findings support its application as a minimally-invasive method for marking amphibians.

Individual hosts differ extensively in their competence for parasites, but traditional research has discounted this variation, partly because modeling such heterogeneity is difficult. This discounting has diminished as tools have improved and recognition has grown that some hosts, the extremely competent, can have exceptional impacts on disease dynamics. Most prominent among these hosts are the superspreaders, but other forms of extreme competence (EC) exist and others await discovery; each with potentially strong but distinct implications for disease emergence and spread. Here, we propose a framework for the study and discovery of EC, suitable for different host–parasite systems, which we hope enhances our understanding of how parasites circulate and evolve in host communities., Highlights A few members of host populations, so-called superspreaders, have disproportionate impacts on the risk of infectious disease emergence and spread. Several other forms of EC exist; some of which might be exceptionally protective. To discover and understand forms of EC, it is imperative to describe the distribution of, and covariation among, traits of individual hosts that mediate the many stages of host–parasite interactions. Here, we provide a framework to do so, emphasizing how interplay among host traits related to parasite exposure behavior, susceptibility, replicability of parasites on/in hosts, and transmissibility, comprise host competence. We hope this framework helps reveal new forms of EC and informs and improves management of disease risk.

Asiatic black bears (Ursus thibetanus) face chronic stress in bile farms. In this study, we investigated whether bile-farmed bears show significantly high levels of stress at rescue and whether stress levels reduce over time in a bear sanctuary where the bears are supported with environmental enrichment and veterinary care to improve animal welfare. We measured stress hormone levels using faecal cortisol metabolites (FCM) in 16 Asiatic black bears freshly rescued from bile farms in Vietnam. Fresh faeces were collected from each bear on the rescue truck and on a weekly basis for a 22-week study period at a bear sanctuary in Vietnam. Results showed that for all 16 rescued bears (with one exception) individual FCM levels from truck samples were above mean baseline FCMs of bears previously rehabilitated to a bear sanctuary. This suggested the majority of the rescued bears were still capable of showing a stress endocrine response during the rescue operation despite being exposed to conditions causing chronic stress in bears on bile farms. Results showed that mean FCM levels of the rescued bears differed significantly between time-periods (higher at the rescue [on truck samples] compared to week 22 samples) and mean FCM levels showed an overall decline over the first 22 weeks after they arrived at the bear sanctuary. The bears also demonstrated acute FCM stress responses to management interventions at the sanctuary, such as veterinary health checks and transportation. In conclusion, rescued bears tend to modulate their stress endocrine response after rehoming at the bear sanctuary. This is an important result, indicating that the rescue effort and rehabilitation of bile-farm bears is effective. Whether this also coincides with behavioural adjustments in rehabilitating bears (eg lessening of stereotypic behaviour) warrants further investigation.

Direct and indirect anthropogenic factors play a massive role in driving wildlife species towards extinction. Longitudinal retrospective studies identify key ‘factors’ responsible for the decline in numbers of wildlife, however, lack the reasoning behind the events leading to mortality. The overarching aim of this study was to categorize these ‘factors’ into different stressor categories faced by reptiles to understand its impact on an individual, and to compare how each stressor category influences the survival of an individual. The results from this study indicated that almost half of the number of reptiles being hospitalized were due to exposure to preliminary stressors such as lawn mowing incidents and pet attack. Primary and secondary admissions were fairly equal in number, however the mortality rate for secondary admissions was drastically high (~80%). The discussion integrates species’ ecology and stress physiology which can prove to have multi-faceted benefits across the fields of ecology and animal welfare. Ecologists can use the results from this study to comprehend species’ activity patterns to better plan reptilian conservation programs, whereas, for wildlife clinicians and rehabilitators, assignment of stressor categories could be a beneficial tool for bolstering the welfare monitoring program for small native reptiles in clinical settings.

Stress impacts on native wildlife because of added pressure from human-induced environmental change. The intrinsic variation in physiological stress responses [e.g. glucocorticoids (GCs)] can be evaluated using the emergency life history stage (ELHS), which has an adaptive value. The temporal and spatial variation in stress responses can also be evaluated using the reactive scope model (RSM). Using the blue-tongue lizard and the Eastern bearded dragon as examples we showed how short- and long-season breeding patterns impacted the adaptive value of the ELHS. Further, with the availability of field endocrinology data (e.g. plasma or faecal GCs), researchers can scope their study species’ stress responses using the RSM to guide conservation and management programs., Glucocorticoids (GCs) are used as biomarkers of physiological stress response in reptiles. Fundamental stress physiology tools including the emergency life history stage (ELHS) and the reactive scope model (RSM) can be useful to determine how individual variation of stress responses shape population ecology. In this perspective, we applied the RSM and ELHS into the context of two urban-dwelling small native Australian reptile species to compare the stress-response patterns in short- and long-breeding lizards. Firstly, by drawing inferences from the ELHS, we presented hypothetical scenarios using sample GC data for a short-breeding species (e.g. common blue-tongue lizard). We showed that activation of the physiological stress response would be non-adaptive due to the consequences of stress on reproduction. Therefore, blue-tongue lizards may become exposed to acute and chronic environmental stressors (e.g. human disturbance and habitat clearance) during the breeding season as they prefer not to activate their hypothalamo-pituitary interrenal (HPI) axis in support of their short-breeding season. On the contrary, long-breeding lizards (e.g. bearded dragons), which have multiple breeding opportunities and are accustomed to living around humans and altered urban environments, tended to operate above the critical adaptive value of the ELHS during their breeding period. This suggests that any future changes to the dynamics of habitat availability and breeding opportunities may favour the dragon differently over the blue-tongue lizard. To further capture the dynamics of stress responses along spatial and temporal scales, we suggested that researchers should collect field data (e.g. blood plasma or faecal GCs) and then use the ELHS and RSM to understand how the environment is shaping the animal’s stress physiology. The application of field stress monitoring and data visualization using the ELHS and RSM could guide environmental monitoring and conservation programs of native wildlife species.

Simple Summary Stress evaluation in wildlife is valuable tool for rehabilitation and injury prevention. This pilot study investigated categories of stress in rescued birds. We determined three categories of stressors (preliminary, primary and secondary) using clinical data of rescued birds from Adelaide, South Australia. It was discovered that birds are highly susceptible to impact injuries (e.g., flying into a building window) and vehicle-related injuries as preliminary stressors, which often result in hospitalisation of birds. Immobility and abnormal behaviour represented the most common primary stressor, while the most common secondary stressors included trauma and fracture. Furthermore, the most common outcome in clinics due to exposure of birds to these three stressor categories was euthanasia. Abstract Urbanisation exposes avian wildlife to an array of environmental stressors that result in clinical admission and hospitalisation. The aim of this pilot study was to conduct a retrospective analysis of clinical data and characterise this based on categories of stress experienced by avian wildlife patients. The results from this study indicated that impact injuries (n = 33, 25%) and vehicle-related injuries (n = 33, 25%) were the most common occurring preliminary stressors that resulted in the hospitalisation of avian wildlife. The most common outcome of avian patients that suffered from vehicle-related injuries was euthanasia (n = 15, 45%), as was avian patients that suffered from impact injuries (n = 16, 48%). Immobility (n = 105, 61%) and abnormal behaviour (n = 24, 14%) were the most commonly occurring primary stressors of avian patients. Finally, trauma (n = 51, 32%) and fractures (n = 44, 27%) were the most common occurring secondary stressors in avian patients. The most common outcome of all these stressors was euthanasia. This study provided further evidence towards the notion that human- and urbanisation-related stressors are the main causes of hospitalisation of avian wildlife, but also indicated that birds admitted as a result of human-related stressors are more likely to be euthanised than released. This study also provided a categorisation system for the stressors identified in avian wildlife patients (preliminary, primary and secondary) that may be used to monitor the stress categories of wildlife patients and gain a deeper understanding of the complex notion of stress.

The koala (Phascolactos cinereus) is currently listed by both the IUCN and the Australian Governments’ Threatened Species Scientific Committee as vulnerable to extinction with a decreasing population trend. This listing can be attributed to climate change and its impact on ecosystems, and anthropomorphic environmental change due to extensive land clearing and habitat fragmentation. These have both been proven to induce stress, which influences the onset of disease. This novel study performed a retrospective analysis whereby records for 12,543 wild, rescued koalas in New South Wales (NSW), Australia were studied in order to determine trends in koala sightings, clinical admissions and injury diagnoses over a period of 29 years (1989-2018). Results indicated that between all three study locations (Port Stephens, Port Macquarie and Lismore), the most common reason koalas were admitted into care was because of disease, the most common disease diagnosed was signs of chlamydia, and the most common outcome for koalas admitted into care was released. At Port Stephens, mature and female koalas were diagnosed with a disease more than any other age or sex, while juvenile and male koalas were released (back into the wild) more than any other age or sex. Additionally, there were fewer koalas with a disease and fewer koalas released in Port Stephens as each year progressed. At Port Macquarie, mature and male koalas were diagnosed with a disease more than any other age or sex, while juvenile and female koalas were released more than any other age or sex. Additionally, there were more koalas with a disease and fewer koalas released in Port Macquarie as each year progressed. At Lismore, adult and female koalas were diagnosed with a disease more than any other age or sex, while joey and male koalas were released more than any other age or sex. Additionally, there were more koalas with a disease and fewer koalas released in Lismore as each year progressed. Determining trends in clinical admissions and diagnosis over such a substantial period of time is an important factor in preventing the continuing decline of koalas throughout Australia, and in particular NSW. It is important to note that there are cultural differences between koala rescue groups in the three study locations (Port Stephens, Port Macquarie and Lismore). These differences may be reflected in the outcomes of koala patients as each group are driven by their own management team. It is essential that any further decline of koala populations is prevented, however this can only be achieved through informed recommendations through research studies such as these. These recommendations should lead to government legislation which can provide stronger protection to koala habitat.