Comparison of animal welfare assessment tools and methodologies: need for an effective approach for captive elephants in Asia.

Welfare is a fundamental aspect of animal management and conservation. In light of growing public awareness and welfare concerns about captive elephants, there is an urgent need for comprehensive, globally coordinated efforts for Asian elephants (Elephas maximus) that participate in religious, logging, or tourist activities in range countries where the majority reside, and where welfare issues have been identified but not addressed. This review provides a comparative analysis of available animal assessment tools. Each offers distinct features for assessment that allow institutions to select criteria for specific needs and available resources. Most are applied to general animal welfare assessments, although some are tailored to particular species, including elephants. The tools span diverse formats, from digital to primarily paper-based assessments. Assessments operate at individual and institutional levels and across multiple welfare domains. Methodologies rely on keeper ratings or expert evaluations, incorporate numerical scoring and Likert scales for welfare grading, and encompass inputs including behaviors, health, and physiological indicators. For tourist camp elephants, one challenge is that the tools were developed in zoos, which may or may not have application to non-zoological settings. Digital tools and assessment methodologies such as keeper ratings face logistical challenges when applied across tourist venues. As with any tool, reliability, validity, and repeatability are essential and must address the unique welfare challenges of diverse captive settings. We propose that a holistic, context-specific, evidence-based, and practical tool be developed to ensure high elephant welfare standards in non-zoological facilities throughout Asia.

Non-invasive assessment of ovarian activity in free-ranging eastern black rhinoceros (Diceros bicornis michaeli) in Kenya.

Eastern black rhinos (Diceros bicornis michaeli) are a critically endangered species living in diverse habitats across Africa. In Kenya, once threatened with extinction due to massive poaching pressures, increased protection has resulted in losses being less than 1% annually today. Still, some populations have failed to achieve desired population growth targets. At Ol Jogi Wildlife Conservancy, some individuals are experiencing sub-optimal reproduction based on historical calving records and long inter-calving intervals (>3 years). Hormones drive the reproductive process, so non-invasive assessments of endocrine patterns can be useful indicators of individual reproductive health. In this study, we analysed longitudinal fecal progestagen metabolite (fPM) concentrations in all breeding female eastern black rhinos at Ol Jogi (n = 17) and compared the prevalence of irregular estrous cycles (longer or shorter than 20-40 days) and anestrous periods (interluteal period more than twice the length of a normal follicular phase, i.e. > 10 days) between optimal (inter-calving interval < 3 years) and sub-optimal (>3 years) reproducing individuals. Ten rhinos were pregnant during at least part of the study period. A total of 12 complete cycles were observed in seven females with an average length of 36 ± 3 days and equal numbers of regular and irregular cycles. Single anestrous periods averaging 67 ± 13 days were observed in five females. Surprisingly, a majority of cycles in optimal reproducing individuals were categorized as irregular based on fPM profiles. Overall, results suggest that irregular ovarian activity and isolated bouts of anestrus do not have negative impacts on reproductive performance in this subpopulation at Ol Jogi. A high priority is to continue using noninvasive hormone monitoring to evaluate how ecological or other variables influence reproductive success in this and other eastern black rhino subpopulations in Kenya.

Characterization of Longitudinal Testosterone, Cortisol, and Musth in Male Asian Elephants (Elephas maximus), Effects of Aging, and Adrenal Responses to Social Changes and Health Events.

The conservation of endangered species and sustainability of managed populations requires considerations to ensure the health and welfare of individuals. Male elephants experience a biological phenomenon called "musth", which is characterized by increased testosterone production, temporal gland secretion and urine dribbling, heightened aggression and sexual behavior, and therefore can pose unique challenges for human safety and animal welfare. This study characterized longitudinal (9 to 22 years) patterns of circulating testosterone and cortisol in relation to musth in four adult Asian elephant bulls spanning ages from 12 to 54 years. Age-related effects on musth activity and adrenal responses to social changes and clinical health events were also examined. All bulls exhibited regular annual musth cycles. Circulating cortisol covaried positively with testosterone and musth, highlighting intrinsic patterns that should be considered when evaluating the impact of social, health, and environmental changes on adrenal glucocorticoid activity. Except for an end-of-life cortisol increase in one bull, there was no clear evidence of chronically elevated cortisol secretion outside of musth in any individual. Testosterone decreased with age in sexually mature bulls, whereas age-related changes in cortisol varied across individuals, with the three older bulls showing the greatest rate of change during musth versus inter-musth periods. In contrast to physiological factors, there was no evidence of social factors, such as addition of a new male and death of male herdmates, impacting adrenal glucocorticoid activity in these bulls in the short term. Changes in cortisol were associated with treatment for Mycobacterium tuberculosis (M. tb) in two bulls, increasing after start of treatment and decreasing with cessation of treatment, but were not clearly associated with activation of disease. This study highlights the importance of longitudinal hormone monitoring to track changes in physiological function and responses to social, health, and environmental change in elephant bulls, which is important for making more informed decisions on how to manage male elephants under varying degrees of human care to ensure welfare and safety.

Perspectives on medical assistance in dying amongst persons with traumatic spinal cord injury: a qualitative study.

STUDY DESIGN: Qualitative. OBJECTIVE: To explore the perspectives of persons living with chronic (>5 y) traumatic spinal cord injuries (tSCI) concerning medical assistance in dying (MAID). SETTING: Saskatchewan, Canada. METHODS: Qualitative exploratory using an interpretative description methodology. Individual semi-structured interviews were held with online video conferencing. Interviews were analyzed using codebook thematic analysis. RESULTS: Ten persons with a chronic (>5 y) tSCI participated (seven males, three females; age 20s-50s; spinal cord level C2-T10). Participants expressed support for persons living with SCI having access to MAID, specifically in the context of preserving autonomy and respecting equality when compared with able-bodied individuals. However, some were concerned about the implications of access to MAID solely based on the diagnosis of a SCI. Participants also emphasized the variability in the initial personal response to injury and expressed concern about MAID being immediately available after injury. They explained the significance and necessity of reflecting on the injury in what can be a lengthy process of adjustment and acceptance. Participants also discussed the importance of hope and highlighted the irreplaceable value of peer support in shaping hope throughout rehabilitation and the need for compassionate, comprehensive, and continuous care to facilitate reintegration and support independence. CONCLUSIONS: Access to MAID provides a greater range of choices for individuals with tSCI but could increase deaths occurring before optimal adaptation and adjustment. The involvement of inter-professional teams, mental health specialists and peers is important to frame hope and understand the potential of a fulfilling life with a tSCI.

Care Considerations in a Patient- and Family-Centered Medical Assistance in Dying Program.

Objective: Medical Assistance in Dying (MAID) became legal in Canada in June 2016. As part of a project designed to improve end-of-life care for those requesting MAID, qualitative data from patients, families, and providers were used to assess opportunities to enhance patient-and family-centered care (PFCC) in this program. Methods: Thirty interviews were conducted with patients, families, and healthcare providers. Five patients who requested an assessment for MAID, 11 family members, and 14 healthcare providers were interviewed about their experiences in 2017. Comparative coding and thematic analysis were completed with the support of NVivo12. Results: Emotional PFCC considerations included: exploring and validating the emotional journey, navigating the uncertain, judgmental experiences, and the emotional impact on families and the care team. Physical PFCC considerations included: sensitivity in eligibility assessments, weaving in interdisciplinary care, provision of anticipatory guidance, and death location. Spiritual PFCC considerations included: honoring choice, listening to life stories, supporting spiritual needs, and acknowledging loss. Relational PFCC considerations included: defining the circle of support, supporting the circle, and relational investments. Conclusion: Fundamental to a PFCC MAID program, practitioners must be afforded time to provide holistic care. Program-related suggestions include incorporating interdisciplinary care early, and throughout the illness trajectory, consistency in care providers, appropriate anticipatory guidance, and bereavement supports for family, and dedicate space for MAID provisions. Patients and families must be included in the ongoing development and re-evaluation of MAID programs to ensure continued focus on quality end-of-life care.

Costs of seasonality at a southern latitude: Behavioral endocrinology of female baboons in the Cape Peninsula of South Africa.

Environmental challenges in the form of temperature extremes and unusual precipitation, which may lead to prolonged periods outside the thermoneutral zone, can be detrimental to animal physiology. Chacma baboons in the Cape Peninsula of South Africa, one of the highest latitudes at which nonhuman primates are found, experience extremes of both temperature and rainfall, as well as seasonal differences in day length that require animals to condense their daily routine into dramatically reduced daylight hours. Here we examine the effects of these climatic factors on the behavior (activity budgets and foraging patterns) and physiology (fecal glucocorticoid concentrations) of adult females (N = 33) in three groups of chacma baboons (Papio ursinus) inhabiting the Cape Peninsula, where temperatures ranged from 7 to 39 °C, monthly rainfall ranged from 2 to 158 mm, and day length varied by 4.5 h across seasons. Climatic variables showed a clear relationship to female baboon glucocorticoid concentrations, which significantly increased with lower temperatures, higher rainfall and shorter day lengths. Activity budgets also differed between summer and winter, with females generally spending less time socializing, moving and resting in the winter compared to summer, with some differences between troops in their feeding-related activities. Cold temperatures accompanied by rainfall and short day lengths may thus represent an ecological constraint for this population. This study highlights the potential impact of anthropogenic climate change on the physiology, behavior, and, ultimately, survival of wildlife populations.

"I Am Okay With It, But I Am Not Going to Do It": The Exogenous Factors Influencing Non-Participation in Medical Assistance in Dying.

Medical assistance in dying (MAID) processes are complex, shaped by legislated directives, and influenced by the discourse regarding its emergence as an end-of-life care option. Physicians and nurse practitioners (NPs) are essential in determining the patient's eligibility and conducting MAID provisions. This research explored the exogenous factors influencing physicians' and NPs' non-participation in formal MAID processes. Using an interpretive description methodology, we interviewed 17 physicians and 18 NPs in Saskatchewan, Canada, who identified as non-participators in MAID. The non-participation factors were related to (a) the health care system they work within, (b) the communities where they live, (c) their current practice context, (d) how their participation choices were visible to others, (e) the risks of participation to themselves and others, (f) time factors, (g) the impact of participation on the patient's family, and (h) patient-HCP relationship, and contextual factors. Practice considerations to support the evolving social contact of care were identified.

"What Is Right for Me, Is Not Necessarily Right for You": The Endogenous Factors Influencing Nonparticipation in Medical Assistance in Dying.

Access to medical assistance in dying (MAID) is influenced by legislation, health care providers (HCPs), the number of patient requests, and the patients' locations. This research explored the factors that influenced HCPs' nonparticipation in formal MAID processes and their needs to support this emerging practice area. Using an interpretive description methodology, we interviewed 17 physicians and 18 nurse practitioners who identified as non-participators in formal MAID processes. Nonparticipation was influenced by their (a) previous personal and professional experiences, (b) comfort with death, (c) conceptualization of duty, (d) preferred end-of-life care approaches, (e) faith or spirituality beliefs, (f) self-accountability, (g) consideration of emotional labor, and (h) future emotional impact. They identified a need for clear care pathways and safe passage. Two separate yet overlapping concepts were identified, conscientious objection to and nonparticipation in MAID, and we discussed options to support the social contract of care between HCPs and patients.

Utilizing Camera Traps, Closed Circuit Cameras and Behavior Observation Software to Monitor Activity Budgets, Habitat Use, and Social Interactions of Zoo-Housed Asian Elephants (Elephus maximus).

Accredited zoos and aquariums value superior animal husbandry and strive to ensure that the physical, psychological, and social needs of animals are met. In North America, the Association of Zoos and Aquariums (AZA) relies on species-specific standards to ensure facilities provide the best care for collection animals. The AZA also makes explicit recommendations for long-term monitoring of welfare. Data collected through behavioral observations can be used to modify management as animals respond over time to social, environmental, or physical changes. In long-lived, social species like elephants, it is particularly important to document herd dynamics, calf development, geriatric health, and social bonds throughout their lifetimes. The Smithsonian's National Zoological Park housed one male and six female Asian elephants in dynamic social groupings. Behavioral observations were conducted on all elephants for two years using two methods involving ZooMonitor, closed circuit cameras, and camera traps. The goal was to compare how these two methods function to provide individual activity budgets, habitat use, and social interactions. Methodologies such as these, alone or in combination, have the potential to produce valuable data about potential changes in welfare over time in a zoological setting and can be performed either by staff or volunteers with high reliability.

Serum Health Biomarkers in African and Asian Elephants: Value Ranges and Clinical Values Indicative of the Immune Response.

Serum biomarkers indicative of inflammation and disease can provide useful information regarding host immune processes, responses to treatment and prognosis. The aims of this study were to assess the use of commercially available anti-equine reagents for the quantification of cytokines (tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukins (IL) 2, 6, and 10) in African (Loxodonta africana, n = 125) and Asian (Elephas maximus, n = 104) elephants, and alongside previously validated anti-human reagents for acute-phase proteins (serum amyloid A and haptoglobin), calculate species-specific biomarker value ranges. In addition, we used opportunistically collected samples to investigate the concentrations of each biomarker during identified clinical cases of illness or injury, as a first step to understanding what biomarkers may be useful to managing elephant health. Immune biomarkers were each elevated above the calculated species-specific value ranges in at least one clinical case, but due to variability in both clinical and non-clinical samples, only serum amyloid A was significantly higher in clinical compared to non-clinical paired samples, with tendencies for higher TNF-α and IL-10. We also detected increased secretion of serum amyloid A and all five cytokines following routine vaccination of a single Asian elephant, indicating that these biomarkers can be beneficial for studying normal immune processes as well as pathology. This study indicates that assays developed with commercial reagents can be used to quantify health biomarkers in wildlife species and identifies several that warrant further investigation to elucidate immune responses to various pathologies.

Welfare Assessment and Activities of Captive Elephants in Thailand.

Thailand is the epicenter of elephant tourism and visiting an elephant camp is a popular activity according to the Tourist Authority of Thailand. However, the welfare of these elephants has been questioned by animal activist groups, international tour operators, and the public. Conclusions that the vast majority of captive elephants are abused often are based on anecdotal evidence and not solid science. So, it is difficult to tease apart emotion, opinion, and fact with regard to what practices are good or bad for elephant welfare. The aim of this paper was to: 1) describe the unique status of captive elephants in Thailand and associated regulations, 2) summarize current issues and challenges facing elephant tourism, 3) review studies conducted on welfare of tourist elephants in Thailand, and 4) offer recommendations for how elephants can be properly cared for under captive conditions in tourist camps. We conclude there are many ways to manage these elephants, and that not all tourist activities are bad for welfare. However, it is essential they be managed in a way that meets physical, physiological and psychological needs, and that management decisions are based on objective data.

Medical Assistance in Dying: Patients', Families', and Health Care Providers' Perspectives on Access and Care Delivery.

Background: Medical assistance in dying (MAID) became legal in Canada in 2016. Although the legislation is federal, each province is responsible for establishing quality care. Objective: To explore patient, family, and health care provider (HCP) perspectives on MAID access and care delivery and improve regional MAID care delivery. Design: Qualitative exploratory. Setting/Subjects: We interviewed 5 patients (4 met the legislated MAID criteria and 1 did not), 11 family members (4 spouses, 5 children, 1 sibling, and 1 friend), and 14 HCP (3 physicians, 4 social workers, and 7 nurses) from June to August 2017. Measurement: Semistructured interviews, content analysis, and thematic summary. Results: Patients, families, and HCPs highlighted access and delivery concerns regarding program sustainability, care pathway ambiguity, lack of support for care choices, institutional conscientious objection (CO), navigating care in institutions with a CO, and postdeath documentation. Patients and families expressed additional concerns regarding lack of ability to provide advanced MAID consent, and the requirement of independent witnesses on MAID request forms and consent immediately before MAID administration. HCPs were additionally uncertain about professional roles and responsibilities. Ten recommendations to improve regional MAID care and the resultant practice change are presented. Conclusion: Quality improvement (QI) processes are essential to devise an accessible dignified patient- and family-centered MAID program. Ensuring patient and family perspectives are integrated into QI initiatives will assist programs in ensuring the needs of all are considered in structuring and staffing a program that is accessible, easy to navigate, and provides dignified end-of-life care in supportive and respectful work environments.

Metabolic health assessment of zoo elephants: Management factors predicting leptin levels and the glucose-to-insulin ratio and their associations with health parameters.

Screening for metabolic-related health problems can enhance animal welfare, so the purpose of this study was to conduct the first metabolic health assessment of zoo elephants and use epidemiological methods to determine how factors in the captive environment were associated with metabolic hormone concentrations. In addition, we examined relationships between metabolic status and several fitness parameters: foot health, musculoskeletal health, reproductive cyclicity, and body condition. Two blood samples were collected 2 weeks apart from 87 Asian (Elephas maximus) and 105 African (Loxodonta africana) elephants managed by zoos accredited by the Association of Zoos and Aquariums for analysis of serum leptin, insulin, glucose and the glucose-to-insulin ratio (G:I). In females, mean (± SD) leptin concentrations and the G:I were lower (P<0.05) in Asian (3.93 ± 2.21 ng/ml and 110 ± 86 units) compared to African (4.37 ± 2.89 ng/ml and 208 ± 133 units) elephants, respectively. For males, mean leptin and the G:I were 4.99 ± 3.61 ng/ml and 253 ± 181 units for Asian, and 3.72 ± 2.00 ng/ml and 326 ± 231 units for African elephants, respectively, with no differences between species (P>0.05). As mean leptin concentration increased there was an increase in the odds of a female being non-cycling (P = 0.0083). The G:I was associated inversely with body condition (P = 0.0002); as the G:I increased there was a decreased risk of BCS = 4 or 5 as compared to the ideal, or BCS = 3. Neither leptin nor G:I were predictive of foot or musculoskeletal health scores. Factors related to walking and feeding practices were most influential in predicting metabolic status, whereas social and housing factors showed smaller, but significant effects. The metabolic health benefits of walking were detected if the time spent in staff-directed walking was 7 hours or more per week. The most protective feeding practices included implementing a random rather than predictable feeding schedule and limiting the number of methods presentation methods. Results indicate that leptin levels and G:I can be used as predictors of both ovarian cycle function and body condition, and are affected by zoo management in elephants.

Reproductive Health Assessment of Female Elephants in North American Zoos and Association of Husbandry Practices with Reproductive Dysfunction in African Elephants (Loxodonta africana).

As part of a multi-institutional study of zoo elephant welfare, we evaluated female elephants managed by zoos accredited by the Association of Zoos and Aquariums and applied epidemiological methods to determine what factors in the zoo environment are associated with reproductive problems, including ovarian acyclicity and hyperprolactinemia. Bi-weekly blood samples were collected from 95 African (Loxodonta africana) and 75 Asian (Elephas maximus) (8-55 years of age) elephants over a 12-month period for analysis of serum progestogens and prolactin. Females were categorized as normal cycling (regular 13- to 17-week cycles), irregular cycling (cycles longer or shorter than normal) or acyclic (baseline progestogens, <0.1 ng/ml throughout), and having Low/Normal (<14 or 18 ng/ml) or High (≥14 or 18 ng/ml) prolactin for Asian and African elephants, respectively. Rates of normal cycling, acyclicity and irregular cycling were 73.2, 22.5 and 4.2% for Asian, and 48.4, 37.9 and 13.7% for African elephants, respectively, all of which differed between species (P < 0.05). For African elephants, univariate assessment found that social isolation decreased and higher enrichment diversity increased the chance a female would cycle normally. The strongest multi-variable models included Age (positive) and Enrichment Diversity (negative) as important factors of acyclicity among African elephants. The Asian elephant data set was not robust enough to support multi-variable analyses of cyclicity status. Additionally, only 3% of Asian elephants were found to be hyperprolactinemic as compared to 28% of Africans, so predictive analyses of prolactin status were conducted on African elephants only. The strongest multi-variable model included Age (positive), Enrichment Diversity (negative), Alternate Feeding Methods (negative) and Social Group Contact (positive) as predictors of hyperprolactinemia. In summary, the incidence of ovarian cycle problems and hyperprolactinemia predominantly affects African elephants, and increases in social stability and feeding and enrichment diversity may have positive influences on hormone status.

Assessment of Body Condition in African (Loxodonta africana) and Asian (Elephas maximus) Elephants in North American Zoos and Management Practices Associated with High Body Condition Scores.

Obesity has a negative effect on health and welfare of many species, and has been speculated to be a problem for zoo elephants. To address this concern, we assessed the body condition of 240 elephants housed in North American zoos based on a set of standardized photographs using a 5-point Body Condition Score index (1 = thinnest; 5 = fattest). A multi-variable regression analysis was then used to determine how demographic, management, housing, and social factors were associated with an elevated body condition score in 132 African (Loxodonta africana) and 108 Asian (Elephas maximus) elephants. The highest BCS of 5, suggestive of obesity, was observed in 34% of zoo elephants. In both species, the majority of elephants had elevated BCS, with 74% in the BCS 4 (40%) and 5 (34%) categories. Only 22% of elephants had BCS 3, and less than 5% of the population was assigned the lowest BCS categories (BCS 1 and 2). The strongest multi-variable model demonstrated that staff-directed walking exercise of 14 hours or more per week and highly unpredictable feeding schedules were associated with decreased risk of BCS 4 or 5, while increased diversity in feeding methods and being female was associated with increased risk of BCS 4 or 5. Our data suggest that high body condition is prevalent among North American zoo elephants, and management strategies that help prevent and mitigate obesity may lead to improvements in welfare of zoo elephants.

Assessment of faecal glucocorticoid metabolite excretion in captive female fishing cats (Prionailurus viverinus) in Thailand.

There is little information on the endocrinology of fishing cats (Prionailurus viverinus), an endangered species in Southeast Asia, especially that pertaining to adrenal function. This study characterized faecal glucocorticoid metabolites in female fishing cats housed at Chiang Mai Night Safari to investigate seasonal and age relationships in hormone patterns. Faecal samples were collected 3 days/week for 1 year from seven females ranging in age from 4.5 to 9.6 years. A corticosterone enzyme immunoassay was validated for fishing cats by showing increases (∼60%) in faecal glucocorticoid immunoactivity above pre-treatment baseline levels within 1-2 days after an adrenocorticotrophic hormone injection. Faecal glucocorticoid metabolite concentrations were not related to age (P > 0.05), but there was a seasonal effect, with concentrations being higher (P < 0.05) during the winter (1.54 ±â€…0.04 µg/g) and rainy season (1.43 ±â€…0.04 µg/g) compared with the summer (1.22 ±â€…0.05 µg/g). Significant relationships were found between faecal glucocorticoids and rainfall (positive) and day length (negative), but not a temperature-humidity index. This is the first study to assess adrenal steroidogenic activity in female fishing cats, and we found that glucocorticoid metabolite production was influenced by seasonal factors, but not by age. We conclude that weather patterns should be taken into consideration in future studies of glucocorticoid activity in this endangered species, especially those studies aimed at improving captive management to create self-sustaining and healthy populations.

Assessment of ovarian activity in captive goral (Naemorhedus griseus) using noninvasive fecal steroid monitoring.

To date, there is no information on gonadal steroidogenic activity of female goral (Naemorhedus griseus), a threatened species of Thailand. Captive goral populations have been established to produce animals for ex situ conservation and reintroduction, but as yet none are self-sustaining. The objectives of the present study were to (1) determine the influence of season on ovarian steriodogenic function; and (2) examine the relationship between gonadal hormone excretion and sexual behaviors throughout the year. Fecal samples were collected 5 to 7 days/wk for 15 months from 8 adult females housed at Omkoi Wildlife Breeding Center in Thailand and analyzed for ovarian steroid metabolites using validated enzyme immunoassays. Observations of sexual behaviors and mating were conducted each morning for 30 min/session. Based on fecal estrogen and progestagen metabolite concentrations, the overall estrous cycle length was about 21 days, with a 2- to 3-day follicular phase and an 18- to 20-day luteal phase. Sexual behaviors, most notably tail-up, increased for 2 to 3 days during the time estrogens were elevated during mating. Fecal progestagens were elevated during luteal phases and increased further during gestation, which lasted approximately 7 months. The lactation period was 5 months, and females were anestrus for 2 to 5 of those months, with the exception of one that cycled continuously throughout. Two females conceived around 2 months postpartum and so were pregnant during lactation. Birth records over the past 21 years indicated young are born throughout the year. This combined with the hormonal data suggests that female gorals are not strongly seasonal, at least in captivity, although there was considerable variation among females in estrogen and progestagen patterns. In conclusion, fecal steroid metabolite monitoring is an effective means of assessing ovarian function in this species and will be a useful tool for breeding management and planned development of assisted reproductive techniques such as artificial insemination and embryo transfer.

An epidemiological approach to welfare research in zoos: the Elephant Welfare Project.

Multi-institutional studies of welfare have proven to be valuable in zoos but are hampered by limited sample sizes and difficulty in evaluating more than just a few welfare indicators. To more clearly understand how interactions of husbandry factors influence the interrelationships among welfare outcomes, epidemiological approaches are needed as well as multifactorial assessments of welfare. Many questions have been raised about the housing and care of elephants in zoos and whether their environmental and social needs are being met in a manner that promotes good welfare. This article describes the background and rationale for a large-scale study of elephant welfare in North American zoos funded by the (U.S.) Institute of Museum and Library Services. The goals of this project are to document the prevalence of positive and negative welfare states in 291 elephants exhibited in 72 Association of Zoos and Aquariums zoos and then determine the environmental, management, and husbandry factors that impact elephant welfare. This research is the largest scale nonhuman animal welfare project ever undertaken by the zoo community, and the scope of environmental variables and welfare outcomes measured is unprecedented.

Development of a versatile enzyme immunoassay for non-invasive assessment of glucocorticoid metabolites in a diversity of taxonomic species.

Endocrinology is a useful tool for conservation biologists and animal managers, and measuring glucocorticoids can help understand biological mechanisms associated with species decline and animal welfare. The current study describes the development and optimization of a glucocorticoid enzyme immunoassay (EIA) to non-invasively assess adrenal activity in a variety of taxa. The antiserum (CJM006) was raised in rabbits to a corticosterone-3-CMO-BSA immunogen and used in a standard competitive EIA system. However, the EIA initially produced results with unacceptably high inter-assay variation, attributed to consistent patterns observed within the optical density of developing plates. To determine the cause of this variability, a number of factors were examined using synthetic corticosterone standard and endogenous faecal extract, including: plate type (Nunc MaxiSorp® II versus Immulon IB plates); the use of non-specific secondary antibody; type (artificial versus natural) and presence (light versus dark) of light during incubation; plate loading temperature (4°C versus room temperature); and substrate reagent temperature (4°C versus room temperature). Results indicated that variability was associated with plate location effects, which were not initially detected because control samples were always run in the same positions across plates. Light and temperature were the two major factors that affected EIA reliability. For this assay, the standard protocol required slight modification, with the optimal protocol using Nunc MaxiSorp® plates, room temperature substrate reagents and dark incubation conditions. Following optimization, this EIA was then validated biochemically for 38 species, through parallel displacement curves and interference assessment tests of faecal and urine samples. Additionally, biological validation was performed opportunistically in a subset of species, with use of this EIA demonstrating significant elevations in faecal glucocorticoid metabolites following potentially challenging events. In summary, this glucocorticoid EIA cross-reacts with excreted glucocorticoid metabolites across a wide range of taxa, including ungulates, primates, felids, birds, rodents and amphibians. We conclude that when used with optimal reagent and incubation conditions, this EIA will be useful for non-invasive monitoring of adrenal activity in a wide range of wildlife species.

Sex, scarring, and stress: understanding seasonal costs in a cryptic marine mammal.

We investigated variation in adrenal activity in the dugong, a vulnerable cryptic marine mammal species, in a population inhabiting the lower thermal limit of their range. We sampled 319 live wild dugongs and examined faecal glucocorticoid (fGC) concentrations in relationship to sex, reproductive state (immature, sexually mature, or pregnant), and season, as well as associations with body condition, male-induced aggressive conspecific interactions, and environmental temperature. Physiological validations with apparently healthy and unhealthy animals indicated that fGC levels reliably reflect adrenal activation in dugongs. Glucocorticoid levels and body condition varied seasonally, with individuals expressing lowest fGC levels and an improving body condition over summer and autumn. Best body condition was observed in winter, which may have enhanced the ability of the animals to cope with thermal and nutritional challenges and helped to sustain the subsequent costs of reproduction (including mating activity in spring). Immature dugongs, especially those recently estranged from their mothers, may be particularly vulnerable to stress during winter, and also at risk of injury from adult conspecifics. During the spring mating period, mature and sexually active males with large erupted tusks had high fGC and poorer body condition compared with all dugongs, except pregnant females, in all other seasons. This finding was consistent with high levels of body scarring on adults of both sexes, resulting from agonistic behaviour by mature males. Competitive reproductive behaviour, inherent in a promiscuous mating system, may have stressful consequences for male dugongs, especially given that strenuous mating activity follows unfavourable thermal and nutritional conditions. Overall, fGC levels appear to be a good proxy for stressors in dugongs associated with season and/or temperature, intra-specific aggression, and pregnancy, but not lactation and/or maternal care. Understanding baseline fGC levels will allow wildlife managers to identify additional and/or unusual stressors that may threaten wild dugong populations.