American Zoos Research Articles

Primary Infection May Be an Underlying Factor Contributing to Lethal Hemorrhagic Disease Caused by Elephant Endotheliotropic Herpesvirus 3 in African Elephants (Loxodonta africana).

ABSTRACTDistinct but related species of elephant endotheliotropic herpesviruses (EEHVs) circulate within Asian and African elephant populations. Primary infection with EEHVs endemic among Asian elephants can cause clinical illness and lethal EEHV hemorrhagic disease (EEHV-HD). The degree to which this occurs among African elephants has not been fully established. Recent cases of EEHV-HD caused by the EEHV3 species in African elephants housed in North American zoos has heightened concern about the susceptibility of this elephant species to EEHV-HD. In this study, we utilize the luciferase immunoprecipitation system (LIPS) to generate a serological assay specific for EEHV3 in African elephants by detecting antibodies against the EEHV3 E34 protein. The results showed that the majority of tested elephants from four separate and genetically unrelated herds, including five elephants that survived clinical illness associated with EEHV3, were positive for prior infection with EEHV3. However, African elephants who succumbed to EEHV3-HD were seronegative for EEHV3 prior to lethal infection. This supports the hypothesis that fatal EEHV-HD caused by EEHV3 is associated with primary infection rather than reactivation of latent virus. Lastly, we observed that African elephants, like Asian elephants, acquire abundant anti-EEHV antibodies prenatally and that anti-EEHV3 specific antibodies were either never detected or declined to undetectable levels in those animals that died from lethal disease following EEHV3 infection.IMPORTANCE Prior to 2019, only five cases of clinical disease from EEHV infection among African elephants had been documented. Since 2019, there have been at least seven EEHV-HD cases in North American zoos, resulting in three fatalities, all associated with EEHV3. Evidence is accumulating to suggest that EEHV-associated clinical illness and death among Asian elephants is due to primary infection and may be associated with waning anti-EEHV antibody levels in young elephants. The development of the EEHV3 serological test described in this study enabled us to confirm that similar dynamics may be contributing to EEHV-HD in African elephants. The ability to screen for EEHV immune status in African elephant calves will have a major impact on managing captive African elephant herds and will provide new tools for investigating and understanding EEHV in wild populations.

Skinks in Zoos: A global approach on distribution patterns of threatened Scincidae in zoological institutions

To manage populations of threatened species according to the IUCN’s One Plan Approach, knowledge about both in situ and ex situ populations is required. To enhance the conservation of threatened skinks and to gain an overview which skink species are kept in zoos, and thus already have an ex situ conservation component, we analysed data from the Zoological Information Management System (ZIMS): their individual numbers, breeding success, and the number of holding institutions. We categorised species as threatened or non-threatened based on IUCN Red List assessments. Only 92 (~5%) of 1727 recognized skink species are held in ZIMS institutions worldwide, mostly in Australia, Europe, and North America. 77% of the species kept globally are classified as non-threatened and ~23% (21 species) are threatened. Only 28% of the species kept have successfully bred in the last year, mostly in one zoo each. Of these seven species were threatened. All threatened species are kept by four zoos at most, generally only in one. Half of the skink species kept are represented by less than 10 individuals. Mainly Australian skink species were kept. To improve the conservation of threatened skinks, a shift towards keeping threatened species should be considered within captive management programmes. European and North American zoos offer capacities and expertise for skink conservation but are outside skink species richness hotspots. Cooperative projects with institutions and stations in such hotspots could greatly benefit the conservation of skinks. Thus, according to the One Plan Approach, the ex situ populations could directly contribute to in situ protection.

Genetic assessment of captive red pandas (Ailurus fulgens) in American zoos to address management separation by putative subspecies.

Red pandas (Ailurus fulgens) are small charismatic mammals native across montane southern Asia, now endangered by human impacts. They are considered "living fossils" as the sole member of a distinct family, warranting higher conservation priority. Therefore, ex situ breeding programs were initiated to prevent extinction and act as genetic reservoirs for reintroduction, although complicated by apparent taxonomic subdivision. This study investigated whether the separation of captive red pandas in the North American Species Survival Plan® by putative subspecies was justified. A 383-bp segment of mitochondrial DNA control region was therefore sequenced from 67 members representing matriarchal lineages of both groups. A network analysis placed the 11 haplotypes found into separate but closely connected clusters, with one group more strongly related than the other. Statistical analyses and diversity indices corroborated differentiation between the two management units. Phylogenetic analyses employing multiple outgroups confirmed, although not robustly, reciprocal monophyly of the four- and seven-haplotype clades representing putative subspecies Ailurus fulgens fulgens and Ailurus fulgens styani, respectively. These empirical results are adequate to justify continued independent management of these zoo subpopulations, but cannot be definitive for taxonomic classification due to limited sampling from their native range. They will, however, be useful in evaluating long-term genetic diversity changes, focusing management efforts on newly revealed evolutionary limitations, and comparing with an assessment of wild red pandas to determine how representative zoo populations are for reintroduction purposes. Maintaining genetic diversity and population structure of endangered species is essential to protect evolutionary potential and adaptations for long-term sustainability.

IDLY INFECTED: A REVIEW OF INFECTIOUS AGENTS IN POPULATIONS OF TWO- AND THREE-TOED SLOTHS (CHOLOEPUS SPECIES AND BRADYPUS SPECIES).

Two- and three-toed sloths (Choloepus spp. and Bradypus spp.) have become popular animals in American culture and in American zoos, where they are frequently used as animal ambassadors. Despite the increased focus on sloth species, the prevalence of infectious diseases in sloth populations and the associated clinical consequences are relatively unknown. This study reviewed all published literature from 1809 to 2019 that examined infectious agents in both captive and wild populations of either two- or three-toed sloths. Online databases were electronically searched for relevant manuscripts using strings of inclusion and exclusion terms, resulting in an initial identification of 5,364 articles. After removing duplications and conducting two relevance screenings, 57 manuscripts were included in the full review. A total of 1,769 individual two-toed sloths and 879 individual three-toed sloths were accounted for in the included studies, with evidence of infection or exposure to infectious agents in 647 (36.6%) and 222 (25.3%) individual two- and three-toed sloths, respectively. Approximately 74% of documented infections were cryptic fungal, parasitic, and viral infections. The remaining 26% of infections represent those that were associated with clinical signs of disease. The infectious agents reported were bacterial (84), parasitic (20), viral (9), and fungal (4). Significant knowledge gaps remain regarding clinical and subclinical infectious disease prevalence and impact in both free-ranging and captive sloths.

Taxonomic Distribution of Neoplasia Among Non-Domestic Felid Species Under Managed Care.

Simple SummaryNeoplasia in nondomestic felids is common, and frequently malignant. However, few studies include large sample sizes of non-Panthera felids. Jaguars are reported to potentially have genetic mutations that predispose them to cancer, but studied populations are limited to North American zoos. By including animals from USA, Mexico, Colombia, Peru, and Brazil, the present study was able to include animals with widely varying genetic backgrounds, diets, and management. This study found that jaguars, lions, tigers and leopards are at a much higher risk of developing cancer than small cats. It also documented that the most common site for neoplasm development is the reproductive tract, followed by the respiratory system and then blood and lymphatic systems. These data provide support for thorough investigation of suspicious lesions to enable early detection of cancer.As evidenced by numerous case reports from zoos, neoplasia in felids is common, but most reports are limited to Panthera species in North America or Europe. In order to obtain a wider epidemiologic understanding of neoplasia distribution, necropsy records at seven facilities (USA, Mexico, Colombia, Peru, and Brazil) were evaluated. In contrast to others, this study population (195 cases, 16 species), included many non-Panthera felids. Overall neoplasia prevalence was 28.2% (55/195). Panthera species had a higher prevalence of neoplasia than non-Panthera species (52.5%; vs. 13.0%). Lions (66.7%), jaguars (55.0%), and tigers (31.3%) had the highest species-specific prevalence of neoplasia. Neoplasms in Panthera species were more frequently malignant than in non-Panthera (86.1% vs. 55.6%). The systems most commonly affected were the reproductive, hematolymphoid, and respiratory. The range of management conditions and more varied genetic backgrounds support a robust taxonomic pattern and suggest that the reported propensity for neoplasia in jaguars may have a genetic basis at a taxonomic level higher than species, as lions and tigers also have high prevalence. Given the high prevalence of neoplasia and high likelihood of malignancy, routine medical exams in all nondomestic felids, but Panthera species in particular, should include thorough assessments of any clinical signs of neoplasia.

Longitudinal Improvements in Zoo-Housed Elephant Welfare: A Case Study at ZSL Whipsnade Zoo.

Simple SummaryZoo elephant welfare has been the topic of much debate over the last two decades, with criticisms made regarding the husbandry and welfare of these species held in European and North American zoos. The aim of this study was to evidence the value of a species-specific behavioural monitoring programme and highlight the positive improvements in elephant welfare that were made in a single collection case study, by the comparison of behavioural activity budgets (time spent performing a particular behaviour) with previous published literature. This study identifies numerous indicators of positive welfare in our collection, including species-appropriate levels of feeding, low engagement in stereotypy (abnormal repetitive behaviour), and proportions of resting behaviour that are consistent with figures published from comparative zoo individuals. Additionally, we show that positive social associations exist between individuals in our study group, with low incidences of agonistic social behaviour and high engagement in positive social interactions. Finally, we acknowledge that improvements are required to further enhance elephant welfare in zoos and we have used the data collected throughout this research programme to adopt an evidence-based approach to the husbandry and management of Asian elephants at Zoological Society of London (ZSL) Whipsnade Zoo.Over the last two decades, criticisms were raised regarding the welfare experienced by elephants in European and North American zoos. Concerns regarding the welfare of zoo-housed elephants in the UK and Europe were consolidated in the publication of several key reports, and media interest peaked. Throughout this study we aim to outline the behavioural measures of welfare observed in the current group of Asian elephants (Elephas maximus) at Zoological Society of London (ZSL) Whipsnade Zoo, using key welfare indicators for this species and comparing them to previous published work. Following the instigation of a species-specific research programme, empirical behavioural data were available to quantify any developments in care and welfare. The collection of behavioural information revealed that individuals in our study group engage in low levels of stereotypic behaviour, have formed and maintain strong associations with one another and display a high proportion of engagement in lying rest. We outline that by applying simple, low-cost methods of behavioural data collection and analysis, it is possible to collect evidence that allows us to evaluate individual level welfare. This facilitates the adoption of an evidence-based approach to zoo management as well as demonstrating compliance with updated legislation for this species.

Specialised for the Swamp, Catered for in Captivity? A Cross-Institutional Evaluation of Captive Husbandry for Two Species of Lechwe.

Simple SummaryLechwe are social antelope adapted to wetland environments that can perform a ritualised courtship display. Although commonly housed in zoos, there is little published information available to guide their management. This study aimed to understand current husbandry practice for southern and Nile lechwe housed in North American and European institutions. A survey was sent to holders of these species, with questions addressing group demographics, enclosure characteristics, diet, enrichment, and occurrence of abnormal behaviours. Results showed that captive lechwe herds consisted of a similar ratio of male to females compared to wild herds, but there may be a limit to the number of male animals available to females at any one time. Lechwe enclosures typically featured wetland areas but these were rarely managed and there were often limited areas of vegetation for cover. The diets provided to lechwe differed when compared to existing husbandry guidelines but did not significantly differ between sampled zoos (in terms of ingredients commonly used). Abnormal behaviours were reported at several zoos but no specific causal factor was identified. This research provides a starting point for further study of the husbandry requirements of these specialised ungulates and considers the role of ecological information to the management of captive wild animals.Lechwe are specialised wetland antelope that can have a strict social hierarchy or perform lekking during breeding. The southern lechwe (Kobus leche) and the Nile lechwe (K. megaceros) are both found in zoos globally, but little research is available to support husbandry decisions. The aim of this research was to investigate current housing and husbandry used for these lechwe across North American and European zoos. A survey was distributed to holders in 2018 and information on 33 herds (18 Nile and 15 southern) was collected. The survey focussed on population demographics, enclosure size, biologically relevant exhibit features, mixed-species holdings, nutrition, use of environmental enrichment and performance of abnormal repetitive behaviours. Results showed that lechwe were housed in herds with similar sex ratios to wild counterparts but with a potential lack of opportunity to lek. Many zoos provided wetland, but this was rarely actively managed, and not all zoos provided cover for hiding and retreat. Current feeding practice differed significantly compared to available antelope husbandry guidelines. No consistency in amounts of pellet, forage or produce provided to lechwe across institutions was found. Abnormal repetitive behaviour was noted by several zoos, but no significant predictor of such behaviour could be identified. Despite some identifiable recognition of ecology informing lechwe management, it is important that evidence-based husbandry decisions are made based on a species’ evolutionary pathway and ecological needs and some fundamental features of lechwe husbandry do not always correlate with the adaptive traits of a specialised wetland ungulate.

Ovarian cyclicity and prolactin status of African elephants (Loxodonta africana) in North American zoos may be influenced by life experience and individual temperament

Hyperprolactinemia is an endocrine disorder associated with infertility in many species, including elephants. In a recent survey of zoos accredited by the Association of Zoos and Aquariums (AZA), over half of African elephant females (N = 101) were not cycling normally, 30% of which exhibited hyperprolactinemia. We examined whether life experience and temperament predict ovarian cyclicity and circulating prolactin status in individual African elephant females. We hypothesized that, similar to humans, acyclicity and hyperprolactinemia in elephants will be associated with an apprehensive or fearful, anxious temperament, and an increased number of potentially challenging life events (transfers, deaths and births). Ninety-five adult African elephant females housed at 37 AZA institutions were included in this study. Blood samples were collected twice a month for 1 year to determine ovarian cycle (cycling, n = 44; irregular, n = 13; non-cycling, n = 38) and prolactin (normal, n = 44; low; n = 23; high; n = 28) status. Keeper ratings on a 6-point scale were obtained on 32 temperament traits in 85 of these elephants. We determined that giving birth and being exposed to herd mates entering the facility were positively associated with normal ovarian cycle and prolactin profiles. By contrast, age, serum cortisol, and an increased number of herd mates leaving a facility were negatively associated with both. Contrary to our hypothesis, hyperprolactinemia was associated with a popular and caring temperament rating, whereas consistently low prolactin was associated with a fearful, apprehensive temperament. These findings indicate that pituitary-ovarian function may be impacted by life history (cyclicity) and temperament (prolactin), which should be taken into consideration when making management decisions.

Commonalities in Management and Husbandry Factors Important for Health and Welfare of Captive Elephants in North America and Thailand.

Simple SummaryThere is considerable concern about the welfare of elephants used for education, research, and entertainment purposes in western zoos and Asian tourist camps, and whether captive venues meet the needs of these highly social and intelligent animals. Therefore, it is important to conduct studies to determine how factors in the captive environment affect animal welfare both positively and negatively. The use of multi-disciplinary approaches to assess welfare has aided improvements in the management of captive elephants in North American zoos and Thailand tourist camps. In this paper, we review the most recent findings of our elephant studies that, using similar methodologies, highlight the importance of proper diets, adequate exercise, natural and stimulating environments, and freedom of movement to welfare. Overall, we found a number of commonalities in how environmental factors affect biological function of elephants managed under vastly different conditions. This type of integrative information is important for establishing welfare guidelines to ensure healthy, sustainable global populations of captive elephants.This review paper is a synthesis of results from multiple studies that we have conducted over the past several years using similar methodologies to identify factors related to welfare of captive populations of elephants in North American zoos and Thailand tourist camps. Using multiple conservation physiology tools, we found that, despite vastly disparate management systems, there are commonalities in how environmental and husbandry factors affect physical and physiological outcomes. Elephants appear to have better welfare, based on fecal glucocorticoid metabolite (FGM) analyses, when housed under conditions that provide a more enriched, stimulating, and less restrictive environment. We also found it is essential to balance diet and exercise for good body condition and metabolic function. In Thailand, use of tools to control elephants, such as the ankus (i.e., guide, hook) and chains, did not equate to poor welfare per se, nor did riding; however, improper uses were associated with higher wound scores and FGM concentrations. Foot health was good overall in both regions, with cracks being the most common problem, and better foot scores were found in elephants kept on softer substrates. Based on these findings, science-based guidelines are being developed in Thailand, while in North America, changes are being incorporated into elephant standards and husbandry resource guides. Management across venues can be improved by encouraging elephant exploration and exercise, establishing socially compatibility groups, ensuring proper use of tools, and providing balanced diets. We contend there is no “one-size-fits-all” management strategy to guarantee good welfare for elephants, but there are essential needs that must be met regardless of where or how they are managed. Future studies are needed to find ways to better socialize elephants; determine how temperament affects coping styles and resilience; study the importance of good handler-elephant relationships; identify more ways for elephants to engage with the environment; and assess the effect of life history on subsequent physiological and psychological well-being.

GREAT APE HEART PROJECT GUIDELINES FOR THE ECHOCARDIOGRAPHIC ASSESSMENT OF GREAT APES.

Cardiovascular disease (CVD) has been identified as a major cause of mortality in all four great ape taxa in zoologic institutions. In an effort to better understand and treat CVD in captive great apes, a program called the Great Ape Heart Project (GAHP), based at Zoo Atlanta, collects and maintains a database of echocardiograms and other relevant medical information relating to the cardiac health status of great apes. Cardiac health assessments have become standard practice among North American zoos that house great apes and are recommended by all four great ape Species Survival Plans (SSP) for the assessment of CVD in captive great apes. As of December 31, 2017, more than 70 ape-holding institutions have submitted approximately 1,100 cardiac examinations of great apes to the GAHP, information from which is stored in the GAHP database. Transthoracic echocardiography is one of the most practical and cost-effective diagnostic imaging techniques for the evaluation of cardiac function in great apes. Standardization of echocardiographic measurements is critical for maximizing the diagnostic value of an echocardiographic exam and for utilization of stored information in comparative studies within and between the great ape taxa. The following manuscript offers suggestions for standardization of nomenclature, imaging technique, echocardiographic measurements, data storage, and reporting of cardiac exams for submission into the GAHP database with the goal of promoting consistency and quality in data collection.

Multi‐institutional survey of causes of maternal rejection in Giraffes Giraffa camelopardalis in North American zoos

Maternal rejection has been attributed to a variety of causes among wild mammals and those in human care, including dystocia, health concerns, primipary, inappropriate maternal care experienced by the mother as a calf and an inability to display natural behaviours, although the specific causes among Giraffes Giraffa camelopardalis have not been isolated. To address this, a survey was designed and disseminated using the Association of Zoos and Aquariums Giraffe Listserv. The primary objectives were to document the circumstances surrounding maternal rejection events and any resulting calf abnormalities. The survey focused specifically on previously documented rejection causes among other ungulate species, as well as commonly suggested causes based on anecdotal evidence. Among the responses from 36 institutions, 12 animals were reported as maternally rejected and nine were maternally separated. Participating institutions frequently cited human presence during labour and low birth weight among rejected calves. Many of the expected causes of maternal rejection, including medical concerns, maternal inexperience and labour duration, were not indicated as contributing factors among participating institutions. Likewise, survey respondents did not note any extreme behavioural differences among Giraffes that receive limited or no maternal care. This survey provides preliminary, descriptive trends; however, future studies should further investigate these relationships. These results may inform husbandry decisions and help avoid future rejection events.

The Andean bear alopecia syndrome may be caused by social housing.

The Andean bear alopecia syndrome is a progressive and chronic condition documented in ex situ populations. Recent advances focus on treating symptoms, not preventing future cases. We therefore explored the epidemiology of this syndrome through an analysis of husbandry and veterinary conditions of 63 Andean bears (26M:37F) housed in North and South American zoos and other ex situ circumstances. We had the most complete information for the North American population and found that 29% of females (n = 24) were affected. No males (n = 26) were affected. An analysis of generalized linear models indicated that three models were competitive in describing the occurrence of the condition (i.e., ΔAICc ≤ 2): the model including only the individual's sex (χ2 = 13.41, df = 1, p < .001), the model including both individual sex and social housing status (χ2 = 1.36, df = 2, p < .001), and the model including both individual sex and the expression of stereotypical behaviors (χ2 = 13.82, df = 2, p = .001). Stereotypical behaviors were common among both males (50%, n = 26) and females (51.9%, n = 27) whether or not they were affected, but the syndrome was seen only in females who had been socially housed. Therefore, we suggest that the Andean bear alopecia syndrome is a symptomatic response to the long-term social housing of bears that would otherwise not live socially. To prevent new cases, we recommend that female Andean bears be housed with adult conspecifics only when females choose to cohabitate.

Relationships between housing and management factors and clinical health events in elephants in North American zoos.

Elephants experience a number of health issues that can contribute to their well-being and survival. In managed populations, housing conditions and management practices can influence individual health, so potential risk factors associated with morbidity or mortality should be identified to ensure the best possible standards of care. The goal of this study was to determine if the number of clinical events experienced could be a useful welfare indicator in zoo elephants, and to determine factors associated with key pathologies. We used an epidemiological approach to investigate how intrinsic (species, sex, age) and extrinsic (housing, management) factors were associated with both the total number of clinical events, and each of the four most prevalent pathology types (gastrointestinal issues, skin lesions, lameness, foot lesions), over a 12-month period. The study included 220 (127 African; 93 Asian) elephants housed at 61 facilities across North America. More than 1100 clinical events were identified. Species and sex differences were apparent in the types of pathology encountered, and unsurprisingly, the number of clinical events was positively correlated with age. Factors relating to housing (percent time with indoor/outdoor choice, space experience inside, number of unique environments an elephant was housed in, percent time on soft substrate) and management (enrichment diversity, spread of feeding opportunities) were also related to the number of clinical events. However, relationships were often counter to our initial hypotheses, highlighting caution in assuming cause and effect from correlational analyses such as these. Other welfare indicators such as serum and fecal glucocorticoids and serum prolactin were also associated with health status, being higher or more variable in individuals with a greater number of events. This approach provides insight into housing and management factors related to the health of these species in zoos, and in some cases, may reflect management changes that have already been made to mitigate existing or anticipated health concerns.

Sex-specific median life expectancies from ex situ populations for 330 animal species

We present life expectancy estimates for hundreds of vertebrate species based on carefully vetted studbook data from North American zoos and aquariums. These data include sex-specific median life expectancies as well as sample size and 95% confidence limits for each estimate. Existing longevity data for animals primarily consist of maximum lifespan values, which are single observations rather than statistically derived estimates of longevity. Moreover, all of our estimates are based on the same type of data and calculated using consistent, standardized methods. To derive these estimates, we conducted Kaplan-Meier survival analyses using individual-level demographic data (i.e., records of birth and mortality events) from studbook records for each ex situ population. Our species set represents a range of vertebrate taxa (primarily mammals, birds, amphibians, and reptiles) and diverse life histories. This dataset will have broad utility, not only for informing comparative demographic and life history studies, but also more broadly for any research or conservation application that requires sex- or species-specific life expectancy information.

Patterns in the long-term viability of North American zoo populations.

Recent concerns about the viability of zoo populations have motivated studies on the historic and current status of animal populations in North American and European zoos. However, these evaluations may not accurately reflect the populations' long-term viability in the decades to come. Here, we assessed the projected future status of North American zoo populations by conducting standardized population viability analyses (PVAs) for 137 cooperative breeding programs. We summarized PVA results to describe patterns in viability across populations, and examined whether viability can be predicted by biological or management-based factors. Under recent management practices and without imports or exports of animals, 64% of populations will decline in size over the next 25 years, and only 18% would retain ≥90% of the founding gene diversity (GD) in 100 years. However, viability would improve if programs can implement management changes (e.g., increasing reproduction, increasing holding space, and importing genetically unique individuals, as appropriate): only 16% of populations would still decline in 25 years, and 49% would retain ≥90% GD in 100 years. Programs with more participating institutions and a "green" Association of Zoos and Aquariums animal program designation were projected to have higher metrics of demographic viability, and those with longer lifespans and lower recent death rates were projected to have higher metrics of genetic viability. Due to the large variation in species life history, management goals, and constraints across programs, our findings suggest there is unlikely to be a single path to long-term viability that would be appropriate for all zoo populations.

Evaluation of management in North American zoos to enhance breeding success of the fishing cat (Prionailurus viverrinus) ex situ population.

The ex situ population of fishing cats (Prionailurus viverrinus) face many challenges to its sustainability such as mate incompatibility, low founder numbers and disease prevalence. The North American population was monitored for a three-year period during institutional transfers and breeding introductions. In total, 26 fishing cats, including 15 different breeding pairs were monitored during 20 transfers. Most institutional transfers occurred in the fall months (September, October and November; 62%; n = 13) and males were transferred more often (62%; n = 13). Breeding success (observed copulations) was recorded in 33% (n = 5) of pairs but only 13% (n = 2 pairs) produced offspring during the study period. Institutions with successful breeding pairs had a greater number of indoor, off-exhibit enclosures (2.67 ± 0.29 enclosures; n = 5) compared to facilities with unsuccessful pairs (1.69 ± 0.25 exhibits; n = 6; p = 0.035). In addition, facilities housing successful pairs performed positive reinforcement training more frequently (14.77 ± 3.27 training days/month) than facilities with unsuccessful pairs (4.00 ± 2.73 days/month; p = 0.035). A binomial generalized linear model showed that friendly vocalizations (p = 0.000) during physical introductions of intended breeding pairs predicted copulation success. Introductions performed when a female was exhibiting estrous behavior (p = 0.020), was also predictive of copulation success. Results from this study are the first comprehensive analyses of captive management in the fishing cat. Environmental factors and management approaches are highlighted that could advance animal welfare and improve reproductive success in this species.

101 Assessment of semen traits in servals (Leptailurus serval) and Canada lynx (Lynx canadensis)

Servals and Canada lynx are managed by species survival plans in North American zoos, but current populations are not sustainable. Increased knowledge of their reproductive biology would benefit breeding management and development of assisted reproductive techniques. The aims of our study were to (1) evaluate effectiveness of urethral catheterization and electroejaculation (EEJ) for semen collection; (2) characterise basal seminal traits; and (3) compare effectiveness of semen cryopreservation methods. Semen was collected from 6 servals and 9 Canada lynx via a urinary catheter (3.5 Fr×22 cm, inserted 15cm into the urethra), followed by EEJ under dexmedetomidine-ketamine anaesthesia. To assess the effect of seasonality on lynx seminal traits, semen was collected before (late January), during (mid-February to mid-March), and after (early April) the peak breeding season. Serval and lynx semen were frozen by conventional slow freezing (i.e. in 0.25-mL straws cooled to 4°C for 2h and frozen in LN vapor) in a soy lecithin-based (SOY) or egg yolk-based (TEY) extender with 4% glycerol and by ultra-rapid freezing (URF; direct pelleting into LN at ≈104°C/min) in SOY medium with 0.2M sucrose. To evaluate post-thaw sperm function in servals, heterologous IVF of domestic cat oocytes was performed, with cleavage rate assessed at 48h post-insemination. Data were analysed by one-way or repeated-measures ANOVA. Data are mean±standard deviation. Sperm recovery by urethral catheterization was negligible in both species, but EEJ allowed sperm collection in all males. Lynx seminal traits were similar during breeding and nonbreeding seasons. Testicular volume (4.81±1.17cm3) and sperm quality (13±11×106 sperm/ejaculate; 49±14% motility; 29±12% normal morphology; 74±13% acrosome integrity) were consistent with previous findings in the lynx genus. Post-thaw sperm quality in lynx has not yet been evaluated. In servals, testes volume was 6.56±2.11cm3 with good sperm quality for most males (46±36×106 sperm/ejaculate; 75±20% motility; 56±36% normal morphology; 84±7% acrosome integrity). Post-thaw, serval sperm acrosome integrity (31±15, 21±13, 24±13% at 0h for TEY, SOY, and URF, respectively; P&amp;gt;0.05) and motility (40±21% at 0h, 20±11% at 6h for TEY; 24±19% at 0h, 6±4% at 6h for SOY; 21±16% at 0h, 3±2% at 6h for URF; treatment: P&amp;gt;0.05; time: P&amp;lt;0.05; interaction: P&amp;gt;0.05) declined substantially. However, thawed sperm could fertilize domestic cat oocytes with no difference among treatments in cleavage success (53±6, 47±4, or 49±14%; TEY, SOY, and URF, respectively; P&amp;gt;0.05), indicating that standard freezing methods are effective in servals. Our findings provide zoos with valuable information about normative reproductive traits in both species. Supported by IMLS and the Roger &amp;amp; Kathy Gross Post-doctoral Fellowship.

SEASONAL VARIATION OF SERUM 25-HYDROXY-VITAMIN D IN TWO CAPTIVE EASTERN BLACK RHINOCEROS ( DICEROS BICORNIS MICHAELI) HOUSED IN A NORTH AMERICAN ZOO.

Black rhinoceros ( Diceros bicornis spp.) are critically endangered species, with less than 65 individual animals housed in captivity within Association of Zoos and Aquariums-accredited zoos within the United States, and an estimated 5,500 individual animals of all subspecies surviving in the wild. Previously published reference values for circulating vitamin D3 (25OHD3; 55.7 ± 34.2 ng/ml) were based upon samples from free-ranging black rhinoceros in Africa. Recent research in human medicine has highlighted the importance of subclinical vitamin D deficiency, with links to increased risks for developing various health conditions. Serum samples collected opportunistically from two captive Eastern black rhinoceros ( Diceros bicornis michaeli) housed with seasonal access outdoors in a North American zoo were tested for 25-hydroxy-vitamin D (25OHD) levels over a 3-yr period. A commercially prepared pelleted diet containing vitamin D3 was fed to both rhinos. This study correlates environmental ultraviolet (UV) index, dietary supplementation, and seasonal serum 25OHD levels to compare with known 25OHD3 levels in free-ranging African black rhinoceros. Results in these two individuals suggest that D. bicornis spp. are dependent upon sunlight or UVB for measurable circulating 25OHD, and that current vitamin D3 supplementation levels may have little effect for Diceros spp. in human care housed in northern latitudes.

Good keeper-elephant relationships in North American zoos are mutually beneficial to welfare

Relationships between animals and their human caretakers can have profound impacts on animal welfare in farms, laboratories and zoos, while human attitudes are important predictors of caretaker behavior towards livestock. In this study, we examined the impact of keeper attitudes about working with elephants on Keeper-Elephant Relationships (KERs) and Bonds (KEBs), and found evidence for reciprocity and welfare benefits to both parties. As part of a large, multi-institutional study of zoo elephant welfare conducted at 60 zoos accredited by the American Zoo and Aquarium Association, blood samples were collected twice monthly for 1 year from 117 African (Loxodont africanus) and 96 Asian (Elephas maximus) elephants for serum cortisol analyses as a measure of well-being. Information was collected via three online questionnaires: 1) a Keeper Survey of 277 elephant keepers about their opinions of and attitudes towards working with elephants and their job satisfaction; 2) an Elephant Behavior Profile Survey where keepers rated a total of 234 elephants on the frequencies of 24 behaviors, and 3) a Keeper-Elephant Bonds Survey in which 209 individual keepers rated the strength of their bond with a specific elephant for a total of 427 keeper-elephant pairings. From the first two surveys, principle components analysis was used to create subscales of keeper attitudes, elephant behaviors and keeper job satisfaction. Component scores were then used as independent variables in epidemiological analyses of elephant mean serum cortisol and mixed model regressions of keeper job satisfaction. For African elephants, risk factors of low serum cortisol included Positive interactions with elephants (p = 0.039), Positive behaviors of elephants (friendly, affiliative) (P = 0.001), and Elephant interacts with public (P = 0.009). Age of the elephant was a small, but significant risk factor for higher cortisol (P = 0.005). For Asian elephants, the risk factors for low cortisol were attitudes indicating social inclusion in elephant groups (Keeper as herdmate, P = 0.039) and Elephant interacts with public (P = 0.006). Latitude of zoo was a predictor of higher cortisol (P = 0.041). Significant predictors of keeper Dissatisfaction with job were weaker Keeper-Elephant Bonds (P = 0.003) and African Species (P = 0.038). Species differences in KERs and KEBs are discussed in terms of differing elephant management factors in zoos. The results provide evidence of the reciprocity of KERs and the mutual benefits of KEBs to both elephant and keeper. These results are relevant for zoo animal management and staff training.

Development of a 12-plex of new microsatellite markers using a novel universal primer method to evaluate the genetic diversity of jaguars (Panthera onca) from North American zoological institutions

Maintenance of genetic diversity in both zoo-managed and wild animals is critical to population health. Jaguars are considered Near Threatened throughout their geographic range, and are Endangered in the United States. Genetic diversity of zoo-managed jaguar populations in North America using microsatellite markers has not been reported. To evaluate genetic diversity in jaguars housed in North American zoological institutions compared to wild and other zoo jaguar populations, we created a 12-plex of ten tetranucleotide (n = 10) and two pentanucleotide (n = 2) autosomal microsatellite loci that have not previously been described for use in jaguars, using a universal primer approach which minimizes cost and DNA quantity requirements. A subset of jaguars from North American zoological institutions (n = 35, approximately 28% of population) were genotyped using DNA extracted from whole blood. Microsatellite loci were polymorphic with a range of 3–10 alleles (average 6.42 alleles per locus), and expected heterozygosity (He) between 0.47 and 0.85. Overall population He was 0.73. Probability of identity of individuals (PI) and siblings (PIsibs) were 6.7 × 10− 13 and 2.1 × 10− 5, respectively, confirming that this panel can be used reliably to identify individuals. Using this novel multiplex microsatellite panel, we concluded that jaguars from the North American zoo population have high genetic diversity, comparable to reported wild and other zoo jaguar populations.