Fecal Glucocorticoid patterns among felids during quarantine and animal transfers Helen Clawitter1, Carol Fieseler2, Corinne Kozlowski1, Steve Bircher3, Julie Hartell-DeNardo3, David Powell1 1Research Department; 2Animal Health Department; 3Carnivore Department; Saint Louis Zoo

Introduction The impact of animal quarantine and transfer on animal well-being has been pondered; however, few studies have quantified how animals respond and adjust to these experiences despite the complete change in the animals’ life. In this study, we document the glucocorticoid responses of five felids transferred to the Saint Louis Zoo (two cheetahs, two snow leopards, one jaguar) preceding transfer, throughout quarantine, and following introduction to a new habitat. Our objectives were to assess the impact of institutional transfer on glucocorticoid production and to determine whether specific behaviors were Figure 2: (A) Cheetah (photo by Gretchen Freund); (B) Snow Leopard associated with increased or reduced glucocorticoid levels during quarantine. (photo by Chuck Dresner); and (C) Jaguar (photo by Christopher Carter) at the Saint Louis Zoo. Material and Methods Animals Results Table 1: Five felids transferred to Saint Louis Zoo and included in study

Figure 4: The percentage of surveys that recorded (A) pacing; (B) eating overnight; and positive, negative or no reactions to (C) training Sample collection and hormone analysis activities; (D) the presentation Fecal samples were collected daily for at least twenty days before institutional of enrichment; and (E) the transfer, during a quarantine period at the Saint Louis Zoo Veterinary Hospital, presence of keepers. and for at least thirty days after transfer to a permanent habitat. Sample collection occurred from October 2015 to March 2016. Samples were collected within 16 hours of defecation and frozen at -80°C until extraction for steroid hormones. In total, 328 fecal samples were collected from five individuals. Results and Findings All animals experienced a 2-3 fold increase in FGM in the first week of Fecal glucocorticoid levels were quantified using a radioimmunoassay quarantine, compared to concentrations prior to transfer. Baseline FGM for (Corticosterone DA 125I Kit, ICN MP Biomedicals). most individuals were around 150 ng/g or less with periodic values in the mid- to upper hundreds. Quarantine Behavior Surveys Quarantine staff completed daily surveys at completion of the work day, noting if Temporal patterns of FGM concentrations differed among individuals. an animal ate overnight, was observed pacing, and the reactions to enrichment, The jaguar and Cheetah 1 demonstrated increased FGM levels throughout training, and care staff. For enrichment, the felids were offered objects (plastic quarantine, while concentrations for cheetah 2 declined each week. balls, barrels, etc.), food (rabbit foot, bone), and other sensory stimulating Concentrations for both snow leopards decreased over time, but items (radio, fresh snow, hay used by another felid), and staff noted if the Snow Leopard 1 showed elevated FGM during its final week in quarantine. reaction was positive (animal interacted with item), no reaction (ignored item), or negative (fear or flight response). The training in quarantine included chute, When moved to their permanent habitat, four individuals experienced scale, target, crate, and hand injection. Staff noted the reactions to training as another increase in FGM. Concentrations during this first week were positive (completed the training with no aggression), no reaction (no participation), substantially greater than levels during the first week in quarantine. Snow and negative (aggression with no participation). Reactions to care staff were also Leopard 2 demonstrated reduced FGM concentrations upon moving to noted as positive (approached staff with little to no aggression as expected for the her habitat. species), no reaction (animal was passive), and negative (animal demonstrated aggression) at the culmination of the work day. Finally, staff noted any special Behavior surveys indicated that cheetahs were less likely to demonstrate events or procedures each day. positive reactions to training and keeper staff. Cheetah 1 had the largest percent increase in FGM concentrations during quarantine, compared to pre-transfer values, and he was also most often recorded as responding Figure 3: Fecal glucocorticoid metabolite (FGM) profiles for negatively to keeper staff. The percentage of days individuals were (A) Cheetah A; (B) Cheetah B; (C) Snow leopard D; (D) Snow Leopard E; documented as eating overnight, pacing, and responses to enrichment and (F) Jaguar C preceding transfer, after transfer during a 30-day did not differ across cats. quarantine period, and after relocation to a permanent habitat. Day 0 indicates the day of arrival at the Saint Louis Zoo, the red asterisk denoted day of quarantine exam with assumed 24 hour delay in corresponding glucocorticoid passage through G.I. tract, and the red Conclusions arrow denoted the day of transfer to a permanent habitat. (F) Mean fecal • While transfer and quarantine may initially be a “stressful” experience, glucocorticoid metabolite (FGM) concentrations (±S.E) prior to transfer, individuals vary substantially in their responses and some appear to during quarantine and after relocation to a permanent habitat. acclimate soon after arrival. • Reactions to keepers may help staff assess individual stress levels during quarantine, whereas responses to training, enrichment and Acknowledgements other behavior may not be informative. Figure 1: (A) Cheetah training for The authors would like to thank the Saint Louis Zoo’s Quarantine hand injection during quarantine staff, Mallory Carmean and Kim Olson, and Carnivore staff for • Transfer to permanent housing may also be a “stressful” experience, (B) Jaguar in quarantine; (C) Snow sample collection, as well as staff from pre-shipment institutions as animals are often introduced to new conspecifics, care staff, and zoo Leopard during crate training in for collection of samples. visitors. Procedures that help animals cope during this time may quarantine. (Photos by Carol improve well-being. Fieseler and Mallory Carmean)