POUCH PRESENTATION OF THE MACROPUS RUFUS RED KANGAROO FOR CYTOLOGICAL ANALYSIS Beatrice DeMarco Thesis Advisor, Chair of the Defense Committee: Dr. Christopher Corbo Committee Member: Dr. Brian Palestis

INTRODUCTION ● Understanding the normal flora that exists in the pouch of marsupials is crucial for the sustainability of species in regards to pathogens and the treatment of disease. ● Developing a basis of what prokaryotic and eukaryotic cells exist in the pouch provide further information about what cells are necessary for healthy developmental growth of the fetal organism from the gestation period. ● Previous research: ○ Tammar wallabies (Macropus eugenii): ■ Actinobacteria, Corynebacterium ■ DNA sequencing (NextGen sequencing - massively parallel sequencing) ○ Koalas (Phascolarctos cinereus): ■ Pouch sampling technique ■ Pseudomonas aeruginosa, Serratia marcescens, Enterococcus faecalis ■ Antimicrobial secretion from pouch epithelial cells. ● Behavioral plasticity of kangaroos from free-ranging to captive conditions. ● Training for voluntary administration of oral medication and injections. ● Behavioral training techniques designed for animals in captivity is crucial for sample collection to be used in both veterinary and research fields. OBJECTIVES

● This experiment is separated into a behavioral and microbial portion curated to determine a catalog of normal flora from the female red kangaroo (Macropus rufus). ● Utilize behavioral techniques and procedures to obtain pouch samples from the pouch of female red kangaroos (Macropus rufus). ● Design behavioral shaping plans under the Association of Zoos & Aquariums (AZA) guidelines to develop training procedures. ● Apply the Diff-Quik and gram stain procedures to identify the cells of the pouch samples. ● Light microscopy attached to a Light Camera System to obtain picture results utilizing oil immersion at different magnifications. “ADELAIDE” AZA Identification Number: A002191 MATERIALS & METHODS: BEHAVIORAL MATERIALS & METHODS: MICROBIAL RESULTS: BEHAVIORAL AD LIBITUM BEHAVIORAL SAMPLING RESULTS: BEHAVIORAL Scan Sampling Technique RESULTS: BEHAVIORAL AZA DESIGNED BEHAVIORAL SHAPING PLAN RESULTS: BEHAVIORAL AZA DESIGNED BEHAVIORAL SHAPING PLAN RESULTS: MICROBIAL RESULTS: MICROBIAL RESULTS: MICROBIAL RESULTS: MICROBIAL RESULTS: MICROBIAL RESULTS: MICROBIAL CONCLUSIONS ● There is both eukaryotic and prokaryotic cells that makeup the normal flora of the red kangaroo (macropus rufus) pouch. ● As per each sample collection and comprised catalogue, there is a greater presence of epithelial cells compared to prokaryotic cells. ● Most of the prokaryotic cells found were gram positive as indicated by the purple stain. ● The incorporation of reinforcers and lag time between training sets within a session proved to have the highest efficiency rate in attaining a behavioral shaping goal. Acknowledgments

● I would like to thank Dr. Sarrah Kaye, the head curator and veterinarian of the Staten Island Zoo. Without her guidance, endless knowledge, and love for these animals this project would not have been possible. ● Additionally, I would like to thank Dr. Christopher Corbo for giving me the ability to conduct this research and helping continuously throughout the process. ● I would like to extend endless gratitude for Dr. Brian Palestis for being constantly available to answer any and all questions I had during the project without his being on campus directly.