This observational study assessed hand hygiene practices among visitors to a petting zoo to evaluate behaviors that reduce zoonotic disease transmission. Of 218 individuals who had animal contact, 49% performed some form of hand hygiene, but only 8.3% properly washed their hands before leaving. Factors such as age, time spent in the exhibit, and presence of staff were associated with hygiene behavior. Proper hand washing was primarily linked to time spent in the exhibit. Findings highlight low compliance and support interventions such as improved signage, facility design, and staff engagement to promote effective hygiene practices.