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Amebiasis caused by Entamoeba invadens is an important disease in reptile collections, causing severe morbidity and mortality. Surveillance of the parasite at the Singapore Zoo was carried out over a 4-yr period by PCR testing on reptiles that presented with lethargy and enteritis for disease investigation. Asymptomatic reptiles sharing the same enclosures as positive individuals were also tested as part of outbreak investigation. Animals in the collection that tested positive for the parasite were treated with metronidazole at various doses, with the addition of paromomycin for two cases, until a negative PCR test result was obtained at the end of the treatment course. A total of 97 samples from 49 individuals across 19 species of reptiles were obtained, of which 24 samples (24.7%) from 19 animals were positive for E. invadens . Of these positive samples, 11 samples were for disease investigation, eight samples for outbreak surveillance, and five samples for treatment monitoring. Treatment was initiated for 10 animals, four of which were showing clinical signs of disease. The parasite was cleared in nine of these 10 animals (90%), with eight animals receiving metronidazole as a sole therapeutic agent. A total of nine animals died of the disease, four of which (44.4%) presented dead or died within 24 h of presentation. Necrotizing enteritis was a consistent postmortem finding resulting in gastrointestinal perforation in two cases, and coelomic adhesions and hepatic trophozoites were each seen in five animals. The results suggest that the management of Entamoeba epizootics in the collection requires prompt outbreak investigation. Diagnosis of the disease with advanced diagnostic tools like PCR, endoscopy, and ultrasonography and treatment with metronidazole in both symptomatic and asymptomatic animals may reduce mortalities during an outbreak.