The adoptability of an animal from a shelter largely depends upon its socialization and friendliness towards humans. For kittens, habituation and proper socialization is an important part of ensuring that the adult cat it will be able to interact properly with humans, thus reducing its chance of being relinquished in the future. In addition, kittens that have been relinquished or placed into a shelter are often subject to several stressors that may impact not only the well-being of the kitten but impair further socialization attempts. The objective of this study was to determine if the kittens’ responses to handling tests improved over a period of 5 consecutive days. This study was conducted at the Animal Rescue League of Iowa (ARL-IA), and involved 14 neonate kittens of mixed sex and breed, between 6 and 8 weeks of age. The treatment was five consecutive days of handling. During treatment kittens were exposed to several handling tests. Data will be presented descriptively. Kittens over the five days scored on average a 1 for the majority of handling tests which indicates a calm kitten. On days four and five, kittens allowed for their rear paws to be held for the maximum 10-s. For front paws on day 4 kittens allowed their paws to be held 9-s but by day 5 this had dropped to 6- s (front left) and 7-s (front right) s respectively. In conclusion, though kittens did not tolerate their front paws being handled as long as rear paws by day five. Overall handling tests conducted on the kittens did not result in any aversive reaction from the kitten to the handler. This data could be useful in further developing socialization and acclimatization programs for kittens in shelters, thereby increasing their adoptability and overall well-being, both in the present and the future.