Digital literacy is one of the important skills that must be mastered by Biology Teacher. An effort has been applied to integrate the learning of Animal Systematics with the training of digital literacy for pre-service biology teachers. This research aimed to evaluate the digital literacy of pre-service teachers that programming the Animal Systematics Course as well as evaluate the need for the course related to the implementation of digital literacy training. This research used mixed method, involving one group shoot case study. The samples were 25 pre-service biology teachers programming Animal Systematics Course. The course implemented projectbased tasks, namely animal systematics mini-research, scientific writing, and phylogenetics reconstruction. The digital literacy of pre-service teachers was evaluated based-on four indicators, namely internet searching, hypertextual navigation, content evaluation, and knowledge assembly. The data of the digital literacy score was analyzed descriptive-qualitatively. The result revealed that literacy digital of pre-service biology teacher can be enhanced through the learning process of Animal Systematics Course. The scores of internet searching, hypertextual navigation, content evaluation, and knowledge assembly rose significantly, reached a score of 85 on average. It can be concluded that digital literacy can be trained during the learning process of animal systematics. This research also recommends the need for an electronic book of animal systematics as supporting learning resources.