本片讲述的是蝙蝠侠（罗杰·克雷格·史密斯配音）以及罗宾联合正义联盟成员闪电侠以及绿箭侠一起对抗被称为Animilitia的犯罪组织。该犯罪组织都是由一些动物性的成员组成，包括银背大猩猩、猎豹女、杀手鳄鱼等，他们在高谭市作恶多端，一般警察对他们束手无策。而且该组织背后的主谋却指向蝙蝠侠的宿敌，同时也是高谭市叱吒风云的黑道大亨企鹅人（DanaSnyder配音）。除了Animilitia组织，蝙蝠侠他们这次还要面对奥斯瓦德公司的商人新发明的邪恶网络动物机器人，一群犯罪恶势力再次聚集在高谭市。
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