Clostridium difficile infection (CDI) has risen in prominence in Australia recently. We conducted laboratory-based surveillance of CDI to examine C. difficile circulating in Australia in October/November 2012. We collected 542 isolates from all States and Territories of Australia except the Northern Territory. The most common ribotypes (RTs) were RTs 014/020 (25.5%), 002 (10.5%), 056 (5.9%) and 070 (4.2%). The survey results were compared with results from a similar Australian survey conducted in 2010. Proportions of RTs 014/020 and 002 remained similar, while RTs 056 (5.9%), 015 (4.1%), 017 (3.3%) and 244 (2.4%) increased in prevalence. Basic clinical and demographic data were available for 338 cases. The majority were healthcare facility-associated (HCFA-CDI, 51.5%) while 17.5% were community-associated (CA-CDI). While no RTs were associated with CA-CDI, RTs 056 and 126 were recently found in Australian production animals, indicating a possible community health threat in Australia.