The goal of this study was to undertake a relatively broad microbiological investigation at historic areas of Cape Evans and Ross Island. The two phylogenetically diverse targets were Bacillus anthracis, the causative agent for anthrax disease, and filamentous micro-fungi associated food products. The human activities were presumed to have played a significant contribution to the introduction of non-indigenous Bacillus anthracis and many filamentous micro-fungi at the historic areas on Ross Island. Bacillus anthracis was suspected to be present at Cape Evans based on a circumstantial clinical analysis of the death of the member in Captain Robert F Scott's Terra Nova Expedition in 1912. Detection methods based on polymerase chain reaction (PCR) targeting B. anthracis specific genes on chromosome and two plasmids were employed. DNA extraction was performed by a bead-beating technique from 74 environmental samples. PCR efficiency was compromised probably due to inhibitors in DNA extracts, but improved with higher concentrations of Taq polymerase. Initially a total of 74 environmental samples were screened with one set of primers before positively tested 19 samples were rigorously investigated with seven sets of primers. Nested PCR also increased the target specificity and detection levels. Sequence analyses of the several positive samples from PCR reactions were characteristic to B. anthracis. A diverse range of filamentous micro-fungi were isolated on three different media at two different temperatures, 15 °C and 25 °C, and identified by classical morphological taxonomy from the foodstuffs and internal environmental samples of Captain Robert F Scott's historic hut at Hut Point built in 1901. In total, there were 22 taxa and 14 genera recorded including many cosmopolitan species isolated from the samples, in particular Penicillium species. An extensive literature review of the :filamentous micro-fungi found in Antarctica identified that 7 taxa isolated in the study were not reported previously. Many isolates were obtained at 15 °C while some isolates grew in the presence of antibiotics.