Australian small mammals such as koalas must cope with immense pressure from anthropogenic induced stressors or trauma such as bushfires, vehicle collision impacts and habitat disturbance and land clearance. In addition, they must cope with diseases such as chlamydia. To date, there is no published literature on physiological stress levels in wild koala populations compared with identified environmental stressors. This study investigated physiological stress levels within sub-populations of wild koalas encountering environmental trauma and disease from New South Wales (NSW), Queensland (QLD) and South Australia (SA). Physiological stress was determined using a faecal glucocorticoid (or cortisol) metabolites (FGMs) enzyme-immunoassay (EIA) from 291 fresh faecal samples collected from wild koalas at the point of rescue. A healthy breeding sub-population from a forest reserve in QLD acted as a control group. Clearance of prime Eucalyptus habitat had the largest impact on FGMs, followed by bushfire related factors (e.g. flat demeanour, dehydration and burns injury). Koalas with other sources of physical injury (dog-attacks and vehicle collisions) and disease (chlamydia) also had higher FGMs compared to healthy wild koalas. Healthy wild koalas expressed the lowest median levels of FGMs. Overall, the results highlight that anthropogenic-induced stressors tend to increase physiological stress in wild koalas. Thus, the ultimate stressors such as habitat clearance and bush fire events could increase the incidences of proximate stressors such as dog attacks and vehicle collisions, and increase risks of foliage shortage, diseases and mortality. Therefore, there is need for ecological monitoring, conservation management actions and policy changes to curb the koala population crisis, especially within on-going and future land and road development programs.