The microbiological and physicochemical characteristics of several commercial beef, kudu and springbok biltong samples from South Africa were assessed in this study. Analysis of samples allowed their differentiation into ‘dry’ and ‘moist’ samples. Dry biltong showed low moisture content (21.5–25.3 g/100 g), a low water activity (0.65–0.68) and a high salt content (5.5–7.9 g/100 g), while moist biltong showed a higher moisture content (35.1–42.8 g/100 g), a higher water activity (0.85 to 0.89) and a lower salt content (3.8–5.6 g/100 g). The pH value did not vary significantly between both groups (5.00–6.26). The results showed that dry biltong presented a low total plate count (TPC) content, a high level of lactic acid bacteria (LAB) and a high level of d-lactic acid as compared to moist biltong. These results suggest that dry biltong complies with the standard hygienic quality (TPC < 7 log cfu/g in agreement with the Food Standards Agency), while moist biltong samples generally showed a low content of d-lactic acid and a low ratio LAB/TPC.