Problems arising from conventional tillage (CT) systems (such as soil erosion, decrease of organic matter, environmental damage etc.) have led many farmers to the adoption of no-till (NT) systems that are more effective in improving soil physical, chemical and microbial properties. Results from this study clearly indicated that NT, mulch tillage (MT), and winter rye cover cropping systems increased the activity of phosphatase, β-glucosidase and arylsulfatase at a 0–10 cm soil depth but decreased the activity of these enzymes at 10–20 cm. The increase in enzyme activity was a good indicator of intensive soil microbial activity in different soil management practices. Poultry litter (PL) application under NT, MT, and rye cropping system could be considered as effective management practices due to the improvement in carbon (C) content and the biochemical quality at the soil surface. The activities of the studied enzymes were highly correlated with soil total nitrogen (STN) soil organic carbon (SOC) at the 0–10 cm soil depth, except for acid phosphatase where no correlation was observed. This study revealed that agricultural practices such as tillage, PL, and cover crop cropping system have a noticeable positive effect on soil biochemical activities under cotton production.