With the climate change, tropical regions of Africa are witnessing a biodiversity decline within agricultural landscapes. This loss of biodiversity threatens ecological integrity and the resilience of food production systems, particularly for smallholder farming who depend of a range of food and cash crops for subsistence and local markets. Unfortunately, the species and the ecological functions which support these smallholder agroecosystems remain poorly understood, limiting the implementation of sustainable practices. This study aimed to assess the biodiversity of smallholder agroforests using ants as biological indicators. Study was focused on the land-use types of smallholder agroforests, including fallows, rural forests, cocoa and rubbers orchards from two different agroecological zones in Côte d’Ivoire. We sampled ants using the standardized methods of winklers, pitfall trap and baiting. Results revealed a rich ant fauna with 81 ant species belonging 27 genera and 6 subfamilies. We recorded 53 and 48 ant species for forest-savanna mosaic and rainy forest regions, respectively. The subfamilies identified includes Myrmicinae (57% of species richness), Formicinae (26%), Ponerinae (8%), Dorylinae and Dolichoderinae (4%, respectively), and Pseudomyrmecinae (1 %). Shannon diversity index value was high in Cocoa RF (3.26) followed by Fallows RF (.90), Cocoa FSM (2.47) and low in Ruralforest (2.04) and Rubber FSM (2.02) Results also showed a variation of ant species composition (ANOSIM: R = 66.4, P= 0.0002) between the agroforests of the both agroecological zones. The most abundant ant species was the aboreal dominant weaver ant Oecophylla longinoda. The most prevalent ant species in rainy forest smallholder agroforests were Crematogaster sp. 2, Nylanderia scintilla, Paltothyreus tarsata, Hypoponera dulcis, Oecophylla longinoda, Monomorium sp. 1 and Carebara distincta. In forest-savanna mosaic agroforest, Oecophylla longinoda, Crematogaster sp. 14, Camponotus maculatus, Paltothyreus tarsata, Pheidole sp.6, Crematogaster solenopsides, Monomorium floricola, Pheidole excellens, Carebara thoracica, Pheidole sp.4. Our study demonstrated that ant communities respond differently to land-use in smallholder agroforests across ecological regions. It provides baseline data of valuable importance and emphasises the importance of continued surveys in smallholder agroecosystems to increase knowledges on ants and other groups of insects, including pollinators and by extension biodiversity.