Ostracods and foraminifera are excellent indicators of environmental change and can act as proxies for the presence of seagrass meadows. These proxies have been under-utilized in vulnerable coastal ecosystems in South-east Asia, and the fundamental habitat and environmental parameters required for such application in environmental monitoring have not yet been established. We investigated the habitat preferences of ostracods and foraminiferal species in seagrass and non-seagrass habitats within Sungai Pulai Estuary (Johor, Malaysia), a system currently undergoing major coastal changes. Samples consisted of surficial and downcore sediments collected from two seagrass meadows and a non-seagrass habitat. Multivariate analysis determined the variations in spatial and depth distribution of the meiofauna. Species dominance, abundance and distribution varied between sites, whereas diversity and community structure varied with sediment depth. We found fewer ostracod individuals (N = 1133) than foraminifera (N = 7242). Ostracods were more species-diverse (H′ = 3.34) in the non-vegetated area compared with seagrass areas (H′ = 2.74), whereas foraminifera species were most diverse (H′ = 3.60) in seagrass areas. Opportunistic species, such as Loxoconcha lilljeborgii, Asterorotalia pulchella, Murrayinella globosa, Ammonia tepida and Elphidium neosimplex dominated the meiofaunal assemblages. The presence of Nummulitidae and Paracyprididae in downcore samples provided information related to rare species and families. Salinity, organic matter and percentage of sand explained much of the meiofaunal distribution. Our findings provide new insight into the factors influencing the presence and distribution of ostracods and foraminifera in the estuary, comprising baseline information for understanding the vulnerability of such habitats to anthropogenic changes.