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Macrobenthos are very useful organisms for monitoring marine environmental and widely use in marine ecology research. They are able to monitor the difference phase in the recovery stage of disturbed sites by appear different species macrobenthos after the cessation of the impact. Univariate and multivariate methods were use to study the macrobenthos community within Sungai Pulai estuary, Johor, Malaysia. Five sub-samples were taken at each sampling sites by using 10 cm diameter corer. Crustaceans were the most abundant at Tanjung Adang (St. 1) and the station of non-seagrass area (St. 2) while polychaetes were the most abundant at Merambong Shoal (St. 3). Higher density of macrobenthos was found at St.3 followed by St. 1 and St. 2. The commonly used population indices such as diversity, richness, evenness and dominance were employed to determine the differences in diversity and abundance of macrobenthos. The diversity, richness and evenness index values showed slight increment from Station 1 to Station 3, while the dominance index decreasing trend from Station 1 to Station 3. A total 21 polychaete families were collected in Sungai Pulai estuary, which was dominated by the Spionidae, Capitellidae and Glyceridae. Cluster (Bray-Curtis similarities) analyses revealed that the Tanjung Adang and Merambong Shoal population were clearly separated from the station non-seagrass. For the time being factors that influence the pattern of distribution of the macrobenthos cannot be determined and subjected to further studies.