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The incidence and abundance of Monogononta rotifer species were recorded from lakes and reservoirs in the upper part of the Cambodian Mekong River basin in April and November 2010. One hundred and seven species are reported, 25 of which are new records to Cambodia and 8 taxa were unidentifiable to species level. Species richness at the regional and local scale was not significantly different between habitat types or between seasons, whether it was estimated using incidence or abundance data. Comparison of incidence data also revealed no significant difference from species richness of ponds concurrently sampled in the same region. There appeared to be a high level of diversity among sites that could not be attributed to nestedness or to the 5 environmental variables measured. Each habitat type and season offered substantially different rotifer communities, with the proportion of unshared species between sample sets ranging from 14-49%. Non-metric multidimensional scaling and PERMANOVA analyses also revealed clustering and significant differences among sample sets based on habitat type and season. Therefore, each habitat type and season contributed to the overall rotifer biodiversity. When the incidence data from this study are combined with those in previously reported studies, the overall species richness estimate for Cambodia is 403 species (95% CI = 386-432) and the number of species records has reached 306.