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Given the wide usage of Roundup, a common herbicide, the impacts of its presence in ecological communities are of great interest. Many studies have investigated the effects of glyphosate, the active ingredient in Roundup, on different factions of an ecosystem including on animals, plants, microorganisms, and nutrients. The current study expanded upon these works using Roundup instead of glyphosate to provide a realistic application in which to observe the development of microbial assemblages and nutrient composition in two different habitats. Winogradsky columns were prepared using benthic material from a ditch and a pond. Varying concentrations of Roundup were introduced to the columns at the beginning of the study and microbial growth and nutrient compositions from each column were measured weekly. The results indicate that the presence of Roundup has varying effects on microorganisms and nutrients. While photosynthetic microbes were negatively impacted, a shift in the microbial composition to heterotrophic microbes indicates that these microorganisms were able to utilize some ingredients in Roundup as a nutrient source. Additionally, the temporal analysis of nutrient compositions indicated that microbes metabolize glyphosate starting with the phosphate moiety even when the other compounds in Roundup are present. While these trends were observed in both benthic habitats, the composition of the ecological community can affect its ability to utilize the ingredients in Roundup as a nutrient source.