This study demonstrated the use of high-throughput sequencing to assess the efficacy of an integrated ultrafiltration (UF)–reverse osmosis (RO) desalination pilot plant located at the Arabian Gulf, and to identify potential microbial-associated problems that may arise in this plant. When integrated into the desalination treatment system, the UF membranes were able to serve as a good pretreatment strategy to delay RO fouling by achieving up to 1.96-log removal of cells from the seawater. Consequently, the differential pressure of the RO membrane remained around 1 bar for the entire six-month study, suggesting no significant biofouling performance issue identified for this RO system. Examples of microbial populations effectively removed by the UF membranes from the feed waters included Nitrosoarchaeum limnia and phototrophic eukaryotes. Microbial-associated problems observed in this pilot plant included the presence of Pseudomonas spp. in coexistence with Desulfovibrio spp. These two bacterial populations can reduce sulfate and produce hydrogen sulfide, which would in turn cause corrosion problems or compromise membrane integrities. Chemical-enhanced backwashing (CEB) can be used as an effective strategy to minimize the associated microbial problems by removing bacterial populations including sulfate reducers from the UF membranes.