Analysis of microbe diversity in freshwater resources and nearshore seawater samples of Upolu Island was performed to investigate the distribution of harmful bacteria. For this, 124 samples were collected from 23 river systems, two volcanic lakes, and 45 locations inside and outside the barrier reef of Upolu Island, Samoa. Physicochemical parameters for general water quality, detection of coliform bacteria and 16S rRNA amplicon sequencing were performed on all samples. Fecal indicator bacteria (FIB) testing indicated a wide distribution of coliform bacteria in all sampled freshwater sites with evidence of fecal coliform in most locations. Importantly, evidence of coliform bacteria was found in most seawater samples inside and along the reef, apart from those samples taken ~20 km offshore. Illumina 16S rRNA amplicon sequencing of the V4 hypervariable region confirmed the presence of various types of harmful bacterial species, namely from the Enterobacteriaceae, Enterococcae, Streptococcaceae, and Vibrionacea families. By combining the sensitivity of FIB testing and next-generation sequencing, we were able to show the extent of potential contaminations in fresh and seawater samples and simultaneously identify the potential pathogenic bacterial genera present. The wide distribution of potential harmful bacteria from river runoff or direct sewage dumping has an impact on human health, leading to many skin and intestinal diseases, and is potentially detrimental to coral reef community health.

Originality-significance statement: Motility and biofilm formation are processes regulated by quorum sensing (QS) in bacteria. Both functions are believed to play an important role in interactions between bacteria and phytoplankton. Here, we show that two bacterial symbionts from the microbial community associated with a ubiquitous diatom switch their motile lifestyle to attached cells while an opportunist bacterium from the same community is incapable of attachment, despite possessing the genetic machinery to do so. Further work indicated that the opportunist lacks QS signal synthases while the symbionts produce three QS signals, one of which is mainly responsible for regulating symbiont colonization of the diatom microenvironment. These findings suggest that QS regulates colonization of diatom surfaces and further work on these model systems will inform our understanding of particle aggregation and bacterial attachment to marine snow and how these processes influence the global carbon cycle. Summary: Interactions between phytoplankton and bacteria play major roles in global biogeochemical cycles and oceanic nutrient fluxes. These interactions occur in the microenvironment surrounding phytoplankton cells, known as the phycosphere. Bacteria in the phycosphere use either chemotaxis or attachment to benefit from algal excretions. Both processes are regulated by quorum sensing (QS), a cell-cell signaling mechanism that uses small infochemicals to coordinate bacterial gene expression. However, the role of QS in regulating bacterial attachment in the phycosphere is not clear. Here, we isolated a Sulfitobacter pseudonitzschiae F5 and a Phaeobacter sp. F10 belonging to the marine Roseobacter group and an Alteromonas macleodii F12 belonging to Alteromonadaceae, from the microbial community of the ubiquitous diatom Asterionellopsis glacialis. We show that only the Roseobacter group isolates (diatom symbionts) can attach to diatom transparent exopolymeric particles. Despite all three bacteria possessing genes involved in motility, chemotaxis, and attachment, only S. pseudonitzschiae F5 and Phaeobacter sp. F10 possessed complete QS systems and could synthesize QS signals. Using UHPLC-MS/MS, we identified three QS molecules produced by both bacteria of which only 3-oxo-C _16:1 -HSL strongly inhibited bacterial motility and stimulated attachment in the phycosphere. These findings suggest that QS signals enable colonization of the phycosphere by algal symbionts.