Interaction among spring phytoplankton succession, water discharge patterns, and hydrogen peroxide dynamics in the Caloosahatchee River in southwest Florida.

Phytoplankton communities are major primary producers in the aquatic realm and are responsible for shaping aquatic ecosystems. The dynamics of algal blooms could be determined by a succession of variable taxonomic groups, which are altered based on complex environmental factors such as nutrient availability and hydraulic factors. In-river structures potentially increase the occurrence of harmful algal blooms (HABs) by increasing water residence time and deteriorating water quality. How flowing water stimulates cell growth and affects the population dynamics of phytoplankton communities is a prioritized question that needs to be addressed for water management tactics. The goal of this study was to determine if an interaction between water flow and water chemistry is present, furthermore, to determine the relationship among phytoplankton community successions in the Caloosahatchee River, a subtropical river strongly influenced by human-controlled water discharge patterns from Lake Okeechobee. Particularly we focused on how phytoplankton community shifts influence the natural abundance of hydrogen peroxide, the most stable reactive oxygen species and a byproduct of oxidative photosynthesis. High-throughput amplicon sequencing using universal primers amplify 23S rRNA gene in cyanobacteria and eukaryotic algal plastids revealed that Synechococcus and Cyanobium were the dominant cyanobacterial genera and their relative abundance ranged between 19.5 and 95.3% of the whole community throughout the monitoring period. Their relative abundance declined when the water discharge increased. On the contrary, the relative abundance of eukaryotic algae sharply increased after water discharge increased. As water temperature increased in May, initially dominant Dolichospermum decreased as Microcystis increased. When Microcystis declined other filamentous cyanobacteria such as Geitlerinema, Pseudanabaena, and Prochlorothreix increased in their relative abundances. Interestingly, a peak of extracellular hydrogen peroxide was observed when Dolichospermum dominance was ended, and M. aeruginosa numbers increased. Overall, phytoplankton communities were strongly impacted by human-induced water discharge patterns.

QT-AMP: Sequencing PCR amplicons from Quanti-Tray wells to analyze enterococci communities.

Enterococcus is ubiquitous in human feces and has been adopted as a useful indicator of human fecal pollution in water. Although regular enterococci monitoring only examines their numbers, identifying human-specific Enterococcus species or genotypes could help discriminate human fecal contamination from other environmental sources. We documented a new approach to characterize enterococci using a high-throughput 16S rRNA gene amplicon sequencing platform from Quanti Trays after following the counting of the most probable numbers of enterococci. We named this method QT-AMP (Quanti-Tray-based amplicon sequencing). We tested surface water samples collected from three rivers in southwest Florida. We detected 11 Enterococcus species from 45 samples in 1.1 million sequence reads. The method detected three rare species and eight cosmopolitan species (Enterococcus faecalis, E. faecium, E. casseliflavus, E. hirae, E. mundtii, E. gallinarum, E. avium, and E. durans) which have been commonly documented in previous studies. The approximate detection level of QT-AMP was four orders of magnitude higher than regular 16S rRNA gene amplicon sequencing. The current Enterolert MPN method only provides quantitative information but now we can look into the relative abundance of Enterococci species composition by accompanying Illumina sequencing. This QT-AMP could be a useful tool to streamline the quantification and identification of enterococci and could be used in various water management projects and human health risk assessment.