Microbial diversity, genomics, and phage-host interactions of cyanobacterial harmful algal blooms.

The occurrence of cyanobacterial harmful algal blooms (cyanoHABs) is related to their physical and chemical environment. However, less is known about their associated microbial interactions and processes. In this study, cyanoHABs were analyzed as a microbial ecosystem, using 1 year of 16S rRNA sequencing and 70 metagenomes collected during the bloom season from Lake Okeechobee (Florida, USA). Biogeographical patterns observed in microbial community composition and function reflected ecological zones distinct in their physical and chemical parameters that resulted in bloom "hotspots" near major lake inflows. Changes in relative abundances of taxa within multiple phyla followed increasing bloom severity. Functional pathways that correlated with increasing bloom severity encoded organic nitrogen and phosphorus utilization, storage of nutrients, exchange of genetic material, phage defense, and protection against oxidative stress, suggesting that microbial interactions may promote cyanoHAB resilience. Cyanobacterial communities were highly diverse, with picocyanobacteria ubiquitous and oftentimes most abundant, especially in the absence of blooms. The identification of novel bloom-forming cyanobacteria and genomic comparisons indicated a functionally diverse cyanobacterial community with differences in its capability to store nitrogen using cyanophycin and to defend against phage using CRISPR and restriction-modification systems. Considering blooms in the context of a microbial ecosystem and their interactions in nature, physiologies and interactions supporting the proliferation and stability of cyanoHABs are proposed, including a role for phage infection of picocyanobacteria. This study displayed the power of "-omics" to reveal important biological processes that could support the effective management and prediction of cyanoHABs. IMPORTANCE: Cyanobacterial harmful algal blooms pose a significant threat to aquatic ecosystems and human health. Although physical and chemical conditions in aquatic systems that facilitate bloom development are well studied, there are fundamental gaps in the biological understanding of the microbial ecosystem that makes a cyanobacterial bloom. High-throughput sequencing was used to determine the drivers of cyanobacteria blooms in nature. Multiple functions and interactions important to consider in cyanobacterial bloom ecology were identified. The microbial biodiversity of blooms revealed microbial functions, genomic characteristics, and interactions between cyanobacterial populations that could be involved in bloom stability and more coherently define cyanobacteria blooms. Our results highlight the importance of considering cyanobacterial blooms as a microbial ecosystem to predict, prevent, and mitigate them.

Periphyton as an indicator of saltwater intrusion into freshwater wetlands: insights from experimental manipulations.

Saltwater intrusion has particularly large impacts on karstic wetlands of the Caribbean Basin due to their porous, carbonate bedrock and low elevation. Increases in salinity and phosphorus (P) accompanying saltwater intrusion into these freshwater, P-limited wetlands are expected to alter biogeochemical cycles along with the structure and function of plant and algal communities. Calcareous periphyton is a characteristic feature of karstic wetlands and plays a central role in trophic dynamics, carbon storage, and nutrient cycling. Periphyton is extremely sensitive to water quality and quantity, but the effects of saltwater intrusion on these microbial mats remain to be understood. We conducted an ex situ mesocosm experiment to test the independent and combined effects of elevated salinity and P on the productivity, nutrient content, and diatom composition of calcareous periphyton from the Florida Everglades. We measured periphyton total carbon, nitrogen, and P concentrations and used settlement plates to measure periphyton accumulation rates and diatom species composition. The light and dark bottle method was used to measure periphyton productivity and respiration. We found that exposure to ~1 g P·m-2 ·yr-1 significantly increased periphyton mat total P concentrations, but had no effect on any other response variable. Mats exposed to elevated salinity (~22 kg salt·m-2 ·yr-1 ) had significantly lower total carbon and tended to have lower biomass and reduced productivity and respiration rates; however, mats exposed to salinity and P simultaneously had greater gross and net productivity. We found strong diatom species dissimilarity between fresh- and saltwater-treated periphyton, while P additions only elicited compositional changes in periphyton also treated with saltwater. This study contributes to our understanding of how the ecologically important calcareous periphyton mats unique to karstic, freshwater wetlands respond to increased salinity and P caused saltwater intrusion and provides a guide to diatom indicator taxa for these two important environmental drivers.

Phosphorus alleviation of salinity stress: effects of saltwater intrusion on an Everglades freshwater peat marsh.

Saltwater intrusion and salinization of coastal wetlands around the world are becoming a pressing issue due to sea level rise. Here, we assessed how a freshwater coastal wetland ecosystem responds to saltwater intrusion. In wetland mesocosms, we continuously exposed Cladium jamaicense Crantz (sawgrass) plants and their peat soil collected from a freshwater marsh to two factors associated with saltwater intrusion in karstic ecosystems: elevated loading of salinity and phosphorus (P) inputs. We took repeated measures using a 2 × 2 factorial experimental design (n = 6) with treatments composed of elevated salinity (~9 ppt), P loading (14.66 µmol P/d), or a combination of both. We measured changes in water physicochemistry, ecosystem productivity, and plant biomass change over two years to assess monthly and two-year responses to saltwater intrusion. In the short-term, plants exhibited positive growth responses with simulated saltwater intrusion (salinity + P), driven by increased P availability. Despite relatively high salinity levels for a freshwater marsh (~9 ppt), gross ecosystem productivity (GEP), net ecosystem productivity (NEP), and aboveground biomass were significantly higher in the elevated salinity + P treated monoliths compared to the freshwater controls. Salinity stress became evident after extended exposure. Although still higher than freshwater controls, GEP and NEP were significantly lower in the elevated salinity + P treatment than the +P treatment after two years. However, elevated salinity decreased live root biomass regardless of whether P was added. Our results suggest that saltwater intrusion into karstic freshwater wetlands may initially act as a subsidy by stimulating aboveground primary productivity of marsh plants. However, chronic exposure to elevated salinity results in plant stress, negatively impacting belowground peat soil structure and stability through a reduction in plant roots.

Salinity pulses interact with seasonal dry-down to increase ecosystem carbon loss in marshes of the Florida Everglades.

Coastal wetlands are globally important sinks of organic carbon (C). However, to what extent wetland C cycling will be affected by accelerated sea-level rise (SLR) and saltwater intrusion is unknown, especially in coastal peat marshes where water flow is highly managed. Our objective was to determine how the ecosystem C balance in coastal peat marshes is influenced by elevated salinity. For two years, we made monthly in situ manipulations of elevated salinity in freshwater (FW) and brackish water (BW) sites within Everglades National Park, Florida, USA. Salinity pulses interacted with marsh-specific variability in seasonal hydroperiods whereby effects of elevated pulsed salinity on gross ecosystem productivity (GEP), ecosystem respiration (ER), and net ecosystem productivity (NEP) were dependent on marsh inundation level. We found little effect of elevated salinity on C cycling when both marsh sites were inundated, but when water levels receded below the soil surface, the BW marsh shifted from a C sink to a C source. During these exposed periods, we observed an approximately threefold increase in CO2 efflux from the marsh as a result of elevated salinity. Initially, elevated salinity pulses did not affect Cladium jamaicense biomass, but aboveground biomass began to be significantly decreased in the saltwater amended plots after two years of exposure at the BW site. We found a 65% (FW) and 72% (BW) reduction in live root biomass in the soil after two years of exposure to elevated salinity pulses. Regardless of salinity treatment, the FW site was C neutral while the BW site was a strong C source (-334 to -454 g C·m-2 ·yr-1 ), particularly during dry-down events. A loss of live roots coupled with annual net CO2 losses as marshes transition from FW to BW likely contributes to the collapse of peat soils observed in the coastal Everglades. As SLR increases the rate of saltwater intrusion into coastal wetlands globally, understanding how water management influences C gains and losses from these systems is crucial. Under current Everglades' water management, drought lengthens marsh dry-down periods, which, coupled with saltwater intrusion, accelerates CO2 loss from the marsh.