Microbial community composition, extracellular enzymatic activities, and structure-function relationships in the central Arctic Ocean, a high-latitude fjord, and the North Atlantic Ocean

Due to their abundance, diversity, and capabilities to transform and metabolize diverse compounds, microbial communities regulate biogeochemical cycles on micro-, regional, and global scales. The activities of microbial communities affect the flow of matter, energy sources of other organisms, and human health, as well as other aspects of life. Yet, the composition, diversity, and ecological roles of microbes in parts of the global oceans—from the high latitudes to the deep water column—remain underexplored. Drawing from microbiological, oceanographic, and ecological concepts, this dissertation explores several fundamental topics: 1) the manner in which hydrographic conditions influence microbial community composition; 2) the ability of these microbial communities across environmental and depth gradients to hydrolyze organic compounds; and 3) microbial structure-function relationships in different habitats and under altered environmental conditions. In the central Arctic Ocean, the composition and enzymatic function of pelagic, particle associated, and benthic bacterial communities varied with depth and region, in parallel with specific hydrographic features. The microbial structure-function relationship in the pelagic realm indicated functional redundancy, suggesting that bacterial compositional shifts—in response to the changing Arctic—may have complex and less predictable functional consequences than previously anticipated. In Tyrolerfjord-Young Sound, northeast Greenland, microbial enzymatic activity patterns were investigated in rivers and within the fjord. Activity patterns correlated with the composition of bacterial communities and dissolved organic matter in the same waters, suggesting that factors extrinsic (organic matter supply) and intrinsic (composition) to microbial communities may, in concert, influence their heterotrophic activities. Finally, functional consequences of differences in community composition were further explored in the North Atlantic. Enriched with high molecular weight organic matter, compositionally-distinct microbial communities exhibited convergent and divergent successional patterns. While convergent features were driven by several initially rare taxa, overarching successional differences in microbial community composition and enzymatic profiles provide evidence for the functional significance of community structure. The integration of community compositional analyses and enzymatic activity measurements has provided valuable information on the identity, ecological roles, and environmental sensitivity of microbial communities in previously underexplored oceanic regions and depths. These insights can be used to evaluate the potential for environmental changes to alter marine microbial community structure and function.Doctor of Philosoph

Impacts of a large storm disturbance on microbial community composition in the Tar River of North Carolina

Microbial community response to environmental disturbances is a crucial topic to investigate as changes in community composition can have widespread consequences on an ecosystem. Microbes are generally sensitive to disturbances, but can exhibit resilience, capable of returning to pre-disturbance composition or achieving an alternative stable state. Here, I investigate microbial community composition in the Tar River of North Carolina from November 2010 to November 2011, capturing the landfall of Hurricane Irene on August 27, 2011. Using 16S ribosomal RNA sequence analyses, I investigated the response of riverine microbial communities to Hurricane Irene and found a likely hurricane-induced change or signature. A finer analysis on two major groups, Betaproteobacteria and Verrucomicrobia, demonstrate the ecophysiological flexibility of their members, persisting and, in some instances, proliferating in the face of disturbance. The alteration in Tar River microbial community composition indicates sensitivity to disturbances, although a larger sampling window is required to assess resilience.Master of Scienc

Depth-related patterns in microbial community responses to complex organic matter in the western North Atlantic Ocean

Oceanic bacterial communities process a major fraction of marine organic carbon. A substantial portion of this carbon transformation occurs in the mesopelagic zone, and a further fraction fuels bacteria in the bathypelagic zone. However, the capabilities and limitations of the diverse microbial communities at these depths to degrade high-molecular-weight (HMW) organic matter are not well constrained. Here, we compared the responses of distinct microbial communities from North Atlantic epipelagic (0–200 m), mesopelagic (200–1000 m), and bathypelagic (1000–4000 m) waters at two open-ocean stations to the same input of diatom-derived HMW particulate and dissolved organic matter. Microbial community composition and functional responses to the input of HMW organic matter – as measured by polysaccharide hydrolase, glucosidase, and peptidase activities – were very similar between the stations, which were separated by 1370 km but showed distinct patterns with depth. Changes in microbial community composition coincided with changes in enzymatic activities: as bacterial community composition changed in response to the addition of HMW organic matter, the rate and spectrum of enzymatic activities increased. In epipelagic mesocosms, the spectrum of peptidase activities became especially broad and glucosidase activities were very high, a pattern not seen at other depths, which, in contrast, were dominated by leucine aminopeptidase and had much lower peptidase and glucosidase rates in general. The spectrum of polysaccharide hydrolase activities was enhanced particularly in epipelagic and mesopelagic mesocosms, with fewer enhancements in rates or spectrum in bathypelagic waters. The timing and magnitude of these distinct functional responses to the same HMW organic matter varied with depth. Our results highlight the importance of residence times at specific depths in determining the nature and quantity of organic matter reaching the deep sea.</p

Riverine Bacterial Communities Reveal Environmental Disturbance Signatures within the Betaproteobacteria and Verrucomicrobia

Riverine bacterial communities play an essential role in the biogeochemical coupling of terrestrial and marine environments, transforming elements and organic matter in their journey from land to sea. However, precisely due to the fact that rivers receive significant terrestrial input, the distinction between resident freshwater taxa vs. land-derived microbes can often become ambiguous. Furthermore, ecosystem perturbations could introduce allochthonous microbial groups and reshape riverine bacterial communities. Using full- and partial-length 16S ribosomal RNA gene sequences, we analyzed the composition of bacterial communities in the Tar River of North Carolina from November 2010 to November 2011, during which a natural perturbation occurred: the inundation of the lower reaches of an otherwise drought-stricken river associated with Hurricane Irene, which passed over eastern North Carolina in late August 2011. This event provided the opportunity to examine the microbiological, hydrological, and geochemical impacts of a disturbance, defined here as the large freshwater influx into the Tar River, superimposed on seasonal changes or other ecosystem variability independent of the hurricane. Our findings demonstrate that downstream communities are more taxonomically diverse and temporally variable than their upstream counterparts. More importantly, pre- vs. post-disturbance taxonomic comparison of the freshwater-dominant Betaproteobacteria class and the phylum Verrucomicrobia reveal a disturbance signature of previously undetected taxa of diverse origins. We use known traits of closely-related taxa to interpret the ecological function of disturbance-associated bacteria, and hypothesize that carbon cycling was enhanced post-disturbance in the Tar River, likely due to the flux of organic carbon into the system associated with the large freshwater pulse. Our analyses demonstrate the importance of geochemical and hydrological alterations in structuring bacterial communities, and illustrate the response of temperate riverine bacteria on fine taxonomic scales to a disturbance

Structure and function of high Arctic pelagic, particle-associated and benthic bacterial communities

Arctic marine microbes are affected by environmental changes that may ultimately influence their functions in carbon cycling. Here, we investigated in concert the structure and enzymatic activities of pelagic, particle-associated and benthic bacterial communities in the central Arctic Ocean, and used these data to evaluate microbial structure–function relationships. Our findings showed influences of hydrographic conditions and particle association on community composition, and sharp pelagic-benthic contrasts. In addition to community compositional differences, regional and depth-related patterns in enzymatic activities were observed. Peptide hydrolysis rates were highest in surface waters, especially at ice-free and first year ice-covered regions, and decreased with depth. While the range of hydrolysed polysaccharides showed varying geographic patterns, particles often showed a wider spectrum of polysaccharide hydrolase activities. Summed benthic peptidase rates differed across stations but showed similar proportions of individual enzyme activities. Analysing for potential linkages between structure and function after subtracting the effect of environmental conditions revealed no direct link, indicating functional redundancy to carry out peptide hydrolysis among pelagic microbes. Thus, while community composition and activities are influenced by environmental conditions, bacterial functional redundancy suggests that compositional shifts – in response to the changing Arctic – may have complex and less predictable functional consequences than previously anticipated

A sea change in microbial enzymes: Heterogeneous latitudinal and depth-related gradients in bulk water and particle-associated enzymatic activities from 30°S to 59°N in the Pacific Ocean

Heterotrophic microbes initiate the degradation of high molecular weight organic matter using extracellular enzymes. Our understanding of differences in microbial enzymatic capabilities, especially among particle-associated taxa and in the deep ocean, is limited by a paucity of hydrolytic enzyme activity measurements. Here, we measured the activities of a broad range of hydrolytic enzymes (glucosidases, peptidases, polysaccharide hydrolases) in epipelagic to bathypelagic bulk water (nonsize-fractionated), and on particles (≥ 3 μm) along a 9800 km latitudinal transect from 30°S in the South Pacific to 59°N in the Bering Sea. Individual enzyme activities showed heterogeneous latitudinal and depth-related patterns, with varying biotic and abiotic correlates. With increasing latitude and decreasing temperature, lower laminarinase activities sharply contrasted with higher leucine aminopeptidase (leu-AMP) and chondroitin sulfate hydrolase activities in bulk water. Endopeptidases (chymotrypsins, trypsins) exhibited patchy spatial patterns, and their activities can exceed rates of the widely measured exopeptidase, leu-AMP. Compared to bulk water, particle-associated enzymatic profiles featured a greater relative importance of endopeptidases, as well as a broader spectrum of polysaccharide hydrolases in some locations, and latitudinal and depth-related trends that are likely consequences of varying particle fluxes. As water depth increased, enzymatic spectra on particles and in bulk water became narrower, and diverged more from one another. These distinct latitudinal and depth-related gradients of enzymatic activities underscore the biogeochemical consequences of emerging global patterns of microbial community structure and function, from surface to deep waters, and among particle-associated taxa

Patterns of environmental variability influence coral-associated bacterial and algal communities on the Mesoamerican Barrier Reef

A coral's capacity to alter its microbial symbionts may enhance its fitness in the face of climate change. Recent work predicts exposure to high environmental variability may increase coral resilience and adaptability to future climate conditions. However, how this heightened environmental variability impacts coral-associated microbial communities remains largely unexplored. Here, we examined the bacterial and algal symbionts associated with two coral species of the genus Siderastrea with distinct life history strategies from three reef sites on the Belize Mesoamerican Barrier Reef System with low or high environmental variability. Our results reveal bacterial community structure, as well as alpha- and beta-diversity patterns, vary by host species. Differences in bacterial communities between host species were partially explained by high abundance of Deltaproteobacteria and Rhodospirillales and high bacterial diversity in Siderastrea radians. Our findings also suggest Siderastrea spp. have dynamic core bacterial communities that likely drive differences observed in the entire bacterial community, which may play a critical role in rapid acclimatization to environmental change. Unlike the bacterial community, Symbiodiniaceae composition was only distinct between host species at high thermal variability sites, suggesting that different factors shape bacterial versus algal communities within the coral holobiont. Our findings shed light on how domain-specific shifts in dynamic microbiomes may allow for unique methods of enhanced host fitness

On Single-Cell Enzyme Assays in Marine Microbial Ecology and Biogeochemistry

Extracellular enzyme activity is a well-established parameter for evaluating microbial biogeochemical roles in marine ecosystems. The presence and activity of extracellular enzymes in seawater provide insights into the quality and quantity of organic matter being processed by the present microorganisms. A key challenge in our understanding of these processes is to decode the extracellular enzyme repertoire and activities of natural communities at the single-cell level. Current measurements are carried out on bulk or size-fractionated samples capturing activities of mixed populations. This approach – even with size-fractionation – cannot be used to trace enzymes back to their producers, nor distinguish the active microbial members, leading to a disconnect between measured activities and the producer cells. By targeting extracellular enzymes and resolving their activities at the single-cell level, we can investigate underlying phenotypic heterogeneity among clonal or closely related organisms, characterize enzyme kinetics under varying environmental conditions, and resolve spatio-temporal distribution of individual enzyme producers within natural communities. In this perspective piece, we discuss state-of-the-art technologies in the fields of microfluidic droplets and functional screening of prokaryotic cells for measuring enzyme activity in marine seawater samples, one cell at a time. We further elaborate on how this single-cell approach can be used to address research questions that cannot be answered with current methods, as pertinent to the enzymatic degradation of organic matter by marine microorganisms

Community structural differences shape microbial responses to high molecular weight organic matter

The extent to which differences in microbial community structure result in variations in organic matter (OM) degradation is not well understood. Here, we tested the hypothesis that distinct marine microbial communities from North Atlantic surface and bottom waters would exhibit varying compositional succession and functional shifts in response to the same pool of complex high molecular weight (HMW-OM). We also hypothesized that microbial communities would produce a broader spectrum of enzymes upon exposure to HMW-OM, indicating a greater potential to degrade these compounds than reflected by initial enzymatic activities. Our results show that community succession in amended mesocosms was congruent with cell growth, increased bacterial production and most notably, with substantial shifts in enzymatic activities. In all amended mesocosms, closely related taxa that were initially rare became dominant at time frames during which a broader spectrum of active enzymes were detected compared to initial timepoints, indicating a similar response among different communities. However, succession on the whole-community level, and the rates, spectra and progression of enzymatic activities, reveal robust differences among distinct communities from discrete water masses. These results underscore the crucial role of rare bacterial taxa in ocean carbon cycling and the importance of bacterial community structure for HMW-OM degradation

Gulf Stream ring water intrusion on the Mid-Atlantic Bight continental shelf break affects microbially driven carbon cycling

Warm core, anticyclonic rings that spin off from the Gulf Stream circulate through the region directly offshore of the Mid-Atlantic Bight. If a warm core ring reaches the continental shelf break, its warm, highly saline water may subduct under cooler, fresher continental shelf surface water, resulting in subsurface waters at the shelf break and over the upper continental slope with high temperatures and salinities and distinct physical and chemical properties characteristic of Gulf Stream water. Such intruding water may also have microbial communities with distinct functional capacities, which may in turn affect the rate and nature of carbon cycling in this coastal/shelf environment. However, the functional capabilities of microbial communities within ring intrusion waters relative to surrounding continental shelf waters are largely unexplored. We investigated microbial community capacity to initiate organic matter remineralization by measuring hydrolysis of a suite of polysaccharide, peptide, and glucose substrates along a transect oriented across the Mid-Atlantic Bight shelf, shelf break, and upper slope. At the outermost sampling site, warm and salty water derived from a Gulf Stream warm core ring was present in the lower portion of the water column. This water exhibited hydrolytic capacities distinct from other sampling sites, and exhibited lower heterotrophic bacterial productivity overall. Warm core rings adjacent to the Mid-Atlantic Bight shelf have increased in frequency and duration in recent years. As the influence of warm core rings on the continental shelf and slope increases in the future, the rate and nature of organic matter remineralization on the continental shelf may also shift