James L. Pinckney

A Mini-review of Microbial Consortia: Their Roles in Aquatic Production and Biogeochemical Cycling

Molecular oxygen (O2) is a potent inhibitor of key microbial processes, including photosynthesis, N2 fixation, denitrification, sulfate reduction, methanogenesis, iron, and metal reduction reactions. Prokaryote survival and proliferation in aquatic environments is often controlled by the ability to tolerate exposure to oxic conditions. Many prokaryotes do not have subcellular organelles for isolating O2-producing from O2-consuming processes and have developed consortial associations with other prokaryotes and eukaryotes that alleviate metabolic constraints of high O2. Nutrient transformations often rely on appropriate cellular and microenvironmental, or microzonal, redox conditions. The spatial and temporal requirements for microenvironmental overlap among microbial groups involved in nutrient transformations necessitates close proximity and diffusional exchange with other biogeochemically distinct, yet complementary, microbial groups. Microbial consortia exist at different levels of community and metabolic complexity, as shown for detrital, microbial mat, biofilm, and planktonic microalgal-bacterial assemblages. To assess the macroscale impacts of consortial interactions, studies should focus on the range of relevant temporal (minutes to hours) and spatial (microns to centimeters) scales controlling microbial production, nutrient exchange, and cycling. In this review, we discuss the utility and application of techniques suitable for determining microscale consortial activity, production, community composition, and interactions in the context of larger scale aquatic ecosystem structure and function.

Microbial indicators of aquatic ecosystem change: current applications to eutrophication studies

Human encroachment on aquatic ecosystems is increasing at an unprecedented rate. The impacts of human pollution and habitat alteration are most evident and of greatest concern at the microbial level, where a bulk of production and nutrient cycling takes place. Aquatic ecosystems are additionally affected by natural perturbations, including droughts, storms, and floods, the frequency and extent of which may be increasing. Distinguishing and integrating the impacts of natural and human stressors is essential for understanding environmentally driven change of microbial diversity and function. Microbial bioindicators play a major role in detecting and characterizing these changes. Complementary use of analytical and molecular indicator tools shows great promise in helping us clarify the processes underlying microbial population, community, and ecosystem change in response to environmental perturbations. This is illustrated in phytoplankton (microalgal and cyanobacterial) and bacterial community changes in a range of US estuarine and coastal ecosystems experiencing increasing development in their water- and airsheds as well as climatic changes (e.g., increasing hurricane frequency). Microbial indicators can be adapted to a range of monitoring programs, including ferries, moored instrumentation, and remote sensing, in order to evaluate environmental controls on microbial community structure and function over ecosystem to global scales.

Phytoplankton Photopigments as Indicators of Estuarine and Coastal Eutrophication

Abstract Human development of coastal watersheds has greatly increased nutrient loading and accelerated estuarine and coastal eutrophication. These waters are also affected by climatic perturbations (e.g., droughts, hurricanes, floods), which may be increasing. The ecological effects of these stressors are often most evident at the microbial level, where the bulk of primary production and biogeochemical cycling occurs. Phytoplankton dominate coastal primary production and thus may be indicative of eutrophication and other major perturbations underlying coastal ecosystem change. Using photopigments that are diagnostic for phytoplankton functional groups (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates), we examined the relative responses of these taxonomic groups to nutrient and hydrologic alterations and evaluated their use as indicators of ecological change in the Neuse River Estuary, North Carolina, and Galveston Bay, Texas. Photopigment indicators can be routinely incorporated in water-quality monitoring programs to assess environmental controls on ecosystem structure and function over varying spatial and temporal scales.

RESPONSES OF THE PHYTOPLANKTON COMMUNITY GROWTH RATE TO NUTRIENT PULSES IN VARIABLE ESTUARINE ENVIRONMENTS

In estuaries, phytoplankton are exposed to rapidly changing conditions that may have profound effects on community structure and function. In these experiments, we evaluated the growth, productivity, and compositional responses of natural phytoplankton communities exposed to limiting nutrient additions and incubation conditions typical of estuarine habitats. Mesocosm bioassays were used to measure the short‐term (2‐day) growth rate, primary productivity, and group‐specific biomass responses of the phytoplankton community in the Neuse River Estuary, North Carolina. A three‐factor (mixing, sediment addition, and nutrient addition) experimental design was applied using 55‐L mesocosm tanks. Growth rates were determined using the 14C photopigment radiolabeling method, and the abundance of algal groups was based on quantification of chemosystematic photopigments by HPLC. For Neuse River Estuary phytoplankton communities, stratified (nonmixed), turbid, and low‐nitrate conditions favored increases in cryptomonad biomass. Mixed, turbid, high‐nitrate conditions were favorable for increased primary productivity and chlorophytes, diatoms, and cyanobacteria. The highest community growth rates occurred under calm, high‐nitrate conditions. This approach provided an assessment of the community‐level phytoplankton responses and insights into the mechanisms driving blooms and bloom species in estuarine waters. The ability to rapidly alter growth rates to capitalize on conditions conducive for growth may play an important role in the timing, extent, and species involved with blooms in estuarine waters. Adaptive growth rate responses of individual species, as well as the community as a whole, further illustrate the sensitivity of estuarine ecosystems to excessive N inputs.

Application of photopigment biomarkers for quantifying microalgal community composition and in situ growth rates

Abstract In estuarine waters, phytoplankton are exposed to rapidly changing conditions that may affect community structure and function. In this study we determined the effects of mixing, turbidity, and limiting nutrient (N) additions on natural phytoplankton growth rates and algal group-specific biomass changes. Mesocosm bioassays were used to quantify the short-term (2–3 day) responses of phytoplankton from the Neuse river estuary, NC. Growth rates were higher under static conditions in N-amended tanks, while biomass of most algal groups was higher under mixed, turbid conditions with N additions. Shifts in community composition did not follow any consistent pattern but each factor influenced phytoplankton growth, biomass, and community composition. Differing growth responses to nutrient additions, mixing, and turbidity resulted in taxonomically-distinct communities. These results highlight the complexity of phytoplankton community structuring processes in estuarine waters. The combination of biomarker quantifications and the radiolabeling method is a useful tool for assessing phytoplankton responses and offers insights into the mechanisms driving blooms and bloom species in estuarine waters.

Salinity control of benthic microbial mat community production in a Bahamian hypersaline lagoon

The purpose of this study was to determine the production and N2 fixation responses of a hypersaline mat community following a reduction in salinity and nutrient enrichment. Cyanobateria-dominated microbial mat samples were collected from hypersaline Storrs Lake and normal seawater salinity Pigeon Creek and preincubated at ambient (90%.) and reduced (45%.) salinities following no nutrient as well as inorganic nutrient (NO3−, PO4−, trace metals) and dissolved organic carbon (DOC, as mannitol) enrichment. CO2 and N2 fixation rates were determined 2 and 4 days later. In addition, DOC (trace concentrations of 3H-labeled glucose and amino acids) uptake was measured in mats under normal and hypersaline conditions. A reduction in salinity from 90 to 45%. significantly enhanced CO2 and N2 fixation rates, but inorganic nutrient and DOC additions did not significantly enhance rates compared with the controls. Dissolved organic carbon/dissolved organic nitrogen (DON) uptake was not influenced over the entire range of salinities (45–90%.) used in this study. When salinity-induced osmotic stress was relieved, mats underwent enhanced primary production and nitrogen fixation. Abiotic stress, induced by hypersaline conditions in Bahamian lagoons, results in lower productivity of the microbial mat communities and this stress may outweigh the typical limiting factors regulating phototrophic community primary production.

COMBINING NEW TECHNOLOGIES FOR DETERMINATION OF PHYTOPLANKTON COMMUNITY STRUCTURE IN THE NORTHERN GULF OF MEXICO 1

In situ analysis of phytoplankton community structure was determined at five stations along the Texas Gulf coast using two instruments, the Fluoroprobe and FlowCAM. Results were compared with traditional methods to determine community structure (pigment analysis and microscopy). Diatoms and small nanoplankton (most likely haptophytes) dominated the phytoplankton community at all stations. Estimated chl concentrations for diatoms+dinoflagellates obtained via the Fluoroprobe were not significantly different for three of the five stations sampled when compared with HPLC‐chemical taxonomy analysis, whereas the concentrations of green algal and cryptophyte chl were overestimated. The FlowCAM estimates of overall nanoplankton and microplankton cell abundance were not significantly different when compared with epifluorescence microscopy, and recorded images of phytoplankton cells provided a representative population of the phytoplankton community at each station. The Fluoroprobe and FlowCAM, when used in tandem, are potentially capable of determining the general characteristics of phytoplankton community structure in situ and could be an important addition to biological observing systems in the coastal ocean.

QUANTIFICATION OF THE RELATIVE ABUNDANCE OF THE TOXIC DINOFLAGELLATE, KARENIA BREVIS (DINOPHYTA), USING UNIQUE PHOTOPIGMENTS1

Diagnostic photopigment analysis is a useful tool for determining the presence and relative abundance of algal groups in natural phytoplankton assemblages. This approach is especially useful when a genus has a unique photopigment composition. The toxic dinoflagellate Karenia brevis (Davis) G. Hansen & Moestrup comb. nov. shares the diagnostic pigment gyroxanthin‐diester with only a few other dinoflagellates and lacks peridinin, one of the major diagnostic pigments of most dinoflagellate species. In this study, measurements of gyroxanthin‐diester and other diagnostic pigments of K. brevis were incorporated into the initial pigment ratio matrix of the chemical taxonomy program (CHEMTAX) to resolve the relative contribution of K. brevis biomass in mixed estuarine phytoplankton assemblages from Florida and Galveston Bay, Texas. The phytoplankton community composition of the bloom in Galveston Bay was calculated based on cell enumerations and biovolumetric measurements in addition to chl a‐specific photopigment estimates of biomass (HPLC and CHEMTAX). The CHEMTAX and biovolume estimates of the phytoplankton community structure were not significantly different and suggest that the HPLC–CHEMTAX approach provides reasonable estimates of K. brevis biomass in natural assemblages. The gyroxanthin‐diester content per cell of K. brevis from Galveston Bay was significantly higher than in K. brevis collected from the west coast of Florida. This pigment‐based approach provides a useful tool for resolving spatiotemporal distributions of phytoplankton in the presence of K. brevis blooms, when an appropriate initial ratio matrix is applied.

Microalgae on seagrass mimics : Does epiphyte community structure differ from live seagrasses ?

The role of seagrasses in regulating epiphytes is a question of central importance for understanding structuring processes in seagrass communities. This study tests the hypothesis that seagrasses are not simply bare substrata for microalgal attachment, but rather influence community composition by altering competitive interactions between different microalgal groups. The effects of wave exposure, depth, seagrass species and seagrass type (mimic vs. live blades) on eelgrass (Zostera marina L.) and shoalgrass (Halodule wrightii Ascher) epiphyte community structure were examined using manipulative field experiments. Relative abundances of major microalgal groups were determined using high-performance liquid chromatographic measurements of diagnostic photopigments. Exposure to wave energy, depth, and seagrass species did not affect epiphyte total biomass. However, epiphyte biomass was significantly greater on live than mimic blades. Diatom biomass was higher under conditions of low wave energy, in deep habitats and on mimic blades. Cyanobacterial biomass was higher in high energy habitats and on live seagrass blades. Although diatoms had a significantly higher biomass on mimic blades, their biomass contribution relative to cyanobacteria was higher on live seagrass blades. The differences in epiphyte community structure on live vs. mimic seagrass blades suggest that competitive interactions between seagrass and epiphytes may result in selection against cyanobacteria or for diatoms. Another possibility is that seagrasses modify the microenvironment on blade surfaces in a way that alters the outcome of competitive interactions between major algal groups (i.e., diatoms and cyanobacteria).

Stimulation of Diesel Fuel Biodegradation by Indigenous Nitrogen Fixing Bacterial Consortia

A bstractSuccessful stimulation of N2 fixation and petroleum hydrocarbon degradation in indigenous microbial consortia may decrease exogenous N requirements and reduce environmental impacts of bioremediation following petroleum pollution. This study explored the biodegradation of petroleum pollution by indigenous N2 fixing marine microbial consortia. Particulate organic carbon (POC) in the form of ground, sterile corn-slash (post-harvest leaves and stems) was added to diesel fuel amended coastal water samples to stimulate biodegradation of petroleum hydrocarbons by native microorganisms capable of supplying a portion of their own N. It was hypothesized that addition of POC to petroleum amended water samples from N-limited coastal waters would promote the growth of N2 fixing consortia and enhance biodegradation of petroleum. Manipulative experiments were conducted using samples from coastal waters (marinas and less polluted control site) to determine the effects of POC amendment on biodegradation of petroleum pollution by native microbial consortia. Structure and function of the microbial consortia were determined by measurement of N2 fixation (acetylene reduction), hydrocarbon biodegradation (14C hexadecane mineralization), bacterial biomass (AODC), number of hydrocarbon degrading bacteria (MPN), and bacterial productivity (3H-thymidine incorporation). Throughout this study there was a consistent enhancement of petroleum hydrocarbon degradation in response to the addition of POC. Stimulation of diesel fuel biodegradation following the addition of POC was likely attributable to increases in bacterial N2 fixation, diesel fuel bioavailability, bacterial biomass, and metabolic activity. Toxicity of the bulk phase water did not appear to be a factor affecting biodegradation of diesel fuel following POC addition. These results indicate that the addition of POC to diesel-fuel-polluted systems stimulated indigenous N2 fixing microbial consortia to degrade petroleum hydrocarbons.