Alan J. Lewitus

Estuaries of the South Atlantic Coast of North America: Their Geographical Signatures

Estuaries of the southeastern Atlantic coastal plain are dominated by shallow meso-tidal bar-built systems interspersed with shallow sounds and both low flow coastal plain and high flow piedmont riverine systems. Three general geographical areas can be discriminated: the sounds of North Carolina; the alternating series of riverine and ocean dominated bar-built systems of South Carolina, Georgia, and northeast Florida, and the subtropical bar-built estuaries of the Florida southeast coast. The regional climate ranges from temperate to subtropical with sea level rise and hurricanes having a major impact on the regions estuaries because of its low and relatively flat geomorphology. Primary production is highest in the central region. Seagrasses are common in the northern and southern most systems, while intertidal salt marshes composed ofSpartina alterniflora reach their greatest extent and productivity in South Carolina and Georgia. Nuisance blooms (cyanobacteria, dinoflagellates, and cryptomonads) occur more frequently in the northern and extreme southern parts of the region. Fishery catches are highest in the North Carolina and Florida areas. Human population growth with its associated urbanization reaches a maximum in Florida and it is thought that the long-term sustainability of the Florida coast for human habitation will be lost within the next 25 years. Tidal flushing appears to play an important role in mitigating anthropogenic inputs in systems of moderate to high tidal range, i.e., the South Carolina and Georgia coasts. The most pressing environmental problems for the estuaries of the southeastern Atlantic coast seem to be nutrient loading and poor land use in North Carolina and high human population density and growth in Florida. The future utilization of these estuarine systems and their services will depend on the development of improved management strategies based on improved data quality.

KLEPTOPLASTIDY IN THE TOXIC DINOFLAGELLATE PFIESTERIA PISCICIDA (DINOPHYCEAE)

The ichthyotoxic dinoflagellate Pfiesteria piscicida Steidinger et Burkholder has a complex life cycle with several heterotrophic flagellated and amoeboid stages. A prevalent flagellated form, the nontoxic zoospore stage, has a proficient grazing ability, especially on cryptophyte prey. Although P. piscicida zoospores lack the genetic capability to synthesize chloroplasts, they can obtain functional chloroplasts from algal prey (i.e. kleptoplastidy), as demonstrated here with a cryptophyte prey. Zoospores grown with Rhodomonas sp. Karsten CCMP757 (Cryptophyceae) grazed the cryptophyte population to minimal densities. After placing the cultures in near darkness where cryptophyte recovery was restricted and further prey ingestion did not occur, the time‐course patterns in growth, prey chloroplast content·zoospore−1, and prey nucleus content·zoospore−1 were followed. Ingested chloroplasts were selectively retained in the dinoflagellate, as indicated by the decline and, ultimately, near absence of cryptophyte nuclei in plastid‐containing zoospores. Chloroplasts retained inside P. piscicida cells for at least a week were photosynthetically active, as indicated by starch accumulation and microscope‐autoradiographic measurements of bicarbonate uptake. Recognition that P. piscicida can function as a phototroph broadens our perspective of the physiological ecology of the dinoflagellate because it suggests that, at least during part of its life cycle, P. piscicida’s growth and survival might be affected by photoregulation and nutritional control of photosynthesis.

Toxicity of Karlodinium micrum (Dinophyceae) associated with a fish kill in a South Carolina brackish retention pond

Abstract A dinoflagellate bloom was found associated with a fish kill event in a South Carolina brackish water retention pond. A multi-analytical approach was used to confirm the identity of the bloom dinoflagellate and evaluate its potential toxicity. Karlodinium micrum was confirmed through light microscopy, pigment profile comparisons, species-specific PCR, and gene sequence data. Necropsy findings on several fish were suggestive of an acute kill event. Toxicity of filtrate from bloom samples was tested by a hemolytic assay using rainbow trout ( Oncorhynchus mykis ) erythrocytes and an ichthyotoxicity assay using larval zebrafish ( Danio rerio ). Hemolytic activity was measurably high (>80% hemolysis) in both whole filtrate and fractionated filtrate (from the 80% MeOH C 18 column elution). This fraction also demonstrated high ichthyotoxic activity as exposed fish experienced rapid death. These results implicate toxic K. micrum as a causative factor in fish death in a non-aquaculture brackish pond associated with a housing development, and extend recent findings linking this species to fish kills in aquaculture ponds.

RAPHIDOPHYCEAE [CHADEFAUD EX SILVA] SYSTEMATICS AND RAPID IDENTIFICATION: SEQUENCE ANALYSES AND REAL-TIME PCR ASSAYS.

Species within the class Raphidophyceae were associated with fish kill events in Japanese, European, Canadian, and U.S. coastal waters. Fish mortality was attributable to gill damage with exposure to reactive oxygen species (peroxide, superoxide, and hydroxide radicals), neurotoxins, physical clogging, and hemolytic substances. Morphological identification of these organisms in environmental water samples is difficult, particularly when fixatives are used. Because of this difficulty and the continued global emergence of these species in coastal estuarine waters, we initiated the development and validation of a suite of real‐time polymerase chain reaction (PCR) assays. Sequencing was used to generate complete data sets for nuclear encoded small‐subunit ribosomal RNA (SSU rRNA; 18S); internal transcribed spacers 1 and 2, 5.8S; and plastid encoded SSU rRNA (16S) for confirmed raphidophyte cultures from various geographic locations. Sequences for several Chattonella species (C. antiqua, C. marina, C. ovata, C. subsalsa, and C. verruculosa), Heterosigma akashiwo, and Fibrocapsa japonica were generated and used to design rapid and specific PCR assays for several species including C. verruculosa Hara et Chihara, C. subsalsa Biecheler, the complex comprised of C. marina Hara et Chihara, C. antiqua Ono and C. ovata, H. akashiwo Ono, and F. japonica Toriumi et Takano using appropriate loci. With this comprehensive data set, we were also able to perform phylogenetic analyses to determine the relationship between these species.

MIXOTROPHY AND NITROGEN UPTAKE BY PFIESTERIA PISCICIDA (DINOPHYCEAE)

The nutritional versatility of dinoflagellates is a complicating factor in identifying potential links between nutrient enrichment and the proliferation of harmful algal blooms. For example, although dinoflagellates associated with harmful algal blooms (e.g. red tides) are generally considered to be phototrophic and use inorganic nutrients such as nitrate or phosphate, many of these species also have pronounced heterotrophic capabilities either as osmotrophs or phagotrophs. Recently, the widespread occurrence of the heterotrophic toxic dinoflagellate, Pfiesteria piscicida Steidinger et Burkholder, has been documented in turbid estuarine waters. Pfiesteria piscicida has a relatively proficient grazing ability, but also has an ability to function as a phototroph by acquiring chloroplasts from algal prey, a process termed kleptoplastidy. We tested the ability of kleptoplastidic P. piscicida to take up 15N‐labeled NH, NO, urea, or glutamate. The photosynthetic activity of these cultures was verified, in part, by use of the fluorochrome, primulin, which indicated a positive relationship between photosynthetic starch production and growth irradiance. All four N substrates were taken up by P. piscicida, and the highest uptake rates were in the range cited for phytoplankton and were similar to N uptake estimates for phagotrophic P. piscicida. The demonstration of direct nutrient acquisition by kleptoplastidic P. piscicida suggests that the response of the dinoflagellate to nutrient enrichment is complex, and that the specific pathway of nutrient stimulation (e.g. indirect stimulation through enhancement of phytoplankton prey abundance vs. direct stimulation by saprotrophic nutrient uptake) may depend on P. piscicida’s nutritional state (phagotrophy vs. phototrophy).

Harmful algal blooms in South Carolina residential and golf course ponds

The South Carolina coastal zone is among the fastest growing areas in the U.S., and population epicenters are marked by dense brackish water pond (lagoon) coverage associated with housing complexes and golf courses. Surveillance efforts in 2001–2002 documented the widespread occurrence of several types of potentially or measurably toxic harmful algal blooms (HABs) in these ponds. These man-made retention ponds were constructed in order to serve as a buffer between developed areas and open estuaries or for aesthetic reasons. However, the combination of restricted tidal flow and nutrient and/or contaminant deposition creates a stimulatory environment for potential HAB formation. These discoveries introduce the need to consider mitigation measures to existing ponds and HAB preventive strategies for future pond construction.

Ecosystem response to bivalve density reduction: management implications

Coastal ecosystems are easily overexploited and changed by physical and biological factors. In this paper, we discuss current ideas and arguments for coastal ecosystem management with an emphasis on systems that have large bivalve filter feeder components. For centuries the species or population approach has been utilized in fisheries management. With the growing knowledge base on specific environmental effects and relationships, it has become increasingly evident that a broad or holistic approach to fisheries management in these systems is usually more appropriate. An ongoing ecosystem scale experiment in which oysters are completely removed from tidal creeks is described and used as a case study. The experimental design takes estimates of the systems carrying capacity into account. Using the population or species approach to monitor the oysters, the only observable change after the experimental manipulation was a slight increase in summer somatic growth and elevated recruitment of oysters in creeks with oyster reefs removed. These data are interpreted as an indication that the creeks with oysters present are below or near carrying capacity. However, when nekton, plankton and water chemistry data are also examined a much more complicated picture emerges. During the summer growing season, nekton biomass in all creeks is often greater than oyster biomass. Also, our calculations show that oysters do not produce enough ammonium to satisfy phytoplankton productivity, but nekton, water column remineralization and sediments can account for most of the deficit. Finally, microflagellates, which are a preferred food for the oysters, dominate the phytoplankton during the summer growing season and diatoms dominate the colder months. The timing of the change in phase of phytoplankton dominance seems to mirror the seasonal arrival and departure times of nekton in the creeks. We argue that dense bivalve reefs and beds are indicative of intense positive feedback loops that make their ecosystems susceptible to dramatic changes in structure. Such changes have not been reported for natural systems, but are found in systems influenced by over-fishing, nutrient loading and pollution. Thus, the management of sustainable fisheries in coastal ecosystems requires an understanding of the ecosystem science and the realization that systems dominated by bivalves exhibit complex responses that are not easily explained by linear dynamics.

LIGHT RESPIRATION IN SIX ESTUARINE PHYTOPLANKTON SPECIES: CONTRASTS UNDER PHOTOAUTOTROPHIC AND MIXOTROPHIC GROWTH CONDITIONS1

The photoresponse of respiration was examined in six estuarine phytoplankton species grown at a limiting irradiance in the presence or absence of glucose. Cellular respiration rates in the dark, at the growth irradiance, and at saturating light levels were higher in glucose‐grown cultures than in photoautotrophic cultures. Glucose uptake also affected the relationship between incident irradiance and gross respiration rate, and the specific respiratory pathways that contributed to the respiration versus irradiance (R vs. I) response. The energy‐conseming Mehler reaction contributed largely to light‐stimulated respiration in photoautotrophic (energy‐limited) cultures. However, the energy‐dissipating alternative pathway was the predominant form of respiration measured in mixotrophic (energy‐sufficient) cultures. Based on these and previous observations, a model of phytoplankton R us. I response is presented, in which the relative contributions of different light respiratory processes (cytochrome pathway respiration, alternative pathway respiration, chlororespiration, Mehler reaction) are dependent on the incident irradiance and cellular energy content.

Relationships among Water-Quality Parameters from the North Inlet–Winyah Bay National Estuarine Research Reserve, South Carolina

Abstract Estuaries integrate atmospheric, watershed, oceanic, and human influences over space and time. Therefore, spatial and temporal patterns in estuarine water-column properties are useful as metrics to evaluate external factors related to internal processes. The National Estuarine Research Reserve monitoring program, including the North Inlet–Winyah Bay complex in South Carolina, provides an ideal setting to track water quality relationships. Our goal was to assess hydrography, chlorophyll a, and particulate and dissolved materials from monitoring data collected at sites from both the salt and estuarine marsh components since 1993–94. Salinity, turbidity, dissolved organic carbon, suspended solids, and chlorophyll a were much greater at the estuarine site, whereas organic nitrogen dominated the total nitrogen pool at both locations. Nitrate was a significant fraction of the total nitrogen pool at the estuarine site but not within the salt marsh. Whereas dissolved organic nitrogen was positively correlated to water temperature, nitrate concentrations were the lowest in the summer. Principal components analysis identified seasonal patterns within the salt marsh for temperature, chlorophyll a, ammonium, suspended solids, and particulate nitrogen. These parameters, grouped together as a primary component, were positively correlated to Spartina alterniflora biomass. In contrast, the estuarine site was more characterized by salinity, pH, and dissolved organic carbon. Although the water-column properties of the salt marsh site reflected a high degree of internal production and remineralization in the summer, patterns at the estuarine site were more likely influenced by seasonal changes in circulation and biogeochemical processing common to coastal plain estuaries.

Kryptoperidinium foliaceum blooms in South Carolina: a multi–analytical approach to identification

Abstract Observations following the discovery of Kryptoperidinium foliaceum blooms in South Carolina (SC), USA, suggest that a multi-analytical approach, using a standard, minimal set of criteria, should be adopted for determining dinoflagellate species identity and taxonomic placement. A combination of morphological, molecular, and biochemical analyses were used to determine the identity of this “red tide” dinoflagellate, first documented in SC waters in the spring of 1998. Results from thecal plate tabulations (based on scanning electron and epifluorescence microscopy), gene sequence data, species-specific PCR probe assays, and microalgal pigment profiles were analyzed and compared to reference cultures of K. foliaceum . Comparative data showed marked inconsistencies among the K. foliaceum reference culture isolates. In addition, the SC bloom isolate was shown to be mononucleate, contrary to previous reports for K. foliaceum , suggesting a more transient endosymbiotic association than previously considered.