Patricia Blackwelder

Biochemical and ultrastructural evidence for the origin of whitings: A biologically induced calcium carbonate precipitation mechanism

We propose that Bahamian whitings, floating patches of lime mud, are in part the result of biological precipitation of calcium carbonate induced by picoplankton and cellular components. The cells may act as nucleation sites for crystallization from seawater supersaturated with CaCO3. Our model represents a new hypothesis concerning Bahamian whiting formation and is based on several lines of evidence. Biochemical data suggest that the macromolecules from carbonate suspended in Bahama Banks whitings are distinct from those found in the lime mud sediment producer Penicillus and from bottom sediment. Direct ultrastructural evidence indicates that mineralization occurs on the surfaces of picoplankton cells and degrading organic cellular components. The organic constituents include, but are not restricted to, whole and fragmented algal cells and composite structures ∼20-30 μm in diameter. The observed calcium carbonate crystals are distinct from skeletal debris. Because of the magnitude of the whiting phenomenon, calculations of carbonate sediment production budgets should include the potential contribution of calcium carbonate produced by epicefular precipitation. Knowledge of carbonate mud genesis is critical to interpretation of ancient occurrences of lime mud as well as to understanding ancient and global carbon cycles.

Thoracosphaera heimii (Lohmann) Kamptner Is a Dinophyte: Observations on Its Morphology and Life Cycle

Abstract Thirty clones of Thoracosphaera heimii (Lohmann) Kamptner have been isolated and cultured from oceanic waters of the western North Atlantic and Gulf of Mexico. Observations on the life cycle and morphology of one clone isolated from the Sargasso Sea are reported herein. The calcareous cell wall is present in the coccoid, vegetative life phase. Reproduction is accomplished by the formation of transitory aplanospore or planospore stages or occasionally by binary fission of weakly calcified cells. The planospore is non-thecate and Gymnodinium-like with an undulating transverse flagellum and a whip-like longitudinal flagellum. All life stages possess chloroplasts and a nucleus with continually condensed chromosomes. The planospore morphology and the dinocaryotic nucleus demonstrate that T. heimii is a dinophyte and not a coccolithophorid. The taxonomic affinity and classification of T. heimii within the Dinophyceae is discussed and a new order Thoracosphaerales Tangen, ord. nov. is proposed for primarily coccoid marine dinoflagellates that possess a calcified cell wall in the vegetative life phase.

Stable isotopic composition of coccoliths

Oxygen isotope analyses of coccoliths from eight coccolithophore species grown in laboratory culture indicate that biological fractionation occurs during the formation of calcium carbonate. Cultures of coccolithophores, including species not previously grown in the laboratory were maintained at various temperatures ranging from 12–28°C. Over this range of temperature, oxygen isotope values for Emiliania huxleyi, Gephyrocapsa oceanica, and Crenalithus sessilis are found to be 1‰ enriched in 18O relative to calcium carbonate precipitated under equilibrium conditions. Coccoliths of Calcidiscus leptoporus, Umbilicosphaera sibogae, U. hulburtiana, Syracosphaera pulchra, and Cricosphaera carterae were found to be 2.5‰ depleted in 18O relative to equilibrium. These findings help clarify previously reported equivocal oxygen isotope data derived from sedimentary calcareous nannofossils and reveal the importance of species-specific isotopic fractionation.

Pyrosequencing of Bacterial Symbionts within Axinella corrugata Sponges: Diversity and Seasonal Variability

Background Marine sponge species are of significant interest to many scientific fields including marine ecology, conservation biology, genetics, host-microbe symbiosis and pharmacology. One of the most intriguing aspects of the sponge “holobiont” system is the unique physiology, interaction with microbes from the marine environment and the development of a complex commensal microbial community. However, intraspecific variability and temporal stability of sponge-associated bacterial symbionts remain relatively unknown. Methodology/Principal Findings We have characterized the bacterial symbiont community biodiversity of seven different individuals of the Caribbean reef sponge Axinella corrugata, from two different Florida reef locations during variable seasons using multiplex 454 pyrosequencing of 16 S rRNA amplicons. Over 265,512 high-quality 16 S rRNA sequences were generated and analyzed. Utilizing versatile bioinformatics methods and analytical software such as the QIIME and CloVR packages, we have identified 9,444 distinct bacterial operational taxonomic units (OTUs). Approximately 65,550 rRNA sequences (24%) could not be matched to bacteria at the class level, and may therefore represent novel taxa. Differentially abundant classes between seasonal Axinella communities included Gammaproteobacteria, Flavobacteria, Alphaproteobacteria, Cyanobacteria, Acidobacter and Nitrospira. Comparisons with a proximal outgroup sponge species (Amphimedon compressa), and the growing sponge symbiont literature, indicate that this study has identified approximately 330 A. corrugata-specific symbiotic OTUs, many of which are related to the sulfur-oxidizing Ectothiorhodospiraceae. This family appeared exclusively within A. corrugata, comprising >34.5% of all sequenced amplicons. Other A. corrugata symbionts such as Deltaproteobacteria, Bdellovibrio, and Thiocystis among many others are described. Conclusions/Significance Slight shifts in several bacterial taxa were observed between communities sampled during spring and fall seasons. New 16 S rDNA sequences and concomitant identifications greatly expand the microbial community profile for this model reef sponge, and will likely be useful as a baseline for any future comparisons regarding sponge microbial community dynamics.

Time-based correlation of biogenic, lithogenic and authigenic sediment components with anthropogenic inputs in the Gulf of Mexico NECOP study area

Hypotheses related to variability in seasonal hypoxic conditions, coastal nutrient enhancement, and off-shelf transport of carbon on the Louisiana continental shelf were tested by characterization of biogenic, lithogenic, and authigenic components from two shelf and one Mississippi Canyon sediment cores. The authigenic-phase glauconite occurs above detection limits only in the core from the hypoxic area. A major increase in glauconite concentration was coincident with the onset (≈1940) of the increased use of commercial fertilizers in the United States. In the same hypoxic-area core, benthic foraminifera species diversity decreases upcore from approximately the turn of the century to the present in a manner concurrent with glauconite and fertilizer increases. A subset of opportunistic benthic foraminifera species, known to become more prominent in stressed environments (i.e., hypoxic), increased upcore from ∼52% of the total population at core bottom to ≈90% at core top. These benthic foraminifera population and diversity changes were not apparent in a “control” core outside the area of documented hypoxia. Seaward of the shelf, in the Mississippi Canyon, coincident increases in sediment accumulation rate, percentages of coarse fraction and of organic carbon at core top indicate increased offshelf transport of carbon and other components. Quartz percentages indicate that episodic down-canyon transport has been active to core bottom (prior to the mid 1800s).

Peptide-Conjugated PAMAM Dendrimer as a Universal DNA Vaccine Platform to Target Antigen-Presenting Cells

DNA-based vaccines hold promise to outperform conventional antigen-based vaccines by virtue of many unique features. However, DNA vaccines have thus far fallen short of expectations, due in part to poor targeting of professional antigen-presenting cells (APC) and low immunogenicity. In this study, we describe a new platform for effective and selective delivery of DNA to APCs in vivo that offers intrinsic immune-enhancing characteristics. This platform is based on conjugation of fifth generation polyamidoamine (G5-PAMAM) dendrimers, a DNA-loading surface, with MHC class II-targeting peptides that can selectively deliver these dendrimers to APCs under conditions that enhance their immune stimulatory potency. DNA conjugated with this platform efficiently transfected murine and human APCs in vitro. Subcutaneous administration of DNA-peptide-dendrimer complexes in vivo preferentially transfected dendritic cells (DC) in the draining lymph nodes, promoted generation of high affinity T cells, and elicited rejection of established tumors. Taken together, our findings show how PAMAM dendrimer complexes can be used for high transfection efficiency and effective targeting of APCs in vivo, conferring properties essential to generate effective DNA vaccines.

Benthic foraminifera from the necop study area impacted by the Mississippi River plume and seasonal hypoxia

Abstract Benthic foraminifera influenced by the Mississippi River plume and seasonal hypoxia were assessed from Louisiana inner-continental shelf sediment samples. Surface foraminifera assemblages were representative of in-situ populations as established by staining techniques. Community diversity and richness/evenness analyses indicate three regimes: high stress (sediment dominated), intermediate stress (hypoxia dominated), and low stress (low sediment accumulation/high oxygen). Epistominella vitrea and Buliminella morgani are useful tracers of rapid sediment accumulation rate and hypoxia. A bottom-water productivity signal west of the Mississippi River plume is indicated by benthic and planktic foraminifera abundance peaks. Surface benthic foraminifera trends are utilized to interpret changes in historical community structure from hypoxic-area sediments deposited since the turn of the century. The hypoxia-tolerant species Buliminella morgani increases markedly upcore, while hypoxia intolerant species decrease or disappear. Diversity and dominance trends temporally correspond to a dramatic increase in U.S. fertilizer application. The results of this study have application to paleoenvironmental research spanning longer geologic timescales. The documented relationships between population structure and stressors in river-dominated marine systems may provide a useful analog for recognition of these conditions in the fossil record.

A Targeted and Adjuvanted Nanocarrier Lowers the Effective Dose of Liposomal Amphotericin B and Enhances Adaptive Immunity in Murine Cutaneous Leishmaniasis

BACKGROUND Amphotericin B (AmB), the most effective drug against leishmaniasis, has serious toxicity. As Leishmania species are obligate intracellular parasites of antigen presenting cells (APC), an immunopotentiating APC-specific AmB nanocarrier would be ideally suited to reduce the drug dosage and regimen requirements in leishmaniasis treatment. Here, we report a nanocarrier that results in effective treatment shortening of cutaneous leishmaniasis in a mouse model, while also enhancing L. major specific T-cell immune responses in the infected host. METHODS We used a Pan-DR-binding epitope (PADRE)-derivatized-dendrimer (PDD), complexed with liposomal amphotericin B (LAmB) in an L. major mouse model and analyzed the therapeutic efficacy of low-dose PDD/LAmB vs full dose LAmB. RESULTS PDD was shown to escort LAmB to APCs in vivo, enhanced the drug efficacy by 83% and drug APC targeting by 10-fold and significantly reduced parasite burden and toxicity. Fortuitously, the PDD immunopotentiating effect significantly enhanced parasite-specific T-cell responses in immunocompetent infected mice. CONCLUSIONS PDD reduced the effective dose and toxicity of LAmB and resulted in elicitation of strong parasite specific T-cell responses. A reduced effective therapeutic dose was achieved by selective LAmB delivery to APC, bypassing bystander cells, reducing toxicity and inducing antiparasite immunity.

Fine-Structural Analysis of Black Band Disease-Infected Coral Reveals Boring Cyanobacteria and Novel Bacteria

Examination of coral fragments infected with black band disease (BBD) at the fine- and ultrastructural levels using scanning (SEM) and transmission electron microscopy (TEM) revealed novel features of the disease. SEM images of the skeleton from the host coral investigated (Montastraea annularis species complex) revealed extensive boring underneath the BBD mat, with cyanobacterial filaments present within some of the bore holes. Cyanobacteria were observed to penetrate into the overlying coral tissue from within the skeleton and were present throughout the mesoglea between tissue layers (coral epidermis and gastrodermis). A population of novel, as yet unidentified, small filamentous bacteria was found at the leading edge of the migrating band. This population increased in number within the band and was present within degrading coral epithelium, suggesting a role in disease etiology. In coral tissue in front of the leading edge of the band, cyanobacterial filaments were observed to be emerging from bundles of sloughed-off epidermal tissue. Degraded gastrodermis that contained actively dividing zooxanthellae was observed using both TEM and SEM. The BBD mat contained cyanobacterial filaments that were twisted, characteristic of negative-tactic responses. Some evidence of boring was found in apparently healthy control coral fragments; however, unlike in BBD-infected fragments, there were no associated cyanobacteria. These results suggest the coral skeleton as a possible source of pathogenic BBD cyanobacteria. Additionally, SEM revealed the presence of a potentially important group of small, filamentous BBD-associated bacteria yet to be identified.

The spatial distribution of silicoflagellates in the region of the Gulf Stream warm-core ring 82B: application to water mass tracer studies

Abstract To delineate potential water mass affinities, we investigated silicoflagellates from the region of Gulf Stream warm-core ring (WCR) 82B in the northwestern Atlantic. Silicoflagellates from 202 samples from N-S and an E-W transects across WCR 82B during late April were analysed. Shelf to Sargasso Sea transects, one completed in early May and the other in June 1982 were also examined. Eight to 11 vertical profiles to 200 m comprised each of the transects. Six taxa of silicoflagellates were found in the samples studied and a total of more than 8000 specimens were encountered. Three major taxa dominated standing stocks: Distephanus speculum, Dictyocha messanensis (intermediate-sized form) and D. mandrai. D. speculum, considered a cold-water taxon in the literature, showed a higher standing stock in the cooler high-velocity region (HVR) of the warm-core ring, continental shelf (SH) and slope (SL) waters. Fewer were present in the wanner ring center (RC), Gulf Stream (GS) and Sargasso Sea (SS). D. mandrai showed a similar distribution to that of D. speculum, but its preference for slightly warmer waters (>~10°C) was noted. In contrast, Dictyocha messanensis (intermediate-sized) and Distephanus pulchra, known to be warm-water taxa, were relatively abundant in the warm ring center. In contrast to standing stock data, ratios between cold- and warm-water taxa correlate well with temperature and salinity in the warm-core ring. Since these ratios are not effected by convective loss, they are excellent water mass tracers in this system. Distribution of the silicoflagellate taxa suggests that WCR82B April had a higher affinity with the Gulf Stream than the Sargasso Sea. Scores derived from factor analysis indicate that silicoflagellate species distributions are highly correlative with water masses. This was evident from correlations with temperature, salinity and with distance from ring center. Nutrients were generally not correlated with species data. This may be due to deep vertical mixing in the ring center of 82B in April that resulted in loss of plankton cells below the euphotic zone which was responsible for low silicoflagellate standing stock. Silicoflagellate productivity, measured as % double skeletons of D. speculum, was approximately the same in all water masses. This result is consistent with the hypothesized plankton population loss due to the convection.