Dennett Mark R.

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Mark R.

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  • Article
    Development and application of a monoclonal-antibody technique for counting Aureococcus anophagefferens, an alga causing recurrent brown tides in the Mid-Atlantic United States
    (American Society for Microbiology, 2003-09) Caron, David A. ; Dennett, Mark R. ; Moran, Dawn M. ; Schaffner, Rebecca A. ; Lonsdale, Darcy J. ; Gobler, Christopher J. ; Nuzzi, Robert ; McLean, Tim I.
    A method was developed for the rapid detection and enumeration of Aureococcus anophagefferens, the cause of harmful algal blooms called "brown tides" in estuaries of the Mid-Atlantic United States. The method employs a monoclonal antibody (MAb) and a colorimetric, enzyme-linked immunosorbent assay format. The MAb obtained exhibits high reactivity with A. anophagefferens and very low cross-reactivities with a phylogenetically diverse array of other protists and bacteria. Standard curves are constructed for each 96-well microtiter plate by using known amounts of a preserved culture of A. anophagefferens. This approach allows estimation of the abundance of the alga in natural samples. The MAb method was compared to an existing method that employs polyclonal antibodies and epifluorescence microscopy and to direct microscopic counts of A. anophagefferens in samples with high abundances of the alga. The MAb method provided increased quantitative accuracy and greatly reduced sample processing time. A spatial survey of several Long Island estuaries in May 2000 using this new approach documented a range of abundances of A. anophagefferens in these bays spanning nearly 3 orders of magnitude.
  • Article
    Victims or vectors : a survey of marine vertebrate zoonoses from coastal waters of the Northwest Atlantic
    (Inter-Research, 2008-08-19) Bogomolni, Andrea L. ; Gast, Rebecca J. ; Ellis, Julie C. ; Dennett, Mark R. ; Pugliares, Katie R. ; Lentell, Betty J. ; Moore, Michael J.
    Surveillance of zoonotic pathogens in marine birds and mammals in the Northwest Atlantic revealed a diversity of zoonotic agents. We found amplicons to sequences from Brucella spp., Leptospira spp., Giardia spp. and Cryptosporidium spp. in both marine mammals and birds. Avian influenza was detected in a harp seal and a herring gull. Routine aerobic and anaerobic culture showed a broad range of bacteria resistant to multiple antibiotics. Of 1460 isolates, 797 were tested for resistance, and 468 were resistant to one or more anti-microbials. 73% (341/468) were resistant to 1–4 drugs and 27% (128/468) resistant to 5–13 drugs. The high prevalence of resistance suggests that many of these isolates could have been acquired from medical and agricultural sources and inter-microbial gene transfer. Combining birds and mammals, 45% (63/141) of stranded and 8% (2/26) of by-caught animals in this study exhibited histopathological and/or gross pathological findings associated with the presence of these pathogens. Our findings indicate that marine mammals and birds in the Northwest Atlantic are reservoirs for potentially zoonotic pathogens, which they may transmit to beachgoers, fishermen and wildlife health personnel. Conversely, zoonotic pathogens found in marine vertebrates may have been acquired via contamination of coastal waters by sewage, run-off and agricultural and medical waste. In either case these animals are not limited by political boundaries and are therefore important indicators of regional and global ocean health.
  • Article
    Characterization of protistan assemblages in the Ross Sea, Antarctica, by denaturing gradient gel electrophoresis
    (American Society for Microbiology, 2004-04) Gast, Rebecca J. ; Dennett, Mark R. ; Caron, David A.
    The diversity of protistan assemblages has traditionally been studied using microscopy and morphological characterization, but these methods are often inadequate for ecological studies of these communities because most small protists inherently lack adequate taxonomic characters to facilitate their identification at the species level and many protistan species also do not preserve well. We have therefore used a culture-independent approach (denaturing gradient gel electrophoresis [DGGE]) to obtain an assessment of the genetic composition and distribution of protists within different microhabitats (seawater, meltwater or slush on sea-ice floes, and ice) of the Ross Sea, Antarctica. Samples of the same type (e.g., water) shared more of the same bands than samples of different types (e.g., ice versus water), despite being collected from different sites. These findings imply that samples from the same environment have a similar protistan species composition and that the type of microenvironment significantly influences the protistan species composition of these Antarctic assemblages. It should be noted that a large number of bands among the samples within each microhabitat were distinct, indicating the potential presence of significant genetic diversity within each microenvironment. Sequence analysis of selected DGGE bands revealed sequences that represent diatoms, dinoflagellates, ciliates, flagellates, and several unidentified eukaryotes.
  • Article
    Upward nitrate transport by phytoplankton in oceanic waters : balancing nutrient budgets in oligotrophic seas
    (PeerJ, 2014-03-13) Villareal, Tracy A. ; Pilskaln, Cynthia H. ; Montoya, Joseph P. ; Dennett, Mark R.
    In oceanic subtropical gyres, primary producers are numerically dominated by small (1–5 µm diameter) pro- and eukaryotic cells that primarily utilize recycled nutrients produced by rapid grazing turnover in a highly efficient microbial loop. Continuous losses of nitrogen (N) to depth by sinking, either as single cells, aggregates or fecal pellets, are balanced by both nitrate inputs at the base of the euphotic zone and N2-fixation. This input of new N to balance export losses (the biological pump) is a fundamental aspect of N cycling and central to understanding carbon fluxes in the ocean. In the Pacific Ocean, detailed N budgets at the time-series station HOT require upward transport of nitrate from the nutricline (80–100 m) into the surface layer (∼0–40 m) to balance productivity and export needs. However, concentration gradients are negligible and cannot support the fluxes. Physical processes can inject nitrate into the base of the euphotic zone, but the mechanisms for transporting this nitrate into the surface layer across many 10s of m in highly stratified systems are unknown. In these seas, vertical migration by the very largest (102–103 µm diameter) phytoplankton is common as a survival strategy to obtain N from sub-euphotic zone depths. This vertical migration is driven by buoyancy changes rather than by flagellated movement and can provide upward N transport as nitrate (mM concentrations) in the cells. However, the contribution of vertical migration to nitrate transport has been difficult to quantify over the required basin scales. In this study, we use towed optical systems and isotopic tracers to show that migrating diatom (Rhizosolenia) mats are widespread in the N. Pacific Ocean from 140°W to 175°E and together with other migrating phytoplankton (Ethmodiscus, Halosphaera, Pyrocystis, and solitary Rhizosolenia) can mediate time-averaged transport of N (235 µmol N m-2 d-1) equivalent to eddy nitrate injections (242 µmol NO3− m-2 d-1). This upward biotic transport can close N budgets in the upper 250 m of the central Pacific Ocean and together with diazotrophy creates a surface zone where biological nutrient inputs rather than physical processes dominate the new N flux. In addition to these numerically rare large migrators, there is evidence in the literature of ascending behavior in small phytoplankton that could contribute to upward flux as well. Although passive downward movement has dominated models of phytoplankton flux, there is now sufficient evidence to require a rethinking of this paradigm. Quantifying these fluxes is a challenge for the future and requires a reexamination of individual phytoplankton sinking rates as well as methods for capturing and enumerating ascending phytoplankton in the sea.
  • Preprint
    Defining DNA-based operational taxonomic units for microbial-eukaryote ecology
    ( 2009-06-19) Caron, David A. ; Countway, Peter D. ; Savai, Pratik ; Gast, Rebecca J. ; Schnetzer, Astrid ; Moorthi, Stefanie D. ; Dennett, Mark R. ; Moran, Dawn M. ; Jones, Adriane C.
    DNA sequence information has been increasingly used in ecological research on microbial eukaryotes. Sequence-based approaches have included studies of the total diversity of selected ecosystems, the autecology of ecologically relevant species, and the identification and enumeration of species of interest to human health. It is still uncommon, however, to delineate protistan species based on their genetic signatures. The reluctance to assign species-level designations based on DNA sequences is partly a consequence of the limited amount of sequence information presently available for many free-living microbial eukaryotes, and partly the problematic nature and debate surrounding the microbial species concept. Despite the difficulties inherent in assigning species names to DNA sequences, there is a growing need to attach meaning to the burgeoning amount of sequence information entering the literature, and a growing desire to apply this information in ecological studies. We describe a computer-based tool that assigns DNA sequences from environmental databases to operational taxonomic units at approximate species-level distinctions. The approach provides a practical method for ecological studies of microbial eukaryotes (primarily protists) by enabling semiautomated analysis of large numbers of samples spanning great taxonomic breadth. Derivation of the algorithm was based on an analysis of complete small subunit ribosomal RNA (18S) gene sequences and partial gene sequences obtained from GenBank for morphologically described protistan species. The program was tested using environmental 18S data sets from two oceanic ecosystems. A total of 388 operational taxonomic units were observed among 2,207 sequences obtained from samples collected in the western North Atlantic and eastern North Pacific.
  • Preprint
    High concentrations of marine snow and diatom algal mats in the North Pacific Subtropical Gyre : implications for carbon and nitrogen cycles in the oligotrophic ocean
    ( 2005-08-02) Pilskaln, Cynthia H. ; Villareal, Tracy A. ; Dennett, Mark R. ; Darkangelo-Wood, C. ; Meadows, G.
    A Video Plankton Recorder (VPR) and remotely operated vehicle (ROV) were utilized on three cruises in the oligotrophic North Pacific Subtropical Gyre (NPSG) between 1995 and 2002 to quantify the size and abundance of marine snow and Rhizosolenia diatom mats within the upper 305 m of the water column. Quantitative image analysis of video collected by the VPR and an ROV-mounted particle imaging system provides the first transect of marine snow size and abundance across the central North Pacific gyre extending from 920 km NW of Oahu to 555 km off Southern California. Snow abundance in the upper 55 m was surprisingly high for this oligotrophic region, with peak values of 6.0-13.0 x 103 aggregates m-3 at the western and eastern-most stations. At stations located in the middle of the transect (farthest from HI and CA), upper water column snow abundance displayed values of ~0.5-1.0 x 103 aggregates m-3. VPR and ROV imagery also provided in-situ documentation of the presence of nitrogen-transporting, vertically migrating Rhizosolenia mats from the surface to >300 m with mat abundances ranging from 0-10 mats m-3. There was clear evidence that Rhizosolenia mats commonly reach sub-nutricline depths. The mats were noted to be a common feature in the North Pacific gyre, with the lower salinity edge of the California Current appearing to be the easternmost extent of their oceanic distribution. Based on ROV observations at depth, flux by large (>1.5 cm) mats is revised upward 4.5 fold, yielding an average value of 40 µmol N m-2 d-1, a value equaling previous estimates that included much smaller mats visible only to towed optical systems. Our results suggest that the occurrence across a broad region of the NPSG of particulate organic matter (POM) production events represented by high concentrations of Rhizosolenia mats, associated mesozooplankton, and abundant detrital marine aggregates may represent significant stochastic components in the overall carbon, nitrogen and silica budgets of the oligotrophic subtropical gyre. Likewise, their presence has important implications for the proposed climate-driven, ecosystem reorganization or domain shift occurring in the NPSG.