The role of heterotrophic microflagellates in plankton communities

Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the Massachusetts Institute of Technology and the Woods Hole Oceanographic Institution June 1984The distribution and feeding behavior of bacterivorous micro flagellates (2-20 μm protozoa) and their ingestion by copepods were examined in an attempt to assess the importance of these protozoa as a trophic link between planktonic bacteria and zooplankton. The abundance of microflagellates relative to other picoplankton (0.2-2.0 μm) and nanoplankton (2-20 μm) populations in water samples in the North Atlantic and in Lake Ontario and on macroaggregates in the North Atlantic was determined using direct microscopical and culture estimation techniques. Seasonal, vertical and geographical changes in the density of microflagellates were generally not greater than one order of magnitude. Microscopical counts of heterotrophic nanoplankton (presumably microflagellates) typically ranged from a few hundred to a few thousand m1-1 for a variety of planktonic environments. They constituted approximately 1/3 to 1/2 of the nanoplankton in the euphotic zone and dominated the nanoplankton in the aphotic zone. Most Probable Number (MPN) estimation of the density of bacterivorous protozoa indicated that microflagellates were, on average, an order of magnitude more abundant than bacterivorous ciliates and amoebae. MPN and direct microscopical counts of microflagellates differed by as much as 104. This discrepancy was smaller in eutrophic environments (e.g. Continental Shelf and Lake Ontario) and on macroscopic detrital aggregates. All microbial populations enumerated were highly concentrated on macroscopic detrital aggregates relative to their abundance in the water surrounding the aggregates. Enrichment factors (the ratio of abundance of a population on a macroaggregate to its abundance in the surrounding water) increased along a eutrophic-to-oligotrophic gradient because of the combined effects of an increased abundance of microorganisms on macroaggregates in oligotrophic environments and a decreased abundance in the surrounding water in these same environments. Average enrichment factors for direct microscopical counts of heterotrophic nanoplankton (range = 17-114) were not as large as enrichment factors observed for MPN estimates of the number of bacterivorous microflagellates (range = 273-18400). Microflagellates numerically dominated the bacterivorous protozoa cultured from macroaggregates by one to two orders of magnitude, but ciliates and amoebae were also highly enriched on macroaggregates. Microenvironments are therefore a potentially important aspect for the ecology of planktonic microorganisms. Observations on the microbial colonization of mucus sloughed by ctenophores and discarded appendicularian houses suggest that these materials may be important sources of macroaggregates. Batch and continuous culture experiments were conducted with clonal cultures of microflagellates to test their ability to grow on various types and densities of bacteria. The doubling time of Monas sp. 1 ranged from 43 hr (when fed the cyanobacterium Synechococcus Strain WH 8101) to 6.9 hr (when fed the heterotrophic bacterium Serratia marinorubra). Cell yields (i.e. the conversion of bacterial biomass into protozoan biomass) of Monas sp. 1 fed two species of heterotrophic bacteria were greater than yields for the microflagellate fed two species chroococcoid cyanobacteria (range = 7-68%). Cell yields of two other species of microflagellates (Monas sp. 2 and Cryptobia maris) were 48% and 61%, respectively, on the bacterium Pseudomonas halodurans. Microflagellates grew in continuous culture at concentrations of bacteria which were lower than bacterial densities required for the growth of ciliates as shown by other investigations. Therefore, microflagellates appear to be well-adapted for grazing bacterioplankton. Microflagellates were also investigated for their ability to graze bacteria attached to particles. Bodo nanorensis and Rhynchomonas nasuta both showed a marked ability to graze attached bacteria and a limited ability to graze unattached cells. These results suggest that microflagellates may also be important consumers of bacteria attached to particles in the plankton and may explain the highly elevated densities of microflagellates on macroaggregates. Grazing experiments performed with the copepod Acartia tonsa indicated that heterotrophic microflagellates were ingested by the copepods at rates comparable to the ingestion of phytoplankton of similar size. The presence of heterotrophic microflagellates did not depress filtration rates of the copepods, and one species (Cryptobia maris) appeared to be selectively grazed. Survival of A. tonsa on a diet of heterotrophic microflagellates was similar to survival on a diet of phytoplankton and was significantly longer than survival of starved Controls or copepods fed only bacteria. Due to their ability to grow at in-situ densities of planktonic bacteria, their relatively high cell yields, and their acceptability as food for zooplankton, it is concluded that bacterivorous microflagellates may constitute an important trophic link between bacteria and zooplankton. This link may provide a mechanism whereby organic material and energy from the detrital food chain can be returned to the classical phytoplankton-copepod-fish food chain.This research was supported by National Science Foundation grants OCE80-2444l and OCE82-l4928 and Ocean Industry Program grant 4473 awarded to Dr. Laurence P. Madin, NSF Doctoral Dissertation grant OCE8l-l299l, the Woods Hole Oceanographic Institution Education Program and the Wood Hole Oceanographic Institution Biology Department

Two-sample Bayesian Nonparametric Hypothesis Testing

In this article we describe Bayesian nonparametric procedures for two-sample hypothesis testing. Namely, given two sets of samples $\mathbf{y}^{\scriptscriptstyle(1)}\;$\stackrel{\scriptscriptstyle{iid}}{\s im}$\;F^{\scriptscriptstyle(1)}$ and $\mathbf{y}^{\scriptscriptstyle(2 )}\;$\stackrel{\scriptscriptstyle{iid}}{\sim}$\;F^{\scriptscriptstyle( 2)}$, with $F^{\scriptscriptstyle(1)},F^{\scriptscriptstyle(2)}$ unknown, we wish to evaluate the evidence for the null hypothesis $H_0:F^{\scriptscriptstyle(1)}\equiv F^{\scriptscriptstyle(2)}$ versus the alternative $H_1:F^{\scriptscriptstyle(1)}\neq F^{\scriptscriptstyle(2)}$. Our method is based upon a nonparametric P\'{o}lya tree prior centered either subjectively or using an empirical procedure. We show that the P\'{o}lya tree prior leads to an analytic expression for the marginal likelihood under the two hypotheses and hence an explicit measure of the probability of the null $\mathrm{Pr}(H_0|\{\mathbf {y}^{\scriptscriptstyle(1)},\mathbf{y}^{\scriptscriptstyle(2)}\}\mathbf{)}$.Comment: Published at http://dx.doi.org/10.1214/14-BA914 in the Bayesian Analysis (http://projecteuclid.org/euclid.ba) by the International Society of Bayesian Analysis (http://bayesian.org/

Nitrogen uptake and NH4+ regeneration by pelagic microplankton and marine snow from the North Atlantic

Comparative rates of nitrogen uptake and NH4+ regeneration by plankton of \u3c153 and \u3c5 μm in size were determined in the Sargasso Sea and Gulf Stream, and by plankton associated with marine snow in the Gulf Stream during May 1982. Rates of total nitrogen uptake of Sargasso Sea phytoplankton exceeded those of the Gulf Stream phytoplankton by factors ranging from 1.8 to 5.6. Rates of microplankton NH4+ regeneration equaled or exceeded rates of NH4+ uptake in the Sargasso Sea, but in the Gulf Stream were negligible in all but one case. Significant rates of NH4+ regeneration were measured for Gulf Stream marine snow, and, in all but one case, exceeded those of NH4+ uptake. Rates of NO3– and urea uptake by the snow were less than half those of NH4+. Protozoan densities were enumerated on aliquots of the same snow particles and compared with previously reported bacterial estimates; enrichment factors of the cultivable ciliates and flagellates were 6500–9000 relative to ambient seawater. These organisms were also grazing and reproducing rapidly. Bacterial densities were also moderately enriched, but their productivity was lower than surrounding seawater bacteria. Thus, the large bacterivorous population associated with marine snow may have accounted for a substantial fraction of the observed NH4+ regeneration

Composition and degradation of salp fecal pellets: Implications for vertical flux in oceanic environments

Changes in the sinking rates, ash-free dry weights, particulate carbon and nitrogen content, and carbon:nitrogen ratios from the fecal pellets of several species of oceanic salps were examined in ten-day decomposition studies. Although bacteria and protozoa became abundant in the incubation vessels, most of the fecal pellets remained physically intact throughout the study. Bacterial activity in the pellets (measured by the rate of uptake of 3H-thymidine) increased, but microbial degradation had little effect on the sinking speeds of most of the fecal pellets. The average losses of ash-free dry weight and carbon and nitrogen content, along with changes in carbon:nitrogen ratio, were small compared to their initial values. We conclude that microbial degradation of large salp fecal pellets would not prevent the vertical flux to the deep ocean of a significant fraction of the particulate organic material contained in the pellets. The fecal pellets of oceanic salps provide a rapid, and potentially important, mechanism for the consolidation and vertical transport of organic and lithogenic material associated with minute particles in the open ocean

In Situ Diazotroph Population Dynamics Under Different Resource Ratios in the North Pacific Subtropical Gyre.

Major advances in understanding the diversity, distribution, and activity of marine N2-fixing microorganisms (diazotrophs) have been made in the past decades, however, large gaps in knowledge remain about the environmental controls on growth and mortality rates. In order to measure diazotroph net growth rates and microzooplankton grazing rates on diazotrophs, nutrient perturbation experiments and dilution grazing experiments were conducted using free-floating in situ incubation arrays in the vicinity of Station ALOHA in March 2016. Net growth rates for targeted diazotroph taxa as well as Prochlorococcus, Synechococcus and photosynthetic picoeukaryotes were determined under high (H) and low (L) nitrate:phosphate (NP) ratio conditions at four depths in the photic zone (25, 45, 75, and 100 m) using quantitative PCR and flow cytometry. Changes in the prokaryote community composition in response to HNP and LNP treatments were characterized using 16S rRNA variable region tag sequencing. Microzooplankton grazing rates on diazotrophs were measured using a modified dilution technique at two depths in the photic zone (15 and 125 m). Net growth rates for most of the targeted diazotrophs after 48 h were not stimulated as expected by LNP conditions, rather enhanced growth rates were often measured in HNP treatments. Interestingly, net growth rates of the uncultivated prymnesiophyte symbiont UCYN-A1 were stimulated in HNP treatments at 75 and 100 m, suggesting that N used for growth was acquired through continuing to fix N2 in the presence of nitrate. Net growth rates for UCYN-A1, UCYN-C, Crocosphaera sp. (UCYN-B) and the diatom symbiont Richelia (associated with Rhizosolenia) were uniformly high at 45 m (up to 1.6 ± 0.5 d-1), implying that all were growing optimally at the onset of the experiment at that depth. Differences in microzooplankton grazing rates on UCYN-A1 and UCYN-C in 15 m waters indicate that the grazer assemblage preyed preferentially on UCYN-A1. Deeper in the water column (125 m), both diazotrophs were grazed at substantial rates, suggesting grazing pressure may increase with depth in the photic zone. Constraining in situ diazotroph growth and mortality rates are important steps for improving parameterization for diazotrophs in global ecosystem models

Paleoproterozoic sterol biosynthesis and the rise of oxygen

Natural products preserved in the geological record can function as ‘molecular fossils’, providing insight into organisms and physiologies that existed in the deep past. One important group of molecular fossils is the steroidal hydrocarbons (steranes), which are the diagenetic remains of sterol lipids. Complex sterols with modified side chains are unique to eukaryotes, although simpler sterols can also be synthesized by a few bacteria. Sterol biosynthesis is an oxygen-intensive process; thus, the presence of complex steranes in ancient rocks not only signals the presence of eukaryotes, but also aerobic metabolic processes. In 1999, steranes were reported in 2.7 billion year (Gyr)-old rocks from the Pilbara Craton in Australia, suggesting a long delay between photosynthetic oxygen production and its accumulation in the atmosphere (also known as the Great Oxidation Event) 2.45–2.32 Gyr ago. However, the recent reappraisal and rejection of these steranes as contaminants pushes the oldest reported steranes forward to around 1.64 Gyr ago (ref. 6). Here we use a molecular clock approach to improve constraints on the evolution of sterol biosynthesis. We infer that stem eukaryotes shared functionally modern sterol biosynthesis genes with bacteria via horizontal gene transfer. Comparing multiple molecular clock analyses, we find that the maximum marginal probability for the divergence time of bacterial and eukaryal sterol biosynthesis genes is around 2.31 Gyr ago, concurrent with the most recent geochemical evidence for the Great Oxidation Event. Our results therefore indicate that simple sterol biosynthesis existed well before the diversification of living eukaryotes, substantially predating the oldest detected sterane biomarkers (approximately 1.64 Gyr ago), and furthermore, that the evolutionary history of sterol biosynthesis is tied to the first widespread availability of molecular oxygen in the ocean–atmosphere system

Enrichment of microbial populations in macroaggregates (marine snow) from surface waters of the North Atlantic

Marine snow particles (macroscopic detrital aggregates) were collected from surface waters throughout the western North Atlantic. Counts of phototrophic and heterotrophic picoplankton, phototrophic and heterotrophic nanoplankton, and phototrophic microplankton were made by epifluorescence microscopy. A Most Probable Number culture technique also was used to estimate the density of bacterivorous protozoa. All microbial populations enumerated were highly enriched on macroaggregates relative to their densities in the surrounding water. The degree of enrichment was greater in open ocean environments because microorganisms in the surrounding water were less abundant in the open ocean than in nearshore waters, and also because microbial density on marine snow was greater in the open ocean than in nearshore environments. Material released by ctenophores and appendicularia is a likely source of marine snow since it supported microbial populations of the same order of magnitude as were observed on SCUBA-collected particles. Heterotrophic nanoflagellates dominated the bacterivorous protozoa cultured from macroaggregates and the surrounding water, but dense populations of ciliates and amoebae also were present on particles. Protozoan populations on marine snow were so dense relative to the surrounding water as to suggest that detrital aggregates are responsible for the planktonic existence of some bacterivorous species

Seasonal analysis of protistan community structure and diversity at the USC Microbial Observatory (San Pedro Channel, North Pacific Ocean)

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/109997/1/lno20105562381.pd

Squeezing in the audio gravitational wave detection band

We demonstrate the generation of broad-band continuous-wave optical squeezing down to 200Hz using a below threshold optical parametric oscillator (OPO). The squeezed state phase was controlled using a noise locking technique. We show that low frequency noise sources, such as seed noise, pump noise and detuning fluctuations, present in optical parametric amplifiers have negligible effect on squeezing produced by a below threshold OPO. This low frequency squeezing is ideal for improving the sensitivity of audio frequency measuring devices such as gravitational wave detectors.Comment: 5 pages, 6 figure

Ultraviolet asymptotics of scalar and pseudoscalar correlators in hot Yang-Mills theory

Inspired by recent lattice measurements, we determine the short-distance (a > omega >> pi T) asymptotics of scalar (trace anomaly) and pseudoscalar (topological charge density) correlators at 2-loop order in hot Yang-Mills theory. The results are expressed in the form of an Operator Product Expansion. We confirm and refine the determination of a number of Wilson coefficients; however some discrepancies with recent literature are detected as well, and employing the correct values might help, on the qualitative level, to understand some of the features observed in the lattice measurements. On the other hand, the Wilson coefficients show slow convergence and it appears uncertain whether this approach can lead to quantitative comparisons with lattice data. Nevertheless, as we outline, our general results might serve as theoretical starting points for a number of perhaps phenomenologically more successful lines of investigation.Comment: 27 pages. v2: minor improvements, published versio