Jarred Swalwell

Genome size differentiates co-occurring populations of the planktonic diatom Ditylum brightwellii (Bacillariophyta)

BackgroundDiatoms are one of the most species-rich groups of eukaryotic microbes known. Diatoms are also the only group of eukaryotic micro-algae with a diplontic life history, suggesting that the ancestral diatom switched to a life history dominated by a duplicated genome. A key mechanism of speciation among diatoms could be a propensity for additional stable genome duplications. Across eukaryotic taxa, genome size is directly correlated to cell size and inversely correlated to physiological rates. Differences in relative genome size, cell size, and acclimated growth rates were analyzed in isolates of the diatom Ditylum brightwellii. Ditylum brightwellii consists of two main populations with identical 18s rDNA sequences; one population is distributed globally at temperate latitudes and the second appears to be localized to the Pacific Northwest coast of the USA. These two populations co-occur within the Puget Sound estuary of WA, USA, although their peak abundances differ depending on local conditions.ResultsAll isolates from the more regionally-localized population (population 2) possessed 1.94 ± 0.74 times the amount of DNA, grew more slowly, and were generally larger than isolates from the more globally distributed population (population 1). The ITS1 sequences, cell sizes, and genome sizes of isolates from New Zealand were the same as population 1 isolates from Puget Sound, but their growth rates were within the range of the slower-growing population 2 isolates. Importantly, the observed genome size difference between isolates from the two populations was stable regardless of time in culture or the changes in cell size that accompany the diatom life history.ConclusionsThe observed two-fold difference in genome size between the D. brightwellii populations suggests that whole genome duplication occurred within cells of population 1 ultimately giving rise to population 2 cells. The apparent regional localization of population 2 is consistent with a recent divergence between the populations, which are likely cryptic species. Genome size variation is known to occur in other diatom genera; we hypothesize that genome duplication may be an active and important mechanism of genetic and physiological diversification and speciation in diatoms.

Ecophysiology of uncultivated marine euryarchaea is linked to particulate organic matter

Particles in aquatic environments host distinct communities of microbes, yet the evolution of particle-specialized taxa and the extent to which specialized microbial metabolism is associated with particles is largely unexplored. Here, we investigate the hypothesis that a widely distributed and uncultivated microbial group—the marine group II euryarchaea (MGII)—interacts with living and detrital particulate organic matter (POM) in the euphotic zone of the central California Current System. Using fluorescent in situ hybridization, we verified the association of euryarchaea with POM. We further quantified the abundance and distribution of MGII 16 S ribosomal RNA genes in size-fractionated seawater samples and compared MGII functional capacity in metagenomes from the same fractions. The abundance of MGII in free-living and >3 μm fractions decreased with increasing distance from the coast, whereas MGII abundance in the 0.8–3 μm fraction remained constant. At several offshore sites, MGII abundance was highest in particle fractions, indicating that particle-attached MGII can outnumber free-living MGII under oligotrophic conditions. Compared with free-living MGII, the genome content of MGII in particle-associated fractions exhibits an increased capacity for surface adhesion, transcriptional regulation and catabolism of high molecular weight substrates. Moreover, MGII populations in POM fractions are phylogenetically distinct from and more diverse than free-living MGII. Eukaryotic phytoplankton additions stimulated MGII growth in bottle incubations, providing the first MGII net growth rate measurements. These ranged from 0.47 to 0.54 d−1. However, MGII were not recovered in whole-genome amplifications of flow-sorted picoeukaryotic phytoplankton and heterotrophic nanoflagellates, suggesting that MGII in particle fractions are not physically attached to living POM. Collectively, our results support a linkage between MGII ecophysiology and POM, implying that marine archaea have a role in elemental cycling through interactions with particles.

Unveiling a phytoplankton hotspot at a narrow boundary between coastal and offshore waters

In terrestrial ecosystems, transitional areas between different plant communities (ecotones) are formed by steep environmental gradients and are commonly characterized by high species diversity and primary productivity, which in turn influences the foodweb structure of these regions. Whether comparable zones of elevated diversity and productivity characterize ecotones in the oceans remains poorly understood. Here we describe a previously hidden hotspot of phytoplankton diversity and productivity in a narrow but seasonally persistent transition zone at the intersection of iron-poor, nitrate-rich offshore waters and iron-rich, nitrate-poor coastal waters of the Northeast Pacific Ocean. Novel continuous measurements of phytoplankton cell abundance and composition identified a complex succession of blooms of five distinct size classes of phytoplankton populations within a 100-km–wide transition zone. The blooms appear to be fueled by natural iron enrichment of offshore communities as they are transported toward the coast. The observed succession of phytoplankton populations is likely driven by spatial gradients in iron availability or time since iron enrichment. Regardless of the underlying mechanism, the resulting communities have a strong impact on the regional biogeochemistry as evidenced by the low partial pressure of CO2 and the nearly complete depletion of nutrients. Enhanced phytoplankton productivity and diversity associated with steep environmental gradients are expected wherever water masses with complementary nutrient compositions mix to create a region more favorable for phytoplankton growth. The ability to detect and track these important but poorly characterized marine ecotones is critical for understanding their impact on productivity and ecosystem structure in the oceans.

Microbubble sizing and shell characterization using flow cytometry

Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled using Mie theory, and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures (from 95 to 333 kPa) by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters.

Light-driven synchrony of Prochlorococcus growth and mortality in the subtropical Pacific gyre

Significance The cyanobacterium Prochlorococcus is the most abundant photosynthetic organism in the oceans, driving marine food webs and biogeochemistry. Estimates of Prochlorococcus cell mortality and cell production are critical to determine how organic matter is transferred into the food web. Using novel high-resolution sampling methods on two winter-time cruises, we show that the daily production of Prochlorococcus cells in surface waters of the subtropical Pacific gyre is consistently balanced by their nightly consumption by other organisms. These synchronized loss processes suggest that Prochlorococcus-derived organic matter stabilizes multiple species interactions, from viruses to grazers. The observed resilience of this synchronized food web dynamic as temperature increased within the gyre suggests that ecosystem stability may persist in future warmer oceans. Theoretical studies predict that competition for limited resources reduces biodiversity to the point of ecological instability, whereas strong predator/prey interactions enhance the number of coexisting species and limit fluctuations in abundances. In open ocean ecosystems, competition for low availability of essential nutrients results in relatively few abundant microbial species. The remarkable stability in overall cell abundance of the dominant photosynthetic cyanobacterium Prochlorococcus is assumed to reflect a simple food web structure strongly controlled by grazers and/or viruses. This hypothesized link between stability and ecological interactions, however, has been difficult to test with open ocean microbes because sampling methods commonly have poor temporal and spatial resolution. Here we use continuous techniques on two different winter-time cruises to show that Prochlorococcus cell production and mortality rates are tightly synchronized to the day/night cycle across the subtropical Pacific Ocean. In warmer waters, we observed harmonic oscillations in cell production and mortality rates, with a peak in mortality rate consistently occurring ∼6 h after the peak in cell production. Essentially no cell mortality was observed during daylight. Our results are best explained as a synchronized two-component trophic interaction with the per-capita rates of Prochlorococcus consumption driven either directly by the day/night cycle or indirectly by Prochlorococcus cell production. Light-driven synchrony of food web dynamics in which most of the newly produced Prochlorococcus cells are consumed each night likely enforces ecosystem stability across vast expanses of the open ocean.

Virtual-core flow cytometry

Traditional flow cytometers use a sheath fluid to position particles or cells for cytometric measurements, but the need for sheath fluid greatly complicates flow cytometric instrumentation. A cytometric detector that is free of the requirements of sheath fluid can simplify the design of flow cytometers and can extend their use into a number of areas. We designed a flow cytometer that uses a combination of three photodetectors to sense the position of a particle in sample stream. The position‐sensitive detectors create a virtual core in the sample stream that eliminates the need for sheath fluid. In this article, we demonstrate the efficacy of a virtual‐core flow cytometer (VCFC) using test particles, immunofluorescently labeled thymocytes, and raw seawater. The VCFC performs accurate measurements that can be used for a number of uses including environmental monitoring or simple immunology tests.

Flow cytometry to characterize microbubbles

Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled with Mie theory and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters.

Single‐bubble sonoluminescence in microgravity

A recent experiment involving single‐bubble sonoluminescence aboard NASA’s parabolic research aircraft will be described. Measurements of the intensity of the light emission were performed during periods of microgravity (near 0 g) and hypergravity (near 2 g). Gravitational effects on the luminescence and extinction pressure thresholds were examined. In addition, the light emission was monitored under constant drive conditions as the gravitational acceleration varied during the parabolas. Measurements show some variability in the sonoluminescence intensity for both thresholds. However, during otherwise constant conditions in which the drive pressure amplitude did not change, the intensity from stable single‐bubble sonoluminescence was observed to increase during periods of microgravity. There was an initial increase in intensity that occurred simultaneous with the decrease in the gravitational acceleration, followed by a slow increase in intensity that appeared to level off near the end of the microgravity...

Dynamics of Teleaulax-like cryptophytes during the decline of a red water bloom in the Columbia River Estuary

The mixotrophic ciliate, Mesodinium rubrum, is a globally distributed ciliate that relies on the acquisition and use of chloroplasts derived from its cryptophyte prey. The ecology and physiology of the cryptophytes is not well known, nor is it clear how their growth influences M. rubrum blooms. A 4-week survey was conducted in the Columbia River estuary in 2013 during the decline of the annual M. rubrum bloom to better understand how environmental factors influence the dynamics of the cryptophyte prey, Teleaulax amphioxeia. Abundances and division rates of freeliving Teleaulax-like cryptophytes were continuously monitored using flow cytometry. Cryptophyte division rates, estimated in situ for the first time using a size-structured division rate model, ranged from 0.2 to 1.5 d, with the highest rates observed in accordance with high abundances. These division rates were positively correlated with concentrations of dissolved inorganic nitrogen and phosphorus, suggesting nutrient availability limited the growth of Teleaulax-like cryptophytes at that time. Assuming a minimum ingestion rate of ~1 cryptophyte ciliate day, the growth of M. rubrum may have been limited by the low abundance of Teleaulax-like cryptophytes during the M. rubrum bloom decline. Our results highlight the importance of prey availability for understanding the dynamics of red water blooms.

Acoustic and optical characterization of ultrasound contrast agents via flow cytometry

Characterizing ultrasound contrast agents (UCAs) involve measuring the size and population distribution. However, these instruments do not allow for characterization of shell properties, which are important for (1) stability to administration and circulation throughout the vasculature; (2) UCA response to ultrasound; and (3) conjugating ligands for molecular imaging. Thus it is critical to understand the physical and rheological properties of shells. We previously developed a light scattering technique to characterize the shell properties of UCAs [Guan and Matula, JASA, Vol 116(5), 2004; Tu, et al., IEEE Trans. Ultrason., Ferroelec., and Freq. Control, vol. 58(5), 2011]. The most recent manifestation involves a flow cytometer modified with a custom square quartz flow cell in place of the standard nozzle and fluid jet. Acoustic coupling to the carrier sheath fluid and UCA samples occurred through a PZT bonded to one side of the flow cell. The PZT-driven UCA oscillations were processed and fitted to the Mar...