The changes in marine ecological conditions brought on by warming and stratification of the oceans have radically shifted many marine environments around the globe. This project aimed to better characterize the aggregation behavior of the abundant picocyanobacterium Prochlorococcus marinus, which is hypothesized to dominate over other phytoplankton as the primary autotroph in increasingly warmer and nutrient poor oceans. This aggregation, believed to be mediated through the secretion of sticky Transparent Exopolymeric Substances (TEP), might be key for Prochlorococcus to sink throughout the ocean and serve as a source of carbon to other communities within its environment. Considering the relatively low concentration of TEP secreted by Prochlorococcus when on its own, this study explored the synergistic effect that heterotrophic bacteria and inorganic minerals in the surrounding seawater may have on the aggregation of P. marinus. This was done by inoculating P. marinus and the model heterotroph Marinobacter adhaerens HP15 individually and mixed in cylindrical roller tanks with the addition of ballasting clay minerals into roller tanks to simulate constant sinking for 7 days. The aggregates which formed after rolling were quantified and their sinking velocities and excess densities were measured. Our results indicate that the most numerous and densest aggregates formed when Prochlorococcus was in the presence of both M. adhaerens and kaolinite clay particles. I will discuss how methodology, particularly cell number, may play a role in the enhanced aggregation that I found when Prochlorococcus was cultured together with the Marinobacter.

Microscopic algae have been investigated extensively by researchers for decades for their ability to bioremediate wastewater and flue gas while producing valuable biomass for use as feed, fuel, fertilizer, nutraceutical, and other specialty products. Reports of the exciting commercial potential of this diverse group of organisms started appearing in the literature as early as the 1940’s. However, nearly 80 years later, relatively few successful commercial microalgae installations exist and algae have not yet reached agricultural commodity status. This dissertation examines three major bottlenecks to commercial microalgae production including lack of an efficient and economical cultivation strategy, poor management of volatile waste nutrients, and costly harvesting and post processing strategies. A chapter is devoted to each of these three areas to gain a better understanding of each bottleneck as well as strategies for overcoming them.

The first chapter demonstrates the capability of two strains of Scenedesmus acutus to grow in ultra-high-density (>10 g L-1 dry weight biomass) cultures in flat panel photobioreactors for year-round production in the desert Southwest with record volumetric biomass productivity. The advantages and efficiency of high-density cultivation are discussed. The second chapter focuses on uptake and utilization of the volatile components of wastewater: ammonia and carbon dioxide. Scenedesmus acutus was cultured on wastewater from both municipal and agricultural origin and was shown to perform significantly better on flue gas as compared to commercial grade CO2 and just as well on waste nutrients as the commonly used BG-11 laboratory culture media, all while producing up to 50% lipids of the dry weight biomass suitable for use in biodiesel. The third chapter evaluates the feasibility of using gravity sedimentation for the harvesting of the difficult-to-separate Scenedesmus acutus green algae biomass followed by microfluidization to disrupt the cells. Lipid-extracted biomass was then studied as a fertilizer for plants and shown to have similar performance to a commercially available 4-6-6 fertilizer. Based on the work from these three chapters, a summary of modifications are suggested to help current and future microalgae companies be more competitive in the marketplace with traditional agricultural commodities.

Coral reefs are diverse marine ecosystems, where reef building corals provide both the structure of the habitat as well as the primary production through their symbiotic algae, and alongside algae living on the reef itself, are the basis of the food web of the reef. In this way, coral reefs are the ocean's "forests" and are estimated to support 25% of all marine species. However, due to the large size of a coral reef, the relative inaccessibility and the reliance on in situ surveying methods, our current understanding of reefs is spatially limited. Understanding coral reefs from a more spatially complete perspective will offer insight into the ecological factors that contribute to coral reef vitality. This has become a priority in recent years due to the rapid decline of coral reefs caused by mass bleaching. Despite this urgency, being able to assess the entirety of a coral reef is physically difficult and this obstacle has not yet been overcome. However, similar difficulties have been addressed in terrestrial ecosystems by using remote sensing methods, which apply hyperspectral imaging to assess large areas of primary producers at high spatial resolutions. Adapting this method of remote spectral sensing to assess coral reefs has been suggested, but in order to quantify primary production via hyper spectral imaging, light-use efficiencies (LUEs) of coral reef communities need to be known. LUEs are estimations of the rate of carbon fixation compared to incident absorbed light. Here, I experimentally determine LUEs and report on several parameters related to LUE, namely net productivity, respiration, and light absorbance for the main primary producers in coral reefs surrounding Bermuda, which consist of algae and coral communities. The derived LUE values fall within typical ranges for LUEs of terrestrial ecosystems, with LUE values for coral averaging 0.022 ± 0.002 mol O2 mol photons-1 day-1 at a water flow rate of 17.5 ± 2 cm s^(-1) and 0.049 ± 0.011 mol O2 mol photons-1 day-1 at a flow rate of 32 ± 4 cm s^(-1) LUE values for algae averaged 0.0335 ± 0.0048 mol O2 mol photons-1 day-1 at a flow rate of 17.5 ± 2 cm s^(-1). These values allow insight into coral reef productivity and opens the door for future remote sensing applications.

Prochlorococcus marinus (MED4), a genus of marine picocyanobacteria that proliferates in open oligotrophic ocean, is one of the most abundant photosynthetic microbes in the world, estimated to contribute up to 10% of the ocean’s primary production. The productivity of these microorganisms is controlled by macronutrient availability in the surface waters. The ratio of macronutrients in the ocean was defined, by Alfred Redfield, as an elemental ratio of 106C:16N:1P. However, the C:N:P ratio varies based on region, season, temperature and irradiance, as well as the composition of the primary producers. In oligotrophic gyres, these nutrient ratios are elevated from the Redfield stoichiometry, but whether this ratio exerts influence on the growth rate of the organism has not been investigated. Elemental stoichiometry of available nutrients can affect the aggregation of organic carbon and exportation of the particles to the ocean depths. The purpose of this study was to investigate the effects of nutrient limitation on aggregation and transparent exopolymeric particle (TEP) production which aids in aggregation. My findings suggested that nutrient limitation reduces TEP production and does not increase aggregate volume concentration. With continued warming, certain regions of the ocean will become more oligotrophic, which further decreases the nutrient supply available for Prochlorococcus. My research shows that this could lead to decreased exportation of organic carbon matter to the depths of the sea.

Safe, readily available, and reliable sources of water are an essential component of any municipality’s infrastructure. Phoenix, Arizona, a southwestern city, has among the highest per capita water use in the United States, making it essential to carefully manage its reservoirs. Generally, municipal water bodies are monitored through field sampling. However, this approach is limited spatially and temporally in addition to being costly. In this study, the application of remotely sensed reflectance data from Landsat 7’s Enhanced Thematic Mapper Plus (ETM+) and Landsat 8’s Operational Land Imager (OLI) along with data generated through field-sampling is used to gain a better understanding of the seasonal development of algal communities and levels of suspended particulates in the three main terminal reservoirs supplying water to the Phoenix metro area: Bartlett Lake, Lake Pleasant, and Saguaro Lake. Algal abundances, particularly the abundance of filamentous cyanobacteria, increased with warmer temperatures in all three reservoirs and reached the highest comparative abundance in Bartlett Lake. Prymnesiophytes (the class of algae to which the toxin-producing golden algae belong) tended to peak between June and August, with one notable peak occurring in Saguaro Lake in August 2017 during which time a fish-kill was observed. In the cooler months algal abundance was comparatively lower in all three lakes, with a more even distribution of abundance across algae classes. In-situ data from March 2017 to March 2018 were compared with algal communities sampled approximately ten years ago in each reservoir to understand any possible long-term changes. The findings show that the algal communities in the reservoirs are relatively stable, particularly those of the filamentous cyanobacteria, chlorophytes, and prymnesiophytes with some notable exceptions, such as the abundance of diatoms, which increased in Bartlett Lake and Lake Pleasant. When in-situ data were compared with Landsat-derived reflectance data, two-band combinations were found to be the best-estimators of chlorophyll-a concentration (as a proxy for algal biomass) and total suspended sediment concentration. The ratio of the reflectance value of the red band and the blue band produced reasonable estimates for the in-situ parameters in Bartlett Lake. The ratio of the reflectance value of the green band and the blue band produced reasonable estimates for the in-situ parameters in Saguaro Lake. However, even the best performing two-band algorithm did not produce any significant correlation between reflectance and in-situ data in Lake Pleasant. Overall, remotely-sensed observations can significantly improve our understanding of the water quality as measured by algae abundance and particulate loading in Arizona Reservoirs, especially when applied over long timescales.

Quagga mussels are an aquatic invasive species capable of causing economic and ecological damage. Despite the quagga mussels’ ability to rapidly spread, two watersheds, the Salt River system and the Verde River system of Arizona, both had no quagga mussel detections for 8 years. The main factor thought to deter quagga mussels was the stratification of the two watersheds during the summer, resulting in high temperatures in the epilimnion and low dissolved oxygen in the hypolimnion. In 2015, Canyon Lake, a reservoir of the Salt River watershed, tested positive for quagga mussel veligers. In this study, I used Landsat 7 and Landsat 8 satellite data to determine if changes in the surface temperature have caused a change to the reservoir allowing quagga mussel contamination. I used a location in the center of the lake with a root mean squared error (RMSE) of 0.80 and a correlation coefficient (R^2) of 0.82, but I did not detect any significant variations in surface temperatures from recent years. I also measured 21 locations on Canyon Lake to determine if the locations in Canyon Lake were able to harbor quagga mussels. I found that summer stratification caused hypolimnion dissolved oxygen levels to drop well below the quagga mussel threshold of 2mg/L. Surface temperatures, however were not high enough throughout the lake to prevent quagga mussels from inhabiting the epilimnion. It is likely that a lack of substrate in the epilimnion have forced any quagga mussel inhabitants in Canyon Lake to specific locations that were not necessarily near the point of quagga veliger detection sampling. The research suggests that while Canyon Lake may have been difficult for quagga mussels to infest, once they become established in the proper locations, where they can survive through the summer, quagga mussels are likely to become more prevalent.

The phytoplankton communities in the open oceans are dominated by picophytoplankton (0.7-2µm) and nanophytoplankton (3-5µm). Studying the community dynamics of these phytoplankton is important to learn about their role in the carbon cycle and food web of the oceans. Dilution experiments were used, along with microscopy and molecular techniques, to determine abundance, biomass and phytoplankton growth and grazing rates in the oligotrophic Sargasso Sea (western North Atlantic subtropical gyre) around the Bermuda Atlantic Time Series Station (BATS) in the summer of 2012. With low biomass and chlorophyll a, the Sargasso Sea appears to be unproductive at first glance, but I found that pico- and nanophytoplankton have high instantaneous growth rates that are balanced by the high grazing rates of microzooplankton.
Mesoscale eddies are important features in the Sargasso Sea that can increase or decrease the available nutrients in the euphotic zone. Two different mesoscale eddies were sampled: an anti-cyclonic eddy and the BATS station which was located at the edge of a cyclonic eddy. The results indicated that BATS had overall higher instantaneous growth (µ between 0.1 d-1 and 3.7 d-1) and grazing rates on pico- and nanophytoplankton, as well as diatoms, compared to the anti-cyclonic eddy (µ between 0.2 d-1 and 3 d-1). I also determined taxon-specific rates using quantitative polymerase chain reaction (qPCR) for the order Mamiellales, one of the smallest representatives of the abundant prasinophytes. This method yielded surprisingly high growth (9.7 d-1 ) and grazing rates (-8.2 d-1) at 80m for BATS. The euphotic zone (~100m) integrated biomass of all phytoplankton did not vary significantly between BATS (379 mg C m-2) and the anti-cyclonic eddy (408 mg C m-2) and the net growth rates at both locations were very close to zero for most of the groups. Although the biomass and net growth rates did not vary greatly between the two locations, the high instantaneous growth and grazing rates of pico- and nano-eukaryotic phytoplankton indicate an increase in the rate of the marine microbial food web, or microbial loop, compared to the anti-cyclonic eddy. This could have been due to the input of new nutrients in the edge of the cyclonic eddy at BATS. Thus, my study suggests that mesoscale variability is of considerable importance for the dynamics of the phytoplankton community and their role in the microbial loop. Much can be learned when using DNA based taxon-specific rates, especially to understand the relative importance and contribution of specific taxa.
More taxon-specific molecular studies will have to be carried out to quantify specific rates of more phytoplankton groups, which will supply a more complete knowledge of phytoplankton community dynamics in the Sargasso Sea. This will increase our understanding of the role of specific groups to the biological carbon dynamics in the euphotic zone into the deep ocean.

It is well known that deficiencies in key chemical elements (such as phosphorus, P) can reduce animal growth; however, recent empirical data have shown that high levels of dietary nutrients can also reduce animal growth. In ecological stoichiometry, this phenomenon is known as the "stoichiometric knife edge," but its underlying mechanisms are not well-known. Previous work has suggested that the crustacean zooplankter Daphnia reduces its feeding rates on phosphorus-rich food, causing low growth due to insufficient C (energy) intake. To test for this mechanism, feeding rates of Daphnia magna on algae (Scenedesmus acutus) differing in C:P ratio (P content) were determined. Overall, there was a significant difference among all treatments for feeding rate (p < 0.05) with generally higher feeding rates on P-rich algae. These data indicate that both high and low food C:P ratio do affect Daphnia feeding rate but are in contradiction with previous work that showed that P-rich food led to strong reductions in feeding rate. Additional experiments are needed to gain further insights.

The giant green sea anemone, Anthopleura xanthogrammica, hosts two different endosymbiotic algae. One is a unicellular chlorophyte, Elliptochloris marina; the other is Symbiodinium muscatinei, a dinoflagellate. Hosting these different symbionts influences the life history strategy of A. xanthogrammica's congener A. elegantissima, directly impacting its reproductive strategy (asexual vs. sexual). My study sought to examine whether the type and density of symbiont also affects the reproductive condition of A. xanthogrammica, which reproduces only sexually. Gonad development was measured in anemones from Slip Point, Clallam Bay, WA and Tongue Point, WA along with symbiont type and density per mg of anemone protein. The results indicate a trend towards brown anemones having more developed gonads, especially in males. This may mean that A. xanthogrammica anemones that host zooxanthellae are more reproductively fit than zoochlorellate anemones. Thus, it may be favorable for anemones to host zooxanthellae. This is especially true in summer months when the high temperatures and mid-day low tides coincide with the period of most rapid gonad development.

Many environmental microorganisms such as marine microbes are un-culturable; hence, they should be analyzed in situ. Even though a few in situ ocean observing instruments have been available to oceanographers, their applications are limited, because these instruments are expensive and power hungry.

In this dissertation project, an inexpensive, portable, low-energy consuming, and highly quantitative microbiological genomic sensor has been developed for in situ ocean-observing systems. A novel real-time colorimetric loop-mediated isothermal amplification (LAMP) technology has been developed for quantitative detection of microbial nucleic acids. This technology was implemented on a chip-level device with an embedded inexpensive imaging device and temperature controller to achieve quantitative detection within one hour. A bubble-free liquid handling approach was introduced to avoid bubble trapping during liquid loading, a common problem in microfluidic devices. An algorithm was developed to reject the effect of bubbles generated during the reaction process, to enable more accurate nucleic acid analysis. This genomic sensor has been validated at gene and gene expression levels using Synechocystis sp. PCC 6803 genomic DNA and total RNA. Results suggest that the detection limits reached 10 copies/μL and 100 fg/μL, respectively. This approach was highly quantitative, with linear standard curves down to 103 copies/μL and 1 pg/μL, respectively. In addition to environmental microbe characterization, this genomic sensor has been employed for viral RNA quantification during an infectious disease outbreak. As the Zika fever was spreading in America, a quantitative detection of Zika virus has been performed. The results show that the genomic sensor is highly quantitative from 10 copies/μL to 105 copies/μL. This suggests that the novel nucleic acid quantification technology is sensitive, quantitative, and robust. It is a promising candidate for rapid microbe detection and quantification in routine laboratories.

In the future, this genomic sensor will be implemented in in situ platforms as a core analytical module with minor modifications, and could be easily accessible by oceanographers. Deployment of this microbial genomic sensor in the field will enable new scientific advances in oceanography and provide a possible solution for infectious disease detection.