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Abundant phytoplankton and bacteria were identified by microscopy and high-throughput 16S rRNA tag Illumina sequencing of samples from water- and ice phases collected during winter at two central European Great Lakes, Balaton and Fertő (Neusiedlersee). Bacterial reads at all sites were dominated (>85%) by Bacteroidetes and Proteobacteria. Amongst phototrophs, microscopy and 16S sequencing revealed that both phytoplankton and cyanobacteria were represented, with a median of 1500 cyanobacterial sequence reads amongst 13 samples analyzed. The sequence analysis compared replicate Balaton and Fertő ice and water samples with an outgroup from three Hungarian soda lakes. In particular, both water and ice from Fertő contained high contributions from cyanobacteria. Two percent of total reads identified to the level of family in water at Fertő were dominated by a single operational taxonomic unit (OTU) of a cyanobacterium within the Rivulariaceae, which was largely absent from ice. Conversely, ice samples from both lakes yielded an abundant OTU assigned to a Flavobacterium sp. known to be associated with freshwater ice. Principal Coordinates Analysis (PCoA) revealed that the ice communities from all sites were similar to one another, and that the water communities did not cluster together. Fluorescence emission spectra obtained at 77 K confirmed the presence of intact cyanobacteria in Fertő water and ice. Photosynthetic characterization of phototrophs resident in water and ice analyzed by assay of acid-stable photosynthetic H14CO3– incorporation showed that communities from both phases were photosynthetically active, thus adding to growing recognition of ice-covered lakes as viable habitat for phototrophs.

The Maumee River is the primary source for nutrients fueling seasonal Microcystis-dominated blooms in western Lake Erie’s open waters though such blooms in the river are infrequent. The river also serves as source water for multiple public water systems and a large food services facility in northwest Ohio, USA. On 20 September 2017, an unprecedented bloom was reported in the Maumee River estuary within the Toledo metropolitan area, which triggered a recreational water advisory. Here we (1) explore physical drivers likely contributing to the bloom’s occurrence, and (2) describe the toxin concentration and bacterioplankton taxonomic composition. A historical analysis using ten-years of seasonal river discharge, water level, and local wind data identified two instances when high-retention conditions occurred over ≥10 days in the Maumee River estuary: in 2016 and during the 2017 bloom. Observation by remote sensing imagery supported the advection of cyanobacterial cells into the estuary from the lake during 2017 and the lack of an estuary bloom in 2016 due to a weak cyanobacterial bloom in the lake. A rapid-response survey during the 2017 bloom determined levels of the cyanotoxins, specifically microcystins, in excess of recreational contact limits at sites within the lower 20 km of the river while amplicon sequencing found these sites were dominated by Microcystis. These results highlight the need to broaden our understanding of physical drivers of cyanobacterial blooms within the interface between riverine and lacustrine systems, particularly as such blooms are expected to become more prominent in response to a changing climate.

This study examined vertically-resolved patterns in microbial community structure across a freshwater trophic gradient extending 1600 km from the oligotrophic waters of Lake Superior to the eutrophic waters of Lake Erie, the most anthropogenically influenced of the Laurentian Great Lakes system. Planktonic bacterial communities clustered by Principal Coordinates Analysis (PCoA) on UniFrac distance matrices into four groups representing the epilimnion and hypolimnion of the upper Great Lakes (Lakes Superior and Huron), Lake Superior's northern bays (Nipigon and Black bays), and Lake Erie. The microbes within the upper Great Lakes hypolimnion were the most divergent of these groups with elevated abundance of Planctomycetes and Chloroflexi compared to the surface mixed layer. Statistical tests of the correlation between distance matrices identified temperature and sample depth as the most influential community structuring parameters, reflecting the strong UniFrac clustering separating mixed-layer and hypolimnetic samples. Analyzing mixed-layer samples alone showed clustering patterns were correlated with nutrient concentrations. Operational taxonomic units (OTU) which were differentially distributed among these conditions often accounted for a large portion of the reads returned. While limited in coverage of temporal variability, this study contributes a detailed description of community variability that can be related to other large freshwater systems characterized by changing trophic state.

Toxic cyanobacterial harmful algal blooms (cyanoHABs) are one of the most significant threats to the security of Earth's surface freshwaters. In the United States, the Federal Water Pollution Control Act of 1972 (i.e., the Clean Water Act) requires that states report any waterbody that fails to meet applicable water quality standards. The problem is that for fresh waters impacted by cyanoHABs, no scientifically-based framework exists for making this designation. This study describes the development of a data-based framework using the Ohio waters of western Lake Erie as an exemplar for large lakes impacted by cyanoHABs. To address this designation for Ohio's open waters, the Ohio Environmental Protection Agency (EPA) assembled a group of academic, state and federal scientists to develop a framework that would determine the criteria for Ohio EPA to consider in deciding on a recreation use impairment designation due to cyanoHAB presence. Typically, the metrics are derived from on-lake monitoring programs, but for large, dynamic lakes such as Lake Erie, using criteria based on discrete samples is problematic. However, significant advances in remote sensing allows for the estimation of cyanoHAB biomass of an entire lake. Through multiple years of validation, we developed a framework to determine lake-specific criteria for designating a waterbody as impaired by cyanoHABs on an annual basis. While the criteria reported in this manuscript are specific to Ohio's open waters, the framework used to determine them can be applied to any large lake where long-term monitoring data and satellite imagery are available.

Ice-nucleating particles (INPs) associated with fresh waters are a neglected, but integral component of the water cycle. Abundant INPs were identified from surface waters of both the Maumee River and Lake Erie with ice nucleus spectra spanning a temperature range from −3 to −15 °C. The majority of river INPs were submicron in size and attributed to biogenic macromolecules, inferred from the denaturation of ice-nucleation activity by heat. In a watershed dominated by row-crop agriculture, higher concentrations of INPs were found in river samples compared to lake samples. Further, ice-nucleating temperatures differed between river and lake samples, which indicated different populations of INPs. Seasonal analysis of INPs that were active at warmer temperatures (≥−10 °C; INP–10) showed their concentration to correlate with river discharge, suggesting a watershed origin of these INPs. A terrestrial origin for INPs in the Maumee River was further supported by a correspondence between the ice-nucleation signatur...

In late May 2016, a cyanobacterial harmful algal bloom (cHAB) was detected in the Maumee River, the largest tributary to Lake Erie, the southernmost lake of the Laurentian Great Lakes system. Testing on 31 May identified Planktothrix agardhii as the dominant cyanobacterium with cell abundance exceeding 1.7×109 cells/L and total microcystins (MC) reaching 19 μg/L MC-LR equivalents, a level over 10-fold higher than the 2015 revised U.S. Environmental Protection Agency (EPA) national health advisory levels for drinking water exposure to adults. Low river discharge coincident with negligible precipitation through the latter half of May coincided with an 80% decline in river turbidity that likely favored bloom formation by a low-light adapted P. agardhii population. Also contributing to the cHAB were high initial nutrient loads and an increase of the river temperature from 13°C to 26°C over this same period. The bloom persisted through 5 June with microcystins exceeding 22 μg/L MC-LR equivalents at the bloom peak. By 6 June, the river had returned to its muddy character following a rain event and sampling on 7 June detected only low levels of toxin (

There is growing recognition that winter is an important season for growth of photoautotrophs in ice-covered freshwater environments. Exemplifying this are expansive under-ice blooms of filamentous diatoms in Lake Erie. Here we describe a metatranscriptome constructed from a phytoplankton community dominated by filamentous diatoms. As expected, a high percentage (> 73%) of the sequences with BLAST hits to nucleotides or proteins in National Center for Biotechnology Information databases were associated with photosynthetic algae of which the majority were diatoms, mainly Aulacoseira spp. and Stephanodiscus spp. which was confirmed by analysis of 18S rRNA gene transcripts and microscopy. Consistent with the winter growth environment, psychrophilic and low-light adaptations were observed. Prominent among adaptations to cold were transcripts for genes involved in biosynthesis of unsaturated fatty acids, which were consistent with expected increased membrane fluidity at low temperatures. Reflecting the combined effect of low winter insolation and high light attenuation were an abundant complement of reads for light-harvesting antennae, mainly genes encoding fucoxanthin chlorophyll a / c proteins. The presence of virulence factors originating from oomycetes offers support for new hypotheses into the eventual decline of Lake Erie's winter diatom bloom. Whereas fungi were identified both through the metatranscriptome and by microscopy, dsRNA viruses of fungi were detected that may indirectly counter against fungal infection. This study demonstrates the utility of an environmental omics approach to yield insights underlying phototrophic life as well as the interactions of the entire microbial community in an extreme environment.

In early August 2014, the municipality of Toledo, OH (USA) issued a ‘do not drink’ advisory on their water supply directly affecting over 400,000 residential customers and hundreds of businesses (Wilson, 2014). This order was attributable to levels of microcystin, a potent liver toxin, which rose to 2.5 μg L−1 in finished drinking water. The Toledo crisis afforded an opportunity to bring together scientists from around the world to share ideas regarding factors that contribute to bloom formation and toxigenicity, bloom and toxin detection as well as prevention and remediation of bloom events. These discussions took place at an NSF- and NOAA-sponsored workshop at Bowling Green State University on April 13 and 14, 2015. In all, more than 100 attendees from six countries and 15 US states gathered together to share their perspectives. The purpose of this review is to present the consensus summary of these issues that emerged from discussions at the Workshop. As additional reports in this special issue provide detailed reviews on many major CHAB species, this paper focuses on the general themes common to all blooms, such as bloom detection, modeling, nutrient loading, and strategies to reduce nutrients.

Oligotrophic Lake Superior and mesotrophic Lake Erie are trophic end members of the hydrologically connected Laurentian Great Lakes system, and as such exhibit different profiles of dissolved nitrogen species. Nitrification in Lake Superior has led to increasing nitrate concentrations over the past century, as opposed to Erie, where nitrate inventories have declined due to denitrification. In this study, we examined the abundance and diversity of nitrifying microbes involved in the oxidation of ammonia to nitrite, and nitrite to nitrate. By in situ hybridization methods, we enumerated the major planktonic ammonia oxidizing bacteria (AOB) and archaea (AOA) during a July 2011 cruise from Lake Superior to Lake Erie. In Lake Superior, AOA dominated compared to AOB, typically exceeding 5 × 103 mL− 1, whereas in Erie, AOB were more abundant than AOA. These data parallel prior work on Lake Superior and Lake Erie sediments, in which AOA are far more abundant in Superior, but AOB dominate in Erie. The lakes were sampled during stratification, and AOA and AOB were largely restricted to the hypolimnion, consistent with the observation that ammonia oxidizers are photoinhibited in surface waters. In Lakes Superior and Erie, we also detected nitrite oxidizing bacteria (NOB) in a pattern paralleling AOA/AOB abundance. Phylogenetic analysis of archaeal 16S rRNA revealed that the planktonic archaea of Lake Superior are members of the ammonia oxidizing Group I.1a Thaumarchaeota most closely related to Nitrosoarchaeum limnia. These AOA are distinct from the Group I.1a AOA in Lake Superior sediments. The major AOB of Lake Erie form a subcluster within the genus Nitrosospira.

Recent global water quality crises point to an urgent need for greater understanding of cyanobacterial harmful algal blooms (cHABs) and their drivers. Nearshore areas of Lake Erie such as Sandusky Bay may become seasonally limited by nitrogen (N) and are characterized by distinct cHAB compositions (i.e., Planktothrix over Microcystis). This study investigated phytoplankton N uptake pathways, determined drivers of N depletion, and characterized the N budget in Sandusky Bay. Nitrate (NO3-) and ammonium (NH4+) uptake, N fixation, and N removal processes were quantified by stable isotopic approaches. Dissimilatory N reduction was a relatively modest N sink, with denitrification, anammox, and N2O production accounting for 84, 14, and 2 % of sediment N removal, respectively. Phytoplankton assimilation was the dominant N uptake mechanism, and NO3- uptake rates were higher than NH4+ uptake rates. Riverine N loading was sometimes insufficient to meet assimilatory and dissimilatory demands, but N fixation alleviated this deficit. N fixation made up 23.7–85.4 % of total phytoplankton N acquisition and indirectly supports Planktothrix blooms. However, N fixation rates were surprisingly uncorrelated with NO3- or NH4+ concentrations. Owing to temporal separation in sources and sinks of N to Lake Erie, Sandusky Bay oscillates between a conduit and a filter of downstream N loading to Lake Erie, delivering extensively recycled forms of N during periods of low export. Drowned river mouths such as Sandusky Bay are mediators of downstream N loading, but climate-change-induced increases in precipitation and N loading will likely intensify N export from these systems.

Sandusky Bay, Lake Erie, receives high nutrient loadings (nitrogen and phosphorus) from the Sandusky River, which drains an agricultural watershed. Eutrophication and cyanobacterial harmful algal blooms (cyanoHABs) persist throughout summer. Planktothrix agardhii is the dominant bloom-forming species and the main producer of microcystins in Sandusky Bay. Non-N2 fixing cyanobacteria, such as Planktothrix and Microcystis, thrive on chemically reduced forms of nitrogen, such as ammonium (NH4+) and urea. Ammonium regeneration and potential uptake rates and total microbial community demand for NH4+ were quantified in Sandusky Bay. Potential NH4+ uptake rates in the light increased from June to August at all stations. Dark uptake rates also increased seasonally and, by the end of August, were on par with light uptake rates. Regeneration rates followed a similar pattern and were significantly higher in August than June. Ammonium uptake kinetics during a Planktothrix-dominated bloom in Sandusky Bay and a Microcystis-dominated bloom in Maumee Bay were also compared. The highest half saturation constant (Km) in Sandusky Bay was measured in June and decreased throughout the season. In contrast, Km values in Maumee Bay were lowest at the beginning of summer and increased in October. A significant increase in Vmax in Sandusky Bay was observed between July and the end of August, reflective of intense competition for depleted NH4+. Metatranscriptome results from Sandusky Bay show a shift from cyanophycin synthetase (luxury NH4+ uptake; cphA1) expression in early summer to cyanophycinase (intracellular N mobilization; cphB/cphA2) expression in August, supporting the interpretation that the microbial community is nitrogen-starved in late summer. Combined, our results show that, in late summer, when nitrogen concentrations are low, cyanoHABs in Sandusky Bay rely on regenerated NH4+ to support growth and toxin production. Increased dark NH4+ uptake late in summer suggests an important heterotrophic contribution to NH4+ depletion in the phycosphere. Kinetic experiments in the two bays suggest a competitive advantage for Planktothrix over Microcystis in Sandusky Bay due to its higher affinity for NH4+ at low concentrations.

Mid-winter limnological surveys of Lake Erie captured extremes in ice extent ranging from expansive ice cover in 2010 and 2011 to nearly ice-free waters in 2012. Consistent with a warming climate, ice cover on the Great Lakes is in decline, thus the ice-free condition encountered may foreshadow the lakes future winter state. Here, we show that pronounced changes in annual ice cover are accompanied by equally important shifts in phytoplankton and bacterial community structure. Expansive ice cover supported phytoplankton blooms of filamentous diatoms. By comparison, ice free conditions promoted the growth of smaller sized cells that attained lower total biomass. We propose that isothermal mixing and elevated turbidity in the absence of ice cover resulted in light limitation of the phytoplankton during winter. Additional insights into microbial community dynamics were gleaned from short 16S rRNA tag (Itag) Illumina sequencing. UniFrac analysis of Itag sequences showed clear separation of microbial communities related to presence or absence of ice cover. Whereas the ecological implications of the changing bacterial community are unclear at this time, it is likely that the observed shift from a phytoplankton community dominated by filamentous diatoms to smaller cells will have far reaching ecosystem effects including food web disruptions.

Recent global water quality crises point to an urgent need for greater understanding of cyanobacterial harmful algal blooms (cHABs) and their drivers. Nearshore areas of Lake Erie such as Sandusky Bay may become seasonally limited by nitrogen (N) and are characterized by distinct cHAB compositions (i.e., Planktothrix over Microcystis). This study investigated phytoplankton N uptake pathways, determined drivers of N depletion, and characterized the N budget in Sandusky Bay. Nitrate (NO3−) and ammonium (NH4+) uptake, N fixation, and N removal processes were quantified by stable isotopic approaches. Dissimilatory N uptake was a relatively modest N sink, with denitrification, anammox, and N2O production accounting for 84, 14, and 2 % of N removal, respectively. Phytoplankton assimilation was the dominant N uptake mechanism, and NO3− uptake rates were higher than NH4+ uptake rates. Riverine DIN loading was sometimes insufficient to meet assimilatory and dissimilatory demands, but N fixation alleviated this deficit. N fixation made up 23.7–85.4 % of total phytoplankton N acquisition and indirectly supports Planktothrix blooms. However, N fixation rates were surprisingly uncorrelated with NO3− or NH4+ concentrations. Owing to temporal separation in sources and sinks of N to Lake Erie, Sandusky Bay pendulums between acting as a strong and weak source of downstream N loading to Lake Erie. Estuarine systems such as Sandusky Bay are mediators of downstream N loading, but climate change-induced increases in precipitation and N loading will likely intensify the swings of the N pendulum in favor of N export.

Pork quality and carcass characteristics are now being integrated into swine breeding objectives because of their economic value. Understanding the genetic basis for these traits is necessary for this to be accomplished. The objective of this study was to estimate phenotypic and genetic parameters for carcass and meat quality traits in 2 Canadian swine populations. Data from a genomic selection study aimed at improving meat quality with a mating system involving hybrid Landrace × Large White and Duroc pigs were used to estimate heritabilities and phenotypic and genetic correlations among them. Data on 2,100 commercial crossbred pigs for meat quality and carcass traits were recorded with pedigrees compromising 9,439 animals over 15 generations. Significant fixed effects (company, sex, and slaughter batch), covariates (cold carcass weight and slaughter age), and random additive and common litter effects were fitted in the models. A series of pairwise bivariate analyses were implemented in ASReml to estimate phenotypic and genetic parameters. Heritability estimates (±SE) for carcass traits were moderate to high and ranged from 0.22 ± 0.08 for longissimus dorsi muscle area to 0.63 ± 0.04 for trimmed ham weight, except for firmness, which was low. Heritability estimates (±SE) for meat quality traits varied from 0.10 ± 0.04 to 0.39 ± 0.06 for the Minolta b* of ham quadriceps femoris muscle and shear force, respectively. Generally, most of the genetic correlations were significant (P < 0.05) and ranged from low (0.18 ± 0.07) to high (-0.97 ± 0.35). There were high negative genetic correlations between drip loss with pH and shear force and a positive correlation with cooking loss. Genetic correlations between carcass weight (both hot and cold) with carcass marbling were highly positive. It was concluded that selection for increasing primal and subprimal cut weights with better pork quality may be possible. Furthermore, the use of pH is confirmed as an indicator for pork water-holding capacity and cooking loss. The heritabilities of carcass and pork quality traits indicated that they can be improved using traditional breeding methods and genomic selection, respectively. The estimated genetic parameters for carcass and meat quality traits can be incorporated into the breeding programs that emphasize product quality in these Canadian swine populations.

Grand Lake St. Marys (Ohio, USA) is a hypereutrophic reservoir prone to persistent toxic cyanobacterial blooms fueled by agricultural nutrient runoff. The pervasive and highly toxic 2010 bloom led to the collapse of the local tourism industry, with microcystin concentrations exceeding 2000 μg mL −1 at some locations during the peak of the bloom. Sampling from a location adjacent to the intake to the Celina Water Treatment Plant, chlorophyll a levels remained above 25 μg L −1 from June to September, reaching a maximum in excess of 100 μg L −1 in July. Molar ratios of dissolved inorganic nitrogen (DIN) to dissolved inorganic phosphorus (DIP) declined below 16 in July and August, suggesting the cyanobacterial population became N-limited during the bloom peak. Cyanobacterial biomass was dominated by Planktothrix spp. throughout late summer 2010, but phylogenetic analysis of mcyA sequences revealed the presence of toxic Microcystis spp. during July that coincided with the highest toxin measurements. August and September samples indicated the community shifted to yield Planktothrix spp. as the major toxic genus. Community shifts of this nature may impede targeted remediation efforts, and therefore a thorough understanding of the taxa involved is necessary prior to implementing strategies to limit bloom formation.

The pelagic regions of Lake Superior and eastern Lake Erie (Laurentian Great Lakes) are typically phosphorus (P)-limited environments, and picocyanobacteria of the genus Synechococcus spp. are prominent primary producers during the summer. As a proxy for their utilization of organic P, the expression of two genes, phnD and phoX , was monitored. The phnD gene encodes the phosphonate binding protein of the ABC-type phosphonate transporter, whereas the phoX gene encodes a calcium-dependent alkaline phosphatase. Furthermore, to assess the ability of freshwater Synechococcus spp. to substitute sulfolipids for phospholipids, sqdX gene (cyanobacterial sulfolipid synthase) expression was examined. We employed PCR primers to detect the presence of all three genes in the endemic Synechococcus spp., and RT-PCR assays of cultured freshwater strains and environmental samples to assess the degree of P-stress in the phytoplankton. We show that the phnD gene was constitutively expressed, suggesting that freshwater picocyanobacteria were metabolizing exogenous phosphonate compounds in Lakes Erie and Superior. By contrast, phoX was regulated by P bioavailability. We also provide evidence that sqdX is expressed during increased growth rates in phosphorus-replete conditions, suggesting that sulfolipid synthesis is not a P conservation mechanism for freshwater Synechococcus spp.

Despite a well-documented rise in nitrate concentration over the past century, Lake Superior has retained an oligotrophic character. In part, this status results from physical attributes of the lake including low temperatures and prolonged isothermy, resulting in deep-mixing and light limitation which constrain primary production. Lake Superior is also phosphorus deficient which limits phytoplankton growth. We conducted large (20 l) volume factorial bioassay experiments to assess the influence of light and nutrients (P, Fe) on nitrate assimilation by a Lake Superior chlorophyte alga. Bioassays seeded with the chlorophyte yielded a strong response to light result- ing in the rapid depletion of nitrate. High light resulted in higher activities of the key N-assimilation enzyme nitrate reductase (NR) and increased algal biomass compared to low light treatments. NR activity was highly correlated with rates of nitrate incorporation in bioassays and field surveys suggesting that NR occupies a critical place in nitrate metabolism. In bioassays, the addition of nutrients (P, Fe) only slightly increased the rate at which nitrate became depleted. Parallel trials using a luminescent cyano- bacterial bioreporter confirmed the lack of response by added nutrients supporting light as an important factor in constraining nitrate assimilation by phytoplankton in the lake.

Pseudomonas aeruginosa is an opportunistic pathogen which is evolving resistance to many currently used antibiotics. While much research has been devoted to the roles of pathogenic P. aeruginosa in cystic fibrosis (CF) patients, less is known of its ecological properties. P. aeruginosa dominates the lungs during chronic infection in CF patients, yet its abundance in some environments is less than that of other diverse groups of pseudomonads. Here, we sought to determine if clinical isolates of P. aeruginosa are vulnerable to environmental pseudomonads that dominate soil and water habitats in one-to-one competitions which may provide a source of inhibitory factors. We isolated a total of 330 pseudomonads from diverse habitats of soil and freshwater ecosystems and competed these strains against one another to determine their capacity for antagonistic activity. Over 900 individual inhibitory events were observed. Extending the analysis to P. aeruginosa isolates revealed that clinical isolates, including ones with increased alginate production, were susceptible to competition by multiple environmental strains. We performed transposon mutagenesis on one isolate and identified an ∼14.8-kb locus involved in antagonistic activity. Only two other environmental isolates were observed to carry the locus, suggesting the presence of additional unique compounds or interactions among other isolates involved in outcompeting P. aeruginosa . This collection of strains represents a source of compounds that are active against multiple pathogenic strains. With the evolution of resistance of P. aeruginosa to currently used antibiotics, these environmental strains provide opportunities for novel compound discovery against drug-resistant clinical strains. IMPORTANCE We demonstrate that clinical CF-derived isolates of P. aeruginosa are susceptible to competition in the presence of environmental pseudomonads. We observed that many diverse environmental strains exhibited varied antagonistic profiles against a panel of clinical P. aeruginosa isolates, suggesting the presence of distinct mechanisms of inhibition among these ecological strains. Understanding the properties of these antagonistic events offers the potential for discoveries of antimicrobial compounds or metabolic pathways important to the development of novel treatments for P. aeruginosa infections.

Recent discoveries have altered prevailing paradigms concerning the conditions under which nitrification takes place and the organisms responsible for nitrification in aquatic ecosystems. In Lake Superior, nitrate (NO { ) concentrations have increased fivefold in the past century. Although previous evidence indicated that most NO { is generated by nitrification within the lake, important questions remain concerning the magnitude and controls of nitrification, and which microbial groups are primarily responsible for this process. We measured water-column nitrification rates in the western basin of Lake Superior during five research cruises from November 2009 to March 2011. Using in situ bottle incubations at 10 depths, we quantified nitrification rates using both the oxidation of 15N-labeled ammonium (NH z ) and the uptake of 14C associated with nitrification. Average rates of NH z oxidation ranged from 18–34 nmol N L21 d21 across the five cruises, similar to values reported for the coastal ocean, and two orders of magnitude lower than values reported from other lakes. Low nitrification rates observed in the epilimnion corresponded to the absence of ammonium-oxidizing archaea and nitrite-oxidizing bacteria. The measured rates of nitrification are . 50-fold greater than the long-term NO { rise in the lake, indicating that N is actively cycling and that long-term change in this ecosystem is mediated by internal dynamics. Nitrification is a key transformation in the nitrogen (N) cycle, important in determining the distribution of N among pools of dissolved inorganic nitrogen (Ward 2011), which in turn can affect rates of primary production (Howarth and Marino 2006) and greenhouse gas emissions

Recent investigations of Lake Erie in the winter have demonstrated the occurrence of substantial phytoplankton communities largely consisting of the diatom Aulacoseira islandica (O. Muller) Simonsen. To assess the activity of this diatom community, multiple measures of production, both general and diatom-specific, were undertaken. We measured oxygen (O 2 ) evolution as proxy for carbon (C)-fixation and 2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl) methoxy)-phenyl)oxazole (PDMPO) incorporation as a measure of silica (Si) deposition. The latter demonstrated conclusively that diatoms were active during winter months and confirmed that diatoms are the primary drivers of winter productivity. The stoichiometric relationship between carbon and silica in the winter Lake Erie phytoplankton assemblage was further compared to the activity of the summer community. Although the winter phytoplankton community was observed to be active, it was less active than the summer community, with lower measured rates of O 2 evolution and Si deposition. These findings provide a new and expanded understanding of the biological carbon production in Lake Erie.

The limnology of offshore Lake Erie during periods of extensive (> 70%) ice cover was examined from ship borne sampling efforts in 2007 to 2010, inclusive. Dense and discrete accumulations of the centric filamentous diatom Aulacoseria islandica (> 10 μg Chl- a /L) were located in the isothermal (

For almost two decades, the western basin of Lake Erie has been plagued with recurring toxic algal blooms dominated by the colonial cyanobacterium, Microcystis spp. Since the Maumee River is a major source of nutrients and sediment inputs into the lake, and Microcystis spp. has been identified as a member of the upstream river algal assemblage, the possibility exists that the river Microcystis species serve as a seed population for the toxic blooms occurring in the lake. Genetic profiling of toxic cyanobacteria using the microcystin synthesis gene, mcyA , clearly indicates that the toxic cyanobacteria of the river are distinct from the toxic Microcystis spp. of Lake Erie. Indeed, mcyA sequences are almost exclusively from toxic Planktothrix spp., similar to what has been documented previously for Sandusky Bay. UniFrac statistical analysis of cyanobacterial community composition by comparison of 16S–23S ITS sequences also show that the Maumee River and Lake Erie communities are distinct. Overall, these data show that despite the importance of nutrient inputs and sediments from the river, the toxic cyanobacterial blooms of Lake Erie do not originate from toxic species endemic to the Maumee River and instead must originate elsewhere, most likely from the lake sediments.

We describe a unique educational collaboration between the U.S. Coast Guard and Bowling Green State University with the goal of engaging Coast Guard crew members in a monitoring program intended to increase both temporal and spatial resolutions of sampling during winter in Lake Erie, a period where extreme weather conditions and safety considerations challenge our ability to sample.

Iron bioavailability in Lake Superior was assessed during field surveys conducted in 2001-2002. Dissolved iron (Fe d ) ranged between 1 and 4 nM at offshore stations and >10 nM at most nearshore sites. Iron availability was assessed using a luminescent Synechococcus bioreporter comprising a luciferase reporter controlled by an iron-responsive promoter isiAB. Bioreporter luminescence was negatively correlated to Fed measured in the samples. Distance from shore was a better predictor of iron bioavailability than was season. Water collected from most offshore stations sampled during spring and summer elicited higher bioreporter luminescence than did nearshore sites. Iron availability did not vary with depth during summer, despite the presence of elevated levels of Fe d in the hypolimnion at most stations. Ultrafiltration (0.02 μm) of Fe d at two offshore sites demonstrated Fed to be present mainly in a colloidal phase, yet the bioreporter responded solely to iron contained in the soluble phase. During spring, a parallel immunochemical assay of diatoms resulted in the detection offerredoxin (Fd) but not flavodoxin (Flvd) at five stations indicating the presence of an iron sufficient assemblage of diatoms at these sites. Whereas neither bioreporter nor immunochemical approaches conducted during spring supported physiological iron deficiency among Lake Superior phytoplankton, the results did point to differences in the availability of iron to prokaryotes and eukaryotes.

The Sea of Okhotsk is known as one of the most biologically productive regions among the world's oceans, and its productivity is supported in part by the discharge of iron (Fe)-rich water from the Amur River. However, little is known about the effect of riverine-derived Fe input on the physiology of the large diatoms which often flourish in surface waters of the productive continental shelf region. We conducted diatom-specific immunochemical ferredoxin (Fd) and flavodoxin (Fld) assays in order to investigate the spatial variability of Fe nutritional status in the microplankton-sized (20–200 μm; hereafter micro-sized) diatoms. The Fd index, defined as the proportion of Fd to the sum of Fd plus Fld accumulations in the cells, was used to assess their Fe nutritional status. Additionally, active chlorophyll fluorescence measurements using pulse–amplitude-modulated (PAM) fluorometry were carried out to obtain the maximum photochemical quantum efficiency (Fv/Fm) of photosystem II for the total micro-sized phytoplankton assemblages including diatoms. During our observations in the summer of 2006, the micro-sized diatoms were relatively abundant (> 10 μg C L−1) in the neritic region, and formed a massive bloom in Sakhalin Bay near the mouth of the Amur River. Values of the Fd index and Fv/Fm were high (>0.9 and >0.65, respectively) near the river mouth, indicating that Fe was sufficient for growth of the diatoms. However, in oceanic waters of the Sea of Okhotsk, the diatom Fd index declined as cellular Fld accumulation increased. These results suggest that there was a distinct gradient in Fe nutritional status in the micro-sized diatoms from near the Amur River mouth to open waters in the Sea of Okhotsk. A significant correlation between dissolved Fe (D-Fe) concentration and the Fd index was found in waters off Sakhalin Island, indicating that D-Fe was a key factor for the photophysiology of this diatom size class. In the vicinity of the Kuril Islands between the Sea of Okhotsk and the Pacific Ocean, micro-sized diatoms only accumulated Fld (i.e., Fd index = 0), despite strong vertical mixing consistent with elevated surface D-Fe levels (>0.4 nM). Since higher Fe quotas are generally required for diatoms growing under low-light conditions, the micro-sized diatoms off the Kuril Islands possibly encountered Fe and light co-limitations. The differential expressions of Fd and Fld in micro-sized diatoms helped us to understand how these organisms respond to Fe availability in the Sea of Okhotsk in connection with the Amur River discharge.

Genetic correlations between performance traits with meat quality and carcass traits were estimated on 6,408 commercial crossbred pigs with performance traits recorded in production systems with 2,100 of them having meat quality and carcass measurements. Significant fixed effects (company, sex and batch), covariates (birth weight, cold carcass weight, and age), random effects (additive, litter and maternal) were fitted in the statistical models. A series of pairwise bivariate analyses were implemented in ASREML to estimate heritability, phenotypic, and genetic correlations between performance traits (n = 9) with meat quality (n = 25) and carcass (n = 19) traits. The animals had a pedigree compromised of 9,439 animals over 15 generations. Performance traits had low-to-moderate heritabilities (±SE), ranged from 0.07±0.13 to 0.45±0.07 for weaning weight, and ultrasound backfat depth, respectively. Genetic correlations between performance and carcass traits were moderate to high. The results indicate that: (a) selection for birth weight may increase drip loss, lightness of longissimus dorsi, and gluteus medius muscles but may reduce fat depth; (b) selection for nursery weight can be valuable for increasing both quantity and quality traits; (c) selection for increased daily gain may increase the carcass weight and most of the primal cuts. These findings suggest that deterioration of pork quality may have occurred over many generations through the selection for less backfat thickness, and feed efficiency, but selection for growth had no adverse effects on pork quality. Low-to-moderate heritabilities for performance traits indicate that they could be improved using traditional selection or genomic selection. The estimated genetic parameters for performance, carcass and meat quality traits may be incorporated into the breeding programs that emphasize product quality in these Canadian swine populations.

Lake St. Clair is the smallest lake in the Laurentian Great Lakes system. MODIS satellite imagery suggests that high algal biomass events have occurred annually along the southern shore during late summer. In this study, we evaluated these events and tested the hypothesis that summer bloom material derived from Lake St. Clair may enter Lake Erie via the Detroit River and represent an overlooked source of potentially toxic Microcystis biomass to the western basin of Lake Erie. We conducted a seasonally and spatially resolved study carried out in the summer of 2013. Our goals were to: 1) track the development of the 2013 summer south-east shore bloom 2) conduct a spatial survey to characterize the extent of toxicity, taxonomic diversity of the total phytoplankton population and the phylogenetic diversity of potential MC-producing cyanobacteria (Microcystis, Planktothrix and Anabaena) during a high biomass event, and 3) compare the strains of potential MC-producers in Lake St. Clair with strains from Lake Erie and Lake Ontario. Our results demonstrated a clear predominance of cyanobacteria during a late August bloom event, primarily dominated by Microcystis, which we traced along the Lake St. Clair coastline downstream to the Detroit River's outflow at Lake Erie. Microcystin levels exceeded the Province of Ontario Drinking Water Quality Standard (1.5 µg L(-1)) for safe drinking water at most sites, reaching up to five times this level in some areas. Microcystis was the predominant microcystin producer, and all toxic Microcystis strains found in Lake St. Clair were genetically similar to toxic Microcystis strains found in lakes Erie and Ontario. These findings suggest extensive genetic connectivity among the three systems.

Flavodoxin is a small electron-transfer protein capable of replacing ferredoxin during periods of Fe deficiency. When evaluating the suitability of flavodoxin as a diagnostic indicator for Fe limitation of phytoplankton growth, we examined its expression in two marine diatoms we cultured using trace-metal-buffered medium. Thalassio-sira weissflogii and Phaeodactylum tricornutum were cultured in ethylenediaminetetraacetic acid-buffered Sargasso Sea water containing from 10 to 1000 nM added Fe. Trace-metal-buffered cultures of each diatom maintained high growth rates across the entire range of Fe additions. Similarly, declines in chlorophyll/cell and in the ratio of photosystem II variable-to-maximum fluorescence were negligible (P. tricornutum) to moderate (T. weissflogii; 54% decline in chlorophyll/cell and 22% decrease in variable-to-maximum fluorescence). Moreover, only minor variations in photosynthetic parameters were observed across the range of additions. In contrast, flavodoxin was expressed to high levels in low-Fe cultures. Despite the inverse relationship between flavodoxin expression and Fe content of the medium, its expression was seemingly independent of any of the indicators of cell physiology that were assayed. It appears that flavodoxin is expressed as an early-stage response to Fe stress and that its accumulation need not be intimately connected to limitations imposed by Fe on the growth rate of these diatoms.

Ammonia oxidation is the first step of nitrification carried out by ammonia-oxidizing Archaea (AOA) and Bacteria (AOB). Lake Superior and Erie are part of the Great Lakes system differing in trophic status with Lake Superior being oligotrophic and Lake Erie meso- to eutrophic. Sediment samples were collected from both lakes and used to characterize abundance and diversity of AOA and AOB based on the ammonia monooxygenase (amoA) gene. Diversity was accessed by a pyro-sequencing approach and the obtained sequences were used to determine the phylogeny and alpha and beta diversity of the AOA and AOB populations. In Lake Erie copy numbers of bacterial amoA genes were in the same order of magnitude or even higher than the copy numbers of the archaeal amoA genes, while in Lake Superior up to 4 orders of magnitude more archaeal than bacterial amoA copies were detected. The AOB detected in the samples from Lake Erie belonged to AOB that are frequently detected in freshwater. Differences were detected between the phylogenetic affiliations of the AOA from the two lakes. Most sequences detected in Lake Erie clustered in the Nitrososphaera cluster (Thaumarchaeal soil group I.1b) where as most of the sequences in Lake Superior were found in the Nitrosopumilus cluster (Thaumarchaeal marine group I.1a) and the Nitrosotalea cluster. Pearson correlations and canonical correspondence analysis (CCA) showed that the differences in abundance and diversity of AOA are very likely related to the sampling location and thereby to the different trophic states of the lakes.

We present evidence for the directed formation of ice by planktonic communities dominated by filamentous diatoms sampled from the ice-covered Laurentian Great Lakes. We hypothesize that ice formation promotes attachment of these non-motile phytoplankton to overlying ice, thereby maintaining a favorable position for the diatoms in the photic zone. However, it is unclear whether the diatoms themselves are responsible for ice nucleation. Scanning electron microscopy revealed associations of bacterial epiphytes with the dominant diatoms of the phytoplankton assemblage, and bacteria isolated from the phytoplankton showed elevated temperatures of crystallization (T-c) as high as -3 degrees C. Ice nucleation-active bacteria were identified as belonging to the genus Pseudomonas, but we could not demonstrate that they were sufficiently abundant to incite the observed freezing. Regardless of the source of ice nucleation activity, the resulting production of frazil ice may provide a means for the diatoms to be recruited to the overlying lake ice, thereby increasing their fitness. Bacterial epiphytes are likewise expected to benefit from their association with the diatoms as recipients of organic carbon excreted by their hosts. This novel mechanism illuminates a previously undescribed stage of the life cycle of the meroplanktonic diatoms that bloom in Lake Erie and other Great Lakes during winter and offers a model relevant to aquatic ecosystems having seasonal ice cover around the world.

Carbon and nutrient cycles in large temperate lakes such as Lake Erie are primarily driven by phototrophic and heterotrophic microorganisms, although our understanding of these is often constrained to late spring through summer due to logistical constraints. During periods of > 90% ice cover in February of 2008, 2009, and 2010, we collected samples from an icebreaker for an examination of bacterial production as well as microbial community structure. In comparison with summer months (August 2002 and 2010), we tested hypotheses concerning seasonal changes in microbial community diversity and production. Bacterial production estimates were c . 2 orders of magnitude higher (volume normalized) in summer relative to winter. Our observations further demonstrate that the microbial community, including single-celled phototrophs, varied in composition between August and February. Sediment traps deployed and collected over a 3 year period (2008–2011) confirmed that carbon export was ongoing and not limiting winter production. The results support the notion that active primary producers in winter months export carbon to the sediments that is not consumed until the warmer seasons. The establishment of this linkage is a critical observation in efforts to understand the extent and severity of annual summertime formations of a zone of regional hypoxia in Lake Erie.

In the Southern Ocean, phytoplankton growth is largely limited by the lack of iron, affecting the biogeochemical cycling not only of iron itself but also of other elements, including nutrients and carbon. It is now recognized that iron limitation affects carbon cycling globally and thus plays a role in Earth’s climate regulation. The bioavailable fraction of iron is the fraction that can effectively interact with phytoplankton to support their iron-dependent metabolic reactions and growth. As such, it is the bioavailable iron pool that shapes phytoplankton communities in most of the Southern Ocean. Despite numerous studies, parameters controlling iron bioavailability to phytoplankton are still poorly understood, probably due to an extremely complex and dynamic interplay between iron chemistry and biology in surface waters. Iron bioavailability depends on chemical and physical speciation and the different uptake strategies of the phyto-and bacterioplankton communities. In the Southern Ocean, 99% of the dissolved iron is complexed by organic ligands, which likely controls its bioavailability. Furthermore, microorganisms also exert feedback on iron chemistry, for instance, by releasing organic iron-binding ligands through production, cell lysis, or degradation of fecal pellets, as well as by reducing iron at the cell surface. Regeneration of iron, through grazing as well as bacterial and viral activities, is another pathway that supplies iron to phytoplankton communities. Field investigations of iron speciation in the Southern Ocean are discussed in conjunction with laboratory assessments of iron speciation and bioavailability using natural assemblages and strains isolated from the Southern Ocean. Methods to measure iron bioavailability and recent developments in mathematical models are also presented.

Blooms of the potentially toxic cyanobacterium Microcystis are common events globally, and as a result significant resources continue to be dedicated to monitoring and con- trolling these events. Recent studies have shown that a significant proportion of total cell-associated phosphorus (P) in marine phytoplankton can be surface adsorbed; as a result studies completed to date do not accurately report the P demands of these organ- isms. In this study we measure the total cell-associated and intracellular P as well as growth rates of two toxic strains of Microcystis aeruginosa Kutz grown under a range of P concentrations. The results show that the intracellular P pool in Microcystis represents a percentage of total cell-associated P (50-90%) similar to what has been reported for actively growing algae in marine systems. Intracellular P concentrations (39-147 fg cell −1 ) generally increased with increasing P concentrations in the growth medium, but growth rate and the ratio of total cell-associated to intracellular P remained generally stable. Intra- cellular P quotas and growth rates in cells grown under the different P treatments illustrate the ability of this organism to successfully respond to changes in ambient P loads, and thus have implications for ecosystem scale productivity models employing P concentrations to predict algal bloom events.

In phosphorus-limited marine environments, picocyanobacteria (Synechococcus and Prochlorococcus spp.) can hydrolyze naturally occurring phosphonates as a P source. Utilization of 2-aminoethylphosphonate (2-AEP) is dependent on expression of the phn genes, encoding functions required for uptake and C-P bond cleavage. Prior work has indicated that expression of picocyanobacterial phnD, encoding the phosphonate binding protein of the phosphonate ABC transporter, is a proxy for the assimilation of phosphonates in natural assemblages of Synechococcus spp. and Prochlorococcus spp (Ilikchyan et al. 2009). In this study, we expand this work to assess seasonal phnD expression in the Sargasso Sea. By RT-PCR, our data confirm that phnD expression is constitutive for the Prochlorococcus spp. detected, but in Synechococcus spp., phnD transcription follows patterns of phosphorus availability in the mixed layer. Specifically, our data suggest that phnD is repressed in the spring when P is bioavailable following deep winter mixing. In the fall, phnD expression follows a depth dependent pattern reflecting depleted P at the surface following summertime drawdown, and elevated P at depth.

RuBisCo activase catalyzes the activation and maintains the activated state of the photosynthetic enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCo, EC 4.1.1.39). We employed antisera prepared against the RuBisCo holoenzyme purified from tobacco and RuBisCo activase isolated from spinach to determine the localization of these proteins in leaves of C3-type higher plants and green algae. In leaves ofVicia faba, both RuBisCo activase and RuBisCo are distributed throughout the chloroplast stroma. In contrast, RuBisCo activase and RuBisCo are predominantly localized to the pyrenoid in the green algaeChlamydomonas reinhardtii andColeochaete scutata. The co-immunolocalization of RuBisCo activase and RuBisCo to the pyrenoid in these two green algal species suggests that pyrenoid-localized RuBisCo is catalytically competent. We conclude that the pyrenoid functions as a unique metabolic compartment of the chloroplast in which the reactions of the photosynthetic carbon reduction pathway are initiated.

[1] An improved knowledge of iron biogeochemistry is needed to better understand key controls on the functioning of high-nitrate low-chlorophyll (HNLC) oceanic regions. Iron budgets for HNLC waters have been constructed using data from disparate sources ranging from laboratory algal cultures to ocean physics. In summer 2003 we conducted FeCycle, a 10-day mesoscale tracer release in HNLC waters SE of New Zealand, and measured concurrently all sources (with the exception of aerosol deposition) to, sinks of iron from, and rates of iron recycling within, the surface mixed layer. A pelagic iron budget (timescale of days) indicated that oceanic supply terms (lateral advection and vertical diffusion) were relatively small compared to the main sink (downward particulate export). Remote sensing and terrestrial monitoring reveal 13 dust or wildfire events in Australia, prior to and during FeCycle, one of which may have deposited iron at the study location. However, iron deposition rates cannot be derived from such observations, illustrating the difficulties in closing iron budgets without quantification of episodic atmospheric supply. Despite the threefold uncertainties reported for rates of aerosol deposition (Duce et al., 1991), published atmospheric iron supply for the New Zealand region is ∼50-fold (i.e., 7- to 150-fold) greater than the oceanic iron supply measured in our budget, and thus was comparable (i.e., a third to threefold) to our estimates of downward export of particulate iron. During FeCycle, the fluxes due to short term (hours) biological iron uptake and regeneration were indicative of rapid recycling and were tenfold greater than for new iron (i.e. estimated atmospheric and measured oceanic supply), giving an “fe” ratio (uptake of new iron/uptake of new + regenerated iron) of 0.17 (i.e., a range of 0.06 to 0.51 due to uncertainties on aerosol iron supply), and an “Fe” ratio (biogenic Fe export/uptake of new + regenerated iron) of 0.09 (i.e., 0.03 to 0.24).

[1] During austral summer 2003, we tracked a patch of surface water infused with the tracer sulfur hexafluoride, but without addition of Fe, through subantarctic waters over 10 days in order to characterize and quantify algal Fe pools and fluxes to construct a detailed biogeochemical budget. Nutrient profiles characterized this patch as a highnitrate, low-silicic acid, low-chlorophyll (HNLSiLC) water mass deficient in dissolved Fe. The low Fe condition was confirmed by several approaches: shipboard iron enrichment experiments and physiological indices of Fe deficiency (Fv/Fm 40% of total chlorophyll. Whereas the picophytoplankton accounted for � 50% of total primary production, they were responsible for the majority of community iron uptake in the mixed layer. Thus ratios of 55 Fe: 14 C uptake were highest for picophytoplankton (median: 17 mmol:mol) and declined to � 5 mmol:mol for the larger algal size fractions. A pelagic Fe budget revealed that picophytoplankton were the largest pool of algal Fe (>90%), which was consistent with the high (� 80%) phytoplankton Fe demand attributed to them. However, Fe regenerated by herbivory satisfied only � 20% of total algal Fe demand. This iron regeneration term increased to 40% of algal Fe demand when we include Fe recycled by bacterivory. As recycled, rather than new, iron dominated the pelagic iron budget (Boyd et al., 2005), it is highly unlikely that the supply of new Fe would redress the imbalance between algal Fe demand and supply. Reasons for this imbalance may include the overestimation of algal iron uptake from radiotracer techniques, or a lack of consideration of other iron regeneration processes. In conclusion, it seems that algal Fe uptake cannot be supported solely by the recycling of algal iron, and may require an Fe ‘‘subsidy’’ from that regenerated by heterotrophic pathways.

Winter research and monitoring of icebound rivers, lakes, and coastal seas to date has usually involved seagoing civilian scientists leading survey efforts. However, because of poor weather conditions and a lack of safe research platforms, scientists collecting data during winter face some difficult and often insurmountable problems. To solve these problems and to further research and environmental monitoring goals, new partnerships can be formed through integrating efforts of the U.S. Coast Guard (USCG) with citizen science initiatives. USCG and a research group at Ohio's Bowling Green State University are entering the third year of such a partnership, in which icebreaking operations in Lake Erie using USCG Cutter Neah Bay support volunteer data collection. With two additional USCG vessels joining the program this winter season, the partnership serves as a timely and useful model for worldwide environmental research and monitoring through citizen science and government collaboration.

The localization of ribulose 1,5-bisphosphate carboxylase/ oxygenase (RuBisCO) in cells ofC. pyrenoidosa grown at varying light intensities was determined by immunoelectron microscopy. Log phase cells grown at photon flux densities of 25 and 75 μEm−2s−1 (light-limiting) and 540 μEm−2s−1 (light-saturating) were fixed in 3% glutaraldehyde and embedded in Lowicryl K4M. Sections were labelled with antiserum to each subunit of RuBisCO followed by protein A-gold. At each light fluence rate, the pyrenoid was heavily labelled by each antibody whereas chloroplast stromal labelling was not above background levels. The apparent absence of stromal RuBisCO at each light level, and hence the lack of enzyme redistribution from pyrenoid to stroma following an increase in light fluence rate, suggests that pyrenoid RuBisCO is functional in vivo.

To address questions regarding the potential impact of elevated total phosphorus (TP) inputs (due to relaxed regulations of TP loading), a series of TP enrichment experiments were con- ducted at pelagic stations in the 3 hydrologically distinct basins of Lake Erie. Results of nutrient assimilation measurements and assays for nutrient bioavailability suggest that the chemical specia- tion, and not concentration, of nitrogenous compounds may influence phytoplankton community structure; this in turn may lead to the selective proliferation of cyanobacteria in the eastern basin of the lake. Assays with cyanobacterial bioluminescent reporter systems for P and N availability as well as Ntot:Ptot assimilation ratios from on-deck incubation experiments support this work. Considered in the context of a microbial food web relative to a grazing food web, the results imply that alterations in current TP loading controls may lead to alterations in the phytoplankton community structure in the different basins of the Lake Erie system.