Water Depth Underpins the Relative Roles and Fates of Nitrogen and Phosphorus in Lakes.

Eutrophication mitigation is an ongoing priority for aquatic ecosystems. However, the current eutrophication control strategies (phosphorus (P) and/or nitrogen (N)) are guided mainly by nutrient addition experiments in small waters without encompassing all in-lake biogeochemical processes that are associated largely with lake morphological characteristics. Here, we use a global lake data set (573 lakes) to show that the relative roles of N vs P in affecting eutrophication are underpinned by water depth. Mean depth and maximum depth relative to mixing depth were used to distinguish shallow (mixing depth > maximum depth), deep (mixing depth < mean depth), and transitional (mean depth ≤ mixing depth ≤ maximum depth) lakes in this study. TN/TP ratio (by mass) was used as an indicator of potential lake nutrient limitation, i.e., N only limitation if N/P < 9, N + P colimitation if 9 ≤ N/P < 22.6, and P only limitation if N/P ≥ 22.6. The results show that eutrophication is favored in shallow lakes, frequently (66.2%) with N limitation while P limitation predominated (94.4%) in most lakes but especially in deep ones. The importance of N limitation increases but P limitation decreases with lake trophic status while N and P colimitation occurs primarily (59.4%) in eutrophic lakes. These results demonstrate that phosphorus reduction can mitigate eutrophication in most large lakes but a dual N and P reduction may be needed in eutrophic lakes, especially in shallow ones (or bays). Our analysis helps clarify the long debate over whether N, P, or both control primary production. While these results imply that more resources be invested in nitrogen management, given the high costs of nitrogen pollution reduction, more comprehensive results from carefully designed experiments at different scales are needed to further verify this modification of the existing eutrophication mitigation paradigm.

Urea dynamics during Lake Taihu cyanobacterial blooms in China.

Lake Taihu, the third largest freshwater lake in China, suffers from harmful cyanobacteria blooms caused by Microcystis spp., which do not fix nitrogen (N). Reduced N (i.e., NH4+, urea and other labile organic N compounds) is an important factor affecting the growth of Microcystis. As the world use of urea as fertilizer has escalated during the past decades, an understanding of how urea cycling relates to blooms of Microcystis is critical to predicting, controlling and alleviating the problem. In this study, the cycling rates of urea-N in Lake Taihu ranged from non-detectable to 1.37 µmol N L-1 h-1 for regeneration, and from 0.042 µmol N L-1 h-1 to 2.27 µmol N L-1 h-1 for potential urea-N removal. The fate of urea-N differed between light and dark incubations. Increased 15NH4+ accumulated and higher quantities of the removed urea-15N remained in the 15NH4+ form were detected in the dark than in the light. A follow-up incubation experiment with 15N-urea confirmed that Microcystis can grow on urea but its effects on urea dynamics were minor, indicating that Microcystis was not the major factor causing the observed fates of urea under different light conditions in Lake Taihu. Bacterial community composition and predicted functional gene data suggested that heterotrophic bacteria metabolized urea, even though Microcystis spp. was the dominant bloom organism.

A retrospective analysis of dental implant survival in HIV patients.

AIM: This 5 years retrospective cohort survival study compared failure rates of dental implants placed in HIV (+) and HIV (-) patients relative to several risk factors. MATERIALS AND METHODS: Between 2006 and 2015, 484 implants placed in HIV (+) patients and 805 implants placed in HIV (-) patients were assessed for survival. The effects of HIV were estimated using propensity weighting. The effects of age, smoking status, diabetes, restoration status, gender, implant type, placement site, hepatitis C status, baseline CD4 count and CD4%, post-placement average CD4%, nadir CD4%, nadir CD4 count and antiviral therapy were analysed. RESULTS: Implants placed in HIV (+) patients and HIV (-) patients had similar failure rates (HR = 1.4, p = 0.34). Increased failure rates were observed in HIV (+) patients with baseline CD4% ≤20 (HR = 2.72, p = 0.04), post-placement CD4% average ≤20% (HR = 2.71, p = 0.04), protease inhibitor administration (HR = 2.74, p = 0.04), smoking (HR = 2.61, p = 0.05) and anterior maxillary placement (HR = 5.82, p < 0.01). Hepatitis C coinfection, viral titre, baseline CD4 count, gender, implant type and restoration type were non-contributory. CONCLUSION: Implants placed in HIV (+) patients had similar survival rates as HIV (-) patients. Failure rates increase significantly when confounding risk factors are present in HIV (+) patients.

Molecular Structure Characterization of Riverine and Coastal Dissolved Organic Matter with Ion Mobility Quadrupole Time-of-Flight LCMS (IM Q-TOF LCMS).

Deciphering molecular structures of dissolved organic matter (DOM) components is key to understanding the formation and transformation of this globally important carbon pool in aquatic environments. Such a task depends on the integrated use of complementary analytical techniques. We characterize the molecular structure of natural DOM using an ion mobility quadrupole time of flight liquid chromatography mass spectrometer (IM Q-TOF LC/MS), which provides multidimensional structural information on DOM molecules. Geometric conformation of DOM molecules is introduced into molecular-level analysis via the ion mobility (IM) in the system, and an actual measurement of isomers is achieved for the first time. Our data show that natural DOM molecules from several south Texas rivers and adjacent coastal waters have smaller geometric conformation compared with standard biomolecules. Furthermore, about 10% of all DOM molecules resolved within the detection limit of IM-MS had at least one but no more than four isomers. With acquired geometric and isomeric information, we established a multidimensional database containing 89 natural DOM compounds. This database provides a foundation to expand further, or compare, with DOM data from different seasons and locations.

Wastewater influences nitrogen dynamics in a coastal catchment during a prolonged drought.

Ecosystem function measurements can enhance our understanding of nitrogen (N) delivery in coastal catchments across river and estuary ecosystems. Here, we contrast patterns of N cycling and export in two rivers, one heavily influenced by wastewater treatment plants (WWTP), in a coastal catchment of south Texas. We measured N export from both rivers to the estuary over 2 yr that encompass a severe drought, along with detailed mechanisms of N cycling in river, tidal river, and two estuary sites during prolonged drought. WWTP nutrient inputs stimulated uptake of N, but denitrification resulting in permanent N removal accounted for only a small proportion of total uptake. During drought periods, WWTP N was the primary source of exported N to the estuary, minimizing the influence of episodic storm-derived nutrients from the WWTP-influenced river to the estuary. In the site without WWTP influence, the river exported very little N during drought, so storm-derived nutrient pulses were important for delivering N loads to the estuary. Overall, N is processed from river to estuary, but sustained WWTP-N loads and periodic floods alter the timing of N delivery and N processing. Research that incorporates empirical measurements of N fluxes from river to estuary can inform management needs in the face of multiple anthropogenic stressors such as demand for freshwater and eutrophication.

Spatial Distribution of Megacopta cribraria (Hemiptera: Plataspidae) Adults, Eggs and Parasitism by Paratelenomus saccharalis (Hymenoptera: Platygastridae) in Soybean.

Since 2014, populations of the kudzu bug, Megacopta cribraria (F.) (Hemiptera: Plataspidae), have declined in the southeastern United States and seldom require treatment. This decline follows the discovery of Paratelenomus saccharalis (Dodd; Hymenoptera: Platygastridae), a non-native egg parasitoid. The objective of this project was to observe the temporal and spatial dynamics of P. saccharalis parasitism of kudzu bug egg masses in commercial soybean fields. Four fields were sampled weekly for kudzu bugs and egg masses at a density of one sample per 0.6 ha. Sampling commenced when soybean reached the R2 maturity stage and continued until no more egg masses were present. Responses including kudzu bugs, egg masses, and parasitism rates were analyzed using ANOVA, Spatial Analysis by Distance Indices (SADIE), and SaTScan spatial analysis software. Egg masses were collected from the field, held in the lab and monitored for emergence of kudzu bug nymphs or P. saccharalis. Kudzu bug populations were generally lower than previously reported in the literature and spatial aggregation was not consistently observed. Egg parasitism was first detected in early July and increased to nearly 40% in mid-August. Significant spatial patterns in parasitism were observed with spatio-temporal clusters being loosely associated with clusters of egg masses. There were no significant differences in parasitism rates between field margins and interiors, suggesting that P. saccharalis is an effective parasitoid of kudzu bug egg masses on a whole-field scale.

Community Biological Ammonium Demand: A Conceptual Model for Cyanobacteria Blooms in Eutrophic Lakes.

Cyanobacterial harmful algal blooms (CyanoHABs) are enhanced by anthropogenic pressures, including excessive nutrient (nitrogen, N, and phosphorus, P) inputs and a warming climate. Severe eutrophication in aquatic systems is often manifested as non-N2-fixing CyanoHABs (e.g., Microcystis spp.), but the biogeochemical relationship between N inputs/dynamics and CyanoHABs needs definition. Community biological ammonium (NH4+) demand (CBAD) relates N dynamics to total microbial productivity and NH4+ deprivation in aquatic systems. A mechanistic conceptual model was constructed by combining nutrient cycling and CBAD observations from a spectrum of lakes to assess N cycling interactions with CyanoHABs. Model predictions were supported with CBAD data from a Microcystis bloom in Maumee Bay, Lake Erie, during summer 2015. Nitrogen compounds are transformed to reduced, more bioavailable forms (e.g., NH4+ and urea) favored by CyanoHABs. During blooms, algal biomass increases faster than internal NH4+ regeneration rates, causing high CBAD values. High turnover rates from cell death and remineralization of labile organic matter consume oxygen and enhance denitrification. These processes drive eutrophic systems to NH4+ limitation or colimitation under warm, shallow conditions and support the need for dual nutrient (N and P) control.

It Takes Two to Tango: When and Where Dual Nutrient (N & P) Reductions Are Needed to Protect Lakes and Downstream Ecosystems.

Preventing harmful algal blooms (HABs) is needed to protect lakes and downstream ecosystems. Traditionally, reducing phosphorus (P) inputs was the prescribed solution for lakes, based on the assumption that P universally limits HAB formation. Reduction of P inputs has decreased HABs in many lakes, but was not successful in others. Thus, the "P-only" paradigm is overgeneralized. Whole-lake experiments indicate that HABs are often stimulated more by combined P and nitrogen (N) enrichment rather than N or P alone, indicating that the dynamics of both nutrients are important for HAB control. The changing paradigm from P-only to consideration of dual nutrient control is supported by studies indicating that (1) biological N fixation cannot always meet lake ecosystem N needs, and (2) that anthropogenic N and P loading has increased dramatically in recent decades. Sediment P accumulation supports long-term internal loading, while N may escape via denitrification, leading to perpetual N deficits. Hence, controlling both N and P inputs will help control HABs in some lakes and also reduce N export to downstream N-sensitive ecosystems. Managers should consider whether balanced control of N and P will most effectively reduce HABs along the freshwater-marine continuum.

Dynamic, mechanistic, molecular-level modelling of cyanobacteria: Anabaena and nitrogen interaction.

Phytoplankton (eutrophication, biogeochemical) models are important tools for ecosystem research and management, but they generally have not been updated to include modern biology. Here, we present a dynamic, mechanistic, molecular-level (i.e. gene, transcript, protein, metabolite) model of Anabaena - nitrogen interaction. The model was developed using the pattern-oriented approach to model definition and parameterization of complex agent-based models. It simulates individual filaments, each with individual cells, each with genes that are expressed to yield transcripts and proteins. Cells metabolize various forms of N, grow and divide, and differentiate heterocysts when fixed N is depleted. The model is informed by observations from 269 laboratory experiments from 55 papers published from 1942 to 2014. Within this database, we identified 331 emerging patterns, and, excluding inconsistencies in observations, the model reproduces 94% of them. To explore a practical application, we used the model to simulate nutrient reduction scenarios for a hypothetical lake. For a 50% N only loading reduction, the model predicts that N fixation increases, but this fixed N does not compensate for the loading reduction, and the chlorophyll a concentration decreases substantially (by 33%). When N is reduced along with P, the model predicts an additional 8% reduction (compared to P only).

Mitigating cyanobacterial harmful algal blooms in aquatic ecosystems impacted by climate change and anthropogenic nutrients.

Mitigating the global expansion of cyanobacterial harmful blooms (CyanoHABs) is a major challenge facing researchers and resource managers. A variety of traditional (e.g., nutrient load reduction) and experimental (e.g., artificial mixing and flushing, omnivorous fish removal) approaches have been used to reduce bloom occurrences. Managers now face the additional effects of climate change on watershed hydrologic and nutrient loading dynamics, lake and estuary temperature, mixing regime, internal nutrient dynamics, and other factors. Those changes favor CyanoHABs over other phytoplankton and could influence the efficacy of control measures. Virtually all mitigation strategies are influenced by climate changes, which may require setting new nutrient input reduction targets and establishing nutrient-bloom thresholds for impacted waters. Physical-forcing mitigation techniques, such as flushing and artificial mixing, will need adjustments to deal with the ramifications of climate change. Here, we examine the suite of current mitigation strategies and the potential options for adapting and optimizing them in a world facing increasing human population pressure and climate change.

A novel membrane inlet mass spectrometer method to measure ¹5NH44⁺ for isotope-enrichment experiments in aquatic ecosystems.

Nitrogen (N) pollution in aquatic ecosystems has attracted much attention over the past decades, but the dynamics of this bioreactive element are difficult to measure in aquatic oxygen-transition environments. Nitrogen-transformation experiments often require measurement of (15)N-ammonium ((15)NH4(+)) ratios in small-volume (15)N-enriched samples. Published methods to determine N isotope ratios of dissolved ammonium require large samples and/or costly equipment and effort. We present a novel ("OX/MIMS") method to determine N isotope ratios for (15)NH4(+) in experimental waters previously enriched with (15)N compounds. Dissolved reduced (15)N (dominated by (15)NH4(+)) is oxidized with hypobromite iodine to nitrogen gas ((29)N2 and/or (30)N2) and analyzed by membrane inlet mass spectrometry (MIMS) to quantify (15)NH4(+) concentrations. The N isotope ratios, obtained by comparing the (15)NH4(+) to total ammonium (via autoanalyzer) concentrations, are compared to the ratios of prepared standards. The OX/MIMS method requires only small sample volumes of water (ca. 12 mL) or sediment slurries and is rapid, convenient, accurate, and precise (R(2) = 0.9994, p < 0.0001) over a range of salinities and (15)N/(14)N ratios. It can provide data needed to quantify rates of ammonium regeneration, potential ammonium uptake, and dissimilatory nitrate reduction to ammonium (DNRA). Isotope ratio results agreed closely (R = 0.998, P = 0.001) with those determined independently by isotope ratio mass spectrometry for DNRA measurements or by ammonium isotope retention time shift liquid chromatography for water-column N-cycling experiments. Application of OX/MIMS should simplify experimental approaches and improve understanding of N-cycling rates and fate in a variety of freshwater and marine environments.

A shift in the archaeal nitrifier community in response to natural and anthropogenic disturbances in the northern Gulf of Mexico.

The Gulf of Mexico is affected by hurricanes and suffers seasonal hypoxia. The Deepwater Horizon oil spill impacted every trophic level in the coastal region. Despite their importance in bioremediation and biogeochemical cycles, it is difficult to predict the responses of microbial communities to physical and anthropogenic disturbances. Here, we quantify sediment ammonia-oxidizing archaeal (AOA) community diversity, resistance and resilience, and important geochemical factors after major hurricanes and the oil spill. Dominant AOA archetypes correlated with different geochemical factors, suggesting that different AOA are constrained by distinct parameters. Diversity was lowest after the hurricanes, showing weak resistance to physical disturbances. However, diversity was highest during the oil spill and coincided with a community shift, suggesting a new alternative stable state sustained for at least 1 year. The new AOA community was not significantly different from that at the spill site 1 year after the spill. This sustained shift in nitrifier community structure may be a result of oil exposure.

Effects of entomopathogenic fungus species, and impact of fertilizers, on biological control of pecan weevil (Coleoptera: Curculionidae).

The pecan weevil, Curculio caryae (Horn), is a key pest of pecan, Carya illinoinensis (Wangenh.) K. Koch. Prior research indicated the potential for use of Hypocreales fungi to suppress C. caryae. We compared the efficacy of two fungal spp., Beauveria bassiana (GHA strain) and Metarhizium brunneum (F52), in their ability to cause C. caryae mortality. The fungus, B. bassiana, was applied to trunks of pecan trees (a method previously shown to be effective in C. caryae suppression) and efficacy was compared with M. brunneum applied to the ground or to the trunk with or without SoyScreen Oil as an ultraviolet protecting agent. Results indicated B. bassiana to be superior to M. brunneum regardless of application method; consequently, the potential for applying B. bassiana to control C. caryae was explored further. Specifically, the impact of different fertilizer regimes (as used by pecan growers) on the persistence of B. bassiana (GHA) in soil was determined. B. bassiana was applied to soil in a pecan orchard after one of several fertilizer treatments--i.e., ammonium nitrate, crimson clover, poultry litter, clover plus poultry litter, and a no-fertilizer control. B. bassiana persistence up to 49 d in 2009 and 2010 was assessed by plating soil onto selective media and determining the number of colony forming units, and by baiting soil with a susceptible host, Galleria mellonella (L.). Fertilizer treatments did not impact B. bassiana persistence. We conclude that standard fertilizers for nitrogen management, when applied according to recommended practices, are unlikely to negatively impact survival of B. bassiana in pecan orchards when the fungus is applied for C. caryae suppression during weevil emergence. Additional research on interactions between entomopathogenic fungi and fertilizer amendments (or other tree nutrition or soil management practices) is merited.

Cumulative impact of a clover cover crop on the persistence and efficacy of Beauveria bassiana in suppressing the pecan weevil (Coleoptera: Curculionidae).

The pecan weevil, Curculio caryae (Horn), is a key pest of pecan. Endemic levels of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin can occur in pecan orchards and contribute to natural control of C. caryae. Commercial formulations of the fungus can also be applied for suppression of C. caryae. We hypothesized that a clover cover crop enhances B. bassiana efficacy and persistence (e.g., by protecting the fungus against abiotic environmental stresses). The hypothesis was tested by conducting field trials in a pecan orchard in Byron, GA, in 2009 and 2010. The study included four treatments arranged in a factorial with two levels of fungus (endemic fungus only, and application of a commercial B. bassiana product), and two levels of clover (white clover, Trifolium repens L., and no clover). Fungal persistence was measured by determining the number of CFUs per gram of soil over time (during 42 d postapplication of B. bassiana in 2009 and 29 d in 2010). Efficacy was measured by capturing naturally emerging C. caryae and subsequently determining mortality and mycosis (over 24 d in 2009 and 17 d in 2010). In 2009, greater prevalence of B. bassiana conidia was detected in plots receiving fungal applications compared with no fungus applications, and no clear effect of clover was observed in plots receiving B. bassiana applications in either year. In 2010, B. bassiana prevalence in the endemic fungus plus clover treatment was higher than fungus without clover, and was similar to plots receiving additional B. bassiana applications. Given that we observed enhanced persistence of endemic B. bassiana in 2010 but not 2009, the impact of clover appears to be a cumulative effect. Mortality of C. caryae (averaged over the sampling periods) ranged between 68-74% in plots receiving B. bassiana applications and 51-56% in plots with endemic fungus only. C. caryae mortality and mycosis data also provided evidence that endemic B. bassiana efficacy was enhanced by clover relative to plots without clover (with no clear clover effect on plots receiving fungus applications). Thus, we conclude that natural control of C. caryae can increase when clover is grown in pecan orchards with endemic populations of B. bassiana.

Controlling harmful cyanobacterial blooms in a hyper-eutrophic lake (Lake Taihu, China): the need for a dual nutrient (N & P) management strategy.

Harmful cyanobacterial blooms, reflecting advanced eutrophication, are spreading globally and threaten the sustainability of freshwater ecosystems. Increasingly, non-nitrogen (N(2))-fixing cyanobacteria (e.g., Microcystis) dominate such blooms, indicating that both excessive nitrogen (N) and phosphorus (P) loads may be responsible for their proliferation. Traditionally, watershed nutrient management efforts to control these blooms have focused on reducing P inputs. However, N loading has increased dramatically in many watersheds, promoting blooms of non-N(2) fixers, and altering lake nutrient budgets and cycling characteristics. We examined this proliferating water quality problem in Lake Taihu, China's 3rd largest freshwater lake. This shallow, hyper-eutrophic lake has changed from bloom-free to bloom-plagued conditions over the past 3 decades. Toxic Microcystis spp. blooms threaten the use of the lake for drinking water, fisheries and recreational purposes. Nutrient addition bioassays indicated that the lake shifts from P limitation in winter-spring to N limitation in cyanobacteria-dominated summer and fall months. Combined N and P additions led to maximum stimulation of growth. Despite summer N limitation and P availability, non-N(2) fixing blooms prevailed. Nitrogen cycling studies, combined with N input estimates, indicate that Microcystis thrives on both newly supplied and previously-loaded N sources to maintain its dominance. Denitrification did not relieve the lake of excessive N inputs. Results point to the need to reduce both N and P inputs for long-term eutrophication and cyanobacterial bloom control in this hyper-eutrophic system.

Comparison of application methods for suppressing the pecan weevil (Coleoptera: Curculionidae) with Beauveria bassiana under field conditions.

The pecan weevil, Curculio caryae (Horn), is a key pest of pecans. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin is pathogenic to C. caryae. One approach to managing C. caryae may be application of B. bassiana directed toward adult weevils as they emerge from the soil to attack nuts in the tree canopy. Our objective was to compare different application methods for suppression of C. caryae adults. Treatments included direct application of B. bassiana (GHA strain) to soil under the tree canopy, soil application followed by cultivation, soil application in conjunction with a cover crop (Sudan grass), direct application to the tree trunk, and application to the trunk with an UV radiation-protecting adjuvant. The study was conducted in a pecan orchard in Byron, GA, in 2005 and 2006. Naturally emerging C. caryae adults, caught after crawling to the trunk, were transported to the laboratory to determine percentage mortality and signs of mycosis. When averaged over the 15-d sampling period, weevil mortality and signs of mycosis were greater in all treatments than in the nontreated control in 2005 and 2006; >75% average mortality was observed with the trunk application both years and in the trunk application with UV protection in 2005. Results indicated trunk applications can produce superior efficacy relative to ground application, particularly if the ground application is followed by cultivation. Efficacy in the cover crop treatment, however, did not differ from other application approaches. Future research should focus on elucidating the causes for treatment differences we observed and the extent to which B. bassiana-induced C. caryae mortality reduces crop damage.

Effect of Soil Moisture and a Surfactant on Entomopathogenic Nematode Suppression of the Pecan Weevil, Curculio caryae.

Our overall goal was to investigate several aspects of pecan weevil, Curculio caryae, suppression with entomopathogenic nematodes. Specifically, our objectives were to: 1) determine optimum moisture levels for larval suppression, 2) determine suppression of adult C. caryae under field conditions, and 3) measure the effects of a surfactant on nematode efficacy. In the laboratory, virulence of Heterorhabditis megidis (UK211) and Steinernema carpocapsae (All) were tested in a loamy sand at gravimetric water contents of negative 0.01, 0.06, 0.3, 1.0, and 15 bars. Curculio caryae larval survival decreased as moisture levels increased. The nematode effect was most pronounced at -0.06 bars. At -0.01 bars, larval survival was

Comparing denitrification estimates for a Texas estuary by using acetylene inhibition and membrane inlet mass spectrometry.

Characterizing denitrification rates in aquatic ecosystems is essential to understanding how systems may respond to increased nutrient loading. Thus, it is important to ensure the precision and accuracy of the methods employed for measuring denitrification rates. The acetylene (C2H2) inhibition method is a simple technique for estimating denitrification. However, potential problems, such as inhibition of nitrification and incomplete inhibition of nitrous oxide reduction, may influence rate estimates. Recently, membrane inlet mass spectrometry (MIMS) has been used to measure denitrification in aquatic systems. Comparable results were obtained with MIMS and C2H2 inhibition methods when chloramphenicol was added to C2H2 inhibition assay mixtures to inhibit new synthesis of denitrifying enzymes. Dissolved-oxygen profiles indicated that surface layers of sediment cores subjected to the MIMS flowthrough incubation remained oxic whereas cores incubated using the C2H2 inhibition methods did not. Analysis of the microbial assemblages before and after incubations indicated significant changes in the sediment surface populations during the long flowthrough incubation for MIMS analysis but not during the shorter incubation used for the C2H2 inhibition method. However, bacterial community changes were also small in MIMS cores at the oxygen transition zone where denitrification occurs. The C2H2 inhibition method with chloramphenicol addition, conducted over short incubation intervals, provides a cost-effective method for estimating denitrification, and rate estimates are comparable to those obtained by the MIMS method.