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Studies to assess natural resource recovery and evaluate monitoring methods for restored bottomland hardwood forests

The Natural Resource Damage Assessment and Restoration process assesses natural resource injury due to oil or chemical spills and calculates the damages to compensate the public for those injuries. Ecological restoration provides a means for recovering resources injured or lost due to contamination from oil or chemical spills by restoring the injured site after remediation, or acquiring or reconstructing equivalent resources off site to replace those lost due to the spill. In the case of restored forests, once restoration is implemented, monitoring of forest ecology helps keep recovery on track, with the maturation of forest vegetation, recovered soil conditions, and development of microbial, fungal, and faunal communities, necessary for ecologically functioning forests. This series of papers focuses on applying methods for monitoring restoration progress in forest vegetation and soils, and amphibian, avian, and mammalian communities, assessing strengths and weaknesses of different methods, and evaluating levels of effort needed to obtain accurate indications of forest ecological condition.

Integrated Environmental Assessment and Management

Microbial diversity, genomics, and phage–host interactions of cyanobacterial harmful algal blooms

The occurrence of cyanobacterial harmful algal blooms (cyanoHABs) is related to their physical and chemical environment. However, less is known about their associated microbial interactions and processes. In this study, cyanoHABs were analyzed as a microbial ecosystem, using 1 year of 16S rRNA sequencing and 70 metagenomes collected during the bloom season from Lake Okeechobee (Florida, USA). Biogeographical patterns observed in microbial community composition and function reflected ecological zones distinct in their physical and chemical parameters that resulted in bloom “hotspots” near major lake inflows. Changes in relative abundances of taxa within multiple phyla followed increasing bloom severity. Functional pathways that correlated with increasing bloom severity encoded organic nitrogen and phosphorus utilization, storage of nutrients, exchange of genetic material, phage defense, and protection against oxidative stress, suggesting that microbial interactions may promote cyanoHAB resilience. Cyanobacterial communities were highly diverse, with picocyanobacteria ubiquitous and oftentimes most abundant, especially in the absence of blooms. The identification of novel bloom-forming cyanobacteria and genomic comparisons indicated a functionally diverse cyanobacterial community with differences in its capability to store nitrogen using cyanophycin and to defend against phage using CRISPR and restriction-modification systems. Considering blooms in the context of a microbial ecosystem and their interactions in nature, physiologies and interactions supporting the proliferation and stability of cyanoHABs are proposed, including a role for phage infection of picocyanobacteria. This study displayed the power of “-omics” to reveal important biological processes that could support the effective management and prediction of cyanoHABs.

Florida

Capturing interactions between nitrogen and hydrological cycles under historical climate and land use: Susquehanna watershed analysis with the GFDL land model LM3-TAN

We developed a process model LM3-TAN to assess the combined effects of direct human influences and climate change on terrestrial and aquatic nitrogen (TAN) cycling. The model was developed by expanding NOAA's Geophysical Fluid Dynamics Laboratory land model LM3V-N of coupled terrestrial carbon and nitrogen (C-N) cycling and including new N cycling processes and inputs such as a soil denitrification, point N sources to streams (i.e., sewage), and stream transport and microbial processes. Because the model integrates ecological, hydrological, and biogeochemical processes, it captures key controls of the transport and fate of N in the vegetation–soil–river system in a comprehensive and consistent framework which is responsive to climatic variations and land-use changes. We applied the model at 1/8° resolution for a study of the Susquehanna River Basin. We simulated with LM3-TAN stream dissolved organic-N, ammonium-N, and nitrate-N loads throughout the river network, and we evaluated the modeled loads for 1986–2005 using data from 16 monitoring stations as well as a reported budget for the entire basin. By accounting for interannual hydrologic variability, the model was able to capture interannual variations of stream N loadings. While the model was calibrated with the stream N loads only at the last downstream Susquehanna River Basin Commission station Marietta (40°02' N, 76°32' W), it captured the N loads well at multiple locations within the basin with different climate regimes, land-use types, and associated N sources and transformations in the sub-basins. Furthermore, the calculated and previously reported N budgets agreed well at the level of the whole Susquehanna watershed. Here we illustrate how point and non-point N sources contributing to the various ecosystems are stored, lost, and exported via the river. Local analysis of six sub-basins showed combined effects of land use and climate on soil denitrification rates, with the highest rates in the Lower Susquehanna Sub-Basin (extensive agriculture; Atlantic coastal climate) and the lowest rates in the West Branch Susquehanna Sub-Basin (mostly forest; Great Lakes and Midwest climate). In the re-growing secondary forests, most of the N from non-point sources was stored in the vegetation and soil, but in the agricultural lands most N inputs were removed by soil denitrification, indicating that anthropogenic N applications could drive substantial increase of N 2 O emission, an intermediate of the denitrification process.

Pennsylvania

Solutions in microbiome engineering: Prioritizing barriers to organism establishment

Microbiome engineering is increasingly being employed as a solution to challenges in health, agriculture, and climate. Often manipulation involves inoculation of new microbes designed to improve function into a preexisting microbial community. Despite, increased efforts in microbiome engineering inoculants frequently fail to establish and/or confer long-lasting modifications on ecosystem function. We posit that one underlying cause of these shortfalls is the failure to consider barriers to organism establishment. This is a key challenge and focus of macroecology research, specifically invasion biology and restoration ecology. We adopt a framework from invasion biology that summarizes establishment barriers in three categories: (1) propagule pressure, (2) environmental filtering, and (3) biotic interactions factors. We suggest that biotic interactions is the most neglected factor in microbiome engineering research, and we recommend a number of actions to accelerate engineering solutions.

The ISME Journal: Multidisciplinary Journal of Mic

The global proliferation of aquatic, benthic Microcoleus : Taxonomy, distribution, toxin production, ecology, and future directions

There have been sporadic reports of aquatic, benthic Microcoleus proliferations in freshwater rivers, lakes, and reservoirs for four decades, with reports increasing in frequency over the last twenty years, suggesting a possible rise in their global distribution, frequency, and intensity. Microcoleus can produce anatoxins which are neurotoxic, and ingestion of toxic mats has caused hundreds of dog fatalities and raised serious human and ecological health concerns. This review synthesizes and evaluates current knowledge on Microcoleus distribution, taxonomy, toxin production, toxicity, ecology, environmental drivers, and biotic interactions. Toxin-producing Microcoleus have been reported in at least 18 countries, though many regions have not conducted toxin testing, suggesting a broader but under-reported distribution. Proliferations occur across diverse habitats, including cobble-bedded streams, large sandy rivers, reservoirs, and lakes. Microcoleus proliferations also occur on macrophytes, both in lakes and rivers. Genomic analyses currently classify anatoxin-producing Microcoleus into distinct species, with all known anatoxin-producers isolated from freshwater ecosystems. Anatoxin concentrations vary widely over space and time, within and among waterbodies. While studies on environmental drivers remain limited, research in cobble-bedded rivers suggests that moderate enrichment of dissolved inorganic nitrogen and low dissolved reactive phosphorus concentrations in the water column promote proliferation. Metagenomic approaches have revealed unique nutrient acquisition and storage strategies used by Microcoleus . Key knowledge gaps remain around the environmental and ecological triggers of proliferation, toxin production, genomic diversity and microbial interactions. Addressing these gaps through coordinated, global studies using robust datasets and consistent methods is critical to improve prediction, monitoring, and mitigation of this increasingly widespread public and ecological health threat.

Water Research

Using biological responses to monitor freshwater post-spill conditions over 3 years in Blacktail Creek, North Dakota, USA

A pipeline carrying unconventional oil and gas (OG) wastewater spilled approximately 11 million liters of wastewater into Blacktail Creek, North Dakota, USA. Flow of the mix of stream water and wastewater down the channel resulted in storage of contaminants in the hyporheic zone and along the banks, providing a long-term source of wastewater constituents to the stream. A multi-level investigation was used to assess the potential effects of oil and brine spills on aquatic life. In this study, we used a combination of experiments using a native fish species, Fathead Minnow ( Pimephales promelas ), field sampling of the microbial community structure, and measures of estrogenicity. The fish investigation included in situ experiments and experiments with collected site water. Estrogenicity was measured in collected site water samples, and microbial community analyses were conducted on collected sediments. During the initial post-spill investigation, February 2015, performing in situ fish bioassays was impossible because of ice conditions. However, microbial community (e.g., the presence of members of the Halomonadaceae, a family that is indicative of elevated salinity) and estrogenicity differences were compared to reference sites and point to early biological effects of the spill. We noted water column effects on in situ fish survival 6 months post-spill during June 2015. At that time, total dissolved ammonium (sum of ammonium and ammonia, TAN) was 4.41 mg NH 4 /L with an associated NH 3 of 1.09 mg/L, a concentration greater than the water quality criteria established to protect aquatic life. Biological measurements in the sediment defined early and long-lasting effects of the spill on aquatic resources. The microbial community structure was affected during all sampling events. Therefore, sediment may act as a sink for constituents spilled and as such provide an indication of continued and cumulative effects post-spill. However, lack of later water column effects may reflect pulse hyporheic flow of ammonia from shallow ground water. Combining fish toxicological, microbial community structure and estrogenicity information provides a complete ecological investigation that defines potential influences of contaminants at organismal, population, and community levels. In general, in situ bioassays have implications for the individual survival and changes at the population level, microbial community structure defines potential changes at the community level, and estrogenicity measurements define changes at the individual and molecular level. By understanding effects at these various levels of biological organization, natural resource managers can interpret how a course of action, especially for remediation/restoration, might affect a larger group of organisms in the system. The current work also reviews potential effects of additional constituents defined during chemistry investigations on aquatic resources.

North Dakota

Microbiome assembly in thawing permafrost and its feedbacks to climate

The physical and chemical changes that accompany permafrost thaw directly influence the microbial communities that mediate the decomposition of formerly frozen organic matter, leading to uncertainty in permafrost–climate feedbacks. Although changes to microbial metabolism and community structure are documented following thaw, the generality of post-thaw assembly patterns across permafrost soils of the world remains uncertain, limiting our ability to predict biogeochemistry and microbial community responses to climate change. Based on our review of the Arctic microbiome, permafrost microbiology, and community ecology, we propose that Assembly Theory provides a framework to better understand thaw-mediated microbiome changes and the implications for community function and climate feedbacks. This framework posits that the prevalence of deterministic or stochastic processes indicates whether the community is well-suited to thrive in changing environmental conditions. We predict that on a short timescale and following high-disturbance thaw (e.g., thermokarst), stochasticity dominates post-thaw microbiome assembly, suggesting that functional predictions will be aided by detailed information about the microbiome. At a longer timescale and lower-intensity disturbance (e.g., active layer deepening), deterministic processes likely dominate, making environmental parameters sufficient for predicting function. We propose that the contribution of stochastic and deterministic processes to post-thaw microbiome assembly depends on the characteristics of the thaw disturbance, as well as characteristics of the microbial community, such as the ecological and phylogenetic breadth of functional guilds, their functional redundancy, and biotic interactions. These propagate across space and time, potentially providing a means for predicting the microbial forcing of greenhouse gas feedbacks to global climate change.

Global Change Biology

A combined microbial and ecosystem metric of carbon retention efficiency explains land cover-dependent soil microbial biodiversity–ecosystem function relationships

While soil organic carbon (C) is the foundation of productive and healthy ecosystems, the impact of the ecology of microorganisms on C-cycling remains unknown. We manipulated the diversity, applied here as species richness, of the microbial community present in similar soils on two contrasting land-covers—an adjacent pasture and forest—and observed the transformations of plant detritus and soil organic matter (SOM) using stable isotope ( 13 C) tracing coupled with a novel nuclear magnetic resonance (NMR) experiment. The amount of detritus-C degraded was not affected by the microbial diversity (p > 0.05), however the fate of detritus- and SOM-C across the diversity gradient was complex and land cover-dependent. For example, in the pasture soil, higher diversity led to lower CO 2 production (p = 0.001), a trend driven solely by SOM-C mineralization. There was no relationship between diversity and detritus-C mineralization or production of new mineral-associations after one year (p > 0.05). In contrast, in the forest soil higher diversity resulted in increased detritus-C (p = 0.01) and SOM-C (p = 0.0008) mineralization and decreased mineral-associated organic matter formation (p = 0.02). In both land cover types, retention efficiency—a measure that integrates both microbial physiology and the ability of the ecosystem to retain C—explained C loss and transformation trends. Overall, this demonstrates that the trajectory of C gained and lost is altered by land management-induced changes to microbial communities, soil structure, and chemical characteristics underlying SOM persistence.

Biogeochemistry Letters

Soil magnetic susceptibility: A quantitative proxy of soil drainage for use in ecological restoration

Flooded, saturated, or poorly drained soils are commonly anaerobic, leading to microbially induced magnetite/maghemite dissolution and decreased soil magnetic susceptibility (MS). Thus, MS is considerably higher in well-drained soils (MS typically 40-80 ?? 10-5 standard international [SI]) compared to poorly drained soils (MS typically 10-25 ?? 10-5 SI) in Illinois, other soil-forming factors being equal. Following calibration to standard soil probings, MS values can be used to rapidly and precisely delineate hydric from nonhydric soils in areas with relatively uniform parent material. Furthermore, soil MS has a moderate to strong association with individual tree species' distribution across soil moisture regimes, correlating inversely with independently reported rankings of a tree species' flood tolerance. Soil MS mapping can thus provide a simple, rapid, and quantitative means for precisely guiding reforestation with respect to plant species' adaptations to soil drainage classes. For instance, in native woodlands of east-central Illinois, Quercus alba , Prunus serotina, and Liriodendron tulipifera predominantly occur in moderately well-drained soils (MS 40-60 ?? 10-5 SI), whereas Acer saccharinum, Carya laciniosa, and Fraxinus pennsylvanica predominantly occur in poorly drained soils (MS <20 ?? 10-5 SI). Using a similar method, an MS contour map was used to guide restoration of mesic, wet mesic, and wet prairie species to pre-settlement distributions at Meadowbrook Park (Urbana, IL, U.S.A.). Through use of soil MS maps calibrated to soil drainage class and native vegetation occurrence, restoration efforts can be conducted more successfully and species distributions more accurately reconstructed at the microecosystem level. ?? 2008 Society for Ecological Restoration International.

Restoration Ecology

Environmental drivers and spatial patterns of antibiotic-resistant, enteric coliforms across a forest–urban riverscape

Antibiotic resistant bacteria are prevalent environmental contaminants in freshwaters, and antibiotic resistance genes circulate throughout the urban water cycle. The increase of antibiotic resistant pathogens threatens public health through direct and indirect exposure, and natural resource managers need information on the spatial patterns of antibiotic resistant bacteria and environmental factors associated with their distribution to improve water quality monitoring and to better assess human, animal, and environmental health risks. We collected water and epilithic biofilm samples and measured physicochemical environmental variables at 29 sites distributed longitudinally in the Green-Duwamish River basin, Washington, USA. We characterized catchment-wide patterns of gram-negative fecal indicator bacteria and hypothesized that the presence of antibiotic resistance would be associated with environmental heterogeneity, bacterial primary ecology, stream compartment, and stream type. Antibiotic resistance was determined by microbial growth on selective media supplemented with 3 different antibiotics (ampicillin, chloramphenicol, or tetracycline). Phenotypic antibiotic resistance was positively associated with disturbance, but resistance to at least 1 antibiotic was also detected in undeveloped river segments, with an 83% overall detection rate (i.e., 24 out of 29 sites, 17 in the mainstem and 7 in tributaries). The most probable number of Escherichia coli was associated with higher levels of antibiotic resistance of non- E. coli coliforms across the basin (ρ = 0.38, p < 0.01) but was not associated with antibiotic resistance of E. coli . Phenotypic resistance was highest among non- E. coli coliforms in the water column of tributaries draining moderately to extensively developed subcatchments. Generalized linear mixed-effects model results showed that 18% of the variance in presence of antibiotic resistance was explained by the fixed effects (summed CV across environmental variables, stream type, primary ecology, and stream compartment), and when a spatial random effect was included, the model explained 27% of the variance. Our study provides new evidence that environmental factors and bacterial primary ecology are important underlying factors associated with spatial patterns of antibiotic resistant enteric coliforms. We used macroecological concepts and a riverscape approach to characterize the distribution of antibiotic resistance with methods applicable to municipalities.

Freshwater Science

Ground-water microbiology and geochemistry

Up-to-date coverage and a unique, multidisciplinary approach The ongoing effort to protect our valuable ground-water resources necessarily involves scientists and engineers from many disciplines. Ground-Water Microbiology and Geochemistry, Second Edition is designed to bridge the historical lack of communication among these disciplines by detailing-in language that cuts across specialties-the impact of microorganisms and microbial processes on ground-water systems. Carefully revised to reflect the many recent discoveries that have been made in the field, the Second Edition begins with an overview of microbiology, ideal for hydrologists and others who may lack formal training in the field. These initial chapters systematically cover the kinds of microorganisms found in subsurface environments, focusing on their growth, metabolism, genetics, and ecology. The second part of the book offers a hydrologic perspective on how microbial processes affect ground-water geochemistry in pristine systems. It also introduces the different classes of ground-water systems, and gives an overview of techniques for sampling subsurface environments. Readers gain an understanding of biogeochemical cycling in ground-water systems-in coverage unique to this book-and how ground-water chemistry can be used to study microbial processes in aquifer systems. The final section of the book deals with the biodegradation of human-introduced contaminants in ground-water systems, with an up-to-date review of the physiology, biochemistry, and redox conditions that favor biodegradation processes. Ground-Water Microbiology and Geochemistry, Second Edition is important reading for geoscientists, hydrologists, and environmental engineers, as well as for water planners and lawyers involved in environmental issues. It also serves as a compelling text for upper-level undergraduate and graduate courses in ground-water chemistry.

Book

Preparation and analysis of nitrogen-bearing compounds in water for stable isotope ratio measurement

This chapter reviews methods for analyzing "natural abundance" nitrogen-bearing compounds in water for δ 15 N, δ 18 O, and δ 17 O. The techniques that are reviewed are Kjeldahl distillation and digestion, diffusion, micro diffusion, ion exchange, and microbial denitrifier methods. It overlaps the contents of the preparation of ecological and biogeochemical samples for isotope analysis and also covers sections on particulate organic nitrogen and biological samples. Nitrogen comprises 78% of the atmosphere, oxygen 21% and argon 1%. There are two stable isotopes of N are 14 N and 15 N and a wide range of oxidation numbers exhibited by nitrogen compounds. Developments of continuous-flow mass spectrometry makes it possible to determine both δ15N and δ18O values of nitrate samples. Such techniques are particularly useful for small-size samples. With decreasing sample size, possible isotopic fractionations associated with extraction from the matrix, contaminants in the reagents, and leaks will become more significant. Due to the interest in distinguishing different species of organic-N compounds, the diffusion method is applied to differentiate types of organic-N.

Book chapter

Energy-related wastewater contamination alters microbial communities of sediment, water, and amphibian skin

To inform responsible energy development, it is important to understand the ecological effects of contamination events. Wastewaters, a common byproduct of oil and gas extraction, often contain high concentrations of sodium chloride (NaCl) and heavy metals (e.g., strontium and vanadium). These constituents can negatively affect aquatic organisms, but there is scarce information for how wastewaters influence potentially distinct microbiomes in wetland ecosystems . Additionally, few studies have concomitantly investigated effects of wastewaters on the habitat (water and sediment) and skin microbiomes of amphibians or relationships among these microbial communities. We sampled microbiomes of water, sediment, and skin of four larval amphibian species across a gradient of chloride contamination (0.04–17,500 mg/L Cl) in the Prairie Pothole Region of North America. We detected 3129 genetic phylotypes and 68 % of those phylotypes were shared among the three sample types. The most common shared phylotypes were Proteobacteria, Firmicutes, and Bacteroidetes. Salinity of wastewaters increased dissimilarity within all three microbial communities, but not the diversity or richness of water and skin microbial communities. Strontium was associated with lower diversity and richness of sediment microbial communities, but not those of water or amphibian skin, likely because metal deposition occurs in sediment when wetlands dry. Based on Bray Curtis distance matrices, sediment microbiomes were similar to those of water, but neither had substantial overlap with amphibian microbiomes. Species identity was the strongest predictor of amphibian microbiomes; frog microbiomes were similar but differed from that of the salamander, whose microbiome had the lowest richness and diversity. Understanding how effects of wastewaters on the dissimilarity, richness, and diversity of microbial communities also influence the ecosystem function of communities will be an important next step. However, our study provides novel insight into the characteristics of, and associations among, different wetland microbial communities and effects of wastewaters from energy production.

Montana, North Dakota

Genotype, soil type, and locale effects on reciprocal transplant vigor, endophyte growth, and microbial functional diversity of a narrow sagebrush hybrid zone in Salt Creek Canyon, Utah

When addressing the nature of ecological adaptation and environmental factors limiting population ranges and contributing to speciation, it is important to consider not only the plant's genotype and its response to the environment, but also any close interactions that it has with other organisms, specifically, symbiotic microorganisms. To investigate this, soils and seedlings were reciprocally transplanted into common gardens of the big sagebrush hybrid zone in Salt Creek Canyon, Utah, to determine location and edaphic effects on the fitness of parental and hybrid plants. Endophytic symbionts and functional microbial diversity of indigenous and transplanted soils and sagebrush plants were also examined. Strong selection occurred against the parental genotypes in the middle hybrid zone garden in middle hybrid zone soil; F1 hybrids had the highest fitness under these conditions. Neither of the parental genotypes had superior fitness in their indigenous soils and habitats; rather F1 hybrids with the nonindigenous maternal parent were superiorly fit. Significant garden-by-soil type interactions indicate adaptation of both plant and soil microorganisms to their indigenous soils and habitats, most notably in the middle hybrid zone garden in middle hybrid zone soil. Contrasting performances of F1 hybrids suggest asymmetrical gene flow with mountain, rather than basin, big sagebrush acting as the maternal parent. We showed that the microbial community impacted the performance of parental and hybrid plants in different soils, likely limiting the ranges of the different genotypes.

Utah

Phenology-based, remote sensing of post-burn disturbance windows in rangelands

Wildland fire activity has increased in many parts of the world in recent decades. Ecological disturbance by fire can accelerate ecosystem degradation processes such as erosion due to combustion of vegetation that otherwise provides protective cover to the soil surface. This study employed a novel ecological indicator based on remote sensing of vegetation greenness dynamics (phenology) to estimate variability in the window of time between fire and the reemergence of green vegetation. The indicator was applied as a proxy for short-term, post-fire disturbance windows in rangelands; where a disturbance window is defined as the time required for an ecological or geomorphic process that is altered to return to pre-disturbance levels. We examined variability in the indicator determined for time series of MODIS and AVHRR NDVI remote sensing data for a database of ∼100 historical wildland fires, with associated post-fire reseeding treatments, that burned 1990–2003 in cold desert shrub steppe of the Great Basin and Columbia Plateau of the western USA. The indicator-based estimates of disturbance window length were examined relative to the day of the year that fires burned and seeding treatments to consider effects of contemporary variability in fire regime and management activities in this environment. A key finding was that contemporary changes of increased length of the annual fire season could have indirect effects on ecosystem degradation, as early season fires appeared to result in longer time that soils remained relatively bare of the protective cover of vegetation after fires. Also important was that reemergence of vegetation did not occur more quickly after fire in sites treated with post-fire seeding, which is a strategy commonly employed to accelerate post-fire vegetation recovery and stabilize soil. Future work with the indicator could examine other ecological factors that are dynamic in space and time following disturbance – such as nutrient cycling, carbon storage, microbial community composition, or soil hydrology – as a function of disturbance windows, possibly using simulation modeling and historical wildfire information.

California;Idaho;Nevada;Oregon;Utah;Washington

Using high-throughput DNA sequencing, genetic fingerprinting, and quantitative PCR as tools for monitoring bloom-forming and toxigenic cyanobacteria in Upper Klamath Lake, Oregon, 2013 and 2014

Monitoring the community structure and metabolic activities of cyanobacterial blooms in Upper Klamath Lake, Oregon, is critical to lake management because these blooms degrade water quality and produce toxic microcystins that are harmful to humans, domestic animals, and wildlife. Genetic tools, such as DNA fingerprinting by terminal restriction fragment length polymorphism (T-RFLP) analysis, high-throughput DNA sequencing (HTS), and real-time, quantitative polymerase chain reaction (qPCR), provide more sensitive and rapid assessments of bloom ecology than traditional techniques. The objectives of this study were (1) to characterize the microbial community at one site in Upper Klamath Lake and determine changes in the cyanobacterial community through time using T-RFLP and HTS in comparison with traditional light microscopy; (2) to determine relative abundances and changes in abundance over time of toxigenic Microcystis using qPCR; and (3) to determine relative abundances and changes in abundance over time of Aphanizomenon , Microcystis , and total cyanobacteria using qPCR. T-RFLP analysis of total cyanobacteria showed a dominance of only one or two distinct genotypes in samples from 2013, but results of HTS in 2013 and 2014 showed more variations in the bloom cycle that fit with the previous understanding of bloom dynamics in Upper Klamath Lake and indicated that potentially toxigenic Microcystis was more prevalent in 2014 than in years prior. The qPCR-estimated copy numbers of all target genes were higher in 2014 than in 2013, when microcystin concentrations also were higher. Total Microcystis density was shown with qPCR to be a better predictor of late-season increases in microcystin concentrations than the relative proportions of potentially toxigenic cells. In addition, qPCR targeting Aphanizomenon at one site in Upper Klamath Lake indicated a moderate bloom of this species (corresponding to chlorophyll a concentrations between approximately 75 and 200 micrograms per liter) from mid-June to mid-August, 2014. After August 18, the Aphanizomenon bloom was overtaken by Microcystis late in the season as microcystin concentrations peaked. Overall, results of this study showed how DNA-based, genetic methods may provide rapid and sensitive diagnoses for the presence of toxigenic cyanobacteria and that they are useful for general monitoring or ecological studies and identification of cyanobacterial community members in complex aquatic habitats. These same methods can also be used to simultaneously address spatial (horizontal and vertical) and temporal variation and under different conditions. Additionally, with some modifications, the same techniques can be applied to different sample types, including water, sediment, and tissue.

Oregon

A survey of microbial community diversity in marine sediments impacted by petroleum hydrocarbons from the Gulf of Mexico and Atlantic shorelines, Texas to Florida

Microbial community genomic DNA was extracted from sediment samples collected along the Gulf of Mexico and Atlantic coasts from Texas to Florida. Sample sites were identified as being ecologically sensitive and (or) as having high potential of being impacted by Macondo-1 (M-1) well oil from the Deepwater Horizon blowout. The diversity within the microbial communities associated with the collected sediments provides a baseline dataset to which microbial community-diversity data from impacted sites could be compared. To determine the microbial community diversity in the samples, genetic fingerprints were generated and compared. Specific sequences within the community genomic DNA were first amplified using the polymerase chain reaction (PCR) with a primer set that provides possible resolution to the species level. A second nested PCR was performed on the primary PCR products using a primer set on which a GC-clamp was attached to one of the primers. The nested PCR products were separated using denaturing-gradient gel electrophoresis (DGGE) that resolves the nested PCR products based on sequence dissimilarities (or similarities), forming a genomic fingerprint of the microbial diversity within the respective samples. Samples with similar fingerprints were grouped and compared to oil-fingerprint data from the same sites (Rosenbauer and others, 2011). The microbial community fingerprints were generally grouped into sites that had been shown to contain background concentrations of non-Deepwater Horizon oil. However, these groupings also included sites where no oil signature was detected. This report represents some of the first information on naturally occurring microbial communities in sediment from shorelines along the Gulf of Mexico and Atlantic coasts from Texas to Florida.

Gulf Of Mexico

Identifying the molecular signatures of agricultural expansion in Amazonian headwater streams

Agricultural impacts on aquatic ecosystems are well-studied, however, most research has focused on temperate regions, whereas the forefront of agricultural expansion is currently in the tropics. At the vanguard of this growth is the boundary between the Amazon and Cerrado biomes in Brazil, driven primarily by expansion of soybean and corn croplands. Here we examine the impacts of cropland expansion on receiving lowland Amazon Basin headwater streams in terms of dissolved organic carbon (DOC) concentration and dissolved organic matter (DOM) composition via ultra-high resolution mass spectrometry. Streams draining croplands had lower DOC concentrations and DOM molecular signatures enriched in N- and S-containing formula in comparison to forested streams. Cropland streams were also enriched in aliphatic, peptide-like and highly unsaturated and phenolic (low O/C) compound categories in comparison to forest streams (enriched in polyphenolics, condensed aromatics and highly unsaturated and phenolic (high O/C) compound categories) indicative of the shifting of sources from organic-rich surface soils and litter layers to autochthonous and more microbial biomass. Distinct molecular assemblages were strongly correlated with cropland and forest catchments, highlighting headwater streams as sentinels for detecting change. On investigation of unique molecular formulae present in only cropland sites, four cropland markers provided the ability to track agricultural impacts in the region. Overall, these patterns indicate reduced organic matter inputs in croplands and greater microbial degradation at these sites leading to declining DOC concentrations, and DOM of more microbial character in receiving streams that is more biolabile, with clear ramifications for downstream ecology and biogeochemical cycles.

Amazon River basin, Tanguro Ranch, Xinga River