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Sample records for aquatic microbial samples

  1. The Sorcerer II Global Ocean Sampling Expedition: Metagenomic Characterization of Viruses within Aquatic Microbial Samples

    PubMed Central

    Williamson, Shannon J.; Rusch, Douglas B.; Yooseph, Shibu; Halpern, Aaron L.; Heidelberg, Karla B.; Glass, John I.; Andrews-Pfannkoch, Cynthia; Fadrosh, Douglas; Miller, Christopher S.; Sutton, Granger; Frazier, Marvin; Venter, J. Craig

    2008-01-01

    Viruses are the most abundant biological entities on our planet. Interactions between viruses and their hosts impact several important biological processes in the world's oceans such as horizontal gene transfer, microbial diversity and biogeochemical cycling. Interrogation of microbial metagenomic sequence data collected as part of the Sorcerer II Global Ocean Expedition (GOS) revealed a high abundance of viral sequences, representing approximately 3% of the total predicted proteins. Cluster analyses of the viral sequences revealed hundreds to thousands of viral genes encoding various metabolic and cellular functions. Quantitative analyses of viral genes of host origin performed on the viral fraction of aquatic samples confirmed the viral nature of these sequences and suggested that significant portions of aquatic viral communities behave as reservoirs of such genetic material. Distributional and phylogenetic analyses of these host-derived viral sequences also suggested that viral acquisition of environmentally relevant genes of host origin is a more abundant and widespread phenomenon than previously appreciated. The predominant viral sequences identified within microbial fractions originated from tailed bacteriophages and exhibited varying global distributions according to viral family. Recruitment of GOS viral sequence fragments against 27 complete aquatic viral genomes revealed that only one reference bacteriophage genome was highly abundant and was closely related, but not identical, to the cyanomyovirus P-SSM4. The co-distribution across all sampling sites of P-SSM4-like sequences with the dominant ecotype of its host, Prochlorococcus supports the classification of the viral sequences as P-SSM4-like and suggests that this virus may influence the abundance, distribution and diversity of one of the most dominant components of picophytoplankton in oligotrophic oceans. In summary, the abundance and broad geographical distribution of viral sequences within

  2. The Sorcerer II Global Ocean Sampling Expedition: metagenomic characterization of viruses within aquatic microbial samples.

    PubMed

    Williamson, Shannon J; Rusch, Douglas B; Yooseph, Shibu; Halpern, Aaron L; Heidelberg, Karla B; Glass, John I; Andrews-Pfannkoch, Cynthia; Fadrosh, Douglas; Miller, Christopher S; Sutton, Granger; Frazier, Marvin; Venter, J Craig

    2008-01-23

    Viruses are the most abundant biological entities on our planet. Interactions between viruses and their hosts impact several important biological processes in the world's oceans such as horizontal gene transfer, microbial diversity and biogeochemical cycling. Interrogation of microbial metagenomic sequence data collected as part of the Sorcerer II Global Ocean Expedition (GOS) revealed a high abundance of viral sequences, representing approximately 3% of the total predicted proteins. Cluster analyses of the viral sequences revealed hundreds to thousands of viral genes encoding various metabolic and cellular functions. Quantitative analyses of viral genes of host origin performed on the viral fraction of aquatic samples confirmed the viral nature of these sequences and suggested that significant portions of aquatic viral communities behave as reservoirs of such genetic material. Distributional and phylogenetic analyses of these host-derived viral sequences also suggested that viral acquisition of environmentally relevant genes of host origin is a more abundant and widespread phenomenon than previously appreciated. The predominant viral sequences identified within microbial fractions originated from tailed bacteriophages and exhibited varying global distributions according to viral family. Recruitment of GOS viral sequence fragments against 27 complete aquatic viral genomes revealed that only one reference bacteriophage genome was highly abundant and was closely related, but not identical, to the cyanomyovirus P-SSM4. The co-distribution across all sampling sites of P-SSM4-like sequences with the dominant ecotype of its host, Prochlorococcus supports the classification of the viral sequences as P-SSM4-like and suggests that this virus may influence the abundance, distribution and diversity of one of the most dominant components of picophytoplankton in oligotrophic oceans. In summary, the abundance and broad geographical distribution of viral sequences within

  3. Autonomous Microbial Sampler (AMS), a device for the uncontaminated collection of multiple microbial samples from submarine hydrothermal vents and other aquatic environments

    NASA Astrophysics Data System (ADS)

    Taylor, Craig D.; Doherty, Kenneth W.; Molyneaux, Stephen J.; Morrison, Archie T.; Billings, John D.; Engstrom, Ivory B.; Pfitsch, Don W.; Honjo, Susumu

    2006-05-01

    An Autonomous Microbial Sampler (AMS) is described that will obtain uncontaminated and exogenous DNA-free microbial samples from most marine, freshwater and hydrothermal ecosystems. Sampling with the AMS may be conducted using manned submersibles, remotely operated vehicles (ROVs), autonomous underwater vehicles (AUVs), or when tethered to a hydrowire during hydrocast operations on research vessels. The modular device consists of a titanium nozzle for sampling in potentially hot environments (>350 °C) and fluid-handling components for the collection of six independent filtered or unfiltered samples. An onboard microcomputer permits sampling to be controlled by the investigator, by external devices (e.g., AUV computer), or by internal programming. Temperature, volume pumped and other parameters are recorded during sampling. Complete protection of samples from microbial contamination was observed in tests simulating deployment of the AMS in coastal seawater, where the sampling nozzle was exposed to seawater containing 1×10 6 cells ml -1 of a red pigmented tracer organism, Serratia marinorubra. Field testing of the AMS at a hydrothermal vent field was successfully undertaken in 2000. Results of DNA destruction studies have revealed that exposure of samples of the Eukaryote Euglena and the bacterium S. marinorubra to 0.5 N sulfuric acid at 23 °C for 1 h was sufficient to remove polymerase chain reaction (PCR) amplifiable DNA. Studies assessing the suitability of hydrogen peroxide as a sterilizing and DNA-destroying agent showed that 20% or 30% hydrogen peroxide sterilized samples of Serratia in 1 h and destroyed the DNA of Serratia in 3 h, but not 1 or 2 h. DNA AWAY™ killed Serratia and destroyed the DNA of both Serratia and the vent microbe (GB-D) of the genus Pyrococcus in 1 h.

  4. Rapid detection of microbial cell abundance in aquatic systems

    DOE PAGES

    Rocha, Andrea M.; Yuan, Quan; Close, Dan M.; ...

    2016-06-01

    The detection and quantification of naturally occurring microbial cellular densities is an essential component of environmental systems monitoring. While there are a number of commonly utilized approaches for monitoring microbial abundance, capacitance-based biosensors represent a promising approach because of their low-cost and label-free detection of microbial cells, but are not as well characterized as more traditional methods. Here, we investigate the applicability of enhanced alternating current electrokinetics (ACEK) capacitive sensing as a new application for rapidly detecting and quantifying microbial cellular densities in cultured and environmentally sourced aquatic samples. ACEK capacitive sensor performance was evaluated using two distinct and dynamicmore » systems the Great Australian Bight and groundwater from the Oak Ridge Reservation in Oak Ridge, TN. Results demonstrate that ACEK capacitance-based sensing can accurately determine microbial cell counts throughout cellular concentrations typically encountered in naturally occurring microbial communities (103 – 106 cells/mL). A linear relationship was observed between cellular density and capacitance change correlations, allowing a simple linear curve fitting equation to be used for determining microbial abundances in unknown samples. As a result, this work provides a foundation for understanding the limits of capacitance-based sensing in natural environmental samples and supports future efforts focusing on evaluating the robustness ACEK capacitance-based within aquatic environments.« less

  5. Rapid detection of microbial cell abundance in aquatic systems.

    PubMed

    Rocha, Andrea M; Yuan, Quan; Close, Dan M; O'Dell, Kaela B; Fortney, Julian L; Wu, Jayne; Hazen, Terry C

    2016-11-15

    The detection and quantification of naturally occurring microbial cellular densities is an essential component of environmental systems monitoring. While there are a number of commonly utilized approaches for monitoring microbial abundance, capacitance-based biosensors represent a promising approach because of their low-cost and label-free detection of microbial cells, but are not as well characterized as more traditional methods. Here, we investigate the applicability of enhanced alternating current electrokinetics (ACEK) capacitive sensing as a new application for rapidly detecting and quantifying microbial cellular densities in cultured and environmentally sourced aquatic samples. ACEK capacitive sensor performance was evaluated using two distinct and dynamic systems - the Great Australian Bight and groundwater from the Oak Ridge Reservation in Oak Ridge, TN. Results demonstrate that ACEK capacitance-based sensing can accurately determine microbial cell counts throughout cellular concentrations typically encountered in naturally occurring microbial communities (10(3)-10(6) cells/mL). A linear relationship was observed between cellular density and capacitance change correlations, allowing a simple linear curve fitting equation to be used for determining microbial abundances in unknown samples. This work provides a foundation for understanding the limits of capacitance-based sensing in natural environmental samples and supports future efforts focusing on evaluating the robustness ACEK capacitance-based within aquatic environments. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Adaptation of Aquatic Microbial Communities to Hg2+ Stress †

    PubMed Central

    Barkay, Tamar

    1987-01-01

    The mechanism of adaptation to Hg2+ in four aquatic habitats was studied by correlating microbially mediated Hg2+ volatilization with the adaptive state of the exposed communities. Community diversity, heterotrophic activity, and Hg2+ resistance measurements indicated that adaptation of all four communities was stimulated by preexposure to Hg2+. In saline water communities, adaptation was associated with rapid volatilization after an initial lag period. This mechanism, however, did not promote adaptation in a freshwater sample, in which Hg2+ was volatilized slowly, regardless of the resistance level of the microbial community. Distribution of the mer operon among representative colonies of the communities was not related to adaptation to Hg2+. Thus, although volatilization enabled some microbial communities to sustain their functions in Hg2+-stressed environments, it was not mediated by the genes that serve as a model system in molecular studies of bacterial resistance to mercurials. PMID:16347488

  7. Estimating Aquatic Insect Populations. Introduction to Sampling.

    ERIC Educational Resources Information Center

    Chihuahuan Desert Research Inst., Alpine, TX.

    This booklet introduces high school and junior high school students to the major groups of aquatic insects and to population sampling techniques. Chapter 1 consists of a short field guide which can be used to identify five separate orders of aquatic insects: odonata (dragonflies and damselflies); ephemeroptera (mayflies); diptera (true flies);…

  8. Estimating Aquatic Insect Populations. Introduction to Sampling.

    ERIC Educational Resources Information Center

    Chihuahuan Desert Research Inst., Alpine, TX.

    This booklet introduces high school and junior high school students to the major groups of aquatic insects and to population sampling techniques. Chapter 1 consists of a short field guide which can be used to identify five separate orders of aquatic insects: odonata (dragonflies and damselflies); ephemeroptera (mayflies); diptera (true flies);…

  9. Selected Nucleic Acid Precursors in Studies of Aquatic Microbial Ecology

    PubMed Central

    Karl, David M.

    1982-01-01

    The use of radiolabeled nucleosides and nucleic acid bases to estimate the rates of RNA and DNA synthesis in naturally occurring microbial assemblages requires numerous assumptions, several of which are evaluated herein. Comparative time series analyses of the uptake and incorporation, labeling specificity, and extent of catabolism of [2-3H]adenine, [methyl-3H]thymidine, and [5-3H]uridine were performed with pure bacterial and algal cultures, as well as with environmental samples. [3H]thymidine yielded the most variable results, especially with regard to the extent of nonspecific macromolecular labeling. The pathways of [3H]thymidine and [3H]adenine metabolism were further evaluated by isotope dilution methods and by comparing incorporation patterns of thymidine labeled at different sites of the molecule. The advantages, uncertainties, and limitations of the use of radiolabeled nucleic acid precursors in studies of aquatic microbial ecology are discussed and a prospectus for future studies presented. PMID:16346114

  10. Effect of pesticides on microbial communities in container aquatic habitats

    PubMed Central

    Muturi, Ephantus J.; Donthu, Ravi Kiran; Fields, Christopher J.; Moise, Imelda K.; Kim, Chang-Hyun

    2017-01-01

    Container aquatic habitats support a specialized community of macroinvertebrates (e.g. mosquitoes) that feed on microbial communities associated with decaying organic matter. These aquatic habitats are often embedded within and around agricultural lands and are frequently exposed to pesticides. We used a microcosm approach to examine the single and combined effects of two herbicides (atrazine, glyphosate), and three insecticides (malathion, carbaryl, permethrin) on microbial communities of container aquatic habitats. MiSeq sequencing of the V4 region of both bacterial and archaeal 16S rRNA gene was used to characterize the microbial communities of indoor microcosms that were either exposed to each pesticide alone, a mix of herbicides, a mix of insecticides, or a mix of all five insecticides. Individual insecticides but not herbicides reduced the microbial diversity and richness and two insecticides, carbaryl and permethrin, also altered the microbial community structure. A mixture of herbicides had no effect on microbial diversity or structure but a mixture of insecticides or all five pesticides reduced microbial diversity and altered the community structure. These findings suggest that exposure of aquatic ecosystems to individual pesticides or their mixtures can disrupt aquatic microbial communities and there is need to decipher how these changes affect resident macroinvertebrate communities. PMID:28300212

  11. Effect of pesticides on microbial communities in container aquatic habitats.

    PubMed

    Muturi, Ephantus J; Donthu, Ravi Kiran; Fields, Christopher J; Moise, Imelda K; Kim, Chang-Hyun

    2017-03-16

    Container aquatic habitats support a specialized community of macroinvertebrates (e.g. mosquitoes) that feed on microbial communities associated with decaying organic matter. These aquatic habitats are often embedded within and around agricultural lands and are frequently exposed to pesticides. We used a microcosm approach to examine the single and combined effects of two herbicides (atrazine, glyphosate), and three insecticides (malathion, carbaryl, permethrin) on microbial communities of container aquatic habitats. MiSeq sequencing of the V4 region of both bacterial and archaeal 16S rRNA gene was used to characterize the microbial communities of indoor microcosms that were either exposed to each pesticide alone, a mix of herbicides, a mix of insecticides, or a mix of all five insecticides. Individual insecticides but not herbicides reduced the microbial diversity and richness and two insecticides, carbaryl and permethrin, also altered the microbial community structure. A mixture of herbicides had no effect on microbial diversity or structure but a mixture of insecticides or all five pesticides reduced microbial diversity and altered the community structure. These findings suggest that exposure of aquatic ecosystems to individual pesticides or their mixtures can disrupt aquatic microbial communities and there is need to decipher how these changes affect resident macroinvertebrate communities.

  12. Two decades of describing the unseen majority of aquatic microbial diversity.

    PubMed

    Zinger, Lucie; Gobet, Angélique; Pommier, Thomas

    2012-04-01

    Aquatic environments harbour large and diverse microbial populations that ensure their functioning and sustainability. In the current context of global change, characterizing microbial diversity has become crucial, and new tools have been developed to overcome the methodological challenges posed by working with microbes in nature. The advent of Sanger sequencing and now next-generation sequencing technologies has enabled the resolution of microbial communities to an unprecedented degree of precision. However, to correctly interpret microbial diversity and its patterns this revolution must also consider conceptual and methodological matters. This review presents advances, gaps and caveats of these recent approaches when considering microorganisms in aquatic ecosystems. We also discuss potentials and limitations of the available methodologies, from water sampling to sequence analysis, and suggest alternative ways to incorporate results in a conceptual and methodological framework. Together, these methods will allow us to gain an unprecedented understanding of microbial diversity in aquatic ecosystems. © 2011 Blackwell Publishing Ltd.

  13. Effect of pesticides on microbial communities in container aquatic habitats

    USDA-ARS?s Scientific Manuscript database

    Mosquitoes develop in a variety of aquatic habitats and feed on microbial communities associated with decaying organic matter. These aquatic habitats are often embedded within and around agricultural lands and are frequently exposed to agricultural chemicals. We used a microcosm approach to examine ...

  14. Minimizing microbial contamination of sperm samples

    USGS Publications Warehouse

    Jenkins, Jill A.; Tiersch, Terrence R.; Green, Christopher C.

    2011-01-01

    Taken from the Methods section: With the collection and translocation of gametes from aquatic species, a potential hazard exists for microbial transfer. Contamination of semen can occur during collection, processing, storage, and transport. Some preventative measures are described below for limiting the spread and amplification of microorganisms such as bacteria, viruses, fungi, mycoplasmas, and parasites. Generally, sanitation during collection is essential. Materials and equipment used to freeze semen should be sterile. Following good practice guidelines for handling and processing samples collected for freezing is especially important for non-domestic animals where disease-free status cannot be guaranteed and unsophisticated technology is used (Russell et al. 1977).

  15. Distinct Factors Shape Aquatic and Sedimentary Microbial Community Structures in the Lakes of Western China

    PubMed Central

    Yang, Jian; Jiang, Hongchen; Wu, Geng; Liu, Wen; Zhang, Guojing

    2016-01-01

    Little is known about the relative importance of spatial and environmental factors to structuring aquatic and sedimentary microbial biogeography in lakes. Here, we investigated the microbial community composition (MCC) of the water (n = 35) and sediment (n = 35) samples from 16 lakes in western China (salinity: freshwater to salt saturation; pairwise geographical distance: 9–2027 km) using high-throughput sequencing and evaluated the relative importance of spatial and environmental factors to microbial (including total, abundant, and rare) distributions. Our results showed that spatial factors were more important than environmental factors in shaping the biogeography of aquatic and sedimentary microbial communities in the studied lakes, and spatial factors on abundant microbial community was stronger than that on the total/rare microbial communities. Moreover, sedimentary rare MCC might be more sensitive to environmental factors than its aquatic counterpart. Such different biogeography responses of total, abundant, and rare communities to environmental and spatial factors could be ascribed to different physiochemical properties between water and sediment. Collectively, this study expands our understanding of factors shaping microbial biogeography of total, abundant, and rare communities between waters and sediments of lakes. PMID:27877171

  16. Understanding aquatic microbial processes using EEM's and in-situ fluorescence sensors

    NASA Astrophysics Data System (ADS)

    Fox, Bethany; Attridge, John; Rushworth, Cathy; Cox, Tim; Anesio, Alexandre; Reynolds, Darren

    2015-04-01

    The diverse origin of dissolved organic matter (DOM) in aquatic systems is well documented within the literature. Previous literature indicates that coloured dissolved organic matter (CDOM) is, in part, transformed by aquatic microbial processes, and that dissolved organic material derived from a microbial origin exhibits tryptophan-like fluorescence. However, this phenomenon is not fully understood and very little data is available within the current literature. The overall aim of our work is to reveal the microbial-CDOM interactions that give rise to the observed tryptophan-like fluorescence. The work reported here investigates the microbial processes that occur within freshwater aquatic samples, as defined by the biochemical oxygen demand (BOD) test, as a function of the T1 peak (λex/em 280/330-370 nm). A series of standard water samples were prepared using glucose, glutamic acid, BOD dilution water and a bacterial seed (Cole-Parmer BOD microbe capsules). Samples were spiked with CDOM (derived from an environmental water body) and subjected to time resolved BOD analysis and as excitation-emission fluorescence spectroscopy. All EEM spectral data was interrogated using parallel factor analysis (PARAFAC) in an attempt to determine the presence and dominance (relative intensities) of the CDOM-related and T1-related fluorophores within the samples. In-situ fluorescence sensors (Chelsea Technologies Group Ltd.) were also used to monitor the T1 fluorescence peak (UviLux Tryptophan) and the CDOM fluorescence peak (UviLux CDOM) during experiments. Tryptophan-like fluorescence was observed (albeit transient) in both spiked and un-spiked standard water samples. By furthering our understanding of aquatic organic matter fluorescence, its origin, transformation, fate and interaction with aquatic microbiological processes, we aim to inform the design of a new generation in-situ fluorescence sensor for the monitoring of aquatic ecosystem health.

  17. Nitrogen transformations in stratified aquatic microbial ecosystems.

    PubMed

    Revsbech, Niels Peter; Risgaard-Petersen, Nils; Schramm, Andreas; Nielsen, Lars Peter

    2006-11-01

    New analytical methods such as advanced molecular techniques and microsensors have resulted in new insights about how nitrogen transformations in stratified microbial systems such as sediments and biofilms are regulated at a microm-mm scale. A large and ever-expanding knowledge base about nitrogen fixation, nitrification, denitrification, and dissimilatory reduction of nitrate to ammonium, and about the microorganisms performing the processes, has been produced by use of these techniques. During the last decade the discovery of anammmox bacteria and migrating, nitrate accumulating bacteria performing dissimilatory reduction of nitrate to ammonium have given new dimensions to the understanding of nitrogen cycling in nature, and the occurrence of these organisms and processes in stratified microbial communities will be described in detail.

  18. Sampling microbial communities in the National Ecological Observatory Network

    NASA Astrophysics Data System (ADS)

    Adams, H. E.; Parnell, J.; Powell, H.

    2012-12-01

    The National Ecological Observatory Network (NEON) is a national-scale research platform to enable the community to assess impacts of climate change, land-use change, and invasive species on ecosystem structure and function at regional and continental scales. The NEON Observatory will collect data on aquatic organisms over 30 years in 36 sites across the United States, including Alaska and Puerto Rico as well as terrestrial organisms at 60 sites including Hawaii. Included in the biological measurements are microbial measurements in terrestrial and aquatic environments, including small, wadeable streams and shallow lakes. Microbial sampling in both aquatic and terrestrial habitats is being planned to coincide with biogeochemical sampling due to similarity of time scale and influence of external drivers. Aquatic sampling is geared toward species diversity and function. Terrestrial sampling aims to collect data on diversity, function, and spatial distribution dynamics. We are in the process of prioritizing data products, so that the most dynamic processes such as enzymatic activity will be measured more frequently and more intensive measures such as metagenome sequence data will be measured on a periodic basis. Here we present our initial microbial sampling strategy and invite the community to provide comment on the design and learn about microbial data products from the Observatory.

  19. Microbial detoxification of metalaxyl in aquatic system.

    PubMed

    Massoud, Ahmed H; Derbalah, Aly S; Belal, El-Sayed B

    2008-01-01

    Four microorganisms, Pseudomonas sp. (ER2), Aspergillus niger (ER6), Cladosporium herbarum (ER4) and Penicilluim sp. (ER3), were isolated from cucumber leaves previously treated with metalaxyl using enrichment technique. These isolates were evaluated for detoxification of metalaxyl at the recommended dose level in aquatic system. The effect of pH and temperature on the growth ability of the tested isolates was also investigated by measuring the intracellular protein and mycelia dry weight for bacterial and fungal isolates, respectively. Moreover, the toxicity of metalaxyl after 28 d of treatment with the tested isolates was evaluated to confirm the complete removal of any toxic materials (metalaxyl and its metabolites). The results showed that the optimum degree pH for the growth of metalaxyl degrading isolates (bacterial and fungal isolates) was 7. The temperature 30 degrees C appeared to be the optimum degree for the growth of either fungal or bacterial isolates. The results showed that Pseudomonas sp. (ER2) was the most effective isolate in metalaxyl degradation followed by Aspergillus niger (ER6), Cladosporium herbarum (ER4) and Penicilluim sp. (ER3), respectively. There is no toxicity of metalaxyl detected in the supernatant after 28 d of treatment with Pseudomonas sp. (ER2). The results suggest that bioremediation by Pseudomonas sp. (ER2) isolate was considered to be effective method for detoxification of metalaxyl in aqueous media.

  20. Photos: Sampling for the National Aquatic Resource Surveys

    EPA Pesticide Factsheets

    The National Aquatic Resource Surveys study all types of waters: rivers and streams, lakes and reservoirs, Great Lakes, coastal waters and wetlands. These photos were taken during in-the-field sampling for recent reports.

  1. Map of National Aquatic Resource Surveys Sampling Locations

    EPA Pesticide Factsheets

    This map displays all of the lakes, rivers and streams, wetlands, and coastal waters sampled by the National Aquatic Resource Surveys, a collaborative EPA program that assesses the condition of the nation's waters using statistical designs.

  2. An endangered oasis of aquatic microbial biodiversity in the Chihuahuan desert

    PubMed Central

    Souza, Valeria; Espinosa-Asuar, Laura; Escalante, Ana E.; Eguiarte, Luis E.; Farmer, Jack; Forney, Larry; Lloret, Lourdes; Rodríguez-Martínez, Juan M.; Soberón, Xavier; Dirzo, Rodolfo; Elser, James J.

    2006-01-01

    The Cuatro Cienegas basin in the Chihuahuan desert is a system of springs, streams, and pools. These ecosystems support >70 endemic species and abundant living stromatolites and other microbial communities, representing a desert oasis of high biodiversity. Here, we combine data from molecular microbiology and geology to document the microbial biodiversity of this unique environment. Ten water samples from locations within the Cuatro Cienegas basin and two neighboring valleys as well as three samples of wet sediments were analyzed. The phylogeny of prokaryotic populations in the samples was determined by characterizing cultured organisms and by PCR amplification and sequencing of 16S rRNA genes from total community DNA. The composition of microbial communities was also assessed by determining profiles of terminal restriction site polymorphisms of 16S rRNA genes in total community DNA. There were 250 different phylotypes among the 350 cultivated strains. Ninety-eight partial 16S rRNA gene sequences were obtained and classified. The clones represented 38 unique phylotypes from ten major lineages of Bacteria and one of Archaea. Unexpectedly, 50% of the phylotypes were most closely related to marine taxa, even though these environments have not been in contact with the ocean for tens of millions of years. Furthermore, terminal restriction site polymorphism profiles and geological data suggest that the aquatic ecosystems of Cuatro Cienegas are hydrologically interconnected with adjacent valleys recently targeted for agricultural intensification. The findings underscore the conservation value of desert aquatic ecosystems and the urgent need for study and preservation of freshwater microbial communities. PMID:16618921

  3. An endangered oasis of aquatic microbial biodiversity in the Chihuahuan desert.

    PubMed

    Souza, Valeria; Espinosa-Asuar, Laura; Escalante, Ana E; Eguiarte, Luis E; Farmer, Jack; Forney, Larry; Lloret, Lourdes; Rodríguez-Martínez, Juan M; Soberón, Xavier; Dirzo, Rodolfo; Elser, James J

    2006-04-25

    The Cuatro Cienegas basin in the Chihuahuan desert is a system of springs, streams, and pools. These ecosystems support >70 endemic species and abundant living stromatolites and other microbial communities, representing a desert oasis of high biodiversity. Here, we combine data from molecular microbiology and geology to document the microbial biodiversity of this unique environment. Ten water samples from locations within the Cuatro Cienegas basin and two neighboring valleys as well as three samples of wet sediments were analyzed. The phylogeny of prokaryotic populations in the samples was determined by characterizing cultured organisms and by PCR amplification and sequencing of 16S rRNA genes from total community DNA. The composition of microbial communities was also assessed by determining profiles of terminal restriction site polymorphisms of 16S rRNA genes in total community DNA. There were 250 different phylotypes among the 350 cultivated strains. Ninety-eight partial 16S rRNA gene sequences were obtained and classified. The clones represented 38 unique phylotypes from ten major lineages of Bacteria and one of Archaea. Unexpectedly, 50% of the phylotypes were most closely related to marine taxa, even though these environments have not been in contact with the ocean for tens of millions of years. Furthermore, terminal restriction site polymorphism profiles and geological data suggest that the aquatic ecosystems of Cuatro Cienegas are hydrologically interconnected with adjacent valleys recently targeted for agricultural intensification. The findings underscore the conservation value of desert aquatic ecosystems and the urgent need for study and preservation of freshwater microbial communities.

  4. MICROBIAL INDICATORS OF AQUATIC ECOSYSTEM CHANGE: CURRENT APPLICATIONS TO EUTROPHICATION STUDIES. (R828677C001)

    EPA Science Inventory

    Human encroachment on aquatic ecosystems is increasing at an unprecedented rate. The impacts of human pollution and habitat alteration are most evident and of greatest concern at the microbial level, where a bulk of production and nutrient cycling takes place. Aquatic ecosyste...

  5. MICROBIAL INDICATORS OF AQUATIC ECOSYSTEM CHANGE: CURRENT APPLICATIONS TO EUTROPHICATION STUDIES. (R828677C001)

    EPA Science Inventory

    Human encroachment on aquatic ecosystems is increasing at an unprecedented rate. The impacts of human pollution and habitat alteration are most evident and of greatest concern at the microbial level, where a bulk of production and nutrient cycling takes place. Aquatic ecosyste...

  6. Identifying the origins of microbially derived aquatic DOM using fluorescence spectroscopy.

    NASA Astrophysics Data System (ADS)

    Fox, Bethany; Thorn, Robin; Anesio, Alexandre; Reynolds, Darren

    2016-04-01

    Dissolved organic matter (DOM) in aquatic systems is an essential support of the microbial population and, therefore, of the entire aquatic ecosystem. Aquatic DOM is also key for global biogeochemical cycling of nutrients and connects land processes to the marine environment via hydrological transportation. There have been multiple advances in technological assessments of the characteristics of aquatic DOM, with spectroscopy becoming widely used. The extensive use of benchtop spectroscopic instruments has led to the development of in situ sensors, improving the spatiotemporal scale of data acquisition. Whilst this has greatly improved understanding of DOM characteristics and patterns, there are still unknown variables, parameters and interactions of DOM within the aquatic environment. In particular, the interactions of aquatic DOM with the microbial population is still mostly unidentified. It is generally accepted that certain DOM fluorescence regions are autochthonous and microbially derived, such as "peak T" fluorescence. However, the origins and metabolic pathways involved in the production and release of these fluorescent molecules is, as yet, not definitively known. Our work focuses on the identification of these metabolic pathways from whence this microbially derived DOM originates. The most recent stage of the research has utilised traditional microbiological techniques, such as growth curves and chemostat experiments, alongside DOM fluorescence spectroscopic analysis and flow cytometry. The information gained regarding the microbial production and processing of DOM is central for the development of novel in situ fluorescence technology, the ultimate aim of this project.

  7. Evaluation of seven aquatic sampling methods for amphibians and other aquatic fauna

    USGS Publications Warehouse

    Gunzburger, M.S.

    2007-01-01

    To design effective and efficient research and monitoring programs researchers must have a thorough understanding of the capabilities and limitations of their sampling methods. Few direct comparative studies exist for aquatic sampling methods for amphibians. The objective of this study was to simultaneously employ seven aquatic sampling methods in 10 wetlands to compare amphibian species richness and number of individuals detected with each method. Four sampling methods allowed counts of individuals (metal dipnet, D-frame dipnet, box trap, crayfish trap), whereas the other three methods allowed detection of species (visual encounter, aural, and froglogger). Amphibian species richness was greatest with froglogger, box trap, and aural samples. For anuran species, the sampling methods by which each life stage was detected was related to relative length of larval and breeding periods and tadpole size. Detection probability of amphibians varied across sampling methods. Box trap sampling resulted in the most precise amphibian count, but the precision of all four count-based methods was low (coefficient of variation > 145 for all methods). The efficacy of the four count sampling methods at sampling fish and aquatic invertebrates was also analyzed because these predatory taxa are known to be important predictors of amphibian habitat distribution. Species richness and counts were similar for fish with the four methods, whereas invertebrate species richness and counts were greatest in box traps. An effective wetland amphibian monitoring program in the southeastern United States should include multiple sampling methods to obtain the most accurate assessment of species community composition at each site. The combined use of frogloggers, crayfish traps, and dipnets may be the most efficient and effective amphibian monitoring protocol. ?? 2007 Brill Academic Publishers.

  8. Changes in microbial community during removal of BDE-153 in four types of aquatic sediments.

    PubMed

    Pan, Ying; Chen, Juan; Zhou, Haichao; Tam, Nora F Y

    2017-09-18

    Indigenous microorganisms in sediments could degrade polybrominated diphenyl ethers (PBDEs), but how the microbial communities respond to PBDEs was seldom reported. The effect of BDE-153, a common congener in aquatic environments, on the microbial communities in four types of aquatic sediments was evaluated during the 150days' incubation under an anaerobic condition. The intrinsic potential to remove BDE-153 varied significantly among four sediment types, and the removal rates of mangrove, mudflat, marine and freshwater sediments were 0.013, 0.013, 0.011, and 0.009day(-1), respectively. The observed microbial species, Simpson, Shannon, and Chao1 indices in all sediments were rather stable and were not changed significantly by BDE-153 amendment. However, BDE-153 amendment altered the microbial community compositions in three saline sediments at the end of the incubation period. Distance-based multivariate multiple regression analysis revealed that salinity, total organic carbon (TOC) and BDE-52, the major debromination product of BDE-153, were the three main factors explaining the variations in microbial community compositions in BDE-treated sediments; whereas salinity, TOC and pH were the main contributing factors in control sediments without BDE-153. The daughter congeners generated during anaerobic debromination process need more attention, especially their effect on the microbial communities in aquatic sediments. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. ANALYSIS OF AQUATIC MICROBIAL COMMUNITIES IMPACTED BY LARGE POULTRY FORMS

    EPA Science Inventory

    Microbial communities often respond more rapidly and extensively to environmental change than communities of higher organisms. Thus, characterizing shifts in the structure of native bacterial communities as a response to changes in nutrients, antimicrobials, and invading pathogen...

  10. ANALYSIS OF AQUATIC MICROBIAL COMMUNITIES IMPACTED BY LARGE POULTRY FORMS

    EPA Science Inventory

    Microbial communities often respond more rapidly and extensively to environmental change than communities of higher organisms. Thus, characterizing shifts in the structure of native bacterial communities as a response to changes in nutrients, antimicrobials, and invading pathogen...

  11. Aquatic hazard assessment of a commercial sample of naphthenic acids.

    PubMed

    Swigert, James P; Lee, Carol; Wong, Diana C L; White, Russell; Scarlett, Alan G; West, Charles E; Rowland, Steven J

    2015-04-01

    This paper presents chemical composition and aquatic toxicity characteristics of a commercial sample of naphthenic acids (NAs). Naphthenic acids are derived from the refining of petroleum middle distillates and can contribute to refinery effluent toxicity. NAs are also present in oil sands process-affected water (OSPW), but differences in the NAs compositions from these sources precludes using a common aquatic toxicity dataset to represent the aquatic hazards of NAs from both origins. Our chemical characterization of a commercial sample of NAs showed it to contain in order of abundance, 1-ring>2-ring>acyclic>3-ring acids (∼84%). Also present were monoaromatic acids (7%) and non-acids (9%, polyaromatic hydrocarbons and sulfur heterocyclic compounds). While the acyclic acids were only the third most abundant group, the five most abundant individual compounds were identified as C(10-14) n-acids (n-decanoic acid to n-tetradecanoic acid). Aquatic toxicity testing of fish (Pimephales promelas), invertebrate (Daphnia magna), algae (Pseudokirchneriella subcapitata), and bacteria (Vibrio fischeri) showed P. promelas to be the most sensitive species with 96-h LL50=9.0 mg L(-1) (LC50=5.6 mg L(-1)). Acute EL50 values for the other species ranged 24-46 mg L(-1) (EC50 values ranged 20-30 mg L(-1)). Biomimetic extraction via solid-phase-microextraction (BE-SPME) suggested a nonpolar narcosis mode of toxic action for D. magna, P. subcapitata, and V. fischeri. The BE analysis under-predicted fish toxicity, which indicates that a specific mode of action, besides narcosis, may be a factor for fishes.

  12. Effect of adaptation to phenol on biodegradation of monosubstituted phenols by aquatic microbial communities

    SciTech Connect

    Shimp, R.J.; Pfaender, F.K.

    1987-07-01

    The adaptation of a mixed aquatic microbial community to phenol was examined in microcosms receiving phenol as a sole carbon source. Extended exposure (adaptation) to phenol resulted in adaptation of the microbial community to the structurally related aromatic compounds m-cresol, m-aminophenol, and p-chlorophenol. The increased biodegradation potential of the phenol-adapted microbial community was accompanied by a concurrent increase in the number of microorganisms able to degrade the three test compounds. Thus, adaptation to the three test chemicals was likely a growth-related result of extended exposure to phenol. The results indicate that adaptation to a single chemical may increase the assimilative capacity of an aquatic environment for other related chemicals even in the absence of adaptation-inducing levels of those materials.

  13. Adaptation of aquatic microbial communities to pollutant stress

    SciTech Connect

    Barkay, T.; Pritchard, H.

    1988-01-01

    Adaptation to biodegradation of p-nitrophenol and to volatilization of Hg/sup 2 +/ are examples of the role the process plays in removal of environmental pollutants and in maintaining active microbial communities in impacted ecosystems. A molecular mechanism of adaptation to Hg/sup 2 +/ is suggested by the enrichment of mercury resistance (MER) genes in some communities upon exposure to mercury.

  14. Rapid assay for microbially reducible ferric iron in aquatic sediments

    USGS Publications Warehouse

    Lovely, Derek R.; Philips , Elizabeth J.P.

    1987-01-01

    The availability of ferric iron for microbial reduction as directly determined by the activity of iron-reducing organisms was compared with its availability as determined by a newly developed chemical assay for microbially reducible iron. The chemical assay was based on the reduction of poorly crystalline ferric iron by hydroxylamine under acidic conditions. There was a strong correlation between the extent to which hydroxylamine could reduce various synthetic ferric iron forms and the susceptibility of the iron to microbial reduction in an enrichment culture of iron-reducing organisms. When sediments that contained hydroxylamine-reducible ferric iron were incubated under anaerobic conditions, ferrous iron accumulated as the concentration of hydroxylamine-reducible ferric iron declined over time. Ferrous iron production stopped as soon as the hydroxylamine-reducible ferric iron was depleted. In anaerobic incubations of reduced sediments that did not contain hydroxylamine-reducible ferric iron, there was no microbial iron reduction, even though the sediments contained high concentrations of oxalate-extractable ferric iron. A correspondence between the presence of hydroxylamine-reducible ferric iron and the extent of ferric iron reduction in anaerobic incubations was observed in sediments from an aquifer and in fresh- and brackish-water sediments from the Potomac River estuary. The assay is a significant improvement over previously described procedures for the determination of hydroxylamine-reducible ferric iron because it provides a correction for the high concentrations of solid ferrous iron which may also be extracted from sediments with acid. This is a rapid, simple technique to determine whether ferric iron is available for microbial reduction.

  15. Rapid assay for microbially reducible ferric iron in aquatic sediments.

    PubMed

    Lovley, D R; Phillips, E J

    1987-07-01

    The availability of ferric iron for microbial reduction as directly determined by the activity of iron-reducing organisms was compared with its availability as determined by a newly developed chemical assay for microbially reducible iron. The chemical assay was based on the reduction of poorly crystalline ferric iron by hydroxylamine under acidic conditions. There was a strong correlation between the extent to which hydroxylamine could reduce various synthetic ferric iron forms and the susceptibility of the iron to microbial reduction in an enrichment culture of iron-reducing organisms. When sediments that contained hydroxylamine-reducible ferric iron were incubated under anaerobic conditions, ferrous iron accumulated as the concentration of hydroxylamine-reducible ferric iron declined over time. Ferrous iron production stopped as soon as the hydroxylamine-reducible ferric iron was depleted. In anaerobic incubations of reduced sediments that did not contain hydroxylamine-reducible ferric iron, there was no microbial iron reduction, even though the sediments contained high concentrations of oxalate-extractable ferric iron. A correspondence between the presence of hydroxylamine-reducible ferric iron and the extent of ferric iron reduction in anaerobic incubations was observed in sediments from an aquifer and in fresh- and brackish-water sediments from the Potomac River estuary. The assay is a significant improvement over previously described procedures for the determination of hydroxylamine-reducible ferric iron because it provides a correction for the high concentrations of solid ferrous iron which may also be extracted from sediments with acid. This is a rapid, simple technique to determine whether ferric iron is available for microbial reduction.

  16. Aquatic Plant/microbial Filters for Treating Septic Tank Effluent

    NASA Technical Reports Server (NTRS)

    Wolverton, B. C.

    1988-01-01

    The use of natural biological processes for treating many types of wastewater have been developed by NASA at the John C. Stennis Space Center, NSTL, Mississippi, during the past 15 years. The simplest form of this technology involves the use of aquatic plant/marsh filters for treatment of septic tank effluent. Septic tank effluent from single home units can be treated to advanced secondary levels and beyond by using a 37.2 sq m (400 sq ft) surface area washed gravel filter. This filter is generally 0.3 m (1 ft) deep with a surface cover of approximately 0.15 m (6 in.) of gravel. The plants in this filter are usually aesthetic or ornamental such as calla lily (Zantedeschia aethiopica), canna lily (Canna flaccida), elephant ear (Colocasia esculenta), and water iris (Iris pseudacorus).

  17. Microbial Characteristics of Native Aquatic Species of Savannah River Wetlands

    SciTech Connect

    McKinsey, P.C.

    2000-12-12

    In 1974 the Savannah River Site (SRS) was established as a National Environmental Research Park (NERP) in the United States. NERP provided locations for long-term ecological research investigation. Many of the ecological studies that have been conducted in the past mainly focused on the macroscopic view. The Savannah River Site contains wetlands that are home to many diverse organisms. We conducted a preliminary survey of microbial habitats in order to explore the biodiversity of species-specific symbionts. Bacterial surveys included viable counts, direct counts, isolation, identification, and metabolic profiles.

  18. Adaptation of Aquatic Microbial Communities to Quaternary Ammonium Compounds

    PubMed Central

    Ventullo, Roy M.; Larson, Robert J.

    1986-01-01

    The effects of long-chain (C12 to C18) quaternary ammonium compounds (QACs) on the density, heterotrophic activity, and biodegradation capabilities of heterotrophic bacteria were examined in situ in a lake ecosystem. Monoalkyl and dialkyl substituted QACs were tested over a range of concentrations (0.001 to 10 mg/liter) in both acute (3 h) and chronic (21 day) exposures. In general, none of the QACs tested had significant adverse effects on bacterial densities in either acute or chronic studies. However, significant decreases in bacterial heterotrophic activity were noted in acute studies at QAC concentrations from 0.1 to 10 mg/liter. Chronic exposure of lake microbial communities to a specific monoalkyl QAC resulted in an adaptive response and recovery of heterotrophic activity. No-observable-effect level in the adapted populations was >10 mg/liter. Chronic exposure also resulted in significant increases in the number and activity of bacteria capable of biodegrading the material. The increase in biodegradation capability was observed at low (microgram per liter) concentrations which are approximately the same as realistic environmental levels. In general, our studies indicated that exposure of lake microbial communities to QACs results in the development of adapted communities which are less sensitive to potential toxic effects and more active in the biodegradation of these materials. PMID:16346991

  19. Microbial ecology of the salmon necrobiome: evidence salmon carrion decomposition influences aquatic and terrestrial insect microbiomes.

    PubMed

    Pechal, Jennifer L; Benbow, M Eric

    2016-05-01

    Carrion decomposition is driven by complex relationships that affect necrobiome community (i.e. all organisms and their genes associated with a dead animal) interactions, such as insect species arrival time to carrion and microbial succession. Little is understood about how microbial communities interact with invertebrates at the aquatic-terrestrial habitat interface. The first objective of the study was to characterize internal microbial communities using high-throughput sequencing of 16S rRNA gene amplicons for aquatic insects (three mayfly species) in streams with salmon carcasses compared with those in streams without salmon carcasses. The second objective was to assess the epinecrotic microbial communities of decomposing salmon carcasses (Oncorhynchus keta) compared with those of terrestrial necrophagous insects (Calliphora terraenovae larvae and adults) associated with the carcasses. There was a significant difference in the internal microbiomes of mayflies collected in salmon carcass-bearing streams and in non-carcass streams, while the developmental stage of blow flies was the governing factor in structuring necrophagous insect internal microbiota. Furthermore, the necrophagous internal microbiome was influenced by the resource on which the larvae developed, and changes in the adult microbiome varied temporally. Overall, these carrion subsidy-driven networks respond to resource pulses with bottom-up effects on consumer microbial structure, as revealed by shifting communities over space and time.

  20. [Seasonal variation of functional diversity of aquatic microbial community in Apostichopus japonicus cultural pond].

    PubMed

    Yan, Fa-Jun; Tian, Xiang-Li; Dong, Shuang-Lin; Yang, Gang

    2014-05-01

    The functional diversity of aquatic microbial communities in sea cucumber (Apostichopus japonicus) cultural ponds was examined in this paper. The Biolog plate technique and redundancy analysis (RDA) method were used to evaluate seasonal changes and their relationships with environmental factors. The results showed that both total amount and types of carbon sources utilized by microbes in the sea cucumber cultural ponds varied seasonally, and were the highest in summer and lowest in winter, with polymers being the main type of carbon sources. Principal component analysis revealed that the carbon utilization diversity of the microbial communities varied significantly over the seasonal courses. A total of 10 categories of carbon sources were significantly related to the principal component 1, among which were polymers, carbohydrates, carboxylic acids, amino acids, and amines. Significant seasonal changes were detected for all carbon utilization diversity indices of the microbial communities, including Shannon, McIntosh, Simpson, and S-E. However, seasonal variations were different among the microbial diversity indices. RDA analysis revealed that TP, NO(3-)-N, TN, and PO4(3-)-P were the critical environmental factors influencing the seasonal changes in functional diversity of aquatic microbial community in sea cucumber cultural ponds.

  1. Probing the pH dependent optical properties of aquatic, terrestrial and microbial humic substances by sodium borohydride reduction

    USDA-ARS?s Scientific Manuscript database

    Chemically reducing humic (HA) and fulvic acids (FA) provides insight into spectroscopically identifiable structural moieties generating the optical properties of HA/FA from aquatic, microbial and terrestrial sources. Sodium borohydride reduction provides targeted reduction of carbonyl groups. The...

  2. Investigations of Sample Stability in Water Chemistry Samples: Implications for the National Aquatic Resource Surveys

    EPA Science Inventory

    Water samples collected for the EPA's National Aquatic Resource Surveys (NARS) typically arrive at an analytical laboratory 2 or 3 days after collection (longer if collected from a remote location), at which point they are stabilized (filtration and/or acid preservation) until an...

  3. Investigations of Sample Stability in Water Chemistry Samples: Implications for the National Aquatic Resource Surveys

    EPA Science Inventory

    Water samples collected for the EPA's National Aquatic Resource Surveys (NARS) typically arrive at an analytical laboratory 2 or 3 days after collection (longer if collected from a remote location), at which point they are stabilized (filtration and/or acid preservation) until an...

  4. Comparison of aquatic macroinvertebrate samples collected using different field methods

    USGS Publications Warehouse

    Lenz, Bernard N.; Miller, Michael A.

    1996-01-01

    Government agencies, academic institutions, and volunteer monitoring groups in the State of Wisconsin collect aquatic macroinvertebrate data to assess water quality. Sampling methods differ among agencies, reflecting the differences in the sampling objectives of each agency. Lack of infor- mation about data comparability impedes data shar- ing among agencies, which can result in duplicated sampling efforts or the underutilization of avail- able information. To address these concerns, com- parisons were made of macroinvertebrate samples collected from wadeable streams in Wisconsin by personnel from the U.S. Geological Survey- National Water Quality Assessment Program (USGS-NAWQA), the Wisconsin Department of Natural Resources (WDNR), the U.S. Department of Agriculture-Forest Service (USDA-FS), and volunteers from the Water Action Volunteer-Water Quality Monitoring Program (WAV). This project was part of the Intergovernmental Task Force on Monitoring Water Quality (ITFM) Wisconsin Water Resources Coordination Project. The numbers, types, and environmental tolerances of the organ- isms collected were analyzed to determine if the four different field methods that were used by the different agencies and volunteer groups provide comparable results. Additionally, this study com- pared the results of samples taken from different locations and habitats within the same streams.

  5. Nutrient enrichment in water more than in leaves affects aquatic microbial litter processing.

    PubMed

    Biasi, Cristiane; Graça, Manuel A S; Santos, Sandro; Ferreira, Verónica

    2017-06-01

    Nutrient enrichment of soils and water will intensify in the future and has the potential to alter fundamental ecosystem processes, such as litter decomposition. We tested the direct (via water nutrient enrichment) and indirect (via changes in leaf chemistry) effects of nutrient enrichment on microbial activity and decomposability of Quercus robur L. (oak) leaves in laboratory microcosms simulating streams. Senescent leaves of oak trees grown without and with fertilization were incubated under ambient and elevated water nutrient [nitrogen (N) and phosphorus (P)] concentrations for 60 days. Soil fertilization led to an increase in leaf (3.4×) and leaf litter (2.3×) N concentration. Increased water-dissolved nutrients concentrations stimulated microbial activity (N uptake, microbial respiration, fungal biomass buildup and conidia production by aquatic hyphomycetes) that translated into accelerated litter decomposition (2.1× for unfertilized and 1.6× for fertilized trees). Leaves from fertilized trees had higher microbial activity and decomposition rates than leaves from unfertilized trees only at low dissolved nutrient availability. When both litter and water nutrients concentration increased, microbial activity and leaf decomposition were stimulated, but the effects were additive and direct effects from increased dissolved nutrient availability were stronger than those mediated by increases in litter N concentration (indirect effects). Our results suggest that increases in water nutrient availability (within the range used in this study) may exert a stronger control on microbial activity and litter decomposition than litter nutrient enrichment.

  6. Microbial plant litter decomposition in aquatic and terrestrial boreal systems along a natural fertility gradient

    NASA Astrophysics Data System (ADS)

    Soares, A. Margarida P. M.; Kritzberg, Emma S.; Rousk, Johannes

    2017-04-01

    Plant litter decomposition is a global ecosystem process, with a crucial role in carbon and nutrient cycling. The majority of litter processing occurs in terrestrial systems, but an important fraction also takes place in inland waters. Among environmental factors, pH impacts the litter decomposition through its selective influence on microbial decomposers. Fungal communities are less affected by pH than bacteria, possibly owing to a wider pH tolerance by this group. On the other hand, bacterial pH optima are constrained to a narrower range of pH values. The microbial decomposition of litter is universally nutrient limited; but few comparisons exist between terrestrial and aquatic systems. We investigated the microbial colonisation and decomposition of plant litter along a fertility gradient, which varied in both pH and N availability in both soil and adjacent water. To do this we installed litterbags with birch (Betula pendula) in streams and corresponding soils in adjacent riparian areas in a boreal system, in Krycklan, Sweden. During the four months covering the ice-free growth season we monitored the successional dynamics of fungal (acetate incorporation into ergosterol) and bacterial growth (thymidine incorporation), microbial respiration in leaf litter, and quantitative and qualitative changes in litter over time. We observed that bacterial growth rates were initially higher in litter decomposing in streams than those in soils, but differences between terrestrial and aquatic bacterial production converged towards the end of the experiment. In litter bags installed in soils, bacterial growth was lower at sites with more acidic pH and lower N availability, while aquatic bacteria were relatively unaffected by the fertility level. Fungal growth rates were two-fold higher for litter decomposing in streams than in soils. In aquatic systems, fungal growth was initially lower in low fertility sites, but differences gradually disappeared over the time course. Fungal

  7. Comparative analyses in aquatic microbial ecology: how far do they go?

    PubMed

    Gasol; Duarte

    2000-02-01

    Methodological developments in recent years have led to an increase in empirical databases on the abundance and functions of aquatic microbes, now allowing synthesis studies. Most of these studies have adopted a comparative approach, such that comparative analyses are now available for most aspects of aquatic microbial food webs (more than 50 papers published in the last 15 years). Some of these analyses apparently yield conflicting results, introducing confusion and unnecessary disputes in the field. We briefly review the comparative analyses so far produced and we highlight generalities, show that some of the perceived discrepancies largely derive from partial analyses of a general underlying trend and formulate predictions based on these general trends that provide new avenues for research.

  8. Microbial community-level toxicity testing of linear alkylbenzene sulfonates in aquatic microcosms.

    PubMed

    Brandt, Kristian K; Jørgensen, Niels O G; Nielsen, Tommy H; Winding, Anne

    2004-08-01

    Complex microbial communities may serve as ideal and ecologically relevant toxicity indicators. We here report an assessment of frequently used methods in microbial ecology for their feasibility to detect toxic effects of the environmentally important surfactant linear alkylbenzene sulfonate (LAS) on microbial communities in lake water and treated waste water. The two microbial communities were evaluated for changes in community structure and function over a period of 7 weeks in replicated aquatic microcosms amended with various levels of LAS (0, 0.1, 1, 10 or 100 mg l(-1)) and inorganic nutrients. In general, the two communities behaved similarly when challenged with LAS. Following lag periods of 1-3 weeks, LAS was degraded to non-toxic substances. Denaturing gradient gel electrophoresis of 16S rRNA gene fragments and [3H]leucine incorporation were the most sensitive assays with effect levels of 0-1 and 1-10 mg LAS l(-1), respectively. Community-level physiological profiles and pollution-induced community tolerance determinations using Biolog microplates demonstrated less sensitivity with effect levels of 10-100 mg LAS l(-1). Total cell counts and net uptake of inorganic N and P were unaffected even at 100 mg LAS l(-1). Interestingly, different microbial communities developed in some replicate microcosms, indicating the importance of stochastic events for community succession. We conclude that microbial community-level toxicity testing holds great promise and suggest a polyphasic approach involving a range of independent methods targeting both the structure and function of the tested microbial communities.

  9. The feasibility of automated online flow cytometry for in-situ monitoring of microbial dynamics in aquatic ecosystems

    PubMed Central

    Besmer, Michael D.; Weissbrodt, David G.; Kratochvil, Bradley E.; Sigrist, Jürg A.; Weyland, Mathias S.; Hammes, Frederik

    2014-01-01

    Fluorescent staining coupled with flow cytometry (FCM) is often used for the monitoring, quantification and characterization of bacteria in engineered and environmental aquatic ecosystems including seawater, freshwater, drinking water, wastewater, and industrial bioreactors. However, infrequent grab sampling hampers accurate characterization and subsequent understanding of microbial dynamics in all of these ecosystems. A logic technological progression is high throughput and full automation of the sampling, staining, measurement, and data analysis steps. Here we assess the feasibility and applicability of automated FCM by means of actual data sets produced with prototype instrumentation. As proof-of-concept we demonstrate examples of microbial dynamics in (i) flowing tap water from a municipal drinking water supply network and (ii) river water from a small creek subject to two rainfall events. In both cases, automated measurements were done at 15-min intervals during 12–14 consecutive days, yielding more than 1000 individual data points for each ecosystem. The extensive data sets derived from the automated measurements allowed for the establishment of baseline data for each ecosystem, as well as for the recognition of daily variations and specific events that would most likely be missed (or miss-characterized) by infrequent sampling. In addition, the online FCM data from the river water was combined and correlated with online measurements of abiotic parameters, showing considerable potential for a better understanding of cause-and-effect relationships in aquatic ecosystems. Although several challenges remain, the successful operation of an automated online FCM system and the basic interpretation of the resulting data sets represent a breakthrough toward the eventual establishment of fully automated online microbiological monitoring technologies. PMID:24917858

  10. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature.

    PubMed

    Tuorto, Steven J; Brown, Chris M; Bidle, Kay D; McGuinness, Lora R; Kerkhof, Lee J

    2015-01-01

    This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry "field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.

  11. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature

    PubMed Central

    Tuorto, Steven J.; Brown, Chris M.; Bidle, Kay D.; McGuinness, Lora R.; Kerkhof, Lee J.

    2015-01-01

    This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry “field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited. PMID:26710122

  12. A generic, process-based model of microbial pollution in aquatic systems

    NASA Astrophysics Data System (ADS)

    Hipsey, Matthew R.; Antenucci, Jason P.; Brookes, Justin D.

    2008-07-01

    Based on a comprehensive synthesis of data available within the literature, a new process-based model of microbial pollution is presented, which is applicable for surface and coastal waters. The model is based on a generic set of parameterisations that describe the dynamics of most protozoan, bacterial and viral organisms of interest, including pathogens and microbial indicator organisms. The parameterisations dynamically account for the effects of temperature, salinity, pH, dissolved oxygen, sunlight, nutrients and turbidity on the growth and mortality of enteric organisms. Parameters for a range of organisms are also presented which are based on collation of literature data. The model has been implemented within an aquatic ecology model, Computational Aquatic Ecosystem Dynamics Model (CAEDYM), which can couple to multidimensional hydrodynamic models. Without adjustment of the literature derived parameter values, a 3-D implementation is validated against observed data from three freshwater systems that differ in their climatic zone, trophic status and operation. The simulations highlight the spatial and temporal variability that may be encountered by operators. Additionally, large differences in the fate and distribution of different species originate from variable rates of growth, mortality and sedimentation and it is emphasized that the use of surrogates for quantifying risk is problematic. The model can be used to help design targeted monitoring programs, explore differences between species, and to support real-time decision-making. Areas where insufficient understanding and data exist are discussed.

  13. Off-site impacts of agricultural composting: role of terrestrially derived organic matter in structuring aquatic microbial communities and their metabolic potential.

    PubMed

    Pommier, Thomas; Merroune, Asmaa; Bettarel, Yvan; Got, Patrice; Janeau, Jean-Louis; Jouquet, Pascal; Thu, Thuy D; Toan, Tran D; Rochelle-Newall, Emma

    2014-12-01

    While considered as sustainable and low-cost agricultural amendments, the impacts of organic fertilizers on downstream aquatic microbial communities remain poorly documented. We investigated the quantity and quality of the dissolved organic matter leaching from agricultural soil amended with compost, vermicompost or biochar and assessed their effects on lake microbial communities, in terms of viral and bacterial abundances, community structure and metabolic potential. The addition of compost and vermicompost significantly increased the amount of dissolved organic carbon in the leachate compared with soil alone. Leachates from these additions, either with or without biochar, were highly bioavailable to aquatic microbial communities, although reducing the metabolic potential of the community and harbouring more specific communities. Although not affecting bacterial richness or taxonomic distributions, the specific addition of biochar affected the original lake bacterial communities, resulting in a strongly different community. This could be partly explained by viral burst and converging bacterial abundances throughout the samples. These results underline the necessity to include off-site impacts of agricultural amendments when considering their cascading effect on downstream aquatic ecosystems.

  14. Effects of riparian plant diversity loss on aquatic microbial decomposers become more pronounced with increasing time.

    PubMed

    Fernandes, Isabel; Duarte, Sofia; Cássio, Fernanda; Pascoal, Cláudia

    2013-11-01

    We examined the potential long-term impacts of riparian plant diversity loss on diversity and activity of aquatic microbial decomposers. Microbial assemblages were obtained in a mixed-forest stream by immersion of mesh bags containing three leaf species (alder, oak and eucalyptus), commonly found in riparian corridors of Iberian streams. Simulation of species loss was done in microcosms by including a set of all leaf species, retrieved from the stream, and non-colonized leaves of three, two or one leaf species. Leaves were renewed every month throughout six months, and microbial inoculum was ensured by a set of colonized leaves from the previous month. Microbial diversity, leaf mass loss and fungal biomass were assessed at the second and sixth months after plant species loss. Molecular diversity of fungi and bacteria, as the total number of operational taxonomic units per leaf diversity treatment, decreased with leaf diversity loss. Fungal biomass tended to decrease linearly with leaf species loss on oak and eucalyptus, suggesting more pronounced effects of leaf diversity on lower quality leaves. Decomposition of alder and eucalyptus leaves was affected by leaf species identity, mainly after longer times following diversity loss. Leaf decomposition of alder decreased when mixed with eucalyptus, while decomposition of eucalyptus decreased in mixtures with oak. Results suggest that the effects of leaf diversity on microbial decomposers depended on leaf species number and also on which species were lost from the system, especially after longer times. This may have implications for the management of riparian forests to maintain stream ecosystem functioning.

  15. Optimization of microbial detoxification for an aquatic mercury-contaminated environment.

    PubMed

    Figueiredo, Neusa L; Canário, João; Serralheiro, Maria Luísa; Carvalho, Cristina

    2017-08-29

    Mercury (Hg) reduction performed by microorganisms is well recognized as a biological means for remediation of contaminated environment. Recently, studies demonstrated that Hg-resistant microorganisms of Tagus Estuary are involved in metal reduction processes. In the present study, aerobic microbial community isolated from a highly Hg-contaminated area of Tagus Estuary was used to determine the optimization of the reduction process in conditions such as the contaminated ecosystem. Factorial design methodology was employed to examine the influence of glucose, sulfate, iron, and chloride on Hg reduction. In the presence of several concentrations of these elements, microbial community reduced Hg in a range of 37-61% of the initial 0.1 mg/ml Hg(2+) levels. The response prediction through central composite design showed that the increase of sulfate concentration led to an optimal response in Hg reduction by microbial community, while the rise in chloride levels markedly decreased metal reduction. Iron may exert antagonistic effects depending upon the media composition. These results are useful in understanding the persistence of Hg contamination in Tagus Estuary after inactivation of critical industrial units, as well as data might also be beneficial for development of new bioremediation strategies either in Tagus Estuary and/or in other Hg-contaminated aquatic environments.

  16. Metaproteomics of aquatic microbial communities in a deep and stratified estuary.

    PubMed

    Colatriano, David; Ramachandran, Arthi; Yergeau, Etienne; Maranger, Roxane; Gélinas, Yves; Walsh, David A

    2015-10-01

    Here we harnessed the power of metaproteomics to assess the metabolic diversity and function of stratified aquatic microbial communities in the deep and expansive Lower St. Lawrence Estuary, located in eastern Canada. Vertical profiling of the microbial communities through the stratified water column revealed differences in metabolic lifestyles and in carbon and nitrogen processing pathways. In productive surface waters, we identified heterotrophic populations involved in the processing of high and low molecular weight organic matter from both terrestrial (e.g. cellulose and xylose) and marine (e.g. organic compatible osmolytes) sources. In the less productive deep waters, chemosynthetic production coupled to nitrification by MG-I Thaumarchaeota and Nitrospina appeared to be a dominant metabolic strategy. Similar to other studies of the coastal ocean, we identified methanol oxidation proteins originating from the common OM43 marine clade. However, we also identified a novel lineage of methanol-oxidizers specifically in the particle-rich bottom (i.e. nepheloid) layer. Membrane transport proteins assigned to the uncultivated MG-II Euryarchaeota were also specifically detected in the nepheloid layer. In total, these results revealed strong vertical structure of microbial taxa and metabolic activities, as well as the presence of specific "nepheloid" taxa that may contribute significantly to coastal ocean nutrient cycling.

  17. Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments (Final Report)

    EPA Science Inventory

    The Ecological Risk Assessment Support Center (ERASC) announced the release of the final report, Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments. This technical paper provides defensible approximations fo...

  18. Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments (Final Report)

    EPA Science Inventory

    The Ecological Risk Assessment Support Center (ERASC) announced the release of the final report, Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments. This technical paper provides defensible approximations fo...

  19. Microbial impact of Canada geese (Branta canadensis) and whistling swans (Cygnus columbianus columbianus) on aquatic ecosystems.

    PubMed

    Hussong, D; Damaré, J M; Limpert, R J; Sladen, W J; Weiner, R M; Colwell, R R

    1979-01-01

    Quantitative and qualitative analyses of the intestinal bacterial flora of Canada geese and whistling swans were carried out with the finding that wild birds harbor significantly more fecal coliforms than fecal streptococci. The reverse was typical of captive and fasting birds. Neither Salmonella spp. nor Shigella spp. were isolated from 44 migratory waterfowl that were wintering in the Chesapeake Bay region. Enteropathogenic Escherichia coli were detected in seven birds. Geese eliminated 10(7) and swans 10(9) fecal coliforms per day. Results of in situ studies showed that large flocks of waterfowl can cause elevated fecal coliform densities in the water column. From the data obtained in this study, it is possible to predict the microbial impact of migratory waterfowl upon aquatic roosting sites.

  20. A synthesis of the effects of pesticides on microbial persistence in aquatic ecosystems.

    PubMed

    Staley, Zachery R; Harwood, Valerie J; Rohr, Jason R

    2015-01-01

    Pesticides have a pervasive presence in aquatic ecosystems throughout the world. While pesticides are intended to control fungi, insects, and other pests, their mechanisms of action are often not specific enough to prevent unintended effects, such as on non-target microbial populations. Microorganisms, including algae and cyanobacteria, protozoa, aquatic fungi, and bacteria, form the basis of many food webs and are responsible for crucial aspects of biogeochemical cycling; therefore, the potential for pesticides to alter microbial community structures must be understood to preserve ecosystem services. This review examines studies that focused on direct population-level effects and indirect community-level effects of pesticides on microorganisms. Generally, insecticides, herbicides, and fungicides were found to have adverse direct effects on algal and fungal species. Insecticides and fungicides also had deleterious direct effects in the majority of studies examining protozoa species, although herbicides were found to have inconsistent direct effects on protozoans. Our synthesis revealed mixed or no direct effects on bacterial species among all pesticide categories, with results highly dependent on the target species, chemical, and concentration used in the study. Examination of community-level, indirect effects revealed that all pesticide categories had a tendency to reduce higher trophic levels, thereby diminishing top-down pressures and favoring lower trophic levels. Often, indirect effects exerted greater influence than direct effects. However, few studies have been conducted to specifically address community-level effects of pesticides on microorganisms, and further research is necessary to better understand and predict the net effects of pesticides on ecosystem health.

  1. A Synthesis of the Effects of Pesticides on Microbial Persistence in Aquatic Ecosystems

    PubMed Central

    Staley, Zachery R.; Harwood, Valerie J.; Rohr, Jason R.

    2016-01-01

    Pesticides are a pervasive presence in aquatic ecosystems throughout the world. While pesticides are intended to control fungi, insects, and other pests, their mechanisms of action are often not specific enough to prevent unintended effects, such as on non-target microbial populations. Microorganisms, including algae and cyanobacteria, protozoa, aquatic fungi, and bacteria, form the basis of many food webs and are responsible for crucial aspects of biogeochemical cycling; therefore, the potential for pesticides to alter microbial community structures must be understood to preserve ecosystem services. This review examines studies that focused on direct population-level effects and indirect community-level effects of pesticides on microorganisms. Generally, insecticides, herbicides, and fungicides were found to have adverse direct effects on algal and fungal species. Insecticides and fungicides also had deleterious direct effects in the majority of studies examining protozoa species, although herbicides were found to have inconsistent direct effects on protozoans. Our synthesis revealed mixed or no direct effects on bacterial species among all pesticide categories, with results highly dependent on the target species, chemical, and concentration used in the study. Examination of community-level, indirect effects revealed that all pesticide categories had a tendency to reduce higher trophic levels, thereby diminishing top-down pressures and favoring lower trophic levels. Often, indirect effects exerted greater influence than direct effects. However, few studies have been conducted to specifically address community-level effects of pesticides on microorganisms and further research is necessary to better understand and predict the net effects of pesticides on ecosystem health. PMID:26565685

  2. Microbial Condition of Water Samples from Foreign Fuel Storage Facilities

    SciTech Connect

    Berry, C.J.; Fliermans, C.B.; Santo Domingo, J.

    1997-10-30

    In order to assess the microbial condition of foreign nuclear fuel storage facilities, fourteen different water samples were received from facilities outside the United States that have sent spent nuclear fuel to SRS for wet storage. Each water sample was analyzed for microbial content and activity as determined by total bacteria, viable aerobic bacteria, viable anaerobic bacteria, viable sulfate- reducing bacteria, viable acid-producing bacteria and enzyme diversity. The results for each water sample were then compared to other foreign samples and to data from the receiving basin for off- site fuel (RBOF) at SRS.

  3. Microbial Activity in Aquatic Environments Measured by Dimethyl Sulfoxide Reduction and Intercomparison with Commonly Used Methods

    PubMed Central

    Griebler, Christian; Slezak, Doris

    2001-01-01

    A new method to determine microbial (bacterial and fungal) activity in various freshwater habitats is described. Based on microbial reduction of dimethyl sulfoxide (DMSO) to dimethyl sulfide (DMS), our DMSO reduction method allows measurement of the respiratory activity in interstitial water, as well as in the water column. DMSO is added to water samples at a concentration (0.75% [vol/vol] or 106 mM) high enough to compete with other naturally occurring electron acceptors, as determined with oxygen and nitrate, without stimulating or inhibiting microbial activity. Addition of NaN3, KCN, and formaldehyde, as well as autoclaving, inhibited the production of DMS, which proves that the reduction of DMSO is a biotic process. DMSO reduction is readily detectable via the formation of DMS even at low microbial activities. All water samples showed significant DMSO reduction over several hours. Microbially reduced DMSO is recovered in the form of DMS from water samples by a purge and trap system and is quantified by gas chromatography and detection with a flame photometric detector. The DMSO reduction method was compared with other methods commonly used for assessment of microbial activity. DMSO reduction activity correlated well with bacterial production in predator-free batch cultures. Cell-production-specific DMSO reduction rates did not differ significantly in batch cultures with different nutrient regimes but were different in different growth phases. Overall, a cell-production-specific DMSO reduction rate of 1.26 × 10−17 ± 0.12 × 10−17 mol of DMS per produced cell (mean ± standard error; R2 = 0.78) was calculated. We suggest that the relationship of DMSO reduction rates to thymidine and leucine incorporation is linear (the R2 values ranged from 0.783 to 0.944), whereas there is an exponential relationship between DMSO reduction rates and glucose uptake, as well as incorporation (the R2 values ranged from 0.821 to 0.931). Based on our results, we conclude that

  4. [A membrane filter sampling method for determining microbial air pollution].

    PubMed

    Cherneva, P; Kiranova, A

    1996-01-01

    The method is a contribution in the evaluation of the exposition and the control of the standards for organic powders. The method concerns the sample-taking procedure and the analysis-making technique for determining of the concentration of the microbial pollution of the air. It is based on filtering of some amount of air through a membrane filter which is then processed for cultivating of microbial colonies on its surface. The results are obtained in number of microbial colonies per unit of air. The method presents opportunity to select and vary the filtered volume of air, to determine the respirable fraction, to determine the personal exposition, as well as for the simultaneous determining of the microbial pollution together with other important parameters of the particle pollutants of the air (metal, fibre and others).

  5. Long-term environmental trends: selection of sampling locations in a reactor-aquatic cooling system.

    PubMed

    Revsin, B K; Watson, J E

    1993-02-01

    The study objective was to determine whether environmental radionuclide accumulations were occurring in an aquatic system with a 13-y history of supplying a power plant with reactor-cooling water as well as receiving plant discharge. The aquatic system consisted of the following: 1) a reactor-cooling lake; 2) a secondary lake approximately 8 km downstream; and 3) a small stream that interfaced with the two lakes. Gamma-emitting radionuclides were identified and quantified in samples of benthic sediments obtained from representative areas of the aquatic system. This study demonstrated that in a reactor-aquatic cooling system, the component of the aquatic system most likely to experience radionuclide accumulation will not necessarily be the reactor-cooling lake, but will be that component of the aquatic system whose benthic sediments contain the highest concentrations of organic matter. Further, it was shown that the quantity of oxidizable organic matter present in a sediment is a good predictor or marker for potential sites of radionuclide accumulation (i.e., 60Co and 137Cs).

  6. When is the best time to sample aquatic macroinvertebrates in ponds for biodiversity assessment?

    PubMed

    Hill, M J; Sayer, C D; Wood, P J

    2016-03-01

    Ponds are sites of high biodiversity and conservation value, yet there is little or no statutory monitoring of them across most of Europe. There are clear and standardised protocols for sampling aquatic macroinvertebrate communities in ponds, but the most suitable time(s) to undertake the survey(s) remains poorly specified. This paper examined the aquatic macroinvertebrate communities from 95 ponds within different land use types over three seasons (spring, summer and autumn) to determine the most appropriate time to undertake sampling to characterise biodiversity. The combined samples from all three seasons provided the most comprehensive record of the aquatic macroinvertebrate taxa recorded within ponds (alpha and gamma diversity). Samples collected during the autumn survey yielded significantly greater macroinvertebrate richness (76% of the total diversity) than either spring or summer surveys. Macroinvertebrate diversity was greatest during autumn in meadow and agricultural ponds, but taxon richness among forest and urban ponds did not differ significantly temporally. The autumn survey provided the highest measures of richness for Coleoptera, Hemiptera and Odonata. However, richness of the aquatic insect order Trichoptera was highest in spring and lowest in autumn. The results illustrate that multiple surveys, covering more than one season, provide the most comprehensive representation of macroinvertebrate biodiversity. When sampling can only be undertaken on one occasion, the most appropriate time to undertake surveys to characterise the macroinvertebrate community biodiversity is during autumn, although this may need to be modified if other floral and faunal groups need to be incorporated into the sampling programme.

  7. Defining Established and Emerging Microbial Risks in the Aquatic Environment: Current Knowledge, Implications, and Outlooks

    PubMed Central

    Rowan, Neil J.

    2011-01-01

    This timely review primarily addresses important but presently undefined microbial risks to public health and to the natural environment. It specifically focuses on current knowledge, future outlooks and offers some potential alleviation strategies that may reduce or eliminate the risk of problematic microbes in their viable but nonculturable (VBNC) state and Cryptosporidium oocysts in the aquatic environment. As emphasis is placed on water quality, particularly surrounding efficacy of decontamination at the wastewater treatment plant level, this review also touches upon other related emerging issues, namely, the fate and potential ecotoxicological impact of untreated antibiotics and other pharmaceutically active compounds in water. Deciphering best published data has elucidated gaps between science and policy that will help stakeholders work towards the European Union's Water Framework Directive (2000/60/EC), which provides an ambitious legislative framework for water quality improvements within its region and seeks to restore all water bodies to “good ecological status” by 2015. Future effective risk-based assessment and management, post definition of the plethora of dynamic inter-related factors governing the occurrence, persistence and/or control of these presently undefined hazards in water will also demand exploiting and harnessing tangential advances in allied disciplines such as mathematical and computer modeling that will permit efficient data generation and transparent reporting to be undertaken by well-balanced consortia of stakeholders. PMID:20976256

  8. Harmonisation of microbial sampling and testing methods for distillate fuels

    SciTech Connect

    Hill, G.C.; Hill, E.C.

    1995-05-01

    Increased incidence of microbial infection in distillate fuels has led to a demand for organisations such as the Institute of Petroleum to propose standards for microbiological quality, based on numbers of viable microbial colony forming units. Variations in quality requirements, and in the spoilage significance of contaminating microbes plus a tendency for temporal and spatial changes in the distribution of microbes, makes such standards difficult to implement. The problem is compounded by a diversity in the procedures employed for sampling and testing for microbial contamination and in the interpretation of the data obtained. The following paper reviews these problems and describes the efforts of The Institute of Petroleum Microbiology Fuels Group to address these issues and in particular to bring about harmonisation of sampling and testing methods. The benefits and drawbacks of available test methods, both laboratory based and on-site, are discussed.

  9. Nonylphenol and estrogenic activity in aquatic environmental samples

    SciTech Connect

    Tanghe, T.; Devriese, G.; Verstraete, W.

    1999-03-01

    The authors surveyed a series of surface waters and sewage treatment plants in Flanders (north of Belgium) for the presence of estrogenic activity and a xeno-estrogenic compound para-nonylphenol (NP), respectively. The surface waters of rural origin, used for drinking water production were free of significant levels of estrogenic activity and NP. Domestic sewage, after proper treatment, appeared to be no major source of this chemical. Yet, in some industrial effluents and surface waters of highly industrialized regions, NP and/or estrogenic activity was prominent, that is, <1 to 122 {micro}g NP/L and 11 to 42 {micro}g NP/L, respectively. This is because of the ongoing use of NP polyethoxylates in industry. The response of the recombinant yeast estrogen assay to the environmental samples tested was not consistent with the detected concentrations of NP. Standard addition of a natural estrogen, 17{beta}-estradiol, generated no or a reduced response compared to the standard curve concentration. Application of humic acids to standard series of NP and 17{beta}-estradiol resulted in a dose-dependent decrease of the estrogenic response. It appears that this bioassay is subject to considerable interferences due to the complexity of environmental samples. Parallel implementation of extensive chemical screening for xenobiotics and use of the bioassay are needed for adequate assessment of the potential estrogenic hazard to avoid false negative evaluations.

  10. Citrobacter portucalensis sp. nov., isolated from an aquatic sample.

    PubMed

    Ribeiro, Teresa Gonçalves; Gonçalves, Bruno Ribeiro; da Silva, Mickael Santos; Novais, Ângela; Machado, Elisabete; Carriço, João André; Peixe, Luísa

    2017-09-01

    A Gram-stain-negative strain, A60T, isolated from a water well sample in Portugal, was characterized phenotypically, genotypically and phylogenetically. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain A60T belonged to the genus Citrobacter, and recN gene phylogeny revealed one strongly supported clade encompassing strain A60T and 13 other strains from public databases, distinct from currently recognized species of the genus Citrobacter. Furthermore, multilocus sequence analysis (MLSA) based on concatenated partial fusA, leuS, pyrG and rpoB sequences confirmed the classification obtained with the recN sequence. In silico genomic comparisons, including average nucleotide identity (ANI) and the genome-to-genome distance calculator (GGDC), showed 94.6 % and 58.4 % identity to the closest relative Citrobacter freundii ATCC 8090T, respectively. The ability to metabolize different compounds further discriminated strain A60T from other species of the genus Citrobacter. The G+C content of strain A60T is 52.0 %. The results obtained support the description of a novel species within the genus Citrobacter, for which the name Citrobacter portucalensis sp. nov. is proposed, with the type strain A60T (=DSM 104542T=CECT 9236T).

  11. Comparative effects of a genetically engineered insect virus and a growth-regulating insecticide on microbial communities in aquatic microcosms.

    PubMed

    Kreutzweiser, D; England, L; Shepherd, J; Conklin, J; Holmes, S

    2001-01-01

    The effects of a genetically engineered insect baculovirus on indigenous aquatic microbial communities were determined in closed, recirculating aquatic microcosms, and compared with the effects of a natural strain of the virus and of a growth-regulating insecticide, Dimilin. The recombinant virus was a nuclear polyhedrosis virus (NPV) of the spruce budworm (Choristoneura fumiferana (Cf)) with a lacZ marker gene inserted into the egt region of the CfNPV. The natural virus was Ireland strain CfNPV. Microbial measurement endpoints included decomposition activity (mass loss of organic material), respiration on two different substrates (O2 consumption), heterotrophic bacterial abundance (plate counts), and microbial community metabolic profiles (carbon source utilization patterns in Biolog GN microplates). Viral DNA of both the natural strain and the recombinant viruses, detected by polymerase chain reaction techniques, settled out of the microcosm water and accumulated on bottom substrates within 3 days of the microcosm inoculations. The viral DNA persisted in bottom substrates for the duration of the 21-day experimental period, although there was some evidence that the recombinant virus was less stable than the natural strain in particulate organic matter. No significant changes in microbial decomposition or respiration activity, bacterial abundance, or average metabolic responses were detected by a time trend analysis in microcosms inoculated with either the lacZ recombinant virus or the natural Ireland strain CfNPV. Significant effects on microbial decomposition and respiration activity were detected in microcosms treated with the growth-regulating insecticide at, and above, the expected environmental concentrations. Despite significant effects on microbial community functional attributes in Dimilin-treated microcosms, there were no detectable changes in community structure in terms of metabolic profiles or bacterial abundance.

  12. Microbial Lineages in Sarcoidosis: A Metagenomic Analysis Tailored for Low Microbial Content Samples.

    PubMed

    Clarke, Erik L; Lauder, Abigail P; Hofstaedter, Casey E; Hwang, Young; Fitzgerald, Ayannah S; Imai, Ize; Biernat, Wojciech; Rękawiecki, Bartłomiej; Majewska, Hanna; Dubaniewicz, Anna; Litzky, Leslie A; Feldman, Michael D; Bittinger, Kyle; Rossman, Milton D; Patterson, Karen C; Bushman, Frederic D; Collman, Ronald G

    2017-08-28

    The etiology of sarcoidosis is unknown, but microbial agents are suspected as triggers. We sought to identify bacterial, fungal or viral lineages in specimens from sarcoidosis patients enriched relative to controls using metagenomic DNA sequencing. Since DNA from environmental contamination contributes disproportionately to samples with low authentic microbial content, we developed improved methods for filtering environmental contamination. We analyzed specimens from sarcoidosis subjects (n=93), non-sarcoidosis control subjects (n=72) and various environmental controls (n=150). Sarcoidosis specimens consisted of two independent sets of formalin-fixed, paraffin-embedded lymph node biopsies, bronchoalveolar lavage (BAL), Kveim reagent, and fresh granulomatous spleen from a sarcoidosis patient. All specimens were analyzed by bacterial 16S and fungal ITS rRNA gene sequencing. In addition, BAL was analyzed by shotgun sequencing of fractions enriched for viral particles, and Kveim and spleen were subjected to whole-genome shotgun sequencing. In one tissue set, fungi in the Cladosporiaceae family were enriched in sarcoidosis compared to non-sarcoidosis tissues; in the other tissue set, we detected enrichment of several bacterial lineages in sarcoidosis, but not Cladosporiaceae. BAL showed limited enrichment of Aspergillus fungi. Several microbial lineages were detected in Kveim and spleen, including Cladosporium. No microbial lineage was enriched in more than one sample type after correction for multiple comparisons. Metagenomic sequencing revealed enrichment of microbes in single types of sarcoidosis samples, but limited concordance across sample types. Statistical analysis accounting for environmental contamination was essential to avoiding false positives.

  13. Elucidating Microbial Adaptation Dynamics via Autonomous Exposure and Sampling

    NASA Technical Reports Server (NTRS)

    Grace, Joseph M.; Verseux, Cyprien; Gentry, Diana; Moffet, Amy; Thayabaran, Ramanen; Wong, Nathan; Rothschild, Lynn

    2013-01-01

    The adaptation of micro-organisms to their environments is a complex process of interaction between the pressures of the environment and of competition. Reducing this multifactorial process to environmental exposure in the laboratory is a common tool for elucidating individual mechanisms of evolution, such as mutation rates. Although such studies inform fundamental questions about the way adaptation and even speciation occur, they are often limited by labor-intensive manual techniques. Current methods for controlled study of microbial adaptation limit the length of time, the depth of collected data, and the breadth of applied environmental conditions. Small idiosyncrasies in manual techniques can have large effects on outcomes; for example, there are significant variations in induced radiation resistances following similar repeated exposure protocols. We describe here a project under development to allow rapid cycling of multiple types of microbial environmental exposure. The system allows continuous autonomous monitoring and data collection of both single species and sampled communities, independently and concurrently providing multiple types of controlled environmental pressure (temperature, radiation, chemical presence or absence, and so on) to a microbial community in dynamic response to the ecosystem's current status. When combined with DNA sequencing and extraction, such a controlled environment can cast light on microbial functional development, population dynamics, inter- and intra-species competition, and microbe-environment interaction. The project's goal is to allow rapid, repeatable iteration of studies of both natural and artificial microbial adaptation. As an example, the same system can be used both to increase the pH of a wet soil aliquot over time while periodically sampling it for genetic activity analysis, or to repeatedly expose a culture of bacteria to the presence of a toxic metal, automatically adjusting the level of toxicity based on the

  14. Improved Method for Recovery of mRNA from Aquatic Samples and Its Application to Detection of mer Expression

    PubMed Central

    Jeffrey, Wade H.; Nazaret, Sylvie; Von Haven, Robin

    1994-01-01

    Previously described methods for extraction of mRNA from environmental samples may preclude detecting transcripts from genes that were present in low abundance in aquatic bacterial communities. By combining a boiling sodium dodecyl sulfate-diethylpyrocarbonate lysis step with acid-guanidinium extraction, we improved recovery of target mRNA from both pure cultures and environmental samples. The most significant advantage of the new protocol is that it is easily adapted to yield high recovery of mRNA from 142-mm-diameter flat filters and high-capacity cartridge filters. The lysis and extraction procedures are more rapid than previously described methods, and many samples can be handled at once. RNA extracts have been shown to be free of contaminating DNA. The lysis procedure does not damage target mRNA sequences, and mRNA can be detected from fewer than 106 bacterial cells. We used the new method to examine transcripts of genes responsible for detoxification of mercurial compounds. Induction of merA (specifying mercuric reductase) transcripts in stationary-phase Pseudomonas aeruginosa containing Tn501 occurred within 60 s of HgCl2 addition and was proportional to the amount of Hg(II) added. The new technique also allowed the detection of merA transcripts from the microbial community of a mercury-contaminated pond (Reality Lake, Oak Ridge, Tenn.). Significant differences in merA transcript abundance were observed between different locations associated with the lake. The results indicate that the new method is simple and rapid and can be applied to the study of mer gene expression of aquatic communities in their natural habitats. PMID:16349274

  15. Aquatic Environments

    USDA-ARS?s Scientific Manuscript database

    Aquatic microbiology can be defined as the study of microorganisms and microbial communities in water environments. Aquatic environments occupy more than 70% of the earth’s surface including oceans, estuaries, rivers, lakes, wetlands, streams, springs, and aquifers. Water is essential for life and m...

  16. Internal nitrogen removal from sediments by the hybrid system of microbial fuel cells and submerged aquatic plants.

    PubMed

    Xu, Peng; Xiao, En-Rong; Xu, Dan; Zhou, Yin; He, Feng; Liu, Bi-Yun; Zeng, Lei; Wu, Zhen-Bin

    2017-01-01

    Sediment internal nitrogen release is a significant pollution source in the overlying water of aquatic ecosystems. This study aims to remove internal nitrogen in sediment-water microcosms by coupling sediment microbial fuel cells (SMFCs) with submerged aquatic plants. Twelve tanks including four treatments in triplicates were designed: open-circuit (SMFC-o), closed-circuit (SMFC-c), aquatic plants with open-circuit (P-SMFC-o) and aquatic plants with closed-circuit (P-SMFC-c). The changes in the bio-electrochemical characteristics of the nitrogen levels in overlying water, pore water, sediments, and aquatic plants were documented to explain the migration and transformation pathways of internal nitrogen. The results showed that both electrogenesis and aquatic plants could facilitate the mineralization of organic nitrogen in sediments. In SMFC, electrogenesis promoted the release of ammonium from the pore water, followed by the accumulation of ammonium and nitrate in the overlying water. The increased redox potential of sediments due to electrogenesis also contributed to higher levels of nitrate in overlying water when nitrification in pore water was facilitated and denitrification at the sediment-water interface was inhibited. When the aquatic plants were introduced into the closed-circuit SMFC, the internal ammonium assimilation by aquatic plants was advanced by electrogenesis; nitrification in pore water and denitrification in sediments were also promoted. These processes might result in the maximum decrease of internal nitrogen with low nitrogen levels in the overlying water despite the lower power production. The P-SMFC-c reduced 8.1%, 16.2%, 24.7%, and 25.3% of internal total nitrogen compared to SMFC-o on the 55th, 82th, 136th, and 190th days, respectively. The smaller number of Nitrospira and the larger number of Bacillus and Pseudomonas on the anodes via high throughput sequencing may account for strong mineralization and denitrification in the sediments

  17. Internal nitrogen removal from sediments by the hybrid system of microbial fuel cells and submerged aquatic plants

    PubMed Central

    Xu, Peng; Xiao, En-Rong; Xu, Dan; Zhou, Yin; He, Feng; Liu, Bi-Yun; Zeng, Lei; Wu, Zhen-Bin

    2017-01-01

    Sediment internal nitrogen release is a significant pollution source in the overlying water of aquatic ecosystems. This study aims to remove internal nitrogen in sediment-water microcosms by coupling sediment microbial fuel cells (SMFCs) with submerged aquatic plants. Twelve tanks including four treatments in triplicates were designed: open-circuit (SMFC-o), closed-circuit (SMFC-c), aquatic plants with open-circuit (P-SMFC-o) and aquatic plants with closed-circuit (P-SMFC-c). The changes in the bio-electrochemical characteristics of the nitrogen levels in overlying water, pore water, sediments, and aquatic plants were documented to explain the migration and transformation pathways of internal nitrogen. The results showed that both electrogenesis and aquatic plants could facilitate the mineralization of organic nitrogen in sediments. In SMFC, electrogenesis promoted the release of ammonium from the pore water, followed by the accumulation of ammonium and nitrate in the overlying water. The increased redox potential of sediments due to electrogenesis also contributed to higher levels of nitrate in overlying water when nitrification in pore water was facilitated and denitrification at the sediment-water interface was inhibited. When the aquatic plants were introduced into the closed-circuit SMFC, the internal ammonium assimilation by aquatic plants was advanced by electrogenesis; nitrification in pore water and denitrification in sediments were also promoted. These processes might result in the maximum decrease of internal nitrogen with low nitrogen levels in the overlying water despite the lower power production. The P-SMFC-c reduced 8.1%, 16.2%, 24.7%, and 25.3% of internal total nitrogen compared to SMFC-o on the 55th, 82th, 136th, and 190th days, respectively. The smaller number of Nitrospira and the larger number of Bacillus and Pseudomonas on the anodes via high throughput sequencing may account for strong mineralization and denitrification in the sediments

  18. A method for sampling microbial aerosols using high altitude balloons.

    PubMed

    Bryan, N C; Stewart, M; Granger, D; Guzik, T G; Christner, B C

    2014-12-01

    Owing to the challenges posed to microbial aerosol sampling at high altitudes, very little is known about the abundance, diversity, and extent of microbial taxa in the Earth-atmosphere system. To directly address this knowledge gap, we designed, constructed, and tested a system that passively samples aerosols during ascent through the atmosphere while tethered to a helium-filled latex sounding balloon. The sampling payload is ~ 2.7 kg and comprised of an electronics box and three sampling chambers (one serving as a procedural control). Each chamber is sealed with retractable doors that can be commanded to open and close at designated altitudes. The payload is deployed together with radio beacons that transmit GPS coordinates (latitude, longitude and altitude) in real time for tracking and recovery. A cut mechanism separates the payload string from the balloon at any desired altitude, returning all equipment safely to the ground on a parachute. When the chambers are opened, aerosol sampling is performed using the Rotorod® collection method (40 rods per chamber), with each rod passing through 0.035 m3 per km of altitude sampled. Based on quality control measurements, the collection of ~ 100 cells rod(-1) provided a 3-sigma confidence level of detection. The payload system described can be mated with any type of balloon platform and provides a tool for characterizing the vertical distribution of microorganisms in the troposphere and stratosphere. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Bacterial Diversity Assessment in Antarctic Terrestrial and Aquatic Microbial Mats: A Comparison between Bidirectional Pyrosequencing and Cultivation

    PubMed Central

    Tytgat, Bjorn; Verleyen, Elie; Obbels, Dagmar; Peeters, Karolien; De Wever, Aaike; D’hondt, Sofie; De Meyer, Tim; Van Criekinge, Wim; Vyverman, Wim; Willems, Anne

    2014-01-01

    The application of high-throughput sequencing of the 16S rRNA gene has increased the size of microbial diversity datasets by several orders of magnitude, providing improved access to the rare biosphere compared with cultivation-based approaches and more established cultivation-independent techniques. By contrast, cultivation-based approaches allow the retrieval of both common and uncommon bacteria that can grow in the conditions used and provide access to strains for biotechnological applications. We performed bidirectional pyrosequencing of the bacterial 16S rRNA gene diversity in two terrestrial and seven aquatic Antarctic microbial mat samples previously studied by heterotrophic cultivation. While, not unexpectedly, 77.5% of genera recovered by pyrosequencing were not among the isolates, 25.6% of the genera picked up by cultivation were not detected by pyrosequencing. To allow comparison between both techniques, we focused on the five phyla (Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes and Deinococcus-Thermus) recovered by heterotrophic cultivation. Four of these phyla were among the most abundantly recovered by pyrosequencing. Strikingly, there was relatively little overlap between cultivation and the forward and reverse pyrosequencing-based datasets at the genus (17.1–22.2%) and OTU (3.5–3.6%) level (defined on a 97% similarity cut-off level). Comparison of the V1–V2 and V3–V2 datasets of the 16S rRNA gene revealed remarkable differences in number of OTUs and genera recovered. The forward dataset missed 33% of the genera from the reverse dataset despite comprising 50% more OTUs, while the reverse dataset did not contain 40% of the genera of the forward dataset. Similar observations were evident when comparing the forward and reverse cultivation datasets. Our results indicate that the region under consideration can have a large impact on perceived diversity, and should be considered when comparing different datasets. Finally, a high number

  20. Bacterial diversity assessment in Antarctic terrestrial and aquatic microbial mats: a comparison between bidirectional pyrosequencing and cultivation.

    PubMed

    Tytgat, Bjorn; Verleyen, Elie; Obbels, Dagmar; Peeters, Karolien; De Wever, Aaike; D'hondt, Sofie; De Meyer, Tim; Van Criekinge, Wim; Vyverman, Wim; Willems, Anne

    2014-01-01

    The application of high-throughput sequencing of the 16S rRNA gene has increased the size of microbial diversity datasets by several orders of magnitude, providing improved access to the rare biosphere compared with cultivation-based approaches and more established cultivation-independent techniques. By contrast, cultivation-based approaches allow the retrieval of both common and uncommon bacteria that can grow in the conditions used and provide access to strains for biotechnological applications. We performed bidirectional pyrosequencing of the bacterial 16S rRNA gene diversity in two terrestrial and seven aquatic Antarctic microbial mat samples previously studied by heterotrophic cultivation. While, not unexpectedly, 77.5% of genera recovered by pyrosequencing were not among the isolates, 25.6% of the genera picked up by cultivation were not detected by pyrosequencing. To allow comparison between both techniques, we focused on the five phyla (Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes and Deinococcus-Thermus) recovered by heterotrophic cultivation. Four of these phyla were among the most abundantly recovered by pyrosequencing. Strikingly, there was relatively little overlap between cultivation and the forward and reverse pyrosequencing-based datasets at the genus (17.1-22.2%) and OTU (3.5-3.6%) level (defined on a 97% similarity cut-off level). Comparison of the V1-V2 and V3-V2 datasets of the 16S rRNA gene revealed remarkable differences in number of OTUs and genera recovered. The forward dataset missed 33% of the genera from the reverse dataset despite comprising 50% more OTUs, while the reverse dataset did not contain 40% of the genera of the forward dataset. Similar observations were evident when comparing the forward and reverse cultivation datasets. Our results indicate that the region under consideration can have a large impact on perceived diversity, and should be considered when comparing different datasets. Finally, a high number of OTUs

  1. Comparisons of discrete and integrative sampling accuracy in estimating pulsed aquatic exposures.

    PubMed

    Morrison, Shane A; Luttbeg, Barney; Belden, Jason B

    2016-11-01

    Most current-use pesticides have short half-lives in the water column and thus the most relevant exposure scenarios for many aquatic organisms are pulsed exposures. Quantifying exposure using discrete water samples may not be accurate as few studies are able to sample frequently enough to accurately determine time-weighted average (TWA) concentrations of short aquatic exposures. Integrative sampling methods that continuously sample freely dissolved contaminants over time intervals (such as integrative passive samplers) have been demonstrated to be a promising measurement technique. We conducted several modeling scenarios to test the assumption that integrative methods may require many less samples for accurate estimation of peak 96-h TWA concentrations. We compared the accuracies of discrete point samples and integrative samples while varying sampling frequencies and a range of contaminant water half-lives (t50 = 0.5, 2, and 8 d). Differences the predictive accuracy of discrete point samples and integrative samples were greatest at low sampling frequencies. For example, when the half-life was 0.5 d, discrete point samples required 7 sampling events to ensure median values > 50% and no sampling events reporting highly inaccurate results (defined as < 10% of the true 96-h TWA). Across all water half-lives investigated, integrative sampling only required two samples to prevent highly inaccurate results and measurements resulting in median values > 50% of the true concentration. Regardless, the need for integrative sampling diminished as water half-life increased. For an 8-d water half-life, two discrete samples produced accurate estimates and median values greater than those obtained for two integrative samples. Overall, integrative methods are the more accurate method for monitoring contaminants with short water half-lives due to reduced frequency of extreme values, especially with uncertainties around the timing of pulsed events. However, the acceptability

  2. Simultaneous Rates of Ribonucleic Acid and Deoxyribonucleic Acid Syntheses for Estimating Growth and Cell Division of Aquatic Microbial Communities

    PubMed Central

    Karl, David M.

    1981-01-01

    A method for measuring rates of ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) syntheses using a single radioactive precursor has been devised and tested using bacterial cultures and natural assemblages of marine and freshwater microorganisms. The procedure is based upon the uptake and incorporation of exogenous [3H]adenine into cellular adenosine triphosphate and deoxyadenosine triphosphate pools which serve as the immediate precursors for the adenine incorporated into RNA and DNA, respectively. It is proposed that the DNA/RNA rate ratio is correlated with the specific growth rate of microorganisms and can be used as an index for estimating and comparing the productivities of microbial assemblages in nature. This technique can also be used to detect discontinuous growth and cell division processes which frequently occur in surface plankton populations. The DNA/RNA rate ratios measured in a variety of aquatic ecosystems ranged from 3.3 to 31.8% without significant correlation to total microbial biomass. PMID:16345882

  3. Catabolism of tritiated thymidine by aquatic microbial communities and incorporation of tritium into RNA and protein.

    PubMed

    Brittain, A M; Karl, D M

    1990-05-01

    The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. All microbial assemblages tested exhibited significant labeling of RNA and protein (i.e., nonspecific labeling), as determined by differential acid-base hydrolysis. Nonspecific labeling was greatest in sediment samples, for which >/=95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. Four different RNA hydrolysis reagents (KOH, NaOH, piperidine, and enzymes) solubilized tritium from cold trichloroacetic acid precipitates. High-pressure liquid chromatography separation of piperidine hydrolysates followed by measurement of isolated monophosphates confirmed the labeling of RNA and indicated that tritium was recovered primarily in CMP and AMP residues. We also evaluated the specificity of [2-H]adenine incorporation into adenylate residues in both RNA and DNA in parallel with the [H]thymidine experiments and compared the degree of nonspecific labeling by [H]adenine with that derived from [H]thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products by high-pressure liquid chromatography separation of the cell-free medium. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples.

  4. Sample preparation procedures utilized in microbial metabolomics: An overview.

    PubMed

    Patejko, Małgorzata; Jacyna, Julia; Markuszewski, Michał J

    2017-02-01

    Bacteria are remarkably diverse in terms of their size, structure and biochemical properties. Due to this fact, it is hard to develop a universal method for handling bacteria cultures during metabolomic analysis. The choice of suitable processing methods constitutes a key element in any analysis, because only appropriate selection of procedures may provide accurate results, leading to reliable conclusions. Because of that, every analytical experiment concerning bacteria requires individually and very carefully planned research methodology. Although every study varies in terms of sample preparation, there are few general steps to follow while planning experiment, like sampling, separation of cells from growth medium, stopping their metabolism and extraction. As a result of extraction, all intracellular metabolites should be washed out from cell environment. What is more, extraction method utilized cannot cause any chemical decomposition or degradation of the metabolome. Furthermore, chosen extraction method should correlate with analytical technique, so it will not disturb or prolong following sample preparation steps. For those reasons, we observe a need to summarize sample preparation procedures currently utilized in microbial metabolomic studies. In the presented overview, papers concerning analysis of extra- and intracellular metabolites, published over the last decade, have been discussed. Presented work gives some basic guidelines that might be useful while planning experiments in microbial metabolomics. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Elucidating Microbial Adaptation Dynamics via Autonomous Exposure and Sampling

    NASA Astrophysics Data System (ADS)

    Grace, J. M.; Verseux, C.; Gentry, D.; Moffet, A.; Thayabaran, R.; Wong, N.; Rothschild, L.

    2013-12-01

    The adaptation of micro-organisms to their environments is a complex process of interaction between the pressures of the environment and of competition. Reducing this multifactorial process to environmental exposure in the laboratory is a common tool for elucidating individual mechanisms of evolution, such as mutation rates[Wielgoss et al., 2013]. Although such studies inform fundamental questions about the way adaptation and even speciation occur, they are often limited by labor-intensive manual techniques[Wassmann et al., 2010]. Current methods for controlled study of microbial adaptation limit the length of time, the depth of collected data, and the breadth of applied environmental conditions. Small idiosyncrasies in manual techniques can have large effects on outcomes; for example, there are significant variations in induced radiation resistances following similar repeated exposure protocols[Alcántara-Díaz et al., 2004; Goldman and Travisano, 2011]. We describe here a project under development to allow rapid cycling of multiple types of microbial environmental exposure. The system allows continuous autonomous monitoring and data collection of both single species and sampled communities, independently and concurrently providing multiple types of controlled environmental pressure (temperature, radiation, chemical presence or absence, and so on) to a microbial community in dynamic response to the ecosystem's current status. When combined with DNA sequencing and extraction, such a controlled environment can cast light on microbial functional development, population dynamics, inter- and intra-species competition, and microbe-environment interaction. The project's goal is to allow rapid, repeatable iteration of studies of both natural and artificial microbial adaptation. As an example, the same system can be used both to increase the pH of a wet soil aliquot over time while periodically sampling it for genetic activity analysis, or to repeatedly expose a culture of

  6. Catabolism of tritiated thymidine by aquatic microbial communities and incorporation of tritium into RNA and protein

    SciTech Connect

    Brittain, A.M.; Karl, D.M. )

    1990-05-01

    The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. Nonspecific labeling was greatest in sediment samples, for which {>=}95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. We also evaluated the specificity of (2-{sup 3}H) adenine incorporation into adenylate residues in both RNA and DNA in parallel with the ({sup 3}H) thymidine experiments and compared the degree of nonspecific labeling by ({sup 3}H) adenine with that derived from ({sup 3}H)thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples.

  7. N-15 NMR spectra of naturally abundant nitrogen in soil and aquatic natural organic matter samples of the International Humic Substances Society

    SciTech Connect

    Thorn, Kevin A.; Cox, Larry G.

    2009-02-28

    The naturally abundant nitrogen in soil and aquatic NOM samples from the International Humic Substances Society has been characterized by solid state CP/MAS ¹⁵N NMR. Soil samples include humic and fulvic acids from the Elliot soil, Minnesota Waskish peat and Florida Pahokee peat, as well as the Summit Hill soil humic acid and the Leonardite humic acid. Aquatic samples include Suwannee River humic, fulvic and reverse osmosis isolates, Nordic humic and fulvic acids and Pony Lake fulvic acid. Additionally, Nordic and Suwannee River XAD-4 acids and Suwannee River hydrophobic neutral fractions were analyzed. Similar to literature reports, amide/aminoquinone nitrogens comprised the major peaks in the solid state spectra of the soil humic and fulvic acids, along with heterocyclic and amino sugar/terminal amino acid nitrogens. Spectra of aquatic samples, including the XAD-4 acids, contain resolved heterocyclic nitrogen peaks in addition to the amide nitrogens. The spectrum of the nitrogen enriched, microbially derived Pony Lake, Antarctica fulvic acid, appeared to contain resonances in the region of pyrazine, imine and/or pyridine nitrogens, which have not been observed previously in soil or aquatic humic substances by ¹⁵N NMR. Liquid state ¹⁵N NMR experiments were also recorded on the Elliot soil humic acid and Pony Lake fulvic acid, both to examine the feasibility of the techniques, and to determine whether improvements in resolution over the solid state could be realized. For both samples, polarization transfer (DEPT) and indirect detection (¹H–¹⁵N gHSQC) spectra revealed greater resolution among nitrogens directly bonded to protons. The amide/aminoquinone nitrogens could also be observed by direct detection experiments.

  8. N-15 NMR spectra of naturally abundant nitrogen in soil and aquatic natural organic matter samples of the International Humic Substances Society

    USGS Publications Warehouse

    Thorn, K.A.; Cox, L.G.

    2009-01-01

    The naturally abundant nitrogen in soil and aquatic NOM samples from the International Humic Substances Society has been characterized by solid state CP/MAS 15N NMR. Soil samples include humic and fulvic acids from the Elliot soil, Minnesota Waskish peat and Florida Pahokee peat, as well as the Summit Hill soil humic acid and the Leonardite humic acid. Aquatic samples include Suwannee River humic, fulvic and reverse osmosis isolates, Nordic humic and fulvic acids and Pony Lake fulvic acid. Additionally, Nordic and Suwannee River XAD-4 acids and Suwannee River hydrophobic neutral fractions were analyzed. Similar to literature reports, amide/aminoquinone nitrogens comprised the major peaks in the solid state spectra of the soil humic and fulvic acids, along with heterocyclic and amino sugar/terminal amino acid nitrogens. Spectra of aquatic samples, including the XAD-4 acids, contain resolved heterocyclic nitrogen peaks in addition to the amide nitrogens. The spectrum of the nitrogen enriched, microbially derived Pony Lake, Antarctica fulvic acid, appeared to contain resonances in the region of pyrazine, imine and/or pyridine nitrogens, which have not been observed previously in soil or aquatic humic substances by 15N NMR. Liquid state 15N NMR experiments were also recorded on the Elliot soil humic acid and Pony Lake fulvic acid, both to examine the feasibility of the techniques, and to determine whether improvements in resolution over the solid state could be realized. For both samples, polarization transfer (DEPT) and indirect detection (1H-15N gHSQC) spectra revealed greater resolution among nitrogens directly bonded to protons. The amide/aminoquinone nitrogens could also be observed by direct detection experiments.

  9. A test of aquatic macroinvertebrate sub-sampling using a gridded screen

    SciTech Connect

    Cross, S.P.

    1994-12-31

    The Biological Resource Evaluations Team of Los Alamos National Laboratories assessed the reliability of a gridded screen sub-sampling technique to estimate aquatic macroinvertebrates in total samples. Benthic macroinvertebrates were collected by kick sampling three riffles areas in Guaje Canyon, Los Alamos County, New Mexico during July and August, 1994. The study included 4,144 macroinvertebrates from samples consisting of 442 to 1005 individuals. The entire samples were spread onto a gridded screen, and 100 macroinvertebrates were randomly selected for identification. To simplify the results, identified macroinvertebrates were assigned to one of six categories: plecoptera, ephemeroptera, trichoptera, coleoptera, diptera, and non-insects. Three sub-samples were taken from each of six full samples. These counts were used as predicted values, while the total sample counts were used as actual values. Single-factor ANOVA tests showed no significant differences between predicted to actual (PTA) values. However, PTA differences indicated that lab-sorting was a more reliable method than live-sorting without a narcotizing agent. Large samples and large numbers in macroinvertebrate categories were tentatively linked with greater PTA differences. PTA differences were less than 5% in 80% of our trials and less than 10% in 95% of our trials. Despite the relatively small size of sub-samples, sub-samples included 60% of taxa found in the total samples. This sub-sampling technique provides accurate estimates of total sample composition in stream reaches rich enough to easily yield the required 100 individuals.

  10. Preliminary Characterisation Of Proteins In Aquatic Samples: A Key To Understanding The Organic Nitrogen Cycle

    NASA Astrophysics Data System (ADS)

    Jones, V.; Ruddell, C. J.; Wainwright, G.; Jaffe, R.; Wolff, G. A.

    When discussing the nitrogen cycle, the dissolved organic nitrogen pool is often treated as a 'black box', due to the analytical difficulties associated with the character- isation of its components. Proteins contain a significant portion of the organic nitro- gen in aquatic systems and recent studies have suggested that certain protein species present in the aquatic environment are not as labile as it was originally thought (e.g. Suzuki et al., 1999) and may therefore form an important part of the long term nitro- gen cycle. The aim of this work is to apply extremely sensitive techniques, recently developed in the biochemical / biomedical field, to characterise proteins in aquatic samples. Two analytical approaches are followed: sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE) with silver staining and liquid chromatography mass spectrometry (peptide mapping). In this poster, we concentrate on SDS PAGE. Large volume water samples were collected at the Everglades National Park, Florida, which encompasses a wide array of ecosystems, ranging from freshwater canals to a shallow marine bay. Samples were concentrated and desalted by tangential flow filtration and proteins were isolated by repeated trichloroacetic acid precipitation. Leaf extracts of the dominant vegetation at each site were prepared. Application of SDS PAGE revealed a large number of distinct protein bands for all sites, corresponding to approximate molecular weights ranging from 30kDa to 250kDa. Protein distributions varied between sites, although bands corresponding to approximate molecular weights of 37kDa, 41kDa, 45kDa, 58kDa and 145kDa were ubiquitous. The 37kDa band in particular was also observed in all leaf extracts pre- pared, suggesting it may represent a recalcitrant molecule, which originates in the higher plant vegetation.

  11. Periphytic photosynthetic stimulation of extracellular enzyme activity in aquatic microbial communities associated with decaying typha litter.

    PubMed

    Francoeur, Steven N; Schaecher, Mark; Neely, Robert K; Kuehn, Kevin A

    2006-11-01

    We examined the effect of light on extracellular enzyme activities of periphytic/endogenous microbial assemblages associated with decomposing litter of an emergent macrophyte Typha angustifolia within a small inland wetland in southeastern Michigan. Standing-dead Typha leaf litter was collected, placed into floating wire mesh litter baskets, and submerged in a wetland pool. Enzyme saturation assays were conducted on three occasions following litter submergence (days 9, 28, and 44) to generate saturation curves for the individual enzymes tested and to examine potential differences in enzyme saturation kinetics during microbial colonization and development. Experimental light manipulations were conducted on two occasions during microbial development (days 10 and 29). Short-term (30 min) light exposure significantly increased extracellular beta-glucosidase activity of litter-associated microbial communities. Activities of beta-xylosidase and leucine-aminopeptidase were not stimulated, and stimulation of phosphatase activity was variable. The exact mechanism for increased enzyme activity remains unknown, but it may have been increased pH arising from periphytic algal photosynthesis. These results suggest that extracellular enzyme activity in microbial communities colonizing natural organic substrata may be influenced by light/photosynthesis, as has previously been demonstrated for periphyton communities grown on artificial, inert substrata. Thus, light/photosynthetic mediated stimulation of extracellular enzyme activities may be a common occurrence in microbial communities associated with natural decaying plant litter in wetlands and might engender diurnal patterns in other microbial decay processes (e.g., production, organic matter decomposition, and mineralization).

  12. Aquatic microbial habitats within a neotropical rainforest: bromeliads and pH-associated trends in bacterial diversity and composition.

    PubMed

    Goffredi, Shana K; Kantor, Adam H; Woodside, Walter T

    2011-04-01

    Tank-forming bromeliads, suspended in the rainforest canopy, possess foliage arranged in compact rosettes capable of long-term retention of rainwater. This large and unique aquatic habitat is inhabited by microorganisms involved in the important decomposition of impounded material. Moreover, these communities are likely influenced by environmental factors such as pH, oxygen, and light. Bacterial community composition and diversity was determined for the tanks of several bromeliad species (Aechmea and Werauhia) from northern Costa Rica, which span a range of parameters, including tank morphology and pH. These were compared with a nearby forest soil sample, an artificial tank (amber bottle), and a commercially available species (Aechmea). Bacterial community diversity, as measured by 16S rRNA analysis and tRFLP, showed a significant positive correlation with tank pH. A majority of 16S rRNA bacterial phylotypes found in association with acidic bromeliad tanks of pH < 5.1 were affiliated with the Alphaproteobacteria, Acidobacteria, Planctomycetes, and Bacteroidetes, and were similar to those found in acidic peat bogs, yet distinct from the underlying soil community. In contrast, bromeliads with tank pH > 5.3, including the commercial bromeliad with the highest pH (6.7), were dominated by Betaproteobacteria, Firmicutes, and Bacteroidetes. To empirically determine the effect of pH on bacterial community, the tank pH of a specimen of Aechmea was depressed, in the field, from 6.5 to 4.5, for 62 days. The resulting community changed predictably with decreased abundance of Betaproteobacteria and Firmicutes and a concomitant increase in Alphaproteobacteria and Acidobacteria. Collectively, these results suggest that bromeliad tanks provide important habitats for a diverse microbial community, distinct from the surrounding environment, which are influenced greatly by acid-base conditions. Additionally, total organic carbon (∼46%) and nitrogen (∼2%) of bromeliad

  13. Determination of the heavy metal binding capacity of aquatic samples using MetPLATE: a preliminary study.

    PubMed

    Huang, F; Bitton, G; Kong, I C

    1999-08-30

    MetPLATE, a microbial toxicity test which is specific for heavy metal toxicity, was used to rapidly determine the heavy metal binding capacity (HMBC) of a wide range of surface waters from Florida and Georgia. HMBC determines the impact of physicochemical factors on metal bioavailability and toxicity. The new developed protocol, using MetPLATE as the toxicity assay, showed that HMBC varied from 1.7 to 39.2 for Cd whereas the ranges for Cu and Ag were < 1-11.9 and < 1-2.1, respectively. The effect of seasons on HMBC was determined using samples from the Hogtown Creek, Gainesville, FL, and from the St John's river in Jacksonville, FL. Both surface waters displayed the highest HMBC during the Fall season. Preliminary examination of the limited data set confirms that HMBC may be influenced by the concentration of dissolved organic carbon (DOC). These preliminary data show that the impact of physical and chemical parameters on the toxicity of metals in aquatic environments can be rapidly assessed using rapid and low-cost microbiotests.

  14. Characterization of Microbial Population Shifts during Sample Storage.

    PubMed

    Mills, Heath J; Reese, Brandi Kiel; Peter, Cruz St

    2012-01-01

    The objective of this study was to determine shifts in the microbial community structure and potential function based on standard Integrated Ocean Drilling Program (IODP) storage procedures for sediment cores. Standard long-term storage protocols maintain sediment temperature at 4°C for mineralogy, geochemical, and/or geotechnical analysis whereas standard microbiological sampling immediately preserves sediments at -80°C. Storage at 4°C does not take into account populations may remain active over geologic time scales at temperatures similar to storage conditions. Identification of active populations within the stored core would suggest geochemical and geophysical conditions within the core change over time. To test this potential, the metabolically active fraction of the total microbial community was characterized from IODP Expedition 325 Great Barrier Reef sediment cores prior to and following a 3-month storage period. Total RNA was extracted from complementary 2, 20, and 40 m below sea floor sediment samples, reverse transcribed to complementary DNA and then sequenced using 454 FLX sequencing technology, yielding over 14,800 sequences from the six samples. Interestingly, 97.3% of the sequences detected were associated with lineages that changed in detection frequency during the storage period including key biogeochemically relevant lineages associated with nitrogen, iron, and sulfur cycling. These lineages have the potential to permanently alter the physical and chemical characteristics of the sediment promoting misleading conclusions about the in situ biogeochemical environment. In addition, the detection of new lineages after storage increases the potential for a wider range of viable lineages within the subsurface that may be underestimated during standard community characterizations.

  15. Characterization of Microbial Population Shifts during Sample Storage

    PubMed Central

    Mills, Heath J.; Reese, Brandi Kiel; Peter, Cruz St.

    2011-01-01

    The objective of this study was to determine shifts in the microbial community structure and potential function based on standard Integrated Ocean Drilling Program (IODP) storage procedures for sediment cores. Standard long-term storage protocols maintain sediment temperature at 4°C for mineralogy, geochemical, and/or geotechnical analysis whereas standard microbiological sampling immediately preserves sediments at −80°C. Storage at 4°C does not take into account populations may remain active over geologic time scales at temperatures similar to storage conditions. Identification of active populations within the stored core would suggest geochemical and geophysical conditions within the core change over time. To test this potential, the metabolically active fraction of the total microbial community was characterized from IODP Expedition 325 Great Barrier Reef sediment cores prior to and following a 3-month storage period. Total RNA was extracted from complementary 2, 20, and 40 m below sea floor sediment samples, reverse transcribed to complementary DNA and then sequenced using 454 FLX sequencing technology, yielding over 14,800 sequences from the six samples. Interestingly, 97.3% of the sequences detected were associated with lineages that changed in detection frequency during the storage period including key biogeochemically relevant lineages associated with nitrogen, iron, and sulfur cycling. These lineages have the potential to permanently alter the physical and chemical characteristics of the sediment promoting misleading conclusions about the in situ biogeochemical environment. In addition, the detection of new lineages after storage increases the potential for a wider range of viable lineages within the subsurface that may be underestimated during standard community characterizations. PMID:22363327

  16. Microbial Groundwater Sampling Protocol for Fecal-Rich Environments

    PubMed Central

    Harter, Thomas; Watanabe, Naoko; Li, Xunde; Atwill, Edward R; Samuels, William

    2014-01-01

    Inherently, confined animal farming operations (CAFOs) and other intense fecal-rich environments are potential sources of groundwater contamination by enteric pathogens. The ubiquity of microbial matter poses unique technical challenges in addition to economic constraints when sampling wells in such environments. In this paper, we evaluate a groundwater sampling protocol that relies on extended purging with a portable submersible stainless steel pump and Teflon® tubing as an alternative to equipment sterilization. The protocol allows for collecting a large number of samples quickly, relatively inexpensively, and under field conditions with limited access to capacity for sterilizing equipment. The protocol is tested on CAFO monitoring wells and considers three cross-contamination sources: equipment, wellbore, and ambient air. For the assessment, we use Enterococcus, a ubiquitous fecal indicator bacterium (FIB), in laboratory and field tests with spiked and blank samples, and in an extensive, multi-year field sampling campaign on 17 wells within 2 CAFOs. The assessment shows that extended purging can successfully control for equipment cross-contamination, but also controls for significant contamination of the well-head, within the well casing and within the immediate aquifer vicinity of the well-screen. Importantly, our tests further indicate that Enterococcus is frequently entrained in water samples when exposed to ambient air at a CAFO during sample collection. Wellbore and air contamination pose separate challenges in the design of groundwater monitoring strategies on CAFOs that are not addressed by equipment sterilization, but require adequate QA/QC procedures and can be addressed by the proposed sampling strategy. PMID:24903186

  17. Microbial groundwater sampling protocol for fecal-rich environments.

    PubMed

    Harter, Thomas; Watanabe, Naoko; Li, Xunde; Atwill, Edward R; Samuels, William

    2014-09-01

    Inherently, confined animal farming operations (CAFOs) and other intense fecal-rich environments are potential sources of groundwater contamination by enteric pathogens. The ubiquity of microbial matter poses unique technical challenges in addition to economic constraints when sampling wells in such environments. In this paper, we evaluate a groundwater sampling protocol that relies on extended purging with a portable submersible stainless steel pump and Teflon(®) tubing as an alternative to equipment sterilization. The protocol allows for collecting a large number of samples quickly, relatively inexpensively, and under field conditions with limited access to capacity for sterilizing equipment. The protocol is tested on CAFO monitoring wells and considers three cross-contamination sources: equipment, wellbore, and ambient air. For the assessment, we use Enterococcus, a ubiquitous fecal indicator bacterium (FIB), in laboratory and field tests with spiked and blank samples, and in an extensive, multi-year field sampling campaign on 17 wells within 2 CAFOs. The assessment shows that extended purging can successfully control for equipment cross-contamination, but also controls for significant contamination of the well-head, within the well casing and within the immediate aquifer vicinity of the well-screen. Importantly, our tests further indicate that Enterococcus is frequently entrained in water samples when exposed to ambient air at a CAFO during sample collection. Wellbore and air contamination pose separate challenges in the design of groundwater monitoring strategies on CAFOs that are not addressed by equipment sterilization, but require adequate QA/QC procedures and can be addressed by the proposed sampling strategy. © 2014 The Authors. Groundwater published by Wiley Periodicals, Inc. on behalf of National Ground Water Association.

  18. Microbial Community Analysis in the Roots of Aquatic Plants and Isolation of Novel Microbes Including an Organism of the Candidate Phylum OP10

    PubMed Central

    Tanaka, Yasuhiro; Tamaki, Hideyuki; Matsuzawa, Hiroaki; Nigaya, Masahiro; Mori, Kazuhiro; Kamagata, Yoichi

    2012-01-01

    A number of molecular ecological studies have revealed complex and unique microbial communities in various terrestrial plant roots; however, little is known about the microbial communities of aquatic plant roots in spite of their potential use for water quality improvement in aquatic environments (e.g. floating treatment wetland system). Here, we report the microbial communities inhabiting the roots of emerged plants, reed (Phragmites australis) and Japanese loosestrife (Lythrum anceps), collected from a floating treatment wetland in a pond by both culture-independent and culture-dependent approaches. Culture-independent analysis based on 16S rRNA gene sequences revealed that the microbial compositions between the two aquatic plant roots were clearly different (e.g. the predominant microbe was Betaproteobacteria for reed and Alphaproteobacteria for Japanese loosestrife). In comparisons of microbial communities between the plant roots and pond water taken from near the plants, the microbial diversity in the plant roots (e.g. 4.40–4.26 Shannon-Weiner index) were higher than that of pond water (e.g. 3.15 Shannon-Weiner index). Furthermore, the plant roots harbored 2.5–3.5 times more phylogenetically novel clone phylotypes than pond water. The culture-dependent approach also revealed differences in the microbial composition and diversity among the two plant roots and pond water. More importantly, compared to pond water, we succeeded in isolating approximately two times more novel isolate phylotypes, including a bacterium of candidate phylum OP10 (recently named Armatimonadetes) from the plant roots. These findings suggest that aquatic plants roots are significant sources for a variety of novel organisms. PMID:22791047

  19. Microbial community analysis in the roots of aquatic plants and isolation of novel microbes including an organism of the candidate phylum OP10.

    PubMed

    Tanaka, Yasuhiro; Tamaki, Hideyuki; Matsuzawa, Hiroaki; Nigaya, Masahiro; Mori, Kazuhiro; Kamagata, Yoichi

    2012-01-01

    A number of molecular ecological studies have revealed complex and unique microbial communities in various terrestrial plant roots; however, little is known about the microbial communities of aquatic plant roots in spite of their potential use for water quality improvement in aquatic environments (e.g. floating treatment wetland system). Here, we report the microbial communities inhabiting the roots of emerged plants, reed (Phragmites australis) and Japanese loosestrife (Lythrum anceps), collected from a floating treatment wetland in a pond by both culture-independent and culture-dependent approaches. Culture-independent analysis based on 16S rRNA gene sequences revealed that the microbial compositions between the two aquatic plant roots were clearly different (e.g. the predominant microbe was Betaproteobacteria for reed and Alphaproteobacteria for Japanese loosestrife). In comparisons of microbial communities between the plant roots and pond water taken from near the plants, the microbial diversity in the plant roots (e.g. 4.40-4.26 Shannon-Weiner index) were higher than that of pond water (e.g. 3.15 Shannon-Weiner index). Furthermore, the plant roots harbored 2.5-3.5 times more phylogenetically novel clone phylotypes than pond water. The culture-dependent approach also revealed differences in the microbial composition and diversity among the two plant roots and pond water. More importantly, compared to pond water, we succeeded in isolating approximately two times more novel isolate phylotypes, including a bacterium of candidate phylum OP10 (recently named Armatimonadetes) from the plant roots. These findings suggest that aquatic plants roots are significant sources for a variety of novel organisms.

  20. Physiological profiling of indigenous aquatic microbial communities to determine toxic effects of metals

    SciTech Connect

    Lehman, R.M.; Colwell, F.S.; Garland, J.L.

    1997-11-01

    Conventional bioassays for environmental assessment frequently rely on nonindigenous single species. The authors employed an assay in which whole environmental samples were distinguished by the ability of the native heterotrophic microbial communities to oxidize 95 different sole carbon sources generating a community-level physiological profile (CLPP). The average metabolic response (AMR) to the 95 variables defining the CLPP was used in laboratory bioassay studies with copper to construct dose-response curves over several different periods of exposure: 1 h (acute), 1 d, 2 d, and 4 d. The acute dose-response of Snake River bacterioplankton communities measured by AMR was compared to the dose-response of Photobacterium phosphoreum (used in the Microtox test) and a proprietary mixed consortia (used in the Polytox test). In laboratory bioassay studies, CLPP, AMR exhibited acute dose-response behavior over a greater range in copper concentrations and with less variability (per dose) than Microtox and Polytox. The acute sensitivity of CLPP AMR to copper was roughly equal to Microtox and much greater than Polytox. After a longer exposure (1 d) to copper, Snake River communities became more sensitive to copper but no additional effect was observed when the exposure was increases to 2 and 4 d. Snake River communities pre-exposed to copper (1 mg/L) for 4 d prior to acute dose-response experiments showed no difference in AMR with respect to doses up to 10 mg/L, indicating the ability of the assay to detect adaptation. Several metal-contaminated streams in Idaho were used to field validate the CLPP approach for detecting impacts of metals in the environment. The response profiles of the bacterioplankton from two downstream sites receiving metal laden mine drainage were compared to those from reference sites upstream and further downstream of the location receiving the mine drainage.

  1. Passive sampling of selected pesticides in aquatic environment using polar organic chemical integrative samplers.

    PubMed

    Thomatou, Alphanna-Akrivi; Zacharias, Ierotheos; Hela, Dimitra; Konstantinou, Ioannis

    2011-08-01

    Polar chemical integrative samplers (POCIS) were examined for their sampling efficiency of 12 pesticides and one metabolite commonly detected in surface waters. Laboratory-based calibration experiments of POCISs were conducted. The determined passive sampling rates were applied for the monitoring of pesticides levels in Lake Amvrakia, Western Greece. Spot sampling was also performed for comparison purposes. Calibration experiments were performed on the basis of static renewal exposure of POCIS under stirred conditions for different time periods of up to 28 days. The analytical procedures were based on the coupling of POCIS and solid phase extraction by Oasis HLB cartridges with gas chromatography-mass spectrometry. The recovery of the target pesticides from the POCIS was generally >79% with relative standard deviation (RSD) <16%. The calibration results revealed an integrative uptake of all pesticides for 28 days and the calculated sampling rates ranged from 0.025 to 0.388 L day(-1) with RSD <29%. Low nanogram/liter levels of pesticides such as diazinon, alachlor, and s-metolachlor were detected during the monitoring campaign using both passive and spot sampling whereas higher concentrations were measured by spot sampling in most cases. Passive sampling by POCIS provides a useful tool for the monitoring of pesticides in aquatic systems since integrative sampling at rates sufficient for analytical quantitation of ambient levels was observed. Calibration data are in demand for a greater number of compounds in order to extend the use in environmental monitoring.

  2. Environmental DNA sampling protocol - filtering water to capture DNA from aquatic organisms

    USGS Publications Warehouse

    Laramie, Matthew B.; Pilliod, David S.; Goldberg, Caren S.; Strickler, Katherine M.

    2015-09-29

    Environmental DNA (eDNA) analysis is an effective method of determining the presence of aquatic organisms such as fish, amphibians, and other taxa. This publication is meant to guide researchers and managers in the collection, concentration, and preservation of eDNA samples from lentic and lotic systems. A sampling workflow diagram and three sampling protocols are included as well as a list of suggested supplies. Protocols include filter and pump assembly using: (1) a hand-driven vacuum pump, ideal for sample collection in remote sampling locations where no electricity is available and when equipment weight is a primary concern; (2) a peristaltic pump powered by a rechargeable battery-operated driver/drill, suitable for remote sampling locations when weight consideration is less of a concern; (3) a 120-volt alternating current (AC) powered peristaltic pump suitable for any location where 120-volt AC power is accessible, or for roadside sampling locations. Images and detailed descriptions are provided for each step in the sampling and preservation process.

  3. Reconnaissance samplings and characterization of aquatic humic substances at the Yuma Desalting Test Facility, Arizona

    USGS Publications Warehouse

    Malcolm, R.L.; Wershaw, R. L.; Thurman, E.M.; Aiken, G.R.; Pinckney, D.J.; Kaakinen, J.

    1981-01-01

    Smectite clay minerals were found to be the principal compound on the surface of the cellulose-acetate, reverse-osmosis membranes at the Yuma Desalting Test Facility. These clay minerals were not present in the pumped ground water, but were blown into the conveyance canal from adjacent soils. Humic substances from the water and suspended sediments were associated with the clay films on the membrane, but no definitive results concerning their role in fouling were achieved. Microbial fouling is believed to be only a minor aspect of membrane fouling. Chemical and physical changes in humic substances were extensively studied at four points in the water-treatment process. Humic substances accounted for the largest component (over 25 percent) of organic constituents. Humic substances in the canal source water were similar to other aquatic humic substances present in natural waters. During the treatment process, these substances are brominated and decolorized. The effect of these halogenated humic substances on membrane fouling is unclear, but their presence in the reverse-osmosis product water and reverse-osmosis reject brine, along with volatile trihalomethanes, has led to environmental concerns. (USGS)

  4. Unexpected toxicity to aquatic organisms of some aqueous bisphenol A samples treated by advanced oxidation processes.

    PubMed

    Tišler, Tatjana; Erjavec, Boštjan; Kaplan, Renata; Şenilă, Marin; Pintar, Albin

    2015-01-01

    In this study, photocatalytic and catalytic wet-air oxidation (CWAO) processes were used to examine removal efficiency of bisphenol A from aqueous samples over several titanate nanotube-based catalysts. Unexpected toxicity of bisphenol A (BPA) samples treated by means of the CWAO process to some tested species was determined. In addition, the CWAO effluent was recycled five- or 10-fold in order to increase the number of interactions between the liquid phase and catalyst. Consequently, the inductively coupled plasma mass spectrometry (ICP-MS) analysis indicated higher concentrations of some toxic metals like chromium, nickel, molybdenum, silver, and zinc in the recycled samples in comparison to both the single-pass sample and the photocatalytically treated solution. The highest toxicity of five- and 10-fold recycled solutions in the CWAO process was observed in water fleas, which could be correlated to high concentrations of chromium, nickel, and silver detected in tested samples. The obtained results clearly demonstrated that aqueous samples treated by means of advanced oxidation processes should always be analyzed using (i) chemical analyses to assess removal of BPA and total organic carbon from treated aqueous samples, as well as (ii) a battery of aquatic organisms from different taxonomic groups to determine possible toxicity.

  5. A strategy to sample nutrient dynamics across the terrestrial-aquatic interface at NEON sites

    NASA Astrophysics Data System (ADS)

    Hinckley, E. S.; Goodman, K. J.; Roehm, C. L.; Meier, C. L.; Luo, H.; Ayres, E.; Parnell, J.; Krause, K.; Fox, A. M.; SanClements, M.; Fitzgerald, M.; Barnett, D.; Loescher, H. W.; Schimel, D.

    2012-12-01

    The construction of the National Ecological Observatory Network (NEON) across the U.S. creates the opportunity for researchers to investigate biogeochemical transformations and transfers across ecosystems at local-to-continental scales. Here, we examine a subset of NEON sites where atmospheric, terrestrial, and aquatic observations will be collected for 30 years. These sites are located across a range of hydrological regimes, including flashy rain-driven, shallow sub-surface (perched, pipe-flow, etc), and deep groundwater, which likely affect the chemical forms and quantities of reactive elements that are retained and/or mobilized across landscapes. We present a novel spatial and temporal sampling design that enables researchers to evaluate long-term trends in carbon, nitrogen, and phosphorus biogeochemical cycles under these different hydrological regimes. This design focuses on inputs to the terrestrial system (atmospheric deposition, bulk precipitation), transfers (soil-water and groundwater sources/chemistry), and outputs (surface water, and evapotranspiration). We discuss both data that will be collected as part of the current NEON design, as well as how the research community can supplement the NEON design through collaborative efforts, such as providing additional datasets, including soil biogeochemical processes and trace gas emissions, and developing collaborative research networks. Current engagement with the research community working at the terrestrial-aquatic interface is critical to NEON's success as we begin construction, to ensure that high-quality, standardized and useful data are not only made available, but inspire further, cutting-edge research.

  6. Species sensitivity distribution for chlorpyrifos to aquatic organisms: Model choice and sample size.

    PubMed

    Zhao, Jinsong; Chen, Boyu

    2016-03-01

    Species sensitivity distribution (SSD) is a widely used model that extrapolates the ecological risk to ecosystem levels from the ecotoxicity of a chemical to individual organisms. However, model choice and sample size significantly affect the development of the SSD model and the estimation of hazardous concentrations at the 5th centile (HC5). To interpret their effects, the SSD model for chlorpyrifos, a widely used organophosphate pesticide, to aquatic organisms is presented with emphases on model choice and sample size. Three subsets of median effective concentration (EC50) with different sample sizes were obtained from ECOTOX and used to build SSD models based on parametric distribution (normal, logistic, and triangle distribution) and nonparametric bootstrap. The SSD models based on the triangle distribution are superior to the normal and logistic distributions according to several goodness-of-fit techniques. Among all parametric SSD models, the one with the largest sample size based on the triangle distribution gives the most strict HC5 with 0.141μmolL(-1). The HC5 derived from the nonparametric bootstrap is 0.159μmol L(-1). The minimum sample size required to build a stable SSD model is 11 based on parametric distribution and 23 based on nonparametric bootstrap. The study suggests that model choice and sample size are important sources of uncertainty for application of the SSD model. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Quantifying the contribution of single microbial cells to nitrogen assimilation in aquatic environments

    NASA Astrophysics Data System (ADS)

    Musat, N.; Kuypers, M. M. M.

    2009-04-01

    Nitrogen is a primary productivity-limiting nutrient in the ocean. The nitrogen limitation of productivity may be overcome by organisms capable of converting dissolved N2 into fixed nitrogen available to the ecosystem. In many oceanic regions, growth of phytoplankton is nitrogen limited because fixation of N2 cannot make up for the removal of fixed inorganic nitrogen (NH4+, NO2-, NO3-) by anaerobic microbial processes. The amount of available fixed nitrogen in the ocean can be changed by the biological processes of heterotrophic denitrification, anaerobic ammonium oxidation and nitrogen fixation. For a complete understanding of nitrogen cycling in the ocean a link between the microbial and biogeochemical processes at the single cell level and their role in global biogeochemical cycles is essential. Here we report a recently developed method, Halogen In Situ Hybridization-Secondary Ion Mass Spectroscopy (HISH-SIMS) and its potential application to study the nitrogen-cycle processes in the ocean. The method allows simultaneous phylogenetic identification and quantitation of metabolic activities of single microbial cells in the environment. It uses horseradish-peroxidase-labeled oligonucleotide probes and fluorine-containing tyramides for the identification of microorganisms in combination with stable-isotope-labeling experiments for analyzing the metabolic function of single microbial cells. HISH-SIMS was successfully used to study nitrogen assimilation and nitrogen fixation by anaerobic phototrophs in a meromictic alpine lake. The HISH-SIMS method enables studies of the ecophysiology of individual, phylogenetically identified microorganisms involved in the N-cycle and allows us to track the flow of nitrogen within microbial communities.

  8. Modeling Microbial Biogeochemistry from Terrestrial to Aquatic Ecosystems Using Trait-Based Approaches

    NASA Astrophysics Data System (ADS)

    King, E.; Molins, S.; Karaoz, U.; Johnson, J. N.; Bouskill, N.; Hug, L. A.; Thomas, B. C.; Castelle, C. J.; Beller, H. R.; Banfield, J. F.; Steefel, C. I.; Brodie, E.

    2014-12-01

    Currently, there is uncertainty in how climate or land-use-induced changes in hydrology and vegetation will affect subsurface carbon flux, the spatial and temporal distribution of flow and transport, biogeochemical cycling, and microbial metabolic activity. Here we focus on the initial development of a Genome-Enabled Watershed Simulation Capability (GEWaSC), which provides a predictive framework for understanding how genomic information stored in a subsurface microbiome affects biogeochemical watershed functioning, how watershed-scale processes affect microbial function, and how these interactions co-evolve. This multiscale framework builds on a hierarchical approach to multiscale modeling, which considers coupling between defined microscale and macroscale components of a system (e.g., a catchment being defined as macroscale and biogeofacies as microscale). Here, we report our progress in the development of a trait-based modeling approach within a reactive transport framework that simulates coupled guilds of microbes. Guild selection is driven by traits extracted from, and physiological properties inferred from, large-scale assembly of metagenome data. Meta-genomic, -transcriptomic and -proteomic information are also used to complement our existing biogeochemical reaction networks and contributes key reactions where biogeochemical analyses are unequivocal. Our approach models the rate of nutrient uptake and the thermodynamics of coupled electron donors and acceptors for a range of microbial metabolisms including heterotrophs and chemolitho(auto)trophs. Metabolism of exogenous substrates fuels catabolic and anabolic processes, with the proportion of energy used for each based upon dynamic intracellular and environmental conditions. In addition to biomass development, anabolism includes the production of key enzymes, such as nitrogenase for nitrogen fixation or exo-enzymes for the hydrolysis of extracellular polymers. This internal resource partitioning represents a

  9. Methylmercury enters an aquatic food web through acidophilic microbial mats in Yellowstone National Park, Wyoming

    USGS Publications Warehouse

    Boyd, E.S.; King, S.; Tomberlin, J.K.; Nordstrom, D.K.; Krabbenhoft, D.P.; Barkay, T.; Geesey, G.G.

    2009-01-01

    Summary Microbial mats are a visible and abundant life form inhabiting the extreme environments in Yellowstone National Park (YNP), WY, USA. Little is known of their role in food webs that exist in the Park's geothermal habitats. Eukaryotic green algae associated with a phototrophic green/purple Zygogonium microbial mat community that inhabits low-temperature regions of acidic (pH ??? 3.0) thermal springs were found to serve as a food source for stratiomyid (Diptera: Stratiomyidae) larvae. Mercury in spring source water was taken up and concentrated by the mat biomass. Monomethylmercury compounds (MeHg +), while undetectable or near the detection limit (0.025 ng l -1) in the source water of the springs, was present at concentrations of 4-7 ng g-1 dry weight of mat biomass. Detection of MeHg + in tracheal tissue of larvae grazing the mat suggests that MeHg+ enters this geothermal food web through the phototrophic microbial mat community. The concentration of MeHg+ was two to five times higher in larval tissue than mat biomass indicating MeHg+ biomagnification occurred between primary producer and primary consumer trophic levels. The Zygogonium mat community and stratiomyid larvae may also play a role in the transfer of MeHg+ to species in the food web whose range extends beyond a particular geothermal feature of YNP. ?? 2008 The Authors. Journal compilation ?? 2008 Society for Applied Microbiology and Blackwell Publishing Ltd.

  10. Biotransformation of herbicides by aquatic microbial communities associated to submerged leaves.

    PubMed

    Carles, Louis; Rossi, Florent; Joly, Muriel; Besse-Hoggan, Pascale; Batisson, Isabelle; Artigas, Joan

    2017-02-01

    Leaf microbial communities possess a large panel of enzymes permitting the breakdown of leaf polymers as well as the transformation of organic xenobiotic compounds present in stream waters. This study aims to assess the potential of leaf microbial communities, exhibiting different exposure histories to pesticides (upstream versus downstream), to biotransform three maize herbicides (mesotrione, S-metolachlor, and nicosulfuron) in single and cocktail molecule exposures. The results showed a high dissipation of nicosulfuron (sulfonylurea herbicide) (from 29.1 ± 10.8% to 66 ± 16.2%, day 40) in both single and cocktail exposures, respectively, but not of mesotrione and S-metolachlor. The formation of nicosulfuron metabolites such as ASDM (2-(aminosulfonyl)-N,N-dimethyl-3-pyridinecarboxamide) and ADMP (2-amino-4,6-dimethoxypyrimidine) and the weak sorption (<0.4%) on the leaf matrix confirmed the transformation of this molecule by leaf microorganisms. In addition, the downstream communities showed a greater ability to transform nicosulfuron than the upstream communities suggesting that the exposure history to pesticides is an important parameter and can enhance the biotransformation potential of leaf microorganisms. After 40-day single exposure to nicosulfuron, the downstream communities were also those experiencing the greatest shifts in fungal and bacterial community diversity suggesting a potential adaptation of microorganisms to this herbicide. Our study emphasizes the importance of leaf microbial communities for herbicide biotransformation in polluted stream ecosystems where fungi could play a crucial role.

  11. Methylmercury enters an aquatic food web through acidophilic microbial mats in Yellowstone National Park, Wyoming.

    PubMed

    Boyd, Eric S; King, Susan; Tomberlin, Jeffery K; Nordstrom, D Kirk; Krabbenhoft, David P; Barkay, Tamar; Geesey, Gill G

    2009-04-01

    Microbial mats are a visible and abundant life form inhabiting the extreme environments in Yellowstone National Park (YNP), WY, USA. Little is known of their role in food webs that exist in the Park's geothermal habitats. Eukaryotic green algae associated with a phototrophic green/purple Zygogonium microbial mat community that inhabits low-temperature regions of acidic (pH approximately 3.0) thermal springs were found to serve as a food source for stratiomyid (Diptera: Stratiomyidae) larvae. Mercury in spring source water was taken up and concentrated by the mat biomass. Monomethylmercury compounds (MeHg(+)), while undetectable or near the detection limit (0.025 ng l(-1)) in the source water of the springs, was present at concentrations of 4-7 ng g(-1) dry weight of mat biomass. Detection of MeHg(+) in tracheal tissue of larvae grazing the mat suggests that MeHg(+) enters this geothermal food web through the phototrophic microbial mat community. The concentration of MeHg(+) was two to five times higher in larval tissue than mat biomass indicating MeHg(+) biomagnification occurred between primary producer and primary consumer trophic levels. The Zygogonium mat community and stratiomyid larvae may also play a role in the transfer of MeHg(+) to species in the food web whose range extends beyond a particular geothermal feature of YNP.

  12. Efficiency of Different Sampling Tools for Aquatic Macroinvertebrate Collections in Malaysian Streams.

    PubMed

    Ghani, Wan Mohd Hafezul Wan Abdul; Rawi, Che Salmah Md; Hamid, Suhaila Abd; Al-Shami, Salman Abdo

    2016-02-01

    This study analyses the sampling performance of three benthic sampling tools commonly used to collect freshwater macroinvertebrates. Efficiency of qualitative D-frame and square aquatic nets were compared to a quantitative Surber sampler in tropical Malaysian streams. The abundance and diversity of macroinvertebrates collected using each tool evaluated along with their relative variations (RVs). Each tool was used to sample macroinvertebrates from three streams draining different areas: a vegetable farm, a tea plantation and a forest reserve. High macroinvertebrate diversities were recorded using the square net and Surber sampler at the forested stream site; however, very low species abundance was recorded by the Surber sampler. Relatively large variations in the Surber sampler collections (RVs of 36% and 28%) were observed for the vegetable farm and tea plantation streams, respectively. Of the three sampling methods, the square net was the most efficient, collecting a greater diversity of macroinvertebrate taxa and a greater number of specimens (i.e., abundance) overall, particularly from the vegetable farm and the tea plantation streams (RV<25%). Fewer square net sample passes (<8 samples) were sufficient to perform a biological assessment of water quality, but each sample required a slightly longer processing time (±20 min) compared with those gathered via the other samplers. In conclusion, all three apparatuses were suitable for macroinvertebrate collection in Malaysian streams and gathered assemblages that resulted in the determination of similar biological water quality classes using the Family Biotic Index (FBI) and the Biological Monitoring Working Party (BMWP). However, despite a slightly longer processing time, the square net was more efficient (lowest RV) at collecting samples and more suitable for the collection of macroinvertebrates from deep, fast flowing, wadeable streams with coarse substrates.

  13. Efficiency of Different Sampling Tools for Aquatic Macroinvertebrate Collections in Malaysian Streams

    PubMed Central

    Ghani, Wan Mohd Hafezul Wan Abdul; Rawi, Che Salmah Md; Hamid, Suhaila Abd; Al-Shami, Salman Abdo

    2016-01-01

    This study analyses the sampling performance of three benthic sampling tools commonly used to collect freshwater macroinvertebrates. Efficiency of qualitative D-frame and square aquatic nets were compared to a quantitative Surber sampler in tropical Malaysian streams. The abundance and diversity of macroinvertebrates collected using each tool evaluated along with their relative variations (RVs). Each tool was used to sample macroinvertebrates from three streams draining different areas: a vegetable farm, a tea plantation and a forest reserve. High macroinvertebrate diversities were recorded using the square net and Surber sampler at the forested stream site; however, very low species abundance was recorded by the Surber sampler. Relatively large variations in the Surber sampler collections (RVs of 36% and 28%) were observed for the vegetable farm and tea plantation streams, respectively. Of the three sampling methods, the square net was the most efficient, collecting a greater diversity of macroinvertebrate taxa and a greater number of specimens (i.e., abundance) overall, particularly from the vegetable farm and the tea plantation streams (RV<25%). Fewer square net sample passes (<8 samples) were sufficient to perform a biological assessment of water quality, but each sample required a slightly longer processing time (±20 min) compared with those gathered via the other samplers. In conclusion, all three apparatuses were suitable for macroinvertebrate collection in Malaysian streams and gathered assemblages that resulted in the determination of similar biological water quality classes using the Family Biotic Index (FBI) and the Biological Monitoring Working Party (BMWP). However, despite a slightly longer processing time, the square net was more efficient (lowest RV) at collecting samples and more suitable for the collection of macroinvertebrates from deep, fast flowing, wadeable streams with coarse substrates. PMID:27019685

  14. Portable system for microbial sample preparation and oligonucleotide microarray analysis.

    SciTech Connect

    Bavykin, S. G.; Akowski, J. P.; Zakhariev, V. M.; Barsky, V. E.; Mirzabekov, A. D.; Perov, A. N.; Biochip Technology Center; Engelhardt Inst. of Molecular Biology

    2001-02-01

    We have developed a three-component system for microbial identification that consists of (i) a universal syringe-operated silica minicolumn for successive DNA and RNA isolation, fractionation, fragmentation, fluorescent labeling, and removal of excess free label and short oligonucleotides; (ii) microarrays of immobilized oligonucleotide probes for 16S rRNA identification; and (iii) a portable battery-powered device for imaging the hybridization of fluorescently labeled RNA fragments with the arrays. The minicolumn combines a guanidine thiocyanate method of nucleic acid isolation with a newly developed hydroxyl radical-based technique for DNA and RNA labeling and fragmentation. DNA and RNA can also be fractionated through differential binding of double- and single-stranded forms of nucleic acids to the silica. The procedure involves sequential washing of the column with different solutions. No vacuum filtration steps, phenol extraction, or centrifugation is required. After hybridization, the overall fluorescence pattern is captured as a digital image or as a Polaroid photo. This three-component system was used to discriminate Escherichia coli, Bacillus subtilis, Bacillus thuringiensis, and human HL60 cells. The procedure is rapid: beginning with whole cells, it takes approximately 25 min to obtain labeled DNA and RNA samples and an additional 25 min to hybridize and acquire the microarray image using a stationary image analysis system or the portable imager.

  15. Sampled-time control of a microbial fuel cell stack

    NASA Astrophysics Data System (ADS)

    Boghani, Hitesh C.; Dinsdale, Richard M.; Guwy, Alan J.; Premier, Giuliano C.

    2017-07-01

    Research into microbial fuel cells (MFCs) has reached the point where cubic metre-scale systems and stacks are being built and tested. Apart from performance enhancement through catalysis, materials and design, an important research area for industrial applicability is stack control, which can enhance MFCs stack power output. An MFC stack is controlled using a sampled-time digital control strategy, which has the advantage of intermittent operation with consequent power saving, and when used in a hybrid series stack connectivity, can avoid voltage reversals. A MFC stack comprising four tubular MFCs was operated hydraulically in series. Each MFC was connected to an independent controller and the stack was connected electrically in series, creating a hybrid-series connectivity. The voltage of each MFC in the stack was controlled such that the overall series stack voltage generated was the algebraic sum (1.26 V) of the individual MFC voltages (0.32, 0.32, 0.32 and 0.3). The controllers were able to control the individual voltages to the point where 2.52 mA was drawn from the stack at a load of 499.9 Ω (delivering 3.18 mW). The controllers were able to reject the disturbances and perturbations caused by electrical loading, temperature and substrate concentration.

  16. An approach for calculating a confidence interval from a single aquatic sample for monitoring hydrophobic organic contaminants.

    PubMed

    Matzke, Melissa M; Allan, Sarah E; Anderson, Kim A; Waters, Katrina M

    2012-12-01

    The use of passive sampling devices (PSDs) for monitoring hydrophobic organic contaminants in aquatic environments can entail logistical constraints that often limit a comprehensive statistical sampling plan, thus resulting in a restricted number of samples. The present study demonstrates an approach for using the results of a pilot study designed to estimate sampling variability, which in turn can be used as variance estimates for confidence intervals for future n = 1 PSD samples of the same aquatic system. Sets of three to five PSDs were deployed in the Portland Harbor Superfund site for three sampling periods over the course of two years. The PSD filters were extracted and, as a composite sample, analyzed for 33 polycyclic aromatic hydrocarbon compounds. The between-sample and within-sample variances were calculated to characterize sources of variability in the environment and sampling methodology. A method for calculating a statistically reliable and defensible confidence interval for the mean of a single aquatic passive sampler observation (i.e., n = 1) using an estimate of sample variance derived from a pilot study is presented. Coverage probabilities are explored over a range of variance values using a Monte Carlo simulation.

  17. Organochlorine and metal pollution in aquatic organisms sampled in the Donana National Park during the period 1983-1986

    SciTech Connect

    Rico, M.C.; Hernandez, L.M.; Gonzalez, M.J.; Fernandez, M.A.; Montero, M.C.

    1987-12-01

    The study area, Donana National Park, is located in the South South-West of Spain, and this is one of the most important reservation of Europe. Samples of aquatic organism were obtained from the principal waterway of Donana National Park to determine the degree of organochlorine and metal contamination of this environment. The sampling was carried out during the period 1983-1986 in order to collect six aquatic species in four sites along the Brazo de la Torre. An agricultural area in the North-West side of the Park and a working mine at about 40 km from its northern boundary were considered as the likely main polluting sources of organochlorine pesticides, PCBs, and heavy metals respectively. The aquatic organism species chosen for analysis were: American crayfish (Procambarus clarckii), carp (Cyprinus carpio), barbel (Barbus barbus), grey mullet (Mugil capito), eel (Anguilla anguilla), and frog (Rana perezi).

  18. Genomic Sequencing of Single Microbial Cells from Environmental Samples

    SciTech Connect

    Ishoey, Thomas; Woyke, Tanja; Stepanauskas, Ramunas; Novotny, Mark; Lasken, Roger S.

    2008-02-01

    Recently developed techniques allow genomic DNA sequencing from single microbial cells [Lasken RS: Single-cell genomic sequencing using multiple displacement amplification, Curr Opin Microbiol 2007, 10:510-516]. Here, we focus on research strategies for putting these methods into practice in the laboratory setting. An immediate consequence of single-cell sequencing is that it provides an alternative to culturing organisms as a prerequisite for genomic sequencing. The microgram amounts of DNA required as template are amplified from a single bacterium by a method called multiple displacement amplification (MDA) avoiding the need to grow cells. The ability to sequence DNA from individual cells will likely have an immense impact on microbiology considering the vast numbers of novel organisms, which have been inaccessible unless culture-independent methods could be used. However, special approaches have been necessary to work with amplified DNA. MDA may not recover the entire genome from the single copy present in most bacteria. Also, some sequence rearrangements can occur during the DNA amplification reaction. Over the past two years many research groups have begun to use MDA, and some practical approaches to single-cell sequencing have been developed. We review the consensus that is emerging on optimum methods, reliability of amplified template, and the proper interpretation of 'composite' genomes which result from the necessity of combining data from several single-cell MDA reactions in order to complete the assembly. Preferred laboratory methods are considered on the basis of experience at several large sequencing centers where >70% of genomes are now often recovered from single cells. Methods are reviewed for preparation of bacterial fractions from environmental samples, single-cell isolation, DNA amplification by MDA, and DNA sequencing.

  19. Aquatic fulvic acids in microbially based ecosystems: results from two desert lakes in Antarctica

    USGS Publications Warehouse

    McKnight, Diane M.; Aiken, G.R.; Smith, R.L.

    1991-01-01

    These lakes receive very limited input of organic material from the surrounding barren desert, but they sustain algal and bacterial populations under permanent ice cover. One lake has an extensive anoxic zone and high salinities; the other is oxic and has low salinities. Despite these differences, fulvic acids from both lakes had similar elemental compositions, carbon distributions, and amino acid contents, indicating that the chemistry of microbially derived fulvic acvids is not strongly influenced by chemical conditions in the water column. Compared to fulvic acids from other natural waters, these fulvic acids have low C:N atomic ratios (19-25) and low contents of aromatic carbons (5-7% of total carbon atoms); they are most similar to marine fulvic acids. -from Authors

  20. In Situ Treatment of PCBs by Anaerobic Microbial Dechlorination in Aquatic Sediment: Are We There Yet?

    PubMed Central

    Sowers, Kevin R.; May, Harold D.

    2012-01-01

    The remediation of PCBs in soils and sediments remains a particularly difficult problem to solve. The possibility of in situ degradation by microorganisms has been pursued for many years since this approach has the potential to provide a cost-effective and environmentally sustainable alternative to dredging for treatment of PCB impacted sites. Because PCBs are hydrophobic and partition into organic material they accumulate in anoxic environments well poised to support anaerobic dechlorination of highly chlorinated commercial PCBs to congeners that are susceptible to complete aerobic degradation. Laboratory research over the past 25 years is now leading to new microbial technologies that could soon be tested for treatment of PCB impacted soils and sediments in the field. PMID:23102490

  1. Molybdenum limitation of microbial nitrogen assimilation in aquatic ecosystems and pure cultures.

    PubMed

    Glass, Jennifer B; Axler, Richard P; Chandra, Sudeep; Goldman, Charles R

    2012-01-01

    Molybdenum (Mo) is an essential micronutrient for biological assimilation of nitrogen gas and nitrate because it is present in the cofactors of nitrogenase and nitrate reductase enzymes. Although Mo is the most abundant transition metal in seawater (107 nM), it is present in low concentrations in most freshwaters, typically <20 nM. In 1960, it was discovered that primary productivity was limited by Mo scarcity (2-4 nM) in Castle Lake, a small, meso-oligotrophic lake in northern California. Follow up studies demonstrated that Mo also limited primary productivity in lakes in New Zealand, Alaska, and the Sierra Nevada. Research in the 1970s and 1980s showed that Mo limited primary productivity and nitrate uptake in Castle Lake only during periods of the growing season when nitrate concentrations were relatively high because ammonium assimilation does not require Mo. In the years since, research has shifted to investigate whether Mo limitation also occurs in marine and soil environments. Here we review studies of Mo limitation of nitrogen assimilation in natural microbial communities and pure cultures. We also summarize new data showing that the simultaneous addition of Mo and nitrate causes increased activity of proteins involved in nitrogen assimilation in the hypolimnion of Castle Lake when ammonium is scarce. Furthermore, we suggest that meter-scale Mo and oxygen depth profiles from Castle Lake are consistent with the hypothesis that nitrogen-fixing cyanobacteria in freshwater periphyton communities have higher Mo requirements than other microbial communities. Finally, we present topics for future research related to Mo bioavailability through time and with changing oxidation state.

  2. Molybdenum limitation of microbial nitrogen assimilation in aquatic ecosystems and pure cultures

    PubMed Central

    Glass, Jennifer B.; Axler, Richard P.; Chandra, Sudeep; Goldman, Charles R.

    2012-01-01

    Molybdenum (Mo) is an essential micronutrient for biological assimilation of nitrogen gas and nitrate because it is present in the cofactors of nitrogenase and nitrate reductase enzymes. Although Mo is the most abundant transition metal in seawater (107 nM), it is present in low concentrations in most freshwaters, typically <20 nM. In 1960, it was discovered that primary productivity was limited by Mo scarcity (2–4 nM) in Castle Lake, a small, meso-oligotrophic lake in northern California. Follow up studies demonstrated that Mo also limited primary productivity in lakes in New Zealand, Alaska, and the Sierra Nevada. Research in the 1970s and 1980s showed that Mo limited primary productivity and nitrate uptake in Castle Lake only during periods of the growing season when nitrate concentrations were relatively high because ammonium assimilation does not require Mo. In the years since, research has shifted to investigate whether Mo limitation also occurs in marine and soil environments. Here we review studies of Mo limitation of nitrogen assimilation in natural microbial communities and pure cultures. We also summarize new data showing that the simultaneous addition of Mo and nitrate causes increased activity of proteins involved in nitrogen assimilation in the hypolimnion of Castle Lake when ammonium is scarce. Furthermore, we suggest that meter-scale Mo and oxygen depth profiles from Castle Lake are consistent with the hypothesis that nitrogen-fixing cyanobacteria in freshwater periphyton communities have higher Mo requirements than other microbial communities. Finally, we present topics for future research related to Mo bioavailability through time and with changing oxidation state. PMID:22993512

  3. Aquatic invertebrate sampling at selected outfalls in Operable Unit 1082; Technical areas 9, 11, 16 and 22

    SciTech Connect

    Cross, S.

    1995-09-01

    The Ecological Studies Team (EST) of ESH-20 at Los Alamos National Laboratory conducted preliminary aquatic sampling at outfalls within Operable Unit 1082 and nearby natural waterways. Eleven outfalls were sampled a total of eighteen times. Three natural waterways (upper Pajarito Canyon, Starmer`s Gulch, and Bulldog Spring) in the vicinity were sampled a total of six times. At most sites, EST recorded hydrological condition, physico-chemical parameters, wildlife uses, and vegetation. At each outfall with water and each natural waterway, EST collected an aquatic invertebrate sample which was analyzed by taxa composition, Wilhm`s biodiversity index, the community tolerance quotient (CTQ), and density. The physico-chemical parameters at most outfalls and natural waterways fell within the normal range of natural waters in the area. However, the outfalls are characterized by low biodiversity and severely stressed communities composed of a restricted number of taxa. The habitat at the other outfalls could probably support well-developed aquatic communities if sufficient water was available. At present, the hydrology at these outfalls is too slight and/or sporadic to support such a community in the foreseeable future. In contrast to the outfalls, the natural waterways of the area had greater densities of aquatic invertebrates, higher biodiversities, and lower CTQs.

  4. Microbial Community Composition and Ecology of an Acidic Aquatic Environment: The Tinto River, Spain.

    PubMed

    López-Archilla, A.I.; Marin, I.; Amils, R.

    2001-01-01

    We studied the correlation between physicochemical and biological characteristics of an acidic river, the Tinto River, in Southwestern Spain. The Tinto River is an extreme environment characterized by its low pH (mean of 2.2) and high concentrations of heavy metals (Fe 2.3 g/L, Zn 0.22 g/L, Cu 0.11 g/L). These extreme conditions are the product of the metabolic activity of chemolithotrophic microorganisms, including iron- and sulfur-oxidizing bacteria, that can be found in high concentrations in its waters. The food chain in the river is very constrained and exclusively microbial. Primary productivity in the Tinto River is the sum of photosynthetic and chemolithotrophic activity. Heterotrophic bacteria and fungi are the major decomposers and protists are the major predators. A correlation analysis including the physicochemical and biological variables suggested a close relationship between the acidic pH values and abundance of both chemolithotrophic bacteria and filamentous fungi. Chemolithotrophic bacteria correlated with the heavy metals found in the river. A principal component analysis of the biotic and abiotic variables suggested that the Tinto River ecosystem can be described as a function of three main groups of variables: pH values, metal concentrations, and biological productivity.

  5. Aquatic Global Passive Sampling (AQUA-GAPS) Revisited: First Steps toward a Network of Networks for Monitoring Organic Contaminants in the Aquatic Environment.

    PubMed

    Lohmann, Rainer; Muir, Derek; Zeng, Eddy Y; Bao, Lian-Jun; Allan, Ian J; Arinaitwe, Kenneth; Booij, Kees; Helm, Paul; Kaserzon, Sarit; Mueller, Jochen F; Shibata, Yasuyuki; Smedes, Foppe; Tsapakis, Manolis; Wong, Charles S; You, Jing

    2017-02-07

    Organic contaminants, in particular persistent organic pollutants (POPs), adversely affect water quality and aquatic food webs across the globe. As of now, there is no globally consistent information available on concentrations of dissolved POPs in water bodies. The advance of passive sampling techniques has made it possible to establish a global monitoring program for these compounds in the waters of the world, which we call the Aquatic Global Passive Sampling (AQUA-GAPS) network. A recent expert meeting discussed the background, motivations, and strategic approaches of AQUA-GAPS, and its implementation as a network of networks for monitoring organic contaminants (e.g., POPs and others contaminants of concern). Initially, AQUA-GAPS will demonstrate its operating principle via two proof-of-concept studies focused on the detection of legacy and emerging POPs in freshwater and coastal marine sites using both polyethylene and silicone passive samplers. AQUA-GAPS is set up as a decentralized network, which is open to other participants from around the world to participate in deployments and to initiate new studies. In particular, participants are sought to initiate deployments and studies investigating the presence of legacy and emerging POPs in Africa, Central, and South America.

  6. Molecular ecology of aquatic microbes

    SciTech Connect

    1994-12-31

    Abstracts of reports are presented from a meeting on Molecular Ecology of Aquatic Microbes. Topics included: opportunities offered to aquatic ecology by molecular biology; the role of aquatic microbes in biogeochemical cycles; characterization of the microbial community; the effect of the environment on aquatic microbes; and the targeting of specific biological processes.

  7. Alternative model and approach for determining microbial heterotrophic activities in aquatic systems.

    PubMed Central

    Dietz, A S; Albright, L J; Tuominen, T

    1977-01-01

    Increasing amounts of high-specific-activity tritiated organic compounds were added to samples of several natural waters such that in situ substrate concentrations might be approximated. The uptake responses by the native heterotrophic microflora suggested that (i) heterotrophic populations metabolize the added nutrients, but (ii) these responses are not necessarily a reflection of Michaelis-Menten enzyme kinetics. The uptake kinetics appeared to be due to dilution of the naturally occurring metabolite by added radioactive substrate and physiological responses of the microflora to organic enrichment. PMID:326186

  8. Chemical Composition of Aquatic Dissolved Organic Matter in Five Boreal Forest Catchments Sampled in Spring and Fall Seasons

    SciTech Connect

    Schumacher,M.; Christl, I.; Vogt, R.; Barmettler, K.; Jacobsen, C.; Kretzschmar, R.

    2006-01-01

    The chemical composition and carbon isotope signature of aquatic dissolved organic matter (DOM) in five boreal forest catchments in Scandinavia were investigated. The DOM was isolated during spring and fall seasons using a reverse osmosis technique. The DOM samples were analyzed by elemental analysis, FT-IR, solid-state CP-MAS {sup 13}C-NMR, and C-1s NEXAFS spectroscopy. In addition, the relative abundance of carbon isotopes ({sup 12}C, {sup 13}C, {sup 14}C) in the samples was measured. There were no significant differences in the chemical composition or carbon isotope signature of the DOM sampled in spring and fall seasons. Also, differences in DOM composition between the five catchments were minor. Compared to reference peat fulvic and humic acids, all DOM samples were richer in O-alkyl carbon and contained less aromatic and phenolic carbon, as shown by FT-IR, {sup 13}C-NMR, and C-1s NEXAFS spectroscopy. The DOM was clearly enriched in {sup 14}C relative to the NBS oxalic acid standard of 1950, indicating that the aquatic DOM contained considerable amounts of organic carbon younger than about 50 years. The weight-based C:N ratios of 31 {+-} 6 and the {delta}{sup 13}Cvalues of -29 {+-} 2{per_thousand}indicate that the isolated DOM is of terrestrial rather than aquatic origin. We conclude that young, hydrophilic carbon compounds of terrestrial origin are predominant in the samples investigated, and that the composition of the aquatic DOM in the studied boreal forest catchments is rather stable during low to intermediate flow conditions.

  9. Estimating site occupancy rates for aquatic plants using spatial sub-sampling designs when detection probabilities are less than one

    USGS Publications Warehouse

    Nielson, Ryan M.; Gray, Brian R.; McDonald, Lyman L.; Heglund, Patricia J.

    2011-01-01

    Estimation of site occupancy rates when detection probabilities are <1 is well established in wildlife science. Data from multiple visits to a sample of sites are used to estimate detection probabilities and the proportion of sites occupied by focal species. In this article we describe how site occupancy methods can be applied to estimate occupancy rates of plants and other sessile organisms. We illustrate this approach and the pitfalls of ignoring incomplete detection using spatial data for 2 aquatic vascular plants collected under the Upper Mississippi River's Long Term Resource Monitoring Program (LTRMP). Site occupancy models considered include: a naïve model that ignores incomplete detection, a simple site occupancy model assuming a constant occupancy rate and a constant probability of detection across sites, several models that allow site occupancy rates and probabilities of detection to vary with habitat characteristics, and mixture models that allow for unexplained variation in detection probabilities. We used information theoretic methods to rank competing models and bootstrapping to evaluate the goodness-of-fit of the final models. Results of our analysis confirm that ignoring incomplete detection can result in biased estimates of occupancy rates. Estimates of site occupancy rates for 2 aquatic plant species were 19–36% higher compared to naive estimates that ignored probabilities of detection <1. Simulations indicate that final models have little bias when 50 or more sites are sampled, and little gains in precision could be expected for sample sizes >300. We recommend applying site occupancy methods for monitoring presence of aquatic species.

  10. Sampling design for aquatic invasive species early detection in Great Lakes ports

    EPA Science Inventory

    From 2006-2012, we evaluated a pilot aquatic invasive species (AIS) early detection monitoring program in Lake Superior that was designed to detect newly introduced fishes. We established survey protocols for three major ports (Duluth-Superior, Sault Ste. Marie, Thunder Bay) and ...

  11. INNOVATIVE TECHNOLOGY EVALUATION REPORT, SEDIMENT SAMPLING TECHNOLOGY, AQUATIC RESEARCH INSTRUMENTS, RUSSIAN PEAT BORER

    EPA Science Inventory

    The Russian Peat Borer designed and fabricated by Aquatic Research Instruments was demonstrated under the U.S. Environmental Protection Agency (EPA) Superfund Innovative Technology Evaluation Program in April and May 1999 at sites in EPA Regions 1 and 5, respectively. In additio...

  12. INNOVATIVE TECHNOLOGY EVALUATION REPORT, SEDIMENT SAMPLING TECHNOLOGY, AQUATIC RESEARCH INSTRUMENTS, RUSSIAN PEAT BORER

    EPA Science Inventory

    The Russian Peat Borer designed and fabricated by Aquatic Research Instruments was demonstrated under the U.S. Environmental Protection Agency (EPA) Superfund Innovative Technology Evaluation Program in April and May 1999 at sites in EPA Regions 1 and 5, respectively. In additio...

  13. Determination of the Biologically Relevant Sampling Depth for Terrestrial and Aquatic Ecological Risk Assessments (Final Report)

    EPA Science Inventory

    This technical paper provides defensible approximations for what the depth of the biologically active zone, or “biotic zone” is within certain environments. The methods used in this study differ somewhat between Part 1 (Terrestrial Biotic Zone) and Part 2 (Aquatic Biotic Zone). ...

  14. Sampling design for early detection of aquatic invasive species in Great Lakes ports

    EPA Science Inventory

    We evaluated a pilot adaptive monitoring program for aquatic invasive species (AIS) early detection in Lake Superior. The monitoring program is designed to detect newly-introduced fishes, and encompasses the lake’s three major ports (Duluth-Superior, Sault Ste. Marie, Thund...

  15. Sampling design for early detection of aquatic invasive species in Great Lakes ports

    EPA Science Inventory

    We evaluated a pilot adaptive monitoring program for aquatic invasive species (AIS) early detection in Lake Superior. The monitoring program is designed to detect newly-introduced fishes, and encompasses the lake’s three major ports (Duluth-Superior, Sault Ste. Marie, Thund...

  16. Sampling design for aquatic invasive species early detection in Great Lakes ports

    EPA Science Inventory

    From 2006-2012, we evaluated a pilot aquatic invasive species (AIS) early detection monitoring program in Lake Superior that was designed to detect newly introduced fishes. We established survey protocols for three major ports (Duluth-Superior, Sault Ste. Marie, Thunder Bay) and ...

  17. Microbial Habitability in Gale Crater: Sample Analysis at Mars (SAM) Instrument Detection of Microbial Essential Carbon and Nitrogen

    NASA Technical Reports Server (NTRS)

    Sutter, B.; Ming, D. W.; Eigenbrode, J. E.; Steele, A.; Stern, J. C.; Gonzalez, R. N.; McAdam, A. C.; Mahaffy, P. R.

    2016-01-01

    Chemical analyses of Mars soils and sediments from previous landed missions have demonstrated that Mars surface materials possessed major (e.g., P, K, Ca, Mg, S) and minor (e.g., Fe, Mn, Zn, Ni, Cl) elements essential to support microbial life. However, the detection of microbial essential organic-carbon (C) and nitrate have been more elusive until the Mars Science Laboratory (MSL) rover mission. Nitrate and organic-C in Gale Crater, Mars have been detected by the Sample Analysis at Mars (SAM) instrument onboard the MSL Curiosity rover. Eolian fines and drilled sedimentary rock samples were heated in the SAM oven from approximately 30 to 860 degrees Centigrade where evolved gases (e.g., nitrous oxide (NO) and CO2) were released and analyzed by SAM’s quadrupole mass spectrometer (MS). The temperatures of evolved NO was assigned to nitrate while evolved CO2 was assigned to organic-C and carbonate. The CO2 releases in several samples occurred below 450 degrees Centigrade suggesting organic-C dominated in those samples. As much as 7 micromoles NO3-N per gram and 200 micromoles CO2-C per gram have been detected in the Gale Crater materials. These N and C levels coupled with assumed microbial biomass (9 x 10 (sup -7) micrograms per cell) C (0.5 micrograms C per micrograms cell) and N (0.14 micrograms N per micrograms cell) requirements, suggests that less than 1 percent and less than 10 percent of Gale Crater C and N, respectively, would be required if available, to accommodate biomass requirements of 1 by 10 (sup 5) cells per gram sediment. While nitrogen is the limiting nutrient, the potential exists that sufficient N and organic-C were present to support limited heterotrophic microbial populations that may have existed on ancient Mars.

  18. Trends in Selenium Utilization in Marine Microbial World Revealed through the Analysis of the Global Ocean Sampling (GOS) Project

    PubMed Central

    Zhang, Yan; Gladyshev, Vadim N.

    2008-01-01

    Selenium is an important trace element that occurs in proteins in the form of selenocysteine (Sec) and in tRNAs in the form of selenouridine. Recent large-scale metagenomics projects provide an opportunity for understanding global trends in trace element utilization. Herein, we characterized the selenoproteome of the microbial marine community derived from the Global Ocean Sampling (GOS) expedition. More than 3,600 selenoprotein gene sequences belonging to 58 protein families were detected, including sequences representing 7 newly identified selenoprotein families, such as homologs of ferredoxin–thioredoxin reductase and serine protease. In addition, a new eukaryotic selenoprotein family, thiol reductase GILT, was identified. Most GOS selenoprotein families originated from Cys-containing thiol oxidoreductases. In both Pacific and Atlantic microbial communities, SelW-like and SelD were the most widespread selenoproteins. Geographic location had little influence on Sec utilization as measured by selenoprotein variety and the number of selenoprotein genes detected; however, both higher temperature and marine (as opposed to freshwater and other aquatic) environment were associated with increased use of this amino acid. Selenoproteins were also detected with preference for either environment. We identified novel fusion forms of several selenoproteins that highlight redox activities of these proteins. Almost half of Cys-containing SelDs were fused with NADH dehydrogenase, whereas such SelD forms were rare in terrestrial organisms. The selenouridine utilization trait was also analyzed and showed an independent evolutionary relationship with Sec utilization. Overall, our study provides insights into global trends in microbial selenium utilization in marine environments. PMID:18551170

  19. Catabolism of Tritiated Thymidine by Aquatic Microbial Communities and Incorporation of Tritium into RNA and Protein †

    PubMed Central

    Brittain, Andrew M.; Karl, David M.

    1990-01-01

    The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. All microbial assemblages tested exhibited significant labeling of RNA and protein (i.e., nonspecific labeling), as determined by differential acid-base hydrolysis. Nonspecific labeling was greatest in sediment samples, for which ≥95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. Four different RNA hydrolysis reagents (KOH, NaOH, piperidine, and enzymes) solubilized tritium from cold trichloroacetic acid precipitates. High-pressure liquid chromatography separation of piperidine hydrolysates followed by measurement of isolated monophosphates confirmed the labeling of RNA and indicated that tritium was recovered primarily in CMP and AMP residues. We also evaluated the specificity of [2-3H]adenine incorporation into adenylate residues in both RNA and DNA in parallel with the [3H]thymidine experiments and compared the degree of nonspecific labeling by [3H]adenine with that derived from [3H]thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products by high-pressure liquid chromatography separation of the cell-free medium. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples. PMID

  20. Random sampling process leads to overestimation of β-diversity of microbial communities.

    PubMed

    Zhou, Jizhong; Jiang, Yi-Huei; Deng, Ye; Shi, Zhou; Zhou, Benjamin Yamin; Xue, Kai; Wu, Liyou; He, Zhili; Yang, Yunfeng

    2013-06-11

    The site-to-site variability in species composition, known as β-diversity, is crucial to understanding spatiotemporal patterns of species diversity and the mechanisms controlling community composition and structure. However, quantifying β-diversity in microbial ecology using sequencing-based technologies is a great challenge because of a high number of sequencing errors, bias, and poor reproducibility and quantification. Herein, based on general sampling theory, a mathematical framework is first developed for simulating the effects of random sampling processes on quantifying β-diversity when the community size is known or unknown. Also, using an analogous ball example under Poisson sampling with limited sampling efforts, the developed mathematical framework can exactly predict the low reproducibility among technically replicate samples from the same community of a certain species abundance distribution, which provides explicit evidences of random sampling processes as the main factor causing high percentages of technical variations. In addition, the predicted values under Poisson random sampling were highly consistent with the observed low percentages of operational taxonomic unit (OTU) overlap (<30% and <20% for two and three tags, respectively, based on both Jaccard and Bray-Curtis dissimilarity indexes), further supporting the hypothesis that the poor reproducibility among technical replicates is due to the artifacts associated with random sampling processes. Finally, a mathematical framework was developed for predicting sampling efforts to achieve a desired overlap among replicate samples. Our modeling simulations predict that several orders of magnitude more sequencing efforts are needed to achieve desired high technical reproducibility. These results suggest that great caution needs to be taken in quantifying and interpreting β-diversity for microbial community analysis using next-generation sequencing technologies. IMPORTANCE Due to the vast diversity

  1. Non-destructive sampling of rock-dwelling microbial communities using sterile adhesive tape.

    PubMed

    Cutler, Nick A; Oliver, Anna E; Viles, Heather A; Whiteley, Andrew S

    2012-12-01

    Building stone provides a habitat for an array of microorganisms, many of which have been demonstrated to have a deleterious effect on the appearance and/or structural integrity of stone masonry. It is essential to understand the composition and structure of stone-dwelling (lithobiontic) microbial communities if successful stone conservation strategies are to be applied, particularly in the face of global environmental change. Ideally, the techniques used to sample such assemblages should be non-destructive due to the sensitive conservation status of many stone buildings. This paper quantitatively assesses the performance of sterile adhesive tape as a non-destructive sampling technique and compares the results of tape sampling with an alternative, destructive, sampling method. We used DNA fingerprinting (TRFLP) to characterise the algal, fungal and bacterial communities living on a stone slab. Our results demonstrate that tape sampling may be used to collect viable quantities of microbial DNA from environmental samples. This technique is ideally suited to the sampling of microbial biofilms, particularly when these communities are dominated by green algae. It provides a good approximation of total community diversity (i.e. the aggregate diversity of epilithic and endolithic communities). Tape sampling is straightforward, rapid and cost effective. When combined with molecular analytical techniques, this sampling method has the potential to make a major contribution to efforts to understand the structure of lithobiontic microbial communities and our ability to predict the response of such communities to future environmental change.

  2. PhyloChip™ microarray comparison of sampling methods used for coral microbial ecology.

    PubMed

    Kellogg, Christina A; Piceno, Yvette M; Tom, Lauren M; DeSantis, Todd Z; Zawada, David G; Andersen, Gary L

    2012-01-01

    Interest in coral microbial ecology has been increasing steadily over the last decade, yet standardized methods of sample collection still have not been defined. Two methods were compared for their ability to sample coral-associated microbial communities: tissue punches and foam swabs, the latter being less invasive and preferred by reef managers. Four colonies of star coral, Montastraea annularis, were sampled in the Dry Tortugas National Park (two healthy and two with white plague disease). The PhyloChip™ G3 microarray was used to assess microbial community structure of amplified 16S rRNA gene sequences. Samples clustered based on methodology rather than coral colony. Punch samples from healthy and diseased corals were distinct. All swab samples clustered closely together with the seawater control and did not group according to the health state of the corals. Although more microbial taxa were detected by the swab method, there is a much larger overlap between the water control and swab samples than punch samples, suggesting some of the additional diversity is due to contamination from water absorbed by the swab. While swabs are useful for noninvasive studies of the coral surface mucus layer, these results show that they are not optimal for studies of coral disease. Published by Elsevier B.V.

  3. PhyloChip™ microarray comparison of sampling methods used for coral microbial ecology

    USGS Publications Warehouse

    Kellogg, Christina A.; Piceno, Yvette M.; Tom, Lauren M.; DeSantis, Todd Z.; Zawada, David G.; Andersen, Gary L.

    2012-01-01

    Interest in coral microbial ecology has been increasing steadily over the last decade, yet standardized methods of sample collection still have not been defined. Two methods were compared for their ability to sample coral-associated microbial communities: tissue punches and foam swabs, the latter being less invasive and preferred by reef managers. Four colonies of star coral, Montastraea annularis, were sampled in the Dry Tortugas National Park (two healthy and two with white plague disease). The PhyloChip™ G3 microarray was used to assess microbial community structure of amplified 16S rRNA gene sequences. Samples clustered based on methodology rather than coral colony. Punch samples from healthy and diseased corals were distinct. All swab samples clustered closely together with the seawater control and did not group according to the health state of the corals. Although more microbial taxa were detected by the swab method, there is a much larger overlap between the water control and swab samples than punch samples, suggesting some of the additional diversity is due to contamination from water absorbed by the swab. While swabs are useful for noninvasive studies of the coral surface mucus layer, these results show that they are not optimal for studies of coral disease.

  4. Application of the BMWP-Costa Rica biotic index in aquatic biomonitoring: sensitivity to collection method and sampling intensity.

    PubMed

    Gutiérrez-Fonseca, Pablo E; Lorion, Christopher M

    2014-04-01

    The use of aquatic macroinvertebrates as bio-indicators in water quality studies has increased considerably over the last decade in Costa Rica, and standard biomonitoring methods have now been formulated at the national level. Nevertheless, questions remain about the effectiveness of different methods of sampling freshwater benthic assemblages, and how sampling intensity may influence biomonitoring results. In this study, we compared the results of qualitative sampling using commonly applied methods with a more intensive quantitative approach at 12 sites in small, lowland streams on the southern Caribbean slope of Costa Rica. Qualitative samples were collected following the official protocol using a strainer during a set time period and macroinvertebrates were field-picked. Quantitative sampling involved collecting ten replicate Surber samples and picking out macroinvertebrates in the laboratory with a stereomicroscope. The strainer sampling method consistently yielded fewer individuals and families than quantitative samples. As a result, site scores calculated using the Biological Monitoring Working Party-Costa Rica (BMWP-CR) biotic index often differed greatly depending on the sampling method. Site water quality classifications using the BMWP-CR index differed between the two sampling methods for 11 of the 12 sites in 2005, and for 9 of the 12 sites in 2006. Sampling intensity clearly had a strong influence on BMWP-CR index scores, as well as perceived differences between reference and impacted sites. Achieving reliable and consistent biomonitoring results for lowland Costa Rican streams may demand intensive sampling and requires careful consideration of sampling methods.

  5. [Wound microbial sampling methods in surgical practice, imprint techniques].

    PubMed

    Chovanec, Z; Veverková, L; Votava, M; Svoboda, J; Peštál, A; Doležel, J; Jedlička, V; Veselý, M; Wechsler, J; Čapov, I

    2012-12-01

    The wound is a damage of tissue. The process of healing is influenced by many systemic and local factors. The most crucial and the most discussed local factor of wound healing is infection. Surgical site infection in the wound is caused by micro-organisms. This information is known for many years, however the conditions leading to an infection occurrence have not been sufficiently described yet. Correct sampling technique, correct storage, transportation, evaluation, and valid interpretation of these data are very important in clinical practice. There are many methods for microbiological sampling, but the best one has not been yet identified and validated. We aim to discuss the problem with the focus on the imprint technique.

  6. Microbial content of house dust samples determined with qPCR.

    PubMed

    Kaarakainen, Pasi; Rintala, Helena; Vepsäläinen, Asko; Hyvärinen, Anne; Nevalainen, Aino; Meklin, Teija

    2009-08-01

    This study was designed to produce information about microbial concentrations using qPCR and their variation in different seasons and home environments with analyses of two types of house dust samples. Also the correlations between the two types of samples and the reproducibility of the parallel subsamples were studied. Two types of vacuumed house dust samples, rug dust and vacuum cleaner bag dust, were collected in 5 normal urban homes in four different seasons (N=20+20). From all dust samples, five parallel subsamples were subjected to qPCR analyses of 17 microbial species or assay groups of microbes. The highest fungal concentrations were found for the Penicillium/Aspergillus/Paecilomyces variotii group, and for the species Aspergillus penicillioides, Aureobasidium pullulans, Cladosporium cladosporioides and Cladosporium herbarum. These species/groups were present in almost all samples. The two types of dust samples gave similar results for most microbial species or groups analyzed, but in general, concentrations were slightly higher in rug dust than in dust from vacuum cleaner bag. Microbial concentrations varied significantly between different seasons and hence the similarity of samples within home was mainly low. The concentrations varied significantly also between different home environments. The reproducibility of the parallel subsamples was good or moderate for most of the analyzed species or assay groups. However, further studies are needed to fully understand the factors causing variation in these methods. Nevertheless, in order to show actual differences in fungal concentrations between urban homes with no known microbial sources, all dust samples to be compared should be taken during the same season.

  7. Determination of selected cardiovascular active compounds in environmental aquatic samples--Methods and results, a review of global publications from the last 10 years.

    PubMed

    Stankiewicz, Albert; Giebułtowicz, Joanna; Stankiewicz, Urszula; Wroczyński, Piotr; Nałęcz-Jawecki, Grzegorz

    2015-11-01

    In recent years cardiovascular diseases were the second most common cause of death worldwide. Therefore, the consumption of cardiovascular drugs is high, which might result in an increase of them in the environment. The major source of aquatic environmental contamination is still effluents of wastewater treatment plants (WWTPs). Unfortunately removal of cardiovascular active compounds and/or their metabolites in WWTP is still unsatisfactory. Among microbial and abiotic degradation of these compounds during wastewater processes, photolysis and photodegradation of cardiovascular drugs also play an important role. New formed compounds may be more toxic or retain the properties of parent compounds. Thus the main goal of this paper was to provide a detailed and comprehensive review of used analytical methods, coupled to liquid chromatography-tandem mass spectrometry, to determine the presence of cardiovascular compounds in surface waters as well as WTTPs effluents and influents. Exhaustive preparation for mass spectrometry detection and quantitation including samples pre-treatment, and the common problem of the matrix effect are thoroughly explored in this paper. Additionally, the article provides some hints in respect of recently noted problematic issue related to the availability of specific standards for the analysis of drug's metabolites. Furthermore, information concerning the metabolism of cardiovascular active compounds including differences in metabolism within enantiomers is described. This article also touches on the problems associated with environmental risk assessment due to the presence of cardiovasculars in the environment. The paper also tries to explain differences in concentrations among cardiovascular compounds between countries worldwide.

  8. DNA-based determination of microbial biomass suitable for frozen and alkaline soil samples

    NASA Astrophysics Data System (ADS)

    Semenov, Mikhail; Blagodatskaya, Evgeniya; Kogut, Boris; Kuzyakov, Yakov

    2015-04-01

    Microbial biomass is a sensitive indicator of changes due to soil management, long before other basic soil measures such as Corg or Ntot. Improvement of methods for determination of microbial biomass still remains relevant, and these methods should be correctly applicable for the soil samples being in various state. This study was designed to demonstrate the applicability of DNA-based determination of microbial biomass under conditions when the common basic approaches, namely chloroform fumigation-extraction (CFE) and substrate-induced respiration (SIR), are restricted by certain soil properties, experimental designs or research needs, e.g. in frozen, alkaline or carbonaceous soils. We compared microbial biomass determined by CFE, SIR and by DNA approaches in the range of neutral and slightly alkaline Chernozem and alkaline Calcisol of semi-arid climate. The samples of natural and agricultural ecosystems were taken throughout the soil profile from long-term static field experiments in the European part of Russia. Extraction and subsequent quantification of dsDNA revealed a strong agreement with SIR and CFE when analyzing the microbial biomass content in soils with pH below 8. The conversion factors (FDNA) from dsDNA to SIR-Cmic (5.10) and CFE-Cmic (4.41) were obtained by testing a range of the soil samples down to 1.5 m depth and indicated a good reproducibility of DNA-based estimations. In alkaline soils (pH > 8), CO2 retention due to alkaline pH and exchange with carbonates resulted in a strong underestimation of soil microbial biomass by SIR or even in the absence of any CO2 emission, especially at low absolute values of microbial biomass in subsoil. Correction of CO2 efflux by theoretical retention pH-dependent factors caused overestimation of SIR-biomass. In alkaline conditions, DNA extraction proved to be a reliable alternative for microbial biomass determination. Moreover, the DNA-based approach can serve as an excellent alternative enabling correct

  9. Quantitative microbial faecal source tracking with sampling guided by hydrological catchment dynamics

    PubMed Central

    Reischer, G H; Haider, J M; Sommer, R; Stadler, H; Keiblinger, K M; Hornek, R; Zerobin, W; Mach, R L; Farnleitner, A H

    2008-01-01

    The impairment of water quality by faecal pollution is a global public health concern. Microbial source tracking methods help to identify faecal sources but the few recent quantitative microbial source tracking applications disregarded catchment hydrology and pollution dynamics. This quantitative microbial source tracking study, conducted in a large karstic spring catchment potentially influenced by humans and ruminant animals, was based on a tiered sampling approach: a 31-month water quality monitoring (Monitoring) covering seasonal hydrological dynamics and an investigation of flood events (Events) as periods of the strongest pollution. The detection of a ruminant-specific and a human-specific faecal Bacteroidetes marker by quantitative real-time PCR was complemented by standard microbiological and on-line hydrological parameters. Both quantitative microbial source tracking markers were detected in spring water during Monitoring and Events, with preponderance of the ruminant-specific marker. Applying multiparametric analysis of all data allowed linking the ruminant-specific marker to general faecal pollution indicators, especially during Events. Up to 80% of the variation of faecal indicator levels during Events could be explained by ruminant-specific marker levels proving the dominance of ruminant faecal sources in the catchment. Furthermore, soil was ruled out as a source of quantitative microbial source tracking markers. This study demonstrates the applicability of quantitative microbial source tracking methods and highlights the prerequisite of considering hydrological catchment dynamics in source tracking study design. PMID:18564182

  10. Variability of soil microbial properties: effects of sampling, handling and storage.

    PubMed

    Cernohlávková, Jitka; Jarkovský, Jirí; Nesporová, Michala; Hofman, Jakub

    2009-11-01

    We investigated the effect of soil spatial variability within the sampling site scale, the effects of sample sieving (1, 2 and 4mm), and storage conditions up to 32 weeks (wet at 4 degrees C, -20 degrees C and air dried) on microbial biomass C, respiration, ammonification and nitrification activities in arable, grassland and forest soil. In general, all results were dependent on soil type. Arable soil showed the highest spatial variability, followed by grassland and forest soil. Sieving did not cause large differences; however, higher biomass C and respiration activity were observed in the 1mm than in the 4mm fraction. Storage at 4 degrees C seemed to be the most appropriate up to 8 weeks showing only minor changes of microbial parameters. Freezing of soils resulted in large increase of respiration. Dried storage indicated disruption of microbial communities even after 2 weeks.

  11. Expedition Six Flight Engineer Pettit uses a chemical/microbial analysis bag to collect water sample

    NASA Image and Video Library

    2002-12-18

    ISS006-E-08628 (18 December 2002) --- Astronaut Donald R. Pettit, Expedition Six NASA ISS science officer, is pictured in the Zvezda Service Module on the International Space Station (ISS) during the scheduled Week 3 potable water sampling and on-orbit chemical/microbial analysis of the SM environment control and life support system.

  12. Expedition Six Flight Engineer Pettit uses a chemical/microbial analysis bag to collect water sample

    NASA Image and Video Library

    2002-12-18

    ISS006-E-08616 (18 December 2002) --- Astronaut Donald R. Pettit, Expedition Six NASA ISS science officer, is pictured in the Zvezda Service Module on the International Space Station (ISS) during the scheduled Week 3 potable water sampling and on-orbit chemical/microbial analysis of the SM environment control and life support system.

  13. Microbial diversity within Juan de Fuca ridge basement fluids sampled from oceanic borehole observatories

    NASA Astrophysics Data System (ADS)

    Jungbluth, S.; Bowers, R.; Lin, H.; Hsieh, C.; Cowen, J. P.; Rappé, M.

    2012-12-01

    Three generations of sampling and instrumentation platforms known as Circulation Obviation Retrofit Kit (CORK) observatories affixed to Ocean Drilling Program (ODP) and Integrated Ocean Drilling Program (IODP) boreholes are providing unrivaled access to fluids originating from 1.2-3.5 million-years (Myr) old basaltic crust of the eastern flank of the Juan de Fuca ridge. Borehole fluid samples obtained via a custom seafloor fluid pumping and sampling system coupled to CORK continuous fluid delivery lines are yielding critical insights into the biogeochemistry and nature of microbial life inhabiting the sediment-covered basement environment. Direct microscopic enumeration revealed microbial cell abundances that are 2-41% of overlying bottom seawater. Snapshots of basement fluid microbial diversity and community structure have been obtained through small subunit ribosomal RNA (SSU rRNA) gene cloning and sequencing from five boreholes that access a range of basement ages and temperatures at the sediment-basement interface. SSU rRNA gene clones were derived from four different CORK installations (1026B, 1301A, 1362A, and 1362B) accessing relatively warmer (65°C) and older (3.5 Myr) ridge flank, and one location (1025C) accessing relatively cooler (39°C) and younger (1.2 Myr) ridge flank, revealing that warmer basement fluids had higher microbial diversity. A sampling time-series collected from borehole 1301A has revealed a microbial community that is temporally variable, with the dominant lineages changing between years. Each of the five boreholes sampled contained a unique microbial assemblage, however, common members are found from both cultivated and uncultivated lineages within the archaeal and bacterial domains, including meso- and thermophilic microbial lineages involved with sulfur cycling (e.g Thiomicrospira, Sulfurimonas, Desulfocapsa, Desulfobulbus). In addition, borehole fluid environmental gene clones were also closely related to uncultivated lineages

  14. Study to determine the aquatic biological effects on the Solid Rocket Booster (SRB). [technique for monitoring marine microbial fouling

    NASA Technical Reports Server (NTRS)

    Colwell, R. R.; Zachary, A.

    1979-01-01

    The surface of the reusable solid rocket boosters (SRB), which are jettisoned from the Shuttle Orbiter to parachute in the sea, are studied for colonization by marine life. Techniques for monitoring the marine microbial fouling of SRB materials are presented. An assessment of the nature and degree of the biofouling expected on the SRB materials in the recovery zone is reported. A determination of the degree and the effects of seasonal variation occurring on microbial fouling in the retrieval zone waters is made. The susceptibility of the SRB parachute recovery system to microbial fouling and biodeterioration is investigated. The development of scanning electron microscopy and epifluorescence microscopic observation techniques for rapid assessment of microbial fouling is discussed.

  15. Trends in monitoring pharmaceuticals and personal-care products in the aquatic environment by use of passive sampling devices

    USGS Publications Warehouse

    Mills, G.A.; Vrana, B.; Allan, I.; Alvarez, D.A.; Huckins, J.N.; Greenwood, R.

    2007-01-01

    The use of passive sampling in monitoring pharmaceuticals and personal-care products (PPCPs) in the aquatic environment is discussed. The utility of passive sampling methods for monitoring the fraction of heavy metals and the biologically available fraction of non-polar organic priority pollutants is recognized and these technologies are being used in surveys of water quality. These devices are used to measure the dissolved fraction and they can yield information that can be used in the development of risk assessments models. These devices can also be used to locate illegal dumping and to monitor specific sources of input of PPCPs into the environment, or to monitor the effectiveness of water treatment processes in the removal of these compounds from wastewater. These devices can provide representative information at low cost which necessitate a combination of laboratory calibration and field studies for emerging pollutants.

  16. I Environmental DNA sampling is more sensitive than a traditional survey technique for detecting an aquatic invader.

    PubMed

    Smart, Adam S; Tingley, Reid; Weeks, Andrew R; van Rooyen, Anthony R; McCarthy, Michael A

    2015-10-01

    Effective management of alien species requires detecting populations in the early stages of invasion. Environmental DNA (eDNA) sampling can detect aquatic species at relatively low densities, but few studies have directly compared detection probabilities of eDNA sampling with those of traditional sampling methods. We compare the ability of a traditional sampling technique (bottle trapping) and eDNA to detect a recently established invader, the smooth newt Lissotriton vulgaris vulgaris, at seven field sites in Melbourne, Australia. Over a four-month period, per-trap detection probabilities ranged from 0.01 to 0.26 among sites where L. v. vulgaris was detected, whereas per-sample eDNA estimates were much higher (0.29-1.0). Detection probabilities of both methods varied temporally (across days and months), but temporal variation appeared to be uncorrelated between methods. Only estimates of spatial variation were strongly correlated across the two sampling techniques. Environmental variables (water depth, rainfall, ambient temperature) were not clearly correlated with detection probabilities estimated via trapping, whereas eDNA detection probabilities were negatively correlated with water depth, possibly reflecting higher eDNA concentrations at lower water levels. Our findings demonstrate that eDNA sampling can be an order of magnitude more sensitive than traditional methods, and illustrate that traditional- and eDNA-based surveys can provide independent information on species distributions when occupancy surveys are conducted over short timescales.

  17. Searching for signatures across microbial communities: Metagenomic analysis of soil samples from mangrove and other ecosystems.

    PubMed

    Imchen, Madangchanok; Kumavath, Ranjith; Barh, Debmalya; Avezedo, Vasco; Ghosh, Preetam; Viana, Marcus; Wattam, Alice R

    2017-08-18

    In this study, we categorize the microbial community in mangrove sediment samples from four different locations within a vast mangrove system in Kerala, India. We compared this data to other samples taken from the other known mangrove data, a tropical rainforest, and ocean sediment. An examination of the microbial communities from a large mangrove forest that stretches across southwestern India showed strong similarities across the higher taxonomic levels. When ocean sediment and a single isolate from a tropical rain forest were included in the analysis, a strong pattern emerged with Bacteria from the phylum Proteobacteria being the prominent taxon among the forest samples. The ocean samples were predominantly Archaea, with Euryarchaeota as the dominant phylum. Principal component and functional analyses grouped the samples isolated from forests, including those from disparate mangrove forests and the tropical rain forest, from the ocean. Our findings show similar patterns in samples were isolated from forests, and these were distinct from the ocean sediment isolates. The taxonomic structure was maintained to the level of class, and functional analysis of the genes present also displayed these similarities. Our report for the first time shows the richness of microbial diversity in the Kerala coast and its differences with tropical rain forest and ocean microbiome.

  18. Community-Level Physiological Profiling of Microbial Communities in Constructed Wetlands: Effects of Sample Preparation.

    PubMed

    Button, Mark; Weber, Kela; Nivala, Jaime; Aubron, Thomas; Müller, Roland Arno

    2016-03-01

    Community-level physiological profiling (CLPP) using BIOLOG® EcoPlates™ has become a popular method for characterizing and comparing the functional diversity, functional potential, and metabolic activity of heterotrophic microbial communities. The method was originally developed for profiling soil communities; however, its usage has expanded into the fields of ecotoxicology, agronomy, and the monitoring and profiling of microbial communities in various wastewater treatment systems, including constructed wetlands for water pollution control. When performing CLPP on aqueous samples from constructed wetlands, a wide variety of sample characteristics can be encountered and challenges may arise due to excessive solids, color, or turbidity. The aim of this study was to investigate the impacts of different sample preparation methods on CLPP performed on a variety of aqueous samples covering a broad range of physical and chemical characteristics. The results show that using filter paper, centrifugation, or settling helped clarify samples for subsequent CLPP analysis, however did not do so as effectively as dilution for the darkest samples. Dilution was able to provide suitable clarity for the darkest samples; however, 100-fold dilution significantly affected the carbon source utilization patterns (CSUPs), particularly with samples that were already partially or fully clear. Ten-fold dilution also had some effect on the CSUPs of samples which were originally clear; however, the effect was minimal. Based on these findings, for this specific set of samples, a 10-fold dilution provided a good balance between ease of use, sufficient clarity (for dark samples), and limited effect on CSUPs. The process and findings outlined here can hopefully serve future studies looking to utilize CLPP for functional analysis of microbial communities and also assist in comparing data from studies where different sample preparation methods were utilized.

  19. GES-5 among the β-lactamases detected in ubiquitous bacteria isolated from aquatic environment samples.

    PubMed

    Manageiro, Vera; Ferreira, Eugénia; Caniça, Manuela; Manaia, Célia M

    2014-02-01

    In this study, we investigated the β-lactamase-encoding genes responsible for β-lactam resistance phenotypes detected among 56 Gram-negative isolates (Gamma- and Alpha-proteobacteria) recovered from wastewater, urban streams, and drinking water. The β-lactam resistance mechanisms detected in 36 isolates comprised the presence of class A (blaTEM-1 , blaSHV-1 , blaSHV-11 , blaGES-5 ), class B (ImiS, L1), class C (blaCMY-2 , blaCMY-34 , blaCMY-65 , blaCMY-89 , blaCMY-90 , blaACC-5 , blaACT-13 ), and class D (blaOXA-309)β-lactamase-encoding genes, some variants described for the first time here. Notably, the results showed antimicrobial resistance genes related not only to commonly used antibiotics, but also to carbapenems, providing the first description of a GES-5-producing Enterobacteriaceae. The importance of ubiquitous bacteria thriving in aquatic environments as reservoirs or carriers of clinically relevant resistance determinants was confirmed, and the need to monitor water habitats as potential sources for the emergence and/or spread of antibiotic resistance in the environment was highlighted.

  20. Microbial processes and communities in sediment samples along a transect across the Lusi mud volcano, Indonesia

    NASA Astrophysics Data System (ADS)

    Krueger, Martin; Straaten, Nontje; Mazzini, Adriano

    2015-04-01

    The Lusi eruption represents one of the largest ongoing sedimentary hosted geothermal systems. This eruption started in 2006 following to a 6.3 M earthquake that stroke Java Island. Since then it has been spewing boiling mud from a central crater with peaks reaching 180.000 m3 per day. Today an area of about 8 km2 is covered by locally dried mud breccia where a network of hundreds of satellite seeping pools is active. Numerous investigations focused on the study of offshore microbial colonies that commonly thrive at offshore methane seeps and mud volcanoes, however very little has been done for onshore seeping structures. Lusi represents a unique opportunity to complete a comprehensive study of onshore microbial communities fed by the seepage of CH4 and CO2 as well as of heavier liquid hydrocarbons originating from several km below the surface. We conducted a sampling campaign at the Lusi site collecting samples of fresh mud close to the erupting crater using a remote controlled drone. In addition we completed a transect towards outer parts of the crater to collect older, weathered samples for comparison. In all samples active microorganisms were present. The highest activities for CO2 and CH4 production as well as for CH4 oxidation and hydrocarbon degradation were observed in medium-age mud samples collected roughly in the middle of the transect. Rates for aerobic methane oxidation were high, as was the potential of the microbial communities to degrade hydrocarbons (oils, alkanes, BTEX tested). The data suggests a transition of microbial populations from an anaerobic, hydrocarbon-driven metabolism in fresher samples from center or from small seeps to more generalistic, aerobic microbial communities in older, more consolidated sediments. Currently, the microbial communities in the different sediment samples are analyzed using quantitative PCR and T-RFLP combined with MiSeq sequencing. This study represents an initial step to better understand onshore seepage

  1. Organic matter loss during hydrochloric acid treatment of aquatic samples: implications for elemental and stable isotopic analyses

    NASA Astrophysics Data System (ADS)

    Levesque, C.; Juniper, K.; Planas, D.

    2009-12-01

    Stable isotope analysis is a standard tool used to investigate organic carbon (Corg) fluxes in the environment. This technique requires the removal of carbonates (particulate inorganic carbon) prior to analysis. A standard approach to remove carbonates consists in treating samples with hydrochloric acid (HCl), followed by rinsing with distilled water to remove acid residues. However, this acid-wash technique has been reported to affect Corg content and isotopic composition. We hypothesized that HCl treatment hydrolyses and dissolves a fraction of the organic matrix, and that the subsequent rinsing of the sample removes this dissolved fraction. We tested the impact of acid treatment on ten types of organic samples from aquatic habitats, including mollusks, crustaceans, fish, diatoms, plants and sediments. Samples were treated with 1N HCl and rinsed with distilled water, and the acid residue was collected. Treated and untreated (control) samples as well as the acid residues were analyzed for carbon content and stable isotopic composition. Between 4.0 and 22.8% of the total Corg was lost as dissolved organic carbon (DOC) during the acid-wash procedure. Samples with low Corg content lost proportionally more Corg as DOC than samples with a higher Corg content. In most cases, DOC loss did not significantly affect sample δ13C values, since the δ13C of DOC was similar to that of the particulate Corg in the sample. In three cases, the DOC lost had a distinct isotopic composition, leading to significant shifts in sample δ13C values (δ13C - 0.4 to + 3.6‰). The commonly-used acid-wash procedure for stable isotope analyses can result in substantial losses of organic carbon and in significant shifts in δ13C.

  2. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community.

    PubMed

    Tapio, Ilma; Shingfield, Kevin J; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J; Wallace, R John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint.

  3. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community

    PubMed Central

    Tapio, Ilma; Shingfield, Kevin J.; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R.; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J.; Wallace, R. John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint. PMID:26986467

  4. The National Ecological Observatory Network Aquatic Sampling: Dissolved Gas Concentrations, Stratification Conditions in Lakes, and Reaeration Rates in Streams

    NASA Astrophysics Data System (ADS)

    Cawley, K.; Goodman, K. J.

    2016-12-01

    The National Ecological Observatory Network (NEON) is deploying instrumentation and collecting samples on a continental spatial scale planned to operate for 30 years starting in 2018. There are five components of NEON: Airborne Observation Platform (AOP), Terrestrial Instrument System (TIS), Terrestrial Observation System (TOS), Aquatic Instrument System (AIS), and Aquatic Observation System (AOS). Collocation of measurements associated with each of these components will allow for linkage and comparison of data related to physical, chemical, and biological parameters. The NEON Aquatic subsystem, comprised of AOS and AIS, will quantify the impacts of climate change, land use, and biological invasions on freshwater populations and processes. Observations including organismal community composition, surface and groundwater chemistry, and habitat structure will be made in addition to deploying instrumentation in and around water bodies. Additionally, data processing is standardized, and quality-controlled data products derived from NEON measurements are freely available through the data portal. Some of the data that will be collected, processed, and published by NEON are particularly relevant to discovering connections between air, land and associated freshwaters, which drive the dynamics of carbon in inland waters. As part of the AOS sub-system, samples will be collected bi-weekly from 24 streams and monthly from 7 lakes for analysis of greenhouse gas (GHG) concentrations (CO2, N2O, and CH4). At the same time, depth profiles for temperature, conductivity, and dissolved oxygen will be collected in the lake sites. From these depth profiles, lake stratification conditions can be discerned. At stream sites, reaeration tracer experiments (simultaneous conservative and gas tracer injection) will be performed about 8 times per year. The stream reaeration rates will be related to discharge values to develop a rating curve from which temporally interpolated reaeration rates can

  5. A Novel Analysis Method for Paired-Sample Microbial Ecology Experiments

    PubMed Central

    Vora, Suhani; Techtmann, Stephen M.; Fortney, Julian L.; Bastidas-Oyanedel, Juan R.; Rodríguez, Jorge; Hazen, Terry C.; Alm, Eric J.

    2016-01-01

    Many microbial ecology experiments use sequencing data to measure a community’s response to an experimental treatment. In a common experimental design, two units, one control and one experimental, are sampled before and after the treatment is applied to the experimental unit. The four resulting samples contain information about the dynamics of organisms that respond to the treatment, but there are no analytical methods designed to extract exactly this type of information from this configuration of samples. Here we present an analytical method specifically designed to visualize and generate hypotheses about microbial community dynamics in experiments that have paired samples and few or no replicates. The method is based on the Poisson lognormal distribution, long studied in macroecology, which we found accurately models the abundance distribution of taxa counts from 16S rRNA surveys. To demonstrate the method’s validity and potential, we analyzed an experiment that measured the effect of crude oil on ocean microbial communities in microcosm. Our method identified known oil degraders as well as two clades, Maricurvus and Rhodobacteraceae, that responded to amendment with oil but do not include known oil degraders. Our approach is sensitive to organisms that increased in abundance only in the experimental unit but less sensitive to organisms that increased in both control and experimental units, thus mitigating the role of “bottle effects”. PMID:27152415

  6. A Novel Analysis Method for Paired-Sample Microbial Ecology Experiments.

    PubMed

    Olesen, Scott W; Vora, Suhani; Techtmann, Stephen M; Fortney, Julian L; Bastidas-Oyanedel, Juan R; Rodríguez, Jorge; Hazen, Terry C; Alm, Eric J

    2016-01-01

    Many microbial ecology experiments use sequencing data to measure a community's response to an experimental treatment. In a common experimental design, two units, one control and one experimental, are sampled before and after the treatment is applied to the experimental unit. The four resulting samples contain information about the dynamics of organisms that respond to the treatment, but there are no analytical methods designed to extract exactly this type of information from this configuration of samples. Here we present an analytical method specifically designed to visualize and generate hypotheses about microbial community dynamics in experiments that have paired samples and few or no replicates. The method is based on the Poisson lognormal distribution, long studied in macroecology, which we found accurately models the abundance distribution of taxa counts from 16S rRNA surveys. To demonstrate the method's validity and potential, we analyzed an experiment that measured the effect of crude oil on ocean microbial communities in microcosm. Our method identified known oil degraders as well as two clades, Maricurvus and Rhodobacteraceae, that responded to amendment with oil but do not include known oil degraders. Our approach is sensitive to organisms that increased in abundance only in the experimental unit but less sensitive to organisms that increased in both control and experimental units, thus mitigating the role of "bottle effects".

  7. A novel analysis method for paired-sample microbial ecology experiments

    SciTech Connect

    Olesen, Scott W.; Vora, Suhani; Techtmann, Stephen M.; Fortney, Julian L.; Bastidas-Oyanedel, Juan R.; Rodriguez, Jorge; Hazen, Terry C.; Alm, Eric J.; Humbert, Jean -Francois

    2016-05-06

    Many microbial ecology experiments use sequencing data to measure a community s response to an experimental treatment. In a common experimental design, two units, one control and one experimental, are sampled before and after the treatment is applied to the experimental unit. The four resulting samples contain information about the dynamics of organisms that respond to the treatment, but there are no analytical methods designed to extract exactly this type of information from this configuration of samples. Here we present an analytical method specifically designed to visualize and generate hypotheses about microbial community dynamics in experiments that have paired samples and few or no replicates. The method is based on the Poisson lognormal distribution, long studied in macroecology, which we found accurately models the abundance distribution of taxa counts from 16S rRNA surveys. To demonstrate the method s validity and potential, we analyzed an experiment that measured the effect of crude oil on ocean microbial communities in microcosm. Our method identified known oil degraders as well as two clades, Maricurvus and Rhodobacteraceae, that responded to amendment with oil but do not include known oil degraders. Furthermore, our approach is sensitive to organisms that increased in abundance only in the experimental unit but less sensitive to organisms that increased in both control and experimental units, thus mitigating the role of bottle effects .

  8. Does equilibrium passive sampling reflect actual in situ bioaccumulation of PAHs and petroleum hydrocarbon mixtures in aquatic worms?

    PubMed

    Muijs, Barry; Jonker, Michiel T O

    2012-01-17

    Over the past couple of years, several analytical methods have been developed for assessing the bioavailability of environmental contaminants in sediments and soils. Comparison studies suggest that equilibrium passive sampling methods generally provide the better estimates of internal concentrations in organisms and thus of subsequent risks. However, field studies to validate the potential of passive sampling to predict actual in situ bioaccumulation are scarce and limited information only exists on selected, individual compounds. The present study investigated whether bioaccumulation of PAH and complex petroleum hydrocarbon mixtures in field-exposed aquatic worms could be predicted properly with passive samplers. To this end, in situ bioaccumulation in aquatic worms at 6 PAH-contaminated locations and 8 petroleum hydrocarbon (oil)-contaminated locations was compared with the results of in situ solid phase micro extraction (SPME) applications. For the oil-contaminated sediments, bioaccumulation was also assessed in the lab with polyoxymethylene solid phase extraction (POM-SPE). Actual PAH bioaccumulation was generally predicted within a factor of 4 with in situ SPME, using temperature-adjusted SPME fiber-water partition coefficients and lab-derived bioaccumulation factors (BAFs) for the worm species used, demonstrating the method's potential under field conditions. In situ SPME appeared to be less suitable for predicting bioaccumulation of oil however, in contrast to POM-SPE in the lab, which assessed in situ oil bioaccumulation within a factor of 3, while also closely reflecting the actual distribution of oil boiling point fractions (the hydrocarbon block profile) as accumulated by the worms. All in all, the results indicated that (specific) equilibrium passive samplers, either applied in the field or the lab, have great potential for assessing bioaccumulation of environmental contaminant mixtures from field-contaminated sediments.

  9. Passive Sampling in Regulatory Chemical Monitoring of Nonpolar Organic Compounds in the Aquatic Environment.

    PubMed

    Booij, Kees; Robinson, Craig D; Burgess, Robert M; Mayer, Philipp; Roberts, Cindy A; Ahrens, Lutz; Allan, Ian J; Brant, Jan; Jones, Lisa; Kraus, Uta R; Larsen, Martin M; Lepom, Peter; Petersen, Jördis; Pröfrock, Daniel; Roose, Patrick; Schäfer, Sabine; Smedes, Foppe; Tixier, Céline; Vorkamp, Katrin; Whitehouse, Paul

    2016-01-05

    We reviewed compliance monitoring requirements in the European Union, the United States, and the Oslo-Paris Convention for the protection of the marine environment of the North-East Atlantic, and evaluated if these are met by passive sampling methods for nonpolar compounds. The strengths and shortcomings of passive sampling are assessed for water, sediments, and biota. Passive water sampling is a suitable technique for measuring concentrations of freely dissolved compounds. This method yields results that are incompatible with the EU's quality standard definition in terms of total concentrations in water, but this definition has little scientific basis. Insufficient quality control is a present weakness of passive sampling in water. Laboratory performance studies and the development of standardized methods are needed to improve data quality and to encourage the use of passive sampling by commercial laboratories and monitoring agencies. Successful prediction of bioaccumulation based on passive sampling is well documented for organisms at the lower trophic levels, but requires more research for higher levels. Despite the existence of several knowledge gaps, passive sampling presently is the best available technology for chemical monitoring of nonpolar organic compounds. Key issues to be addressed by scientists and environmental managers are outlined.

  10. Modelling sediment-microbial dynamics in the South Nation River, Ontario, Canada: Towards the prediction of aquatic and human health risk.

    PubMed

    Droppo, I G; Krishnappan, B G; Liss, S N; Marvin, C; Biberhofer, J

    2011-06-01

    Runoff from agricultural watersheds can carry a number of agricultural pollutants and pathogens; often associated with the sediment fraction. Deposition of this sediment can impact water quality and the ecology of the river, and the re-suspension of such sediment can become sources of contamination for reaches downstream. In this paper a modelling framework to predict sediment and associated microbial erosion, transport and deposition is proposed for the South Nation River, Ontario, Canada. The modelling framework is based on empirical relationships (deposition and re-suspension fluxes), derived from laboratory experiments in a rotating circular flume using sediment collected from the river bed. The bed shear stress governing the deposition and re-suspension processes in the stream was predicted using a one dimensional mobile boundary flow model called MOBED. Counts of live bacteria associated with the suspended and bed sediments were used in conjunction with measured suspended sediment concentration at an upstream section to allow for the estimation of sediment associated microbial erosion, transport and deposition within the modelled river reach. Results suggest that the South Nation River is dominated by deposition periods with erosion only occurring at flows above approximately 250 m(3) s(-1) (above this threshold, all sediment (suspended and eroded) with associated bacteria are transported through the modelled reach). As microbes are often associated with sediments, and can survive for extended periods of time, the river bed is shown to be a possible source of pathogenic organisms for erosion and transport downstream during large storm events. It is clear that, shear levels, bacteria concentrations and suspended sediment are interrelated requiring that these parameters be studied together in order to understand aquatic microbial dynamics. It is important that any management strategies and operational assessments for the protection of human and aquatic health

  11. Quantitative reverse sample genome probing of microbial communities and its application to oil field production waters

    SciTech Connect

    Voordouw, G.; Shen, Y.; Harrington, C.S.; Teland, A.J. ); Jack, T.R. ); Westlake, W.S. )

    1993-12-01

    This paper presents a protocol for quantitative analysis of microbial communities by reverse sample genome probing is presented in which (i) whole community DNA is isolated and labeled in the presence of a known amount of an added internal standard and (ii) the resulting spiked reverse genome probe is hybridized with a master filter on which denatured genomic DNAs from bacterial standards isolated from the target environment were spotted in large amounts (up to 1,500 ng) in order to improve detection sensitivity. This protocol allowed reproducible fingerprinting of the microbial community in oil field production waters at 19 sites from which water and biofilm samples were collected. It appeared that selected sulfate-reducing bacteria were significantly enhanced in biofilms covering the metal surfaces in contact with the production waters.

  12. Quantitative Reverse Sample Genome Probing of Microbial Communities and Its Application to Oil Field Production Waters

    PubMed Central

    Voordouw, Gerrit; Shen, Yin; Harrington, Clare S.; Telang, Anita J.; Jack, Thomas R.; Westlake, Donald W. S.

    1993-01-01

    This paper presents a protocol for quantitative analysis of microbial communities by reverse sample genome probing is presented in which (i) whole community DNA is isolated and labeled in the presence of a known amount of an added internal standard and (ii) the resulting spiked reverse genome probe is hybridized with a master filter on which denatured genomic DNAs from bacterial standards isolated from the target environment were spotted in large amounts (up to 1,500 ng) in order to improve detection sensitivity. This protocol allowed reproducible fingerprinting of the microbial community in oil field production waters at 19 sites from which water and biofilm samples were collected. It appeared that selected sulfate-reducing bacteria were significantly enhanced in biofilms covering the metal surfaces in contact with the production waters. Images PMID:16349111

  13. Microbial activity in subsurface samples before and during nitrate-enhanced bioremediation

    SciTech Connect

    Thomas, J.M.; Gordy, V.R.; Bruce, C.L.; Ward, C.H.; Hutchins, S.R.; Sinclair, J.L.

    1995-12-31

    A study was conducted to determine the microbial activity at a site contaminated with JP-4 jet fuel before and during nitrate-enhanced bioremediation. Samples at three depths from six different locations were collected aseptically under anaerobic conditions before and during treatment. Cores were located in or close to the source of contamination, downgradient of the source, or outside the zone of contamination. Parameters for microbial characterization included (1) viable counts of aerobic heterotrophic, JP-4 degrading, and oligotrophic bacteria; (2) the most probable number (MPN) of aerobic and anaerobic protozoa; (3) the MPN of total denitrifiers; and (4) the MPN of denitrifiers in hydrocarbon-amended microcosms. The results indicate that the total number of denitrifiers increased by an order of magnitude during nitrate-enhanced bioremediation in most samples. The number of total heterotrophs and JP-4-degrading microorganisms growing aerobically also increased. In addition, the first anaerobic protozoa associated with hydrocarbon-contaminated subsurface materials were detected.

  14. Gold-FISH: A correlative approach to microscopic imaging of single microbial cells in environmental samples

    NASA Astrophysics Data System (ADS)

    Schmidt, Hannes; Seki, David; Woebken, Dagmar; Eickhorst, Thilo

    2017-04-01

    Fluorescence in situ hybridization (FISH) is routinely used for the phylogenetic identification, detection, and quantification of single microbial cells environmental microbiology. Oligonucleotide probes that match the 16S rRNA sequence of target organisms are generally applied and the resulting signals are visualized via fluorescence microscopy. Consequently, the detection of the microbial cells of interest is limited by the resolution and the sensitivity of light microscopy where objects smaller than 0.2 µm can hardly be represented. Visualizing microbial cells at magnifications beyond light microscopy, however, can provide information on the composition and potential complexity of microbial habitats - the actual sites of nutrient cycling in soil and sediments. We present a recently developed technique that combines (1) the phylogenetic identification and detection of individual microorganisms by epifluorescence microscopy, with (2) the in situ localization of gold-labelled target cells on an ultrastructural level by SEM. Based on 16S rRNA targeted in situ hybridization combined with catalyzed reporter deposition, a streptavidin conjugate labeled with a fluorescent dye and nanogold particles is introduced into whole microbial cells. A two-step visualization process including an autometallographic enhancement of nanogold particles then allows for either fluorescence or electron microscopy, or a correlative application thereof. We will present applications of the Gold-FISH protocol to samples of marine sediments, agricultural soils, and plant roots. The detection and enumeration of bacterial cells in soil and sediment samples was comparable to CARD-FISH applications via fluorescence microscopy. Examples of microbe-surface interaction analysis will be presented on the basis of bacteria colonizing the rhizoplane of rice roots. In principle, Gold-FISH can be performed on any material to give a snapshot of microbe-surface interactions and provides a promising tool for

  15. Microbial Contamination in Next Generation Sequencing: Implications for Sequence-Based Analysis of Clinical Samples

    PubMed Central

    Strong, Michael J.; Xu, Guorong; Morici, Lisa; Splinter Bon-Durant, Sandra; Baddoo, Melody; Lin, Zhen; Fewell, Claire; Taylor, Christopher M.; Flemington, Erik K.

    2014-01-01

    The high level of accuracy and sensitivity of next generation sequencing for quantifying genetic material across organismal boundaries gives it tremendous potential for pathogen discovery and diagnosis in human disease. Despite this promise, substantial bacterial contamination is routinely found in existing human-derived RNA-seq datasets that likely arises from environmental sources. This raises the need for stringent sequencing and analysis protocols for studies investigating sequence-based microbial signatures in clinical samples. PMID:25412476

  16. Fitting distributions to microbial contamination data collected with an unequal probability sampling design.

    PubMed

    Williams, M S; Ebel, E D; Cao, Y

    2013-01-01

    The fitting of statistical distributions to microbial sampling data is a common application in quantitative microbiology and risk assessment applications. An underlying assumption of most fitting techniques is that data are collected with simple random sampling, which is often times not the case. This study develops a weighted maximum likelihood estimation framework that is appropriate for microbiological samples that are collected with unequal probabilities of selection. A weighted maximum likelihood estimation framework is proposed for microbiological samples that are collected with unequal probabilities of selection. Two examples, based on the collection of food samples during processing, are provided to demonstrate the method and highlight the magnitude of biases in the maximum likelihood estimator when data are inappropriately treated as a simple random sample. Failure to properly weight samples to account for how data are collected can introduce substantial biases into inferences drawn from the data. The proposed methodology will reduce or eliminate an important source of bias in inferences drawn from the analysis of microbial data. This will also make comparisons between studies and the combination of results from different studies more reliable, which is important for risk assessment applications. © 2012 No claim to US Government works.

  17. Analysing Microbial Community Composition through Amplicon Sequencing: From Sampling to Hypothesis Testing.

    PubMed

    Hugerth, Luisa W; Andersson, Anders F

    2017-01-01

    Microbial ecology as a scientific field is fundamentally driven by technological advance. The past decade's revolution in DNA sequencing cost and throughput has made it possible for most research groups to map microbial community composition in environments of interest. However, the computational and statistical methodology required to analyse this kind of data is often not part of the biologist training. In this review, we give a historical perspective on the use of sequencing data in microbial ecology and restate the current need for this method; but also highlight the major caveats with standard practices for handling these data, from sample collection and library preparation to statistical analysis. Further, we outline the main new analytical tools that have been developed in the past few years to bypass these caveats, as well as highlight the major requirements of common statistical practices and the extent to which they are applicable to microbial data. Besides delving into the meaning of select alpha- and beta-diversity measures, we give special consideration to techniques for finding the main drivers of community dissimilarity and for interaction network construction. While every project design has specific needs, this review should serve as a starting point for considering what options are available.

  18. Analysing Microbial Community Composition through Amplicon Sequencing: From Sampling to Hypothesis Testing

    PubMed Central

    Hugerth, Luisa W.; Andersson, Anders F.

    2017-01-01

    Microbial ecology as a scientific field is fundamentally driven by technological advance. The past decade's revolution in DNA sequencing cost and throughput has made it possible for most research groups to map microbial community composition in environments of interest. However, the computational and statistical methodology required to analyse this kind of data is often not part of the biologist training. In this review, we give a historical perspective on the use of sequencing data in microbial ecology and restate the current need for this method; but also highlight the major caveats with standard practices for handling these data, from sample collection and library preparation to statistical analysis. Further, we outline the main new analytical tools that have been developed in the past few years to bypass these caveats, as well as highlight the major requirements of common statistical practices and the extent to which they are applicable to microbial data. Besides delving into the meaning of select alpha- and beta-diversity measures, we give special consideration to techniques for finding the main drivers of community dissimilarity and for interaction network construction. While every project design has specific needs, this review should serve as a starting point for considering what options are available. PMID:28928718

  19. A novel analysis method for paired-sample microbial ecology experiments

    DOE PAGES

    Olesen, Scott W.; Vora, Suhani; Techtmann, Stephen M.; ...

    2016-05-06

    Many microbial ecology experiments use sequencing data to measure a community s response to an experimental treatment. In a common experimental design, two units, one control and one experimental, are sampled before and after the treatment is applied to the experimental unit. The four resulting samples contain information about the dynamics of organisms that respond to the treatment, but there are no analytical methods designed to extract exactly this type of information from this configuration of samples. Here we present an analytical method specifically designed to visualize and generate hypotheses about microbial community dynamics in experiments that have paired samplesmore » and few or no replicates. The method is based on the Poisson lognormal distribution, long studied in macroecology, which we found accurately models the abundance distribution of taxa counts from 16S rRNA surveys. To demonstrate the method s validity and potential, we analyzed an experiment that measured the effect of crude oil on ocean microbial communities in microcosm. Our method identified known oil degraders as well as two clades, Maricurvus and Rhodobacteraceae, that responded to amendment with oil but do not include known oil degraders. Furthermore, our approach is sensitive to organisms that increased in abundance only in the experimental unit but less sensitive to organisms that increased in both control and experimental units, thus mitigating the role of bottle effects .« less

  20. Calibration of a passive sampling device for time-integrated sampling of hydrophilic herbicides in aquatic environments.

    PubMed

    Tran, Anh T K; Hyne, Ross V; Doble, P

    2007-03-01

    Two types of solid-phase materials, a styrenedivinylbenzene copolymer sorbent (embedded in a SDB-XC Empore disk) and a styrenedivinylbenzene copolymer sorbent modified with sulfonic acid functional groups (embedded in a SDB-RPS Empore disk), were compared as a receiving phase in a passive sampling device for monitoring polar pesticides. The SDB-XC Empore disk was selected for further evaluation, overlayed with either a polysulfone or a polyethersulfone diffusion membrane. The target herbicides included five nonionized herbicides (simazine, atrazine, diuron, clomazone, and metolachlor) and four phenoxy acid herbicides (dicamba, (2,4-dichlorophenoxy)acetic acid [2,4-D], (4-chloro-2-methylphenoxy)acetic acid [MCPA], and triclopyr) with log octanol/water partition coefficient (log K(OW)) values of less than three in water. Uptake of these herbicides generally was higher into a device constructed of a SDB-XC Empore disk as a receiving phase covered with a polyethersulfone membrane compared to a similar device covered with a polysulfone membrane. Using the device with a SDB-XC Empore disk covered with a polyethersulfone membrane, linear uptake of simazine, atrazine, diuron, clomazone, and metolachlor was observed for up to 21 d, and daily sampling rates of the herbicides from water in a laboratory flow-through system were determined. The uptake rate of each nonionized herbicide by the Empore disk-based passive sampler was linearly proportional to its concentration in the water, and the sampling rate was independent of the water concentrations over the 21-d period. Uptake of the phenoxy acid herbicides (2,4-D, MCPA, and triclopyr) obeyed first-order kinetics and rapidly reached equilibrium in the passive sampler after approximately 12 d of exposure. The Empore disk-based passive sampler displayed isotropic kinetics, with a release half-life for triclopyr of approximately 6 d.

  1. Combining Passive Sampling with Toxicological Characterization of Complex Mixtures of Pollutants from the Aquatic Environment.

    PubMed

    Jahnke, Annika; Witt, Gesine; Schäfer, Sabine; Haase, Nora; Escher, Beate I

    2016-08-04

    The combination of polymer-based passive sampling to collect complex environmental mixtures of pollutants, the transfer of these mixtures into bioassays, and their related toxicological characterization is still in its infancy. However, this approach has considerable potential to improve environmental hazard and risk assessment for two reasons. First, the passive sampler collects a broad range of chemicals representing the fraction of compounds available for diffusion and (bio)uptake, excluding a large part of the matrix; thus, extensive sample cleanup which could discriminate certain compounds can be avoided. Second, the toxicological characterization of samples using bioassays is complementary to chemical (target) analysis within environmental monitoring because it captures all chemicals exerting the same mode of toxic action and acting jointly in mixtures, thus providing a comprehensive picture of their overall combined effects. The scientific literature describes a range of examples from the water phase where passive sampling is usually carried out in the kinetic uptake regime for most chemicals although some may already have reached equilibrium. The composition of the chemical mixture changes from the water phase to the passive sampling material because of kinetic effects and polymer/water partition coefficients which depend on the chemicals' hydrophobicity. In contrast, only a few applications in sediment and biota have been described, but amongst these some pioneering studies have demonstrated the feasibility and potential of this combined approach. This chapter gives an overview of what has been carried out in this research area, focusing on opportunities and challenges, and points out desirable future developments with a focus on the importance of choosing a suitable combination of sampling and dosing to transfer (or re-establish) the environmental mixture into the bioassay.

  2. Passive Sampling in Regulatory Chemical Monitoring of Nonpolar Organic Compounds in the Aquatic Environment

    EPA Science Inventory

    We reviewed compliance monitoring requirements in the European Union (EU), the United States(USA), and the Oslo-Paris Convention for the protection of the marine environment of the North-East Atlantic (OSPAR), and evaluated if these are met by passive sampling methods for nonpola...

  3. Amphibian breeding phenology and reproductive outcome: an examination using terrestrial and aquatic sampling

    Treesearch

    C.H. Greenberg; S.A. Johnson; R. Owen; A. Storfer

    2017-01-01

    Worldwide amphibian declines highlight the need for programs that monitor species presence and track population trends. We sampled larval amphibians with a box trap at 3-week intervals for 23 months in eight wetlands, and concurrently trapped adults and juveniles with drift fences, to examine spatiotemporal patterns of tadpole occurrence; explore relationships between...

  4. Development of a luminescent mutagenicity test for high-throughput screening of aquatic samples.

    PubMed

    Zwart, Nick; Lamoree, Marja; Houtman, Corine; de Boer, Jacob; Kool, Jeroen; Hamers, Timo

    2017-09-06

    The Salmonella reversion based Ames test is the most widely used method for mutagenicity testing. For rapid toxicity assessment of e.g. water samples and for effect-directed analysis, however, the Ames test suffers from lack of throughput and is regarded as a laborious, time consuming method. To achieve faster analysis, with increased throughput, a (downscaled) luminescent derivative of the Ames Salmonella/microsome fluctuation test has been developed through expression of the Photorhabdus luminescens luciferase in the Salmonella TA98 and TA100 strains. The applicability of this test is demonstrated by analysis of environmentally relevant compounds, a suspended particulate matter extract and an industrial effluent sample. Use of the luminescent reporter reduced the required detection time from 48 to 28h with a specificity of 84% for responses reported in the literature to a set of 14 mutagens as compared to 72% in the unmodified fluctuation test. Testing of the same compounds in a downscaled luminescent format resulted in an 88% similarity with the response found in the regular luminescent format. The increase in throughput, faster analysis and potential for real-time bacterial quantification that luminescence provides, allows future application in the high-throughput screening of large numbers of samples or sample fractions, as required in effect-directed analysis in order to accelerate the identification of (novel) mutagens. Copyright © 2017. Published by Elsevier Ltd.

  5. Passive Sampling in Regulatory Chemical Monitoring of Nonpolar Organic Compounds in the Aquatic Environment

    EPA Science Inventory

    We reviewed compliance monitoring requirements in the European Union (EU), the United States(USA), and the Oslo-Paris Convention for the protection of the marine environment of the North-East Atlantic (OSPAR), and evaluated if these are met by passive sampling methods for nonpola...

  6. Optimized Extraction Method To Remove Humic Acid Interferences from Soil Samples Prior to Microbial Proteome Measurements.

    PubMed

    Qian, Chen; Hettich, Robert L

    2017-07-07

    The microbial composition and their activities in soil environments play a critical role in organic matter transformation and nutrient cycling. Liquid chromatography coupled to high-performance mass spectrometry provides a powerful approach to characterize soil microbiomes; however, the limited microbial biomass and the presence of abundant interferences in soil samples present major challenges to proteome extraction and subsequent MS measurement. To this end, we have designed an experimental method to improve microbial proteome measurement by removing the soil-borne humic substances coextraction from soils. Our approach employs an in situ detergent-based microbial lysis/TCA precipitation coupled to an additional cleanup step involving acidified precipitation and filtering at the peptide level to remove most of the humic acid interferences prior to proteolytic peptide measurement. The novelty of this approach is an integration to exploit two different characteristics of humic acids: (1) Humic acids are insoluble in acidic solution but should not be removed at the protein level, as undesirable protein removal may also occur. Rather it is better to leave the humics acids in the samples until the peptide level, at which point the significant differential solubility of humic acids versus peptides at low pH can be exploited very efficiently. (2) Most of the humic acids have larger molecule weights than the peptides. Therefore, filtering a pH 2 to 3 peptide solution with a 10 kDa filter will remove most of the humic acids. This method is easily interfaced with normal proteolytic processing approaches and provides a reliable and straightforward protein extraction method that efficiently removes soil-borne humic substances without inducing proteome sample loss or biasing protein identification in mass spectrometry. In general, this humic acid removal step is universal and can be adopted by any workflow to effectively remove humic acids to avoid them negatively competing

  7. Effect of DNA extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.

    PubMed

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J; Waghorn, Garry C; Janssen, Peter H

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  8. Effect of DNA Extraction Methods and Sampling Techniques on the Apparent Structure of Cow and Sheep Rumen Microbial Communities

    PubMed Central

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J.; Waghorn, Garry C.; Janssen, Peter H.

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  9. Contending with the Sampling Compromise of Time VS. Space: a Case Study in Aquatic Monitoring

    NASA Astrophysics Data System (ADS)

    Currie, W. J.

    2013-12-01

    In every field study, there is always a trade-off between the number of samples that can be analyzed and the temporal or spatial coverage. Project Quinte, is a 40 year collaborative study investigating the biological and physico-chemical changes to the Bay of Quinte (BOQ), Lake Ontario, which was prompted primarily by eutrophication concerns in the late 1960s. Sampling of water chemistry, plankton composition and primary productivity occurs biweekly, but at a small number of stations and only for the period of May-Oct. Spatially limited sites are chosen with the intent of being representative of a larger region, but the validity of this is often left untested. In order to discern the resulting ecological effects on the plankton community, the field data have been used with some success to show changes in nutrients, plankton composition and growth, but are complicated by other factors such as changes in phosphorous controls and the arrival of a series of invasive species (Dreissenid mussels, predatory cladocerans, round goby). The dataset is extensive but has large consistent gaps (late fall - early spring) and spatially limited sampling stations can mask trends. In this study we explore a combination of spatial and temporal analysis techniques that can overcome these limitations. Recently we have begun to use a towed sensor body containing a CTD and Laser Optical Plankton Counter (LOPC) to give both increased spatial resolution and sampling extent in an effort to determine the probability distribution and intermittency of both physical-chemical (e.g. temperature, turbidity) and biological (e.g. zooplankton and phytoplankton biomass) measures. These transects are used to compare the variability in phys-chem and plankton distributions with those collected at long-term stations to determine scales appropriate to the field monitoring program.

  10. Understanding microbial/DOM interactions using fluorescence and flow cytometry

    NASA Astrophysics Data System (ADS)

    Fox, Bethany; Rushworth, Cathy; Attridge, John; Anesio, Alexandre; Cox, Tim; Reynolds, Darren

    2015-04-01

    The transformation and movement of dissolved organic carbon (DOC) within freshwater aquatic systems is an important factor in the global cycling of carbon. DOC within aquatic systems is known to underpin the microbial food web and therefore plays an essential role in supporting and maintaining the aquatic ecosystem. Despite this the interactions between bacteria and dissolved organic matter (DOM) are not well understood, although the literature indicates that the microbial processing of bioavailable DOM is essential during the production of autochthonous, labile, DOM. DOM can be broadly characterised by its fluorescing properties and Coble et al. (2014) define terrestrially derived DOM as exhibiting "peak C" fluorescence, whilst labile microbially derived DOM is defined as showing "peak T" fluorescence. Our work explores the microbial/DOM interactions by analysing aquatic samples using fluorescence excitation and emission matrices (EEMs) in conjunction with microbial consumption of dissolved oxygen. Environmental and synthetic water samples were subjected to fluorescence characterisation using both fluorescence spectroscopy and in situ fluorescence sensors (Chelsea Technologies Group Ltd.). PARAFAC analysis and peak picking were performed on EEMs and compared with flow cytometry data, used to quantify bacterial numbers present within samples. Synthetic samples were created using glucose, glutamic acid, nutrient-rich water and a standard bacterial seed. Synthetic samples were provided with terrestrially derived DOM via the addition of an aliquot of environmental water. Using a closed system approach, samples were incubated over time (up to a maximum of 20 days) and analysed at pre-defined intervals. The main focus of our work is to improve our understanding of microbial/DOM interactions and how these interactions affect both the DOM characteristics and microbial food web in freshwater aquatic systems. The information gained, in relation to the origin, microbial

  11. Microbial quality of frozen Nile crocodile (Crocodylus niloticus) meat samples from three selected farms in Zimbabwe.

    PubMed

    Makanyanga, Tsitsi B; Mutema, Gideon; Mukarati, Norman L; Chikerema, Sylvester M; Makaya, Pious V; Musari, Shuvai; Matope, Gift

    2014-01-17

    Microbial quality of frozen Nile crocodile (Crocodylus niloticus) meat from three farms in Zimbabwe was assessed based on 2051 samples collected for pre-export testing during 2006 to 2011. Data were perused by season and year in terms of aerobic plate (APC), coliform (CC), Escherichia coli (ECC) and Listeria monocytogenes (LMC) counts and the presence of Salmonella spp. The log10-transformed data were compared among the farms and seasons using the Kruskal-Wallis test. Microbial quality of the samples was graded based on the EC No. 2073.2005 criteria for beef. The mean APC and CC for the crocodile meat differed significantly (P=0.000) among the farms with the highest APC (3.2±0.05 log10 cfu/g) and the lowest (2.7±0.05 log10 cfu/g) recorded from farms A and C, respectively. There were no significant differences (P>0.05) in ECC and LMC among the farms, while Salmonella spp. were only isolated from one farm. Although the microbial quality of frozen crocodile meat from these farms was generally within acceptable limits, the isolation of E. coli and Salmonella spp. is of public health concern. Thus, implementing of measures to control the pasteurizing process and to minimize bacterial contamination of crocodile meat after pasteurization need to be carefully considered. © 2013.

  12. Random Sampling Process Leads to Overestimation of β-Diversity of Microbial Communities

    PubMed Central

    Zhou, Jizhong; Jiang, Yi-Huei; Deng, Ye; Shi, Zhou; Zhou, Benjamin Yamin; Xue, Kai; Wu, Liyou; He, Zhili; Yang, Yunfeng

    2013-01-01

    ABSTRACT The site-to-site variability in species composition, known as β-diversity, is crucial to understanding spatiotemporal patterns of species diversity and the mechanisms controlling community composition and structure. However, quantifying β-diversity in microbial ecology using sequencing-based technologies is a great challenge because of a high number of sequencing errors, bias, and poor reproducibility and quantification. Herein, based on general sampling theory, a mathematical framework is first developed for simulating the effects of random sampling processes on quantifying β-diversity when the community size is known or unknown. Also, using an analogous ball example under Poisson sampling with limited sampling efforts, the developed mathematical framework can exactly predict the low reproducibility among technically replicate samples from the same community of a certain species abundance distribution, which provides explicit evidences of random sampling processes as the main factor causing high percentages of technical variations. In addition, the predicted values under Poisson random sampling were highly consistent with the observed low percentages of operational taxonomic unit (OTU) overlap (<30% and <20% for two and three tags, respectively, based on both Jaccard and Bray-Curtis dissimilarity indexes), further supporting the hypothesis that the poor reproducibility among technical replicates is due to the artifacts associated with random sampling processes. Finally, a mathematical framework was developed for predicting sampling efforts to achieve a desired overlap among replicate samples. Our modeling simulations predict that several orders of magnitude more sequencing efforts are needed to achieve desired high technical reproducibility. These results suggest that great caution needs to be taken in quantifying and interpreting β-diversity for microbial community analysis using next-generation sequencing technologies. PMID:23760464

  13. Changes in aquatic microbial responses to C-substrates with stream water and sediment quality related to land use pressures.

    PubMed

    Stutter, Marc I; Cains, Jonathan

    2017-10-01

    The degradation of aquatic ecosystems by multiply-acting stressors is a key threat requiring new approaches to identify impairment processes and restoration targets. Heterotrophic respiration of C-substrates can be an integrative indicator of how aquatic ecosystems respond to pollution. We utilised sixteen C-substrates (simple metabolites to dissolved organic matter (DOM)) to characterise substrate induced respiration (SIR) for sediments from sixteen NE Scotland streams covering a range of land use-related pollution pressures. Pollution explanatory variables were as assessed from GIS-derived land cover areas, dissolved water chemistry (N, P, base cations and trace elements) and sediment trace metals (by aqua-regia digest). Large inter-site variation in dissolved and sediment chemical concentrations was strongly significantly correlated with land cover: dissolved N, Ca, Mg, K, Na positively with agriculture and urban, negatively with semi-natural land areas; sediment As, Ba, Co, Zn with agriculture, Cu, Pb, Zn with urban areas. Simple linear regression modelling was used to explore the influences across land cover, dissolved and sediment chemistry with C-substrate responses, both individually and using principal components-derived SIR 'fingerprints'. The data supported the hypothesis that pollution pressures altered water and sediment quality, in turn affecting sediment microbes and their respiration responses to a range of C-substrates, especially discriminating the DOM respiration. Since headwater DOM is a dominant pool of C driving ecosystem processes downstream then any loss of ability to utilise DOM should be further explored in terms of possible connections to pollution processes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Abundance-weighted phylogenetic diversity measures distinguish microbial community states and are robust to sampling depth.

    PubMed

    McCoy, Connor O; Matsen, Frederick A

    2013-01-01

    In microbial ecology studies, the most commonly used ways of investigating alpha (within-sample) diversity are either to apply non-phylogenetic measures such as Simpson's index to Operational Taxonomic Unit (OTU) groupings, or to use classical phylogenetic diversity (PD), which is not abundance-weighted. Although alpha diversity measures that use abundance information in a phylogenetic framework do exist, they are not widely used within the microbial ecology community. The performance of abundance-weighted phylogenetic diversity measures compared to classical discrete measures has not been explored, and the behavior of these measures under rarefaction (sub-sampling) is not yet clear. In this paper we compare the ability of various alpha diversity measures to distinguish between different community states in the human microbiome for three different datasets. We also present and compare a novel one-parameter family of alpha diversity measures, BWPDθ, that interpolates between classical phylogenetic diversity (PD) and an abundance-weighted extension of PD. Additionally, we examine the sensitivity of these phylogenetic diversity measures to sampling, via computational experiments and by deriving a closed form solution for the expectation of phylogenetic quadratic entropy under re-sampling. On the three datasets, a phylogenetic measure always performed best, and two abundance-weighted phylogenetic diversity measures were the only measures ranking in the top four across all datasets. OTU-based measures, on the other hand, are less effective in distinguishing community types. In addition, abundance-weighted phylogenetic diversity measures are less sensitive to differing sampling intensity than their unweighted counterparts. Based on these results we encourage the use of abundance-weighted phylogenetic diversity measures, especially for cases such as microbial ecology where species delimitation is difficult.

  15. Stabilization of human urine doping control samples: II. microbial degradation of steroids.

    PubMed

    Tsivou, M; Livadara, D; Georgakopoulos, D G; Koupparis, M A; Atta-Politou, J; Georgakopoulos, C G

    2009-05-01

    The transportation of urine samples, collected for doping control analysis, does not always meet ideal conditions of storage and prompt delivery to the World Anti-Doping Agency (WADA) accredited laboratories. Because sample collection is not conducted under sterile conditions, microbial activity may cause changes to the endogenous steroid profiles of samples. In the current work, funded by WADA, a chemical mixture consisting of antibiotics, antimycotic substances and protease inhibitors was applied in urine aliquots fortified with conjugated and deuterated steroids and inoculated with nine representative microorganisms. Aliquots with and without the chemical mixture were incubated at 37 degrees C for 7 days to simulate the transportation period, whereas another series of aliquots was stored at -20 degrees C as reference. Microbial growth was assessed immediately after inoculation and at the end of the incubation period. Variations in pH and specific gravity values were recorded. Gas chromatography-mass spectrometry (GC-MS) analysis was performed for the detection of steroids in the free, glucuronide, and sulfate fractions. The addition of the chemical stabilization mixture to urine samples inhibited microorganism growth and prevented steroid degradation at 37 degrees C. On the other hand, four of the nine microorganisms induced alterations in the steroid profile of the unstabilized samples incubated at 37 degrees C.

  16. Sample processing and cDNA preparation for microbial metatranscriptomics in complex soil communities.

    PubMed

    Carvalhais, Lilia C; Schenk, Peer M

    2013-01-01

    Soil presents one of the most complex environments for microbial communities as it provides many microhabitats that allow coexistence of thousands of species with important ecosystem functions. These include biomass and nutrient cycling, mineralization, and detoxification. Culture-independent DNA-based methods, such as metagenomics, have revealed operational taxonomic units that suggest a high diversity of microbial species and associated functions in soil. An emerging but technically challenging area to profile the functions of microorganisms and their activities is mRNA-based metatranscriptomics. Here, we describe issues and important considerations of soil sample processing and cDNA preparation for metatranscriptomics from bacteria and archaea and provide a set of methods that can be used in the required experimental steps.

  17. Separation and analysis of trace volatile formaldehyde in aquatic products by a MoO₃/polypyrrole intercalative sampling adsorbent with thermal desorption gas chromatography and mass spectrometry.

    PubMed

    Ma, Yunjian; Zhao, Cheng; Zhan, Yisen; Li, Jianbin; Zhang, Zhuomin; Li, Gongke

    2015-05-01

    An in situ embedded synthesis strategy was developed for the preparation of a MoO3 /polypyrrole intercalative sampling adsorbent for the separation and analysis of trace volatile formaldehyde in aquatic products. Structural and morphological characteristics of the MoO3 /polypyrrole intercalative adsorbent were investigated by a series of characterization methods. The MoO3 /polypyrrole sampling adsorbent possessed a higher sampling capacity and selectivity for polar formaldehyde than commonly used commercial adsorbent Tenax TA. Finally, the MoO3 /polypyrrole adsorbent was packed in the thermal desorption tube that was directly coupled to gas chromatography with mass spectrometry for the analysis of trace volatile formaldehyde in aquatic products. Trace volatile formaldehyde from real aquatic products could be selectively sampled and quantified to be 0.43-6.6 mg/kg. The detection limit was achieved as 0.004 μg/L by this method. Good recoveries for spiked aquatic products were achieved in range of 75.0-108% with relative standard deviations of 1.2-9.0%. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Examination of Microbial Proteome Preservation Techniques Applicable to Autonomous Environmental Sample Collection

    PubMed Central

    Saito, Mak A.; Bulygin, Vladimir V.; Moran, Dawn M.; Taylor, Craig; Scholin, Chris

    2011-01-01

    Improvements in temporal and spatial sampling frequency have the potential to open new windows into the understanding of marine microbial dynamics. In recent years, efforts have been made to allow automated samplers to collect microbial biomass for DNA/RNA analyses from moored observatories and autonomous underwater vehicles. Measurements of microbial proteins are also of significant interest given their biogeochemical importance as enzymes that catalyze reactions and transporters that interface with the environment. We examined the influence of five preservatives solutions (SDS-extraction buffer, ethanol, trichloroacetic acid, B-PER, and RNAlater) on the proteome integrity of the marine cyanobacterium Synechococcus WH8102 after 4 weeks of storage at room temperature. Four approaches were used to assess degradation: total protein recovery, band integrity on an SDS detergent polyacrylamide electrophoresis (SDS-PAGE) gel, and number of protein identifications and relative abundances by 1-dimensional LC–MS/MS proteomic analyses. Total protein recoveries from the preserved samples were lower than the frozen control due to processing losses, which could be corrected for with internal standardization. The trichloroacetic acid preserved sample showed significant loss of protein band integrity on the SDS-PAGE gel. The RNAlater preserved sample showed the highest number of protein identifications (103% relative to the control; 520 ± 31 identifications in RNAlater versus 504 ± 4 in the control), equivalent to the frozen control. Relative abundances of individual proteins in the RNAlater treatment were quite similar to that of the frozen control (average ratio of 1.01 ± 0.27 for the 50 most abundant proteins), while the SDS-extraction buffer, ethanol, and B-PER all showed significant decreases in both number of identifications and relative abundances of individual proteins. Based on these findings, RNAlater was an effective proteome preservative, although

  19. Examination of microbial proteome preservation techniques applicable to autonomous environmental sample collection.

    PubMed

    Saito, Mak A; Bulygin, Vladimir V; Moran, Dawn M; Taylor, Craig; Scholin, Chris

    2011-01-01

    Improvements in temporal and spatial sampling frequency have the potential to open new windows into the understanding of marine microbial dynamics. In recent years, efforts have been made to allow automated samplers to collect microbial biomass for DNA/RNA analyses from moored observatories and autonomous underwater vehicles. Measurements of microbial proteins are also of significant interest given their biogeochemical importance as enzymes that catalyze reactions and transporters that interface with the environment. We examined the influence of five preservatives solutions (SDS-extraction buffer, ethanol, trichloroacetic acid, B-PER, and RNAlater) on the proteome integrity of the marine cyanobacterium Synechococcus WH8102 after 4 weeks of storage at room temperature. Four approaches were used to assess degradation: total protein recovery, band integrity on an SDS detergent polyacrylamide electrophoresis (SDS-PAGE) gel, and number of protein identifications and relative abundances by 1-dimensional LC-MS/MS proteomic analyses. Total protein recoveries from the preserved samples were lower than the frozen control due to processing losses, which could be corrected for with internal standardization. The trichloroacetic acid preserved sample showed significant loss of protein band integrity on the SDS-PAGE gel. The RNAlater preserved sample showed the highest number of protein identifications (103% relative to the control; 520 ± 31 identifications in RNAlater versus 504 ± 4 in the control), equivalent to the frozen control. Relative abundances of individual proteins in the RNAlater treatment were quite similar to that of the frozen control (average ratio of 1.01 ± 0.27 for the 50 most abundant proteins), while the SDS-extraction buffer, ethanol, and B-PER all showed significant decreases in both number of identifications and relative abundances of individual proteins. Based on these findings, RNAlater was an effective proteome preservative, although

  20. Effect of short-term room temperature storage on the microbial community in infant fecal samples

    PubMed Central

    Guo, Yong; Li, Sheng-Hui; Kuang, Ya-Shu; He, Jian-Rong; Lu, Jin-Hua; Luo, Bei-Jun; Jiang, Feng-Ju; Liu, Yao-Zhong; Papasian, Christopher J.; Xia, Hui-Min; Deng, Hong-Wen; Qiu, Xiu

    2016-01-01

    Sample storage conditions are important for unbiased analysis of microbial communities in metagenomic studies. Specifically, for infant gut microbiota studies, stool specimens are often exposed to room temperature (RT) conditions prior to analysis. This could lead to variations in structural and quantitative assessment of bacterial communities. To estimate such effects of RT storage, we collected feces from 29 healthy infants (0–3 months) and partitioned each sample into 5 portions to be stored for different lengths of time at RT before freezing at −80 °C. Alpha diversity did not differ between samples with storage time from 0 to 2 hours. The UniFrac distances and microbial composition analysis showed significant differences by testing among individuals, but not by testing between different time points at RT. Changes in the relative abundance of some specific (less common, minor) taxa were still found during storage at room temperature. Our results support previous studies in children and adults, and provided useful information for accurate characterization of infant gut microbiomes. In particular, our study furnished a solid foundation and justification for using fecal samples exposed to RT for less than 2 hours for comparative analyses between various medical conditions. PMID:27226242

  1. Empirical testing of modified Salmonella MLST in aquatic environmental samples by in silico analysis.

    PubMed

    Ho, Ying-Ning; Chou, Ming-Yuan; Tsai, Hsin-Chi; Huang, Tung-Yi; Fan, Cheng-Wei; Hsu, Bing-Mu

    2017-03-01

    Multilocus sequence typing (MLST) is an approach for prediction of Salmonella servoar and eBRUST groups (eBGs) based on seven typing scheme of housekeeping genes. Up to date, >220.000 allelic profiles and 65,973 Salmonella strains have been established in the MLST database. Several studies have modified MLST method with fewer targeted housekeeping genes for the purpose of economy and efficiency. Nevertheless, no study has conducted systematically to evaluate the correlation between the numbers of housekeeping genes targeted and the accuracy of prediction rate. In this study, we aimed to tackle this problem by extracting data from the MLST database as a whole using the software RStudio. Our results indicated that as the numbers of genes in MLST scheme increased, the accuracy of the eBGs prediction rate increased and reached 100% when the gene numbers are greater than or equal to 5. To examine the applicability of the approach, 395 environmental water samples were subjected to this study. A set of 52 Salmonella enterica isolates was initially used to develop MLST targeting seven housekeeping genes. A total of 29 sequence types, including 11 new sequence types were found among the 52 sequenced isolates that differentiated into 19 serotypes. Moreover, two novel sequence types did not belong to current classification. Our results show that the outcome in the three-gene sequence typing (aroC, hisD, and purE) was as accurate as in the seven-gene sequence typing for prediction of environmental Salmonella isolates. Our data suggested that this five-gene and reduced gene-number sequence-typing scheme can serve as an alternative modified MLST when effectiveness and financial management were the concerns.

  2. Biomarker Analysis of Samples Visually Identified as Microbial in the Eocene Green River Formation: An Analogue for Mars.

    PubMed

    Olcott Marshall, Alison; Cestari, Nicholas A

    2015-09-01

    One of the major exploration targets for current and future Mars missions are lithofacies suggestive of biotic activity. Although such lithofacies are not confirmation of biotic activity, they provide a way to identify samples for further analyses. To test the efficacy of this approach, we identified carbonate samples from the Eocene Green River Formation as "microbial" or "non-microbial" based on the macroscale morphology of their laminations. These samples were then crushed and analyzed by gas chromatography/mass spectroscopy (GC/MS) to determine their lipid biomarker composition. GC/MS analysis revealed that carbonates visually identified as "microbial" contained a higher concentration of more diverse biomarkers than those identified as "non-microbial," suggesting that this could be a viable detection strategy for selecting samples for further analysis or caching on Mars.

  3. Linking questions to practices in the study of microbial pathogens: sampling bias and typing methods.

    PubMed

    Gómez-Díaz, Elena

    2009-12-01

    The importance of understanding the population genetics and evolution of microbial pathogens is increasing as a result of the spread and re-emergence of many infectious diseases and their impact for public health. In the last few years, the development of high throughput multi-gene sequence methodologies has opened new opportunities for studying pathogen populations, providing reliable and robust means for both epidemiological and evolutionary investigations. For instance, for many pathogens, multilocus sequence typing has become the "gold standard" in molecular epidemiology, allowing strain identification and discovery. However, there is a huge gap between typing a clinical collection of isolates and making inferences about their evolutionary history and population genetics. Critical issues for studying microbial pathogens such as an adequate sampling design and the appropriate selection of the genetic technique are also required, and will rely on the scale of study and the characteristics of the biological system (e.g., multi- vs. single-host pathogens and vector vs. food or air-borne pathogens). My aim here is to discuss some of these issues in more detail and illustrate how these aspects are often overlooked and easily neglected in the field. Finally, given the rapid accumulation of complete genome sequences and the increasing effort on microbiology research, it is clear that now more than ever integrative approaches bringing together epidemiology and evolutionary biology are needed for understanding the diversity of microbial pathogens.

  4. Direct and indirect effects of temperature on the population dynamics and ecosystem functioning of aquatic microbial ecosystems.

    PubMed

    Beveridge, Oliver S; Petchey, Owen L; Humphries, Stuart

    2010-11-01

    1. While much is known about the direct effect that temperature can have on aquatic communities, less is known about its indirect effect via the temperature dependence of viscosity and temperature-dependent trophic interactions. 2. We manipulated the temperature (5-20 °C) and the viscosity (equivalent to 5-20 °C) of water in laboratory-based bacteria-protist communities. Communities contained food chains with one, two or three trophic levels. Responses measured were population dynamics (consumer carrying capacity and growth rate, average species population density, and the coefficient of variation of population density through time) and ecosystem function (decomposition). 3. Temperature, viscosity and food chain length produced significant responses in population dynamics. Temperature-dependent viscosity had a significant effect on the carrying capacity and growth rates of consumers, as well as the average density of the top predator. Overall, indirect effects of temperature via changes in viscosity were subtle in comparison to the indirect effect of temperature via trophic interactions. 4. Our results highlight the importance of direct and indirect effects of temperature, mediated through trophic interactions and physical changes in the environment, both for population dynamics and ecosystem processes. Future mechanistic modelling of effects of environmental change on species will benefit from distinguishing the different mechanisms of the overall effect of temperature.

  5. GPU-Meta-Storms: computing the structure similarities among massive amount of microbial community samples using GPU.

    PubMed

    Su, Xiaoquan; Wang, Xuetao; Jing, Gongchao; Ning, Kang

    2014-04-01

    The number of microbial community samples is increasing with exponential speed. Data-mining among microbial community samples could facilitate the discovery of valuable biological information that is still hidden in the massive data. However, current methods for the comparison among microbial communities are limited by their ability to process large amount of samples each with complex community structure. We have developed an optimized GPU-based software, GPU-Meta-Storms, to efficiently measure the quantitative phylogenetic similarity among massive amount of microbial community samples. Our results have shown that GPU-Meta-Storms would be able to compute the pair-wise similarity scores for 10 240 samples within 20 min, which gained a speed-up of >17 000 times compared with single-core CPU, and >2600 times compared with 16-core CPU. Therefore, the high-performance of GPU-Meta-Storms could facilitate in-depth data mining among massive microbial community samples, and make the real-time analysis and monitoring of temporal or conditional changes for microbial communities possible. GPU-Meta-Storms is implemented by CUDA (Compute Unified Device Architecture) and C++. Source code is available at http://www.computationalbioenergy.org/meta-storms.html.

  6. Microbial sequences retrieved from environmental samples from seasonal arctic snow and meltwater from Svalbard, Norway.

    PubMed

    Larose, Catherine; Berger, Sibel; Ferrari, Christophe; Navarro, Elisabeth; Dommergue, Aurélien; Schneider, Dominique; Vogel, Timothy M

    2010-03-01

    16S rRNA gene (rrs) clone libraries were constructed from two snow samples (May 11, 2007 and June 7, 2007) and two meltwater samples collected during the spring of 2007 in Svalbard, Norway (79 degrees N). The libraries covered 19 different microbial classes, including Betaproteobacteria (21.3%), Sphingobacteria (16.4%), Flavobacteria (9.0%), Acidobacteria (7.7%) and Alphaproteobacteria (6.5%). Significant differences were detected between the two sets of sample libraries. First, the meltwater libraries had the highest community richness (Chao1: 103.2 and 152.2) and Shannon biodiversity indices (between 3.38 and 3.59), when compared with the snow libraries (Chao1: 14.8 and 59.7; Shannon index: 1.93 and 3.01). Second, integral-LIBSHUFF analyses determined that the bacterial communities in the snow libraries were significantly different from those of the meltwater libraries. Despite these differences, our data also support the theory that a common core group of microbial populations exist within a variety of cryohabitats. Electronic supplementary material The online version of this article (doi:10.1007/s00792-009-0299-2) contains supplementary material, which is available to authorized users.

  7. Vibrational Spectroscopy for Imaging Single Microbial Cells in Complex Biological Samples

    PubMed Central

    Harrison, Jesse P.; Berry, David

    2017-01-01

    Vibrational spectroscopy is increasingly used for the rapid and non-destructive imaging of environmental and medical samples. Both Raman and Fourier-transform infrared (FT-IR) imaging have been applied to obtain detailed information on the chemical composition of biological materials, ranging from single microbial cells to tissues. Due to its compatibility with methods such as stable isotope labeling for the monitoring of cellular activities, vibrational spectroscopy also holds considerable power as a tool in microbial ecology. Chemical imaging of undisturbed biological systems (such as live cells in their native habitats) presents unique challenges due to the physical and chemical complexity of the samples, potential for spectral interference, and frequent need for real-time measurements. This Mini Review provides a critical synthesis of recent applications of Raman and FT-IR spectroscopy for characterizing complex biological samples, with a focus on developments in single-cell imaging. We also discuss how new spectroscopic methods could be used to overcome current limitations of single-cell analyses. Given the inherent complementarity of Raman and FT-IR spectroscopic methods, we discuss how combining these approaches could enable us to obtain new insights into biological activities either in situ or under conditions that simulate selected properties of the natural environment. PMID:28450860

  8. Microbial Air and Surface Monitoring Results from International Space Station Samples

    NASA Technical Reports Server (NTRS)

    Ott, C. Mark; Bruce, Rebekah J.; Castro, Victoria A.; Novikova, Natalia D.; Pierson, D. L.

    2005-01-01

    Over the course of long-duration spaceflight, spacecraft develop a microbial ecology that directly interacts with the crew of the vehicle. While most microorganisms are harmless or beneficial to the inhabitants of the vehicle, the presence of medically significant organisms appearing in this semi-closed environment could adversely affect crew health and performance. The risk of exposure of the crew to medically significant organisms during a mission is estimated using information gathered during nominal and contingency environmental monitoring. Analysis of the air and surface microbiota in the habitable compartments of the International Space Station (ISS) over the last four years indicate a high presence of Staphylococcus species reflecting the human inhabitants of the vehicle. Generally, air and surface microbial concentrations are below system design specifications, suggesting a lower risk of contact infection or biodegradation. An evaluation of sample frequency indicates a decrease in the identification of new species, suggesting a lower potential for unknown microorganisms to be identified. However, the opportunistic pathogen, Staphylococcus aureus, has been identified in 3 of the last 5 air samples and 5 of the last 9 surface samples. In addition, 47% of the coagulase negative Staphylococcus species that were isolated from the crew, ISS, and its hardware were found to be methicillin resistance. In combination, these observations suggest the potential of methicillin resistant infectious agents over time.

  9. Microbial Air and Surface Monitoring Results from International Space Station Samples

    NASA Technical Reports Server (NTRS)

    Ott, C. Mark; Bruce, Rebekah J.; Castro, Victoria A.; Novikova, Natalia D.; Pierson, D. L.

    2005-01-01

    Over the course of long-duration spaceflight, spacecraft develop a microbial ecology that directly interacts with the crew of the vehicle. While most microorganisms are harmless or beneficial to the inhabitants of the vehicle, the presence of medically significant organisms appearing in this semi-closed environment could adversely affect crew health and performance. The risk of exposure of the crew to medically significant organisms during a mission is estimated using information gathered during nominal and contingency environmental monitoring. Analysis of the air and surface microbiota in the habitable compartments of the International Space Station (ISS) over the last four years indicate a high presence of Staphylococcus species reflecting the human inhabitants of the vehicle. Generally, air and surface microbial concentrations are below system design specifications, suggesting a lower risk of contact infection or biodegradation. An evaluation of sample frequency indicates a decrease in the identification of new species, suggesting a lower potential for unknown microorganisms to be identified. However, the opportunistic pathogen, Staphylococcus aureus, has been identified in 3 of the last 5 air samples and 5 of the last 9 surface samples. In addition, 47% of the coagulase negative Staphylococcus species that were isolated from the crew, ISS, and its hardware were found to be methicillin resistance. In combination, these observations suggest the potential of methicillin resistant infectious agents over time.

  10. Vibrational spectroscopy for imaging single microbial cells in complex biological samples

    DOE PAGES

    Harrison, Jesse P.; Berry, David

    2017-04-13

    Here, vibrational spectroscopy is increasingly used for the rapid and non-destructive imaging of environmental and medical samples. Both Raman and Fourier-transform infrared (FT-IR) imaging have been applied to obtain detailed information on the chemical composition of biological materials, ranging from single microbial cells to tissues. Due to its compatibility with methods such as stable isotope labeling for the monitoring of cellular activities, vibrational spectroscopy also holds considerable power as a tool in microbial ecology. Chemical imaging of undisturbed biological systems (such as live cells in their native habitats) presents unique challenges due to the physical and chemical complexity of themore » samples, potential for spectral interference, and frequent need for real-time measurements. This Mini Review provides a critical synthesis of recent applications of Raman and FT-IR spectroscopy for characterizing complex biological samples, with a focus on developments in single-cell imaging. We also discuss how new spectroscopic methods could be used to overcome current limitations of singlecell analyses. Given the inherent complementarity of Raman and FT-IR spectroscopic methods, we discuss how combining these approaches could enable us to obtain new insights into biological activities either in situ or under conditions that simulate selected properties of the natural environment.« less

  11. Adenosine triphosphate bioluminescence analysis for rapid screening of microbial contamination in non-sterile pharmaceutical samples.

    PubMed

    Jimenez, Luis

    2004-01-01

    An Adenosine Triphosphate (ATP) bioluminescence system was compared and validated against standard methods for rapid microbiological monitoring of several non-sterile pharmaceutical formulations such as creams, tablets, and capsules. Results obtained using 1%, 2.5%, and 10% of product suspensions indicated that most samples that did not contain non-microbial ATP neither inhibited the bioluminescence reaction nor did something else. Ten percent product suspensions were inoculated with different concentrations of Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Candida albicans, and Aspergillus niger. Samples were incubated for 24-120 h at 35 degrees C with shaking. Results indicated a strong inhibitory effect of microbial growth, as no microorganisms were detected by using the ATP bioluminescence assay. However, when 1% and 2.5% product suspensions were spiked with the same microorganisms, positive detection was confirmed. After incubation, all microorganisms were detected by the bioluminescence system within 24-72 h. All positive samples were confirmed by using standard plating media. However, to optimize detection of all microorganisms, different enrichment media were developed.

  12. Microbial profiling of South African acid mine water samples using next generation sequencing platform.

    PubMed

    Kamika, I; Azizi, S; Tekere, M

    2016-07-01

    This study monitored changes in bacterial and fungal structure in a mine water in a monthly basis over 4 months. Over the 4-month study period, mine water samples contained more bacteria (91.06 %) compared to fungi (8.94 %). For bacteria, mine water samples were dominated by Proteobacteria (39.14 to 65.06 %) followed by Firmicutes (26.34 to 28.9 %) in summer, and Cyanobacteria (27.05 %) in winter. In the collected samples, 18 % of bacteria could not be assigned to a phylum and remained unclassified suggesting hitherto vast untapped microbial diversity especially during winter. The fungal domain was the sole eukaryotic microorganism found in the mine water samples with unclassified fungi (68.2 to 91 %) as the predominant group, followed by Basidiomycota (6.9 to 27.8 %). The time of collection, which was linked to the weather, had higher impact on bacterial community than fungal community. The bacterial operational taxonomic units (OTUs) ranged from 865 to 4052 over the 4-month sampling period, while fungal OTUs varied from 73 to 249. The diversity indices suggested that the bacterial community inhabiting the mine water samples were more diverse than the fungal community. The canonical correspondence analysis (CCA) results highlighted that the bacterial community variance had the strongest relationship with water temperature, conductivity, pH, and dissolved oxygen (DO) content, as compared to fungi and water characteristics, had the greatest contribution to both bacterial and fungal community variance. The results provided the relationships between microbial community and environmental variables in the studied mining sites.

  13. Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

    PubMed Central

    Boardman, Anna K.; Campbell, Jennifer; Wirz, Holger; Sharon, Andre; Sauer-Budge, Alexis F.

    2015-01-01

    Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample. PMID:25675242

  14. Microbial Metagenomics Mock Scenario-based Sample Simulation (M3S3).

    PubMed

    Motro, Yair; Moran-Gilad, Jacob

    2017-08-12

    Shotgun sequencing in increasingly applied in clinical microbiology for unbiased culture-independent diagnosis. While software solutions for metagenomics proliferate, integration of metagenomics in clinical care, requires method standardisation and validation. Virtual metagenomics samples could underpin validation by substituting real samples and thus we sought to develop a novel solution for simulation of metagenomics samples based on user-defined clinical scenarios. We designed the Microbial Metagenomics Mock Scenario-based Sample Simulation (M3S3) workflow, which allows users to generate virtual samples from raw reads or assemblies. The M3S3 output is a mock sample in FASTQ or FASTA format. M3S3 was tested by generating virtual samples for ten challenging infectious disease scenarios, involving a background matrix 'spiked' in silico with pathogens including mixtures. Replicate samples (seven per scenario) were used to represent different compositional ratios. Virtual samples were analysed using Taxonomer and Kraken db. The ten challenge scenarios were successfully applied, generating 80 samples. For all tested scenarios, the virtual samples showed sequence compositions as predicted from the user input. Spiked pathogen sequences were identified with the majority of the replicates and most exhibited acceptable abundance (deviation between expected and observed abundance of spiked pathogens), with slight differences observed between software tools. Despite demonstrated proof-of-concept, integration of clinical metagenomics in routine microbiology remains a substantial challenge. M3S3 is capable of producing virtual samples on-demand, simulating a spectrum of clinical diagnostic scenarios of varying complexity. The M3S3 tool can therefore support the development and validation of standardised metagenomics applications. Copyright © 2017. Published by Elsevier Ltd.

  15. Sampling natural biofilms: a new route to build efficient microbial anodes.

    PubMed

    Erable, Benjamin; Roncato, Marie-Anne; Achouak, Wafa; Bergel, Alain

    2009-05-01

    Electrochemically active biofilms were constructed on graphite anodes under constant polarization at -0.1V vs saturated calomel reference (SCE) with 10 mM acetate as substrate. The reactors were inoculated with three different microbial samples that were drawn from exactly the same place in a French Atlantic coastal port (i) by scraping the biofilm that had formed naturally on the surface of a floating bridge, (ii) by taking marine sediments just under the floating bridge, and (iii) by taking nearby beach sand. Current densities of 2.0 A/m2 were reached using the biofilm sample as inoculum while only 0.4 A/m2 and 0.8 A/m2 were obtained using the underlying sediments and the beach sand, respectively. The structure of bacterial communities forming biofilms was characterized by denaturing gradient gel electrophoresis (DGGE) analysis, and revealed differences between samples with the increase in relative intensities of some bands and the appearance of others. Bacteria close related to Bacteroidetes, Halomonas, and Marinobacterium were retrieved only from the efficient EA-biofilms formed from natural biofilms, whereas, bacteria close related to Mesoflavibacter were predominant on biofilm formed from sediments. The marine biofilm was selected as the inoculum to further optimize the microbial anode. Epifluorescence microscopy and SEM confirmed that maintaining the electrode under constant polarization promoted rapid settlement of the electrode surface by a bacterial monolayer film. The microbial anode was progressively adapted to the consumption of acetate by three serial additions of substrate, thus improving the Coulombic efficiency of acetate consumption from 31 to 89%. The possible oxidation of sulfide played only a very small part in the current production and the biofilm was not able to oxidize hydrogen. Graphite proved to be more efficient than dimensionally stable anode (DSA) or stainless steel butthis result might be due to differences in the surface roughness

  16. Ancient pathogen DNA in archaeological samples detected with a Microbial Detection Array

    PubMed Central

    Devault, Alison M.; McLoughlin, Kevin; Jaing, Crystal; Gardner, Shea; Porter, Teresita M.; Enk, Jacob M.; Thissen, James; Allen, Jonathan; Borucki, Monica; DeWitte, Sharon N.; Dhody, Anna N.; Poinar, Hendrik N.

    2014-01-01

    Ancient human remains of paleopathological interest typically contain highly degraded DNA in which pathogenic taxa are often minority components, making sequence-based metagenomic characterization costly. Microarrays may hold a potential solution to these challenges, offering a rapid, affordable, and highly informative snapshot of microbial diversity in complex samples without the lengthy analysis and/or high cost associated with high-throughput sequencing. Their versatility is well established for modern clinical specimens, but they have yet to be applied to ancient remains. Here we report bacterial profiles of archaeological and historical human remains using the Lawrence Livermore Microbial Detection Array (LLMDA). The array successfully identified previously-verified bacterial human pathogens, including Vibrio cholerae (cholera) in a 19th century intestinal specimen and Yersinia pestis (“Black Death” plague) in a medieval tooth, which represented only minute fractions (0.03% and 0.08% alignable high-throughput shotgun sequencing reads) of their respective DNA content. This demonstrates that the LLMDA can identify primary and/or co-infecting bacterial pathogens in ancient samples, thereby serving as a rapid and inexpensive paleopathological screening tool to study health across both space and time. PMID:24603850

  17. Ancient pathogen DNA in archaeological samples detected with a Microbial Detection Array.

    PubMed

    Devault, Alison M; McLoughlin, Kevin; Jaing, Crystal; Gardner, Shea; Porter, Teresita M; Enk, Jacob M; Thissen, James; Allen, Jonathan; Borucki, Monica; DeWitte, Sharon N; Dhody, Anna N; Poinar, Hendrik N

    2014-03-06

    Ancient human remains of paleopathological interest typically contain highly degraded DNA in which pathogenic taxa are often minority components, making sequence-based metagenomic characterization costly. Microarrays may hold a potential solution to these challenges, offering a rapid, affordable, and highly informative snapshot of microbial diversity in complex samples without the lengthy analysis and/or high cost associated with high-throughput sequencing. Their versatility is well established for modern clinical specimens, but they have yet to be applied to ancient remains. Here we report bacterial profiles of archaeological and historical human remains using the Lawrence Livermore Microbial Detection Array (LLMDA). The array successfully identified previously-verified bacterial human pathogens, including Vibrio cholerae (cholera) in a 19th century intestinal specimen and Yersinia pestis ("Black Death" plague) in a medieval tooth, which represented only minute fractions (0.03% and 0.08% alignable high-throughput shotgun sequencing reads) of their respective DNA content. This demonstrates that the LLMDA can identify primary and/or co-infecting bacterial pathogens in ancient samples, thereby serving as a rapid and inexpensive paleopathological screening tool to study health across both space and time.

  18. Microbial degradation of gasoline in soil: Effect of season of sampling.

    PubMed

    Turner, D A; Pichtel, J; Rodenas, Y; McKillip, J; Goodpaster, J V

    2015-06-01

    In cases where fire debris contains soil, microorganisms can rapidly and irreversibly alter the chemical composition of any ignitable liquid residue that may be present. In this study, differences in microbial degradation due to the season in which the sample is collected was examined. Soil samples were collected from the same site during Fall, Winter, Spring and Summer and the degradation of gasoline was monitored over 30 days. Predominant viable bacterial populations enumerated using real-time PCR and reverse transcriptase polymerase chain reaction (RT-PCR) enumeration revealed the predominant viable bacterial genera to be Alcaligenes, Bacillus, and Flavobacterium. Overall, the compounds most vulnerable to microbial degradation are the n-alkanes, followed by the mono-substituted alkylbenzenes (e.g., toluene, ethylbenzene, propylbenzene and isopropylbenzene). Benzaldehyde (a degradation product of toluene) was also identified as a marker for the extent of biodegradation. Ultimately, it was determined that soil collected during an unusually hot and dry summer exhibited the least degradation with little to no change in gasoline for up to 4 days, readily detectable n-alkanes for up to 7 days and relatively high levels of resilient compounds such as o-xylene, p-xylene and 1,3,5-trimethylbenzene. These results demonstrate, however, that prompt preservation and/or analysis of soil evidence is required in order to properly classify an ignitable liquid residue. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. Functionality of microbial communities in constructed wetlands used for pesticide remediation: Influence of system design and sampling strategy.

    PubMed

    Lv, Tao; Carvalho, Pedro N; Zhang, Liang; Zhang, Yang; Button, Mark; Arias, Carlos A; Weber, Kela P; Brix, Hans

    2017-03-01

    The objective of this study was to compare the microbial community metabolic function from both unsaturated and saturated constructed wetland mesocosms (CWs) when treating the pesticide tebuconazole. The comparison was performed for both interstitial water and substrate biofilm by community level physiological profiling (CLPP) via BIOLOG™ EcoPlates. For each CW design (saturated or unsaturated), six mesocosms were established including one unplanted and five planted individually with either Juncus effusus, Typha latifolia, Berula erecta, Phragmites australis or Iris pseudacorus. Microbial activity and metabolic richness of interstitial water from unsaturated CWs were significantly lower than that from saturated CWs. However, in general, the opposite result was observed for biofilm samples. Wetland plants promoted significantly higher biofilm microbial activity and metabolic richness than unplanted CWs in both CW designs. Differences in the microbial community functional profiles between plant species were only found for saturated CWs. Biofilm microbial metabolic richness was generally statistically higher than that of interstitial water in both unsaturated (1.4-24 times higher) and saturated (1.2-1.7 times higher) CWs. Carbon source (guild) utilization patterns were generally different between interstitial water and biofilm samples. Functionality of the biofilm microbial community was positively correlated to the removal of all pollutants (TN, NH4(+)-N, TP, TOC and tebuconazole) for both unsaturated and saturated CWs, suggesting the biofilm plays a more important role in pollutant removal than the interstitial water microbial community. Thus, merely observing the interstitial water microbial communities may underestimate the role of the microbial community in CW performance. Interestingly, the ability for the biofilm microbial community to utilize amino acids and amines/amides was positively correlated with tebuconazole removal in all system types. Copyright

  20. Analysis of ethanol in fermentation samples by a robust nanocomposite-based microbial biosensor.

    PubMed

    Sefčovičová, Jana; Filip, Jaroslav; Mastihuba, Vladimír; Gemeiner, Peter; Tkac, Jan

    2012-06-01

    A robust microbial biosensor was constructed from a bionanocomposite prepared by a direct mixing of bacterial cells of Gluconobacter oxydans and carbon nanotubes with ferricyanide employed as a mediator for enhanced sensitivity of ethanol oxidation. A successful integration of the device into flow injection analysis mode of operation provided a high sensitivity of detection of (74 ± 2.7) μA mM(-1) cm(-2), a low detection limit of 5 μM and a linear range from 10 μM up to 1 mM. A short response time of the biosensor allowed a sample throughput of 67 h(-1) at 0.3 ml min(-1). The biosensor exhibited high operational stability with a decrease in the biosensor response of 1.7% during 43 h of continuous operation. The device was used to analyse ethanol in fermentation samples with a good agreement with a HPLC method.

  1. Treatability studies on different refinery wastewater samples using high-throughput microbial electrolysis cells (MECs).

    PubMed

    Ren, Lijiao; Siegert, Michael; Ivanov, Ivan; Pisciotta, John M; Logan, Bruce E

    2013-05-01

    High-throughput microbial electrolysis cells (MECs) were used to perform treatability studies on many different refinery wastewater samples all having appreciably different characteristics, which resulted in large differences in current generation. A de-oiled refinery wastewater sample from one site (DOW1) produced the best results, with 2.1±0.2 A/m(2) (maximum current density), 79% chemical oxygen demand removal, and 82% headspace biological oxygen demand removal. These results were similar to those obtained using domestic wastewater. Two other de-oiled refinery wastewater samples also showed good performance, with a de-oiled oily sewer sample producing less current. A stabilization lagoon sample and a stripped sour wastewater sample failed to produce appreciable current. Electricity production, organics removal, and startup time were improved when the anode was first acclimated to domestic wastewater. These results show mini-MECs are an effective method for evaluating treatability of different wastewaters. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. High-throughput automated microfluidic sample preparation for accurate microbial genomics

    PubMed Central

    Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B.; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P.; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C.

    2017-01-01

    Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications. PMID:28128213

  3. Microbial counts and particulate matter levels in roadside air samples under skytrain stations, Bangkok, Thailand.

    PubMed

    Luksamijarulkul, Pipat; Kongtip, Pornpimol

    2010-05-01

    In conditions with heavy traffic and crowds of people on roadside areas under skytrain stations in Bangkok, the natural air ventilation may be insufficient and air quality may be poor. A study of 350 air samples collected from the roadside, under skytrain stations in Bangkok, was carried out to assess microbial counts (210 air samples) and particulate matter (PM10) levels (140 samples). The results reveal the mean +/- standard deviation bacterial counts and fungal counts were 406.8 +/- 302.7 cfu/m3 and 128.9 +/- 89.7 cfu/m3, respectively. The PM10 level was 186.1 +/- 188.1 microg/m3. When compared to recommended levels, 4.8% of air samples (10/210 samples) had bacterial counts more than recommended levels (> 1,000 cfu/ m3) and 27.1% (38/140 samples) had PM10 levels more than recommended levels (> 120 microg/m3). These may affect human health, especially of street venders who spend most of their working time in these areas.

  4. High-throughput automated microfluidic sample preparation for accurate microbial genomics.

    PubMed

    Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C

    2017-01-27

    Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications.

  5. Estimating Virus Production Rates in Aquatic Systems

    PubMed Central

    Matteson, Audrey R.; Budinoff, Charles R.; Campbell, Claire E.; Buchan, Alison; Wilhelm, Steven W.

    2010-01-01

    Viruses are pervasive components of marine and freshwater systems, and are known to be significant agents of microbial mortality. Developing quantitative estimates of this process is critical as we can then develop better models of microbial community structure and function as well as advance our understanding of how viruses work to alter aquatic biogeochemical cycles. The virus reduction technique allows researchers to estimate the rate at which virus particles are released from the endemic microbial community. In brief, the abundance of free (extracellular) viruses is reduced in a sample while the microbial community is maintained at near ambient concentration. The microbial community is then incubated in the absence of free viruses and the rate at which viruses reoccur in the sample (through the lysis of already infected members of the community) can be quantified by epifluorescence microscopy or, in the case of specific viruses, quantitative PCR. These rates can then be used to estimate the rate of microbial mortality due to virus-mediated cell lysis. PMID:20972392

  6. Microfluidic-based mini-metagenomics enables discovery of novel microbial lineages from complex environmental samples.

    PubMed

    Yu, Feiqiao Brian; Blainey, Paul C; Schulz, Frederik; Woyke, Tanja; Horowitz, Mark A; Quake, Stephen R

    2017-07-05

    Metagenomics and single-cell genomics have enabled genome discovery from unknown branches of life. However, extracting novel genomes from complex mixtures of metagenomic data can still be challenging and represents an ill-posed problem which is generally approached with ad hoc methods. Here we present a microfluidic-based mini-metagenomic method which offers a statistically rigorous approach to extract novel microbial genomes while preserving single-cell resolution. We used this approach to analyze two hot spring samples from Yellowstone National Park and extracted 29 new genomes, including three deeply branching lineages. The single-cell resolution enabled accurate quantification of genome function and abundance, down to 1% in relative abundance. Our analyses of genome level SNP distributions also revealed low to moderate environmental selection. The scale, resolution, and statistical power of microfluidic-based mini-metagenomics make it a powerful tool to dissect the genomic structure of microbial communities while effectively preserving the fundamental unit of biology, the single cell.

  7. Single sample resolution of rare microbial dark matter in a marine invertebrate metagenome

    PubMed Central

    Miller, Ian J.; Weyna, Theodore R.; Fong, Stephen S.; Lim-Fong, Grace E.; Kwan, Jason C.

    2016-01-01

    Direct, untargeted sequencing of environmental samples (metagenomics) and de novo genome assembly enable the study of uncultured and phylogenetically divergent organisms. However, separating individual genomes from a mixed community has often relied on the differential-coverage analysis of multiple, deeply sequenced samples. In the metagenomic investigation of the marine bryozoan Bugula neritina, we uncovered seven bacterial genomes associated with a single B. neritina individual that appeared to be transient associates, two of which were unique to one individual and undetectable using certain “universal” 16S rRNA primers and probes. We recovered high quality genome assemblies for several rare instances of “microbial dark matter,” or phylogenetically divergent bacteria lacking genomes in reference databases, from a single tissue sample that was not subjected to any physical or chemical pre-treatment. One of these rare, divergent organisms has a small (593 kbp), poorly annotated genome with low GC content (20.9%) and a 16S rRNA gene with just 65% sequence similarity to the closest reference sequence. Our findings illustrate the importance of sampling strategy and de novo assembly of metagenomic reads to understand the extent and function of bacterial biodiversity. PMID:27681823

  8. Single sample resolution of rare microbial dark matter in a marine invertebrate metagenome

    DOE PAGES

    Miller, Ian J.; Weyna, Theodore R.; Fong, Stephen S.; ...

    2016-09-29

    Direct, untargeted sequencing of environmental samples (metagenomics) and de novo genome assembly enable the study of uncultured and phylogenetically divergent organisms. However, separating individual genomes from a mixed community has often relied on the differential-coverage analysis of multiple, deeply sequenced samples. In the metagenomic investigation of the marine bryozoan Bugula neritina, we uncovered seven bacterial genomes associated with a single B. neritina individual that appeared to be transient associates, two of which were unique to one individual and undetectable using certain “universal” 16S rRNA primers and probes. We recovered high quality genome assemblies for several rare instances of “microbial darkmore » matter,” or phylogenetically divergent bacteria lacking genomes in reference databases, from a single tissue sample that was not subjected to any physical or chemical pre-treatment. One of these rare, divergent organisms has a small (593 kbp), poorly annotated genome with low GC content (20.9%) and a 16S rRNA gene with just 65% sequence similarity to the closest reference sequence. Lastly, our findings illustrate the importance of sampling strategy and de novo assembly of metagenomic reads to understand the extent and function of bacterial biodiversity.« less

  9. Single sample resolution of rare microbial dark matter in a marine invertebrate metagenome

    SciTech Connect

    Miller, Ian J.; Weyna, Theodore R.; Fong, Stephen S.; Lim-Fong, Grace E.; Kwan, Jason C.

    2016-09-29

    Direct, untargeted sequencing of environmental samples (metagenomics) and de novo genome assembly enable the study of uncultured and phylogenetically divergent organisms. However, separating individual genomes from a mixed community has often relied on the differential-coverage analysis of multiple, deeply sequenced samples. In the metagenomic investigation of the marine bryozoan Bugula neritina, we uncovered seven bacterial genomes associated with a single B. neritina individual that appeared to be transient associates, two of which were unique to one individual and undetectable using certain “universal” 16S rRNA primers and probes. We recovered high quality genome assemblies for several rare instances of “microbial dark matter,” or phylogenetically divergent bacteria lacking genomes in reference databases, from a single tissue sample that was not subjected to any physical or chemical pre-treatment. One of these rare, divergent organisms has a small (593 kbp), poorly annotated genome with low GC content (20.9%) and a 16S rRNA gene with just 65% sequence similarity to the closest reference sequence. Lastly, our findings illustrate the importance of sampling strategy and de novo assembly of metagenomic reads to understand the extent and function of bacterial biodiversity.

  10. Automated Three-Dimensional Microbial Sensing and Recognition Using Digital Holography and Statistical Sampling

    PubMed Central

    Moon, Inkyu; Yi, Faliu; Javidi, Bahram

    2010-01-01

    We overview an approach to providing automated three-dimensional (3D) sensing and recognition of biological micro/nanoorganisms integrating Gabor digital holographic microscopy and statistical sampling methods. For 3D data acquisition of biological specimens, a coherent beam propagates through the specimen and its transversely and longitudinally magnified diffraction pattern observed by the microscope objective is optically recorded with an image sensor array interfaced with a computer. 3D visualization of the biological specimen from the magnified diffraction pattern is accomplished by using the computational Fresnel propagation algorithm. For 3D recognition of the biological specimen, a watershed image segmentation algorithm is applied to automatically remove the unnecessary background parts in the reconstructed holographic image. Statistical estimation and inference algorithms are developed to the automatically segmented holographic image. Overviews of preliminary experimental results illustrate how the holographic image reconstructed from the Gabor digital hologram of biological specimen contains important information for microbial recognition. PMID:22163664

  11. Automated three-dimensional microbial sensing and recognition using digital holography and statistical sampling.

    PubMed

    Moon, Inkyu; Yi, Faliu; Javidi, Bahram

    2010-01-01

    We overview an approach to providing automated three-dimensional (3D) sensing and recognition of biological micro/nanoorganisms integrating Gabor digital holographic microscopy and statistical sampling methods. For 3D data acquisition of biological specimens, a coherent beam propagates through the specimen and its transversely and longitudinally magnified diffraction pattern observed by the microscope objective is optically recorded with an image sensor array interfaced with a computer. 3D visualization of the biological specimen from the magnified diffraction pattern is accomplished by using the computational Fresnel propagation algorithm. For 3D recognition of the biological specimen, a watershed image segmentation algorithm is applied to automatically remove the unnecessary background parts in the reconstructed holographic image. Statistical estimation and inference algorithms are developed to the automatically segmented holographic image. Overviews of preliminary experimental results illustrate how the holographic image reconstructed from the Gabor digital hologram of biological specimen contains important information for microbial recognition.

  12. Microbial quality of bagged baby spinach and romaine lettuce: effects of top versus bottom sampling.

    PubMed

    Kase, Julie A; Borenstein, Stacey; Blodgett, Robert J; Feng, Peter C H

    2012-01-01

    Contamination with Escherichia coli O157:H7 and Salmonella have called into question the safety and microbial quality of bagged ready-to-eat leafy greens. This study expands on previous findings that these goods have high total bacteria counts (TBC) and coliform counts, variation in counts among different lots, that Escherichia coli is present, and disparities in counts when bags are top or bottom sampled. Nearly 100 bags of baby spinach and hearts of romaine lettuce from a single brand were subjected to both top and bottom sampling. Product was blended, and a portion serially diluted and plated to obtain TBC. Total coliform and E. coli levels were estimated by the most-probable-number (MPN) technique with ColiComplete discs. Top-sampled TBC from bags of baby spinach (48 bags, 13 different lots) ranged from 3.9 to 8.1 log CFU/g and bottom-sampled TBC ranged from 4.0 to 8.2 log CFU/g, with 52% of the bags (or 39% of the lots) producing TBC higher in bottom samples. For hearts of romaine (47 bags from 19 different lots), top-sampled bags had TBC ranging from 2.4 to 7.0 log, and bottom-sampled bags had TBC from 3.3 to 7.3 log, with 64% of the bags (or 63% of the lots) showing higher TBC in bottom samples. However, we are unable to reject the hypothesis that the top and bottom samples from either commodity contain the same TBC (P ≥ 0.08). No E. coli was detected and total coliform bacteria counts were, with few exceptions, ≥210 MPN/g, irrespective of TBC. In general, lots with the most number of days before the printed "use-by" date had lower TBC. However, the R(2) values for either baby spinach (0.4085) or hearts of romaine (0.2946) suggest that age might not be a very good predictor of higher TBC. TBC varied widely between lots and even more so within same-lot samples, as indicated by the sum of squares results. This finding, along with higher TBC in bottom samples, suggests further consideration when a microbiological sampling scheme of bagged produce is

  13. Determination of ATP-activity as a useful tool for monitoring microbial load in aqueous humidifier samples.

    PubMed

    Liebers, Verena; Bachmann, Dieter; Franke, Gabriele; Freundt, Susanne; Stubel, Heike; Düser, Maria; Kendzia, Benjamin; Böckler, Margret; Brüning, Thomas; Raulf, Monika

    2015-03-01

    Air humidifier water tanks are potential sources of microbial contaminants. Aerosolization of these contaminants is associated with the development of airway and lung diseases; therefore, implementation of preventive strategies including monitoring of the microbial contamination is recommended. So far, culture-based methods that include measuring colony forming units (CFU) are widely used to monitor microbial load. However, these methods are time consuming and have considerable drawbacks. As a result, alternative methods are needed which provide not only clear and accurate results concerning microbial load in water samples, but are also rapid and easy to use in the field. This paper reports on a rapid test for ATP quantification as an alternative method for microbial monitoring, including its implementation, validation and application in the field. For this purpose, 186 water samples were characterized with different methods, which included ATP analysis, culture-based methods, endotoxin activity (common and rapid test), pyrogenic activity and number of particles. Half of the samples was measured directly in the field and the other half one day later in the laboratory. The results of both tests are highly correlated. Furthermore, to check how representative the result from one sample of a water source is, a second sample of the same water tank were collected and measured. Bioluminescence results of the undiluted samples covered a range between 20 and 25,000 relative light units (RLU) and correlated with the results obtained using the other methods. The highest correlation was found between bioluminescence and endotoxin activity (rs=0.79) as well as pyrogenic activity (rs=0.75). Overall, the results of this study indicate that ATP measurement using bioluminescence is a suitable tool to obtain rapid, reproducible and sensitive information on the microbial load of water samples, and is suitable to use in the field. However, to use ATP measurement as an indicator of

  14. Rapid comparison and correlation analysis among massive number of microbial community samples based on MDV data model

    PubMed Central

    Su, Xiaoquan; Hu, Jianqiang; Huang, Shi; Ning, Kang

    2014-01-01

    The research in microbial communities would potentially impact a vast number of applications in “bio”-related disciplines. Large-scale analyses became a clear trend in microbial community studies, thus it is increasingly important to perform efficient and in-depth data mining for insightful biological principles from large number of samples. However, as microbial communities are from different sources and of different structures, comparison and data-mining from large number of samples become quite difficult. In this work, we have proposed a data model to represent large-scale comparison of microbial community samples, namely the “Multi-Dimensional View” data model (the MDV model) that should at least include 3 aspects: samples profile (S), taxa profile (T) and meta-data profile (V). We have also proposed a method for rapid data analysis based on the MDV model and applied it on the case studies with samples from various environmental conditions. Results have shown that though sampling environments usually define key variables, the analysis could detect bio-makers and even subtle variables based on large number of samples, which might be used to discover novel principles that drive the development of communities. The efficiency and effectiveness of data analysis method based on the MDV model have been validated by the results. PMID:25227622

  15. Rapid comparison and correlation analysis among massive number of microbial community samples based on MDV data model

    NASA Astrophysics Data System (ADS)

    Su, Xiaoquan; Hu, Jianqiang; Huang, Shi; Ning, Kang

    2014-09-01

    The research in microbial communities would potentially impact a vast number of applications in ``bio''-related disciplines. Large-scale analyses became a clear trend in microbial community studies, thus it is increasingly important to perform efficient and in-depth data mining for insightful biological principles from large number of samples. However, as microbial communities are from different sources and of different structures, comparison and data-mining from large number of samples become quite difficult. In this work, we have proposed a data model to represent large-scale comparison of microbial community samples, namely the ``Multi-Dimensional View'' data model (the MDV model) that should at least include 3 aspects: samples profile (S), taxa profile (T) and meta-data profile (V). We have also proposed a method for rapid data analysis based on the MDV model and applied it on the case studies with samples from various environmental conditions. Results have shown that though sampling environments usually define key variables, the analysis could detect bio-makers and even subtle variables based on large number of samples, which might be used to discover novel principles that drive the development of communities. The efficiency and effectiveness of data analysis method based on the MDV model have been validated by the results.

  16. Impact of temperature and time storage on the microbial detection of oral samples by Checkerboard DNA-DNA hybridization method.

    PubMed

    do Nascimento, Cássio; dos Santos, Janine Navarro; Pedrazzi, Vinícius; Pita, Murillo Sucena; Monesi, Nadia; Ribeiro, Ricardo Faria; de Albuquerque, Rubens Ferreira

    2014-01-01

    Molecular diagnosis methods have been largely used in epidemiological or clinical studies to detect and quantify microbial species that may colonize the oral cavity in healthy or disease. The preservation of genetic material from samples remains the major challenge to ensure the feasibility of these methodologies. Long-term storage may compromise the final result. The aim of this study was to evaluate the effect of temperature and time storage on the microbial detection of oral samples by Checkerboard DNA-DNA hybridization. Saliva and supragingival biofilm were taken from 10 healthy subjects, aliquoted (n=364) and processed according to proposed protocols: immediate processing and processed after 2 or 4 weeks, and 6 or 12 months of storage at 4°C, -20°C and -80°C. Either total or individual microbial counts were recorded in lower values for samples processed after 12 months of storage, irrespective of temperatures tested. Samples stored up to 6 months at cold temperatures showed similar counts to those immediately processed. The microbial incidence was also significantly reduced in samples stored during 12 months in all temperatures. Temperature and time of oral samples storage have relevant impact in the detection and quantification of bacterial and fungal species by Checkerboard DNA-DNA hybridization method. Samples should be processed immediately after collection or up to 6 months if conserved at cold temperatures to avoid false-negative results. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Rapid comparison and correlation analysis among massive number of microbial community samples based on MDV data model.

    PubMed

    Su, Xiaoquan; Hu, Jianqiang; Huang, Shi; Ning, Kang

    2014-09-17

    The research in microbial communities would potentially impact a vast number of applications in "bio"-related disciplines. Large-scale analyses became a clear trend in microbial community studies, thus it is increasingly important to perform efficient and in-depth data mining for insightful biological principles from large number of samples. However, as microbial communities are from different sources and of different structures, comparison and data-mining from large number of samples become quite difficult. In this work, we have proposed a data model to represent large-scale comparison of microbial community samples, namely the "Multi-Dimensional View" data model (the MDV model) that should at least include 3 aspects: samples profile (S), taxa profile (T) and meta-data profile (V). We have also proposed a method for rapid data analysis based on the MDV model and applied it on the case studies with samples from various environmental conditions. Results have shown that though sampling environments usually define key variables, the analysis could detect bio-makers and even subtle variables based on large number of samples, which might be used to discover novel principles that drive the development of communities. The efficiency and effectiveness of data analysis method based on the MDV model have been validated by the results.

  18. Screening of new bioactive materials from microbial extracts of soil microorganism (I). Antimicrobial activity from 200 samples using microdilution assay.

    PubMed

    Jung, S O; Kim, J; Chang, I M; Ryu, J C

    1998-06-01

    The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screening of antibiotic susceptibility than those previously reported.

  19. Simple absolute quantification method correcting for quantitative PCR efficiency variations for microbial community samples.

    PubMed

    Brankatschk, Robert; Bodenhausen, Natacha; Zeyer, Josef; Bürgmann, Helmut

    2012-06-01

    Real-time quantitative PCR (qPCR) is a widely used technique in microbial community analysis, allowing the quantification of the number of target genes in a community sample. Currently, the standard-curve (SC) method of absolute quantification is widely employed for these kinds of analysis. However, the SC method assumes that the amplification efficiency (E) is the same for both the standard and the sample target template. We analyzed 19 bacterial strains and nine environmental samples in qPCR assays, targeting the nifH and 16S rRNA genes. The E values of the qPCRs differed significantly, depending on the template. This has major implications for the quantification. If the sample and standard differ in their E values, quantification errors of up to orders of magnitude are possible. To address this problem, we propose and test the one-point calibration (OPC) method for absolute quantification. The OPC method corrects for differences in E and was derived from the ΔΔC(T) method with correction for E, which is commonly used for relative quantification in gene expression studies. The SC and OPC methods were compared by quantifying artificial template mixtures from Geobacter sulfurreducens (DSM 12127) and Nostoc commune (Culture Collection of Algae and Protozoa [CCAP] 1453/33), which differ in their E values. While the SC method deviated from the expected nifH gene copy number by 3- to 5-fold, the OPC method quantified the template mixtures with high accuracy. Moreover, analyzing environmental samples, we show that even small differences in E between the standard and the sample can cause significant differences between the copy numbers calculated by the SC and the OPC methods.

  20. Detection of Microbial Life in Glacial Samples - Laboratories Studies and Development for Field use

    NASA Astrophysics Data System (ADS)

    Barnett, M. J.; Cullen, D. C.; Telling, J.; Wadham, J. L.; Holt, J.; Sims, M.

    2007-12-01

    Adenosine triphosphate (ATP) is frequently used as a proxy for bulk microbial biomass in environmental sciences and, in the food and health industries. Despite successful ATP detection in a variety of ecosystems, very little data are available on ATP levels in the glacial system. In this study, protocols for ATP detection on glacial ice and sediment samples are investigated, in order to aid in the development of a single-use device for in-field life detection, and also to increase the available data on biomass estimates in the cryosphere. ATP detection in two glacial samples reveals concentrations indistinguishable from internal blanks. Therefore, the samples were centrifuged and their particulate loads were subjected to four different extraction processes. Applying these extraction methods resulted in higher ATP concentration than samples with no extraction process; the different techniques increase the ATP detected between 5 and 15 times (also relative to an internal standard). Concurrent with the laboratory based development of extraction protocols is the development of a single-use device for the detection of ATP at the sampling site, in icy environments. The device is microfluidic-based, using commercially available reagents for the detection of ATP by bioluminescence. In order to produce a robust measure of biomass, both laboratory and field based analyses need to be carried out. This work shows the potential of ATP detection in glacial samples and the early development of a device for in situ life detection. The quantification of ATP in microfluidic format is being developed as the preliminary target for an integrated life detection and characterisation device.

  1. Design and Implementation of an Automated Illuminating, Culturing, and Sampling System for Microbial Optogenetic Applications.

    PubMed

    Stewart, Cameron J; McClean, Megan N

    2017-02-19

    Optogenetic systems utilize genetically-encoded proteins that change conformation in response to specific wavelengths of light to alter cellular processes. There is a need for culturing and measuring systems that incorporate programmed illumination and stimulation of optogenetic systems. We present a protocol for building and using a continuous culturing apparatus to illuminate microbial cells with programmed doses of light, and automatically acquire and analyze images of cells in the effluent. The operation of this apparatus as a chemostat allows the growth rate and the cellular environment to be tightly controlled. The effluent of the continuous cell culture is regularly sampled and the cells are imaged by multi-channel microscopy. The culturing, sampling, imaging, and image analysis are fully automated so that dynamic responses in the fluorescence intensity and cellular morphology of cells sampled from the culture effluent are measured over multiple days without user input. We demonstrate the utility of this culturing apparatus by dynamically inducing protein production in a strain of Saccharomyces cerevisiae engineered with an optogenetic system that activates transcription.

  2. Potential microbial contamination during sampling of permafrost soil assessed by tracers.

    PubMed

    Bang-Andreasen, Toke; Schostag, Morten; Priemé, Anders; Elberling, Bo; Jacobsen, Carsten S

    2017-02-23

    Drilling and handling of permanently frozen soil cores without microbial contamination is of concern because contamination e.g. from the active layer above may lead to incorrect interpretation of results in experiments investigating potential and actual microbial activity in these low microbial biomass environments. Here, we present an example of how microbial contamination from active layer soil affected analysis of the potentially active microbial community in permafrost soil. We also present the development and use of two tracers: (1) fluorescent plastic microspheres and (2) Pseudomonas putida genetically tagged with Green Fluorescent Protein production to mimic potential microbial contamination of two permafrost cores. A protocol with special emphasis on avoiding microbial contamination was developed and employed to examine how far microbial contamination can penetrate into permafrost cores. The quantity of tracer elements decreased with depth into the permafrost cores, but the tracers were detected as far as 17 mm from the surface of the cores. The results emphasize that caution should be taken to avoid microbial contamination of permafrost cores and that the application of tracers represents a useful tool to assess penetration of potential microbial contamination into permafrost cores.

  3. Potential microbial contamination during sampling of permafrost soil assessed by tracers

    PubMed Central

    Bang-Andreasen, Toke; Schostag, Morten; Priemé, Anders; Elberling, Bo; Jacobsen, Carsten S.

    2017-01-01

    Drilling and handling of permanently frozen soil cores without microbial contamination is of concern because contamination e.g. from the active layer above may lead to incorrect interpretation of results in experiments investigating potential and actual microbial activity in these low microbial biomass environments. Here, we present an example of how microbial contamination from active layer soil affected analysis of the potentially active microbial community in permafrost soil. We also present the development and use of two tracers: (1) fluorescent plastic microspheres and (2) Pseudomonas putida genetically tagged with Green Fluorescent Protein production to mimic potential microbial contamination of two permafrost cores. A protocol with special emphasis on avoiding microbial contamination was developed and employed to examine how far microbial contamination can penetrate into permafrost cores. The quantity of tracer elements decreased with depth into the permafrost cores, but the tracers were detected as far as 17 mm from the surface of the cores. The results emphasize that caution should be taken to avoid microbial contamination of permafrost cores and that the application of tracers represents a useful tool to assess penetration of potential microbial contamination into permafrost cores. PMID:28230151

  4. Potential microbial contamination during sampling of permafrost soil assessed by tracers

    NASA Astrophysics Data System (ADS)

    Bang-Andreasen, Toke; Schostag, Morten; Priemé, Anders; Elberling, Bo; Jacobsen, Carsten S.

    2017-02-01

    Drilling and handling of permanently frozen soil cores without microbial contamination is of concern because contamination e.g. from the active layer above may lead to incorrect interpretation of results in experiments investigating potential and actual microbial activity in these low microbial biomass environments. Here, we present an example of how microbial contamination from active layer soil affected analysis of the potentially active microbial community in permafrost soil. We also present the development and use of two tracers: (1) fluorescent plastic microspheres and (2) Pseudomonas putida genetically tagged with Green Fluorescent Protein production to mimic potential microbial contamination of two permafrost cores. A protocol with special emphasis on avoiding microbial contamination was developed and employed to examine how far microbial contamination can penetrate into permafrost cores. The quantity of tracer elements decreased with depth into the permafrost cores, but the tracers were detected as far as 17 mm from the surface of the cores. The results emphasize that caution should be taken to avoid microbial contamination of permafrost cores and that the application of tracers represents a useful tool to assess penetration of potential microbial contamination into permafrost cores.

  5. A mud-sampling technique for the study of the ecology of aquatic snails, and its use in the evaluation of the efficacy of molluscicides in field trials*

    PubMed Central

    Crossland, N. O.

    1962-01-01

    The author describes a sampling technique suitable for estimating the population densities of African aquatic snails in the field by the removal of a plug of mud and its subsequent detailed examination. This device has been used in field trials with the molluscicides ICI 24223 and Bayer 73. Estimates of percentage kills were based on counts of live and dead snails in each mud sample before and after molluscicide treatment. Statistical analysis of these counts suggests that most snails killed by the molluscicides were subsequently recovered, thus providing some confirmatory evidence of the validity of these estimates. PMID:13882393

  6. Sampling in the Middle Adriatic Sea - Relations Between Mercury and Microbial Species

    NASA Astrophysics Data System (ADS)

    Kotnik, J.; Zivkovic, I.; Horvat, M.; Solic, M.; Fajon, V.

    2016-02-01

    Chemical and biological processes can transform mercury species. These (bio)chemical reactions are of great importance because they facilitate mercury's entrance into the marine food web where it readily bioaccumulates in the form of methylmercury (MeHg). Microorganisms might represent a critical Hg bioaccumulation step in the entire marine food web. In order to establish relationship between mercury and microbial species, we performed series of samplings. Samplings were conducted during oceanographic cruises aboard the Croatian research vessel Bios Dva from March 2014 to June 2015 in the Middle Adriatic Sea. Our research is constrained in transect from the Bay of Kaštela to the island of Vis. Water profiles of total mercury (THg), dissolved gaseous mercury (DGM), methylmercury (MeHg)1,2 and picoeukaryotes3 in non-filtered seawater samples were created for both coastal and open waters. THg concentrations range from 0.14-1.10 ng/L near the island of Vis, which represents pristine marine environment. The highest values for THg (0.92-5.58 ng/L) are found in the Bay of Kaštela that had been affected by previous contamination from chlor-alkali industrial waste waters. DGM always shows lower values in the pristine environment (22.1-245 pg/L) than in the contaminated area (31.8-351 pg/L). At the Split Channel stations, which is located between pristine and contaminated environment, values for DGM concentrations are in between 23.0-374 pg/L. MeHg concentrations vary, but range from 1.07-34.3 pg/L for all stations, with the highest values found in the Bay of Kaštela. Number of picoeukaryotes is the highest in the central part of the Bay of Kaštela (0.44×103-31.8×103/mL) while the lowest number (0.63×103-19.9×103/mL) is found at the island of Vis. All our results will be presented herein. 1. Horvat, M. et al. Speciation of mercury in surface and deep-sea waters in the Mediterranean Sea. Atmos. Environ. 37, S93-S108 (2003). 2. Kotnik, J. et al. Mercury speciation in

  7. Supercritical fluid extraction and ultra performance liquid chromatography of respiratory quinones for microbial community analysis in environmental and biological samples.

    PubMed

    Hanif, Muhammad; Atsuta, Yoichi; Fujie, Koichi; Daimon, Hiroyuki

    2012-03-05

    Microbial community structure plays a significant role in environmental assessment and animal health management. The development of a superior analytical strategy for the characterization of microbial community structure is an ongoing challenge. In this study, we developed an effective supercritical fluid extraction (SFE) and ultra performance liquid chromatography (UPLC) method for the analysis of bacterial respiratory quinones (RQ) in environmental and biological samples. RQ profile analysis is one of the most widely used culture-independent tools for characterizing microbial community structure. A UPLC equipped with a photo diode array (PDA) detector was successfully applied to the simultaneous determination of ubiquinones (UQ) and menaquinones (MK) without tedious pretreatment. Supercritical carbon dioxide (scCO(2)) extraction with the solid-phase cartridge trap proved to be a more effective and rapid method for extracting respiratory quinones, compared to a conventional organic solvent extraction method. This methodology leads to a successful analytical procedure that involves a significant reduction in the complexity and sample preparation time. Application of the optimized methodology to characterize microbial communities based on the RQ profile was demonstrated for a variety of environmental samples (activated sludge, digested sludge, and compost) and biological samples (swine and Japanese quail feces).

  8. Trace fossils of microbial colonization on Mars: Criteria for search and for sample return

    NASA Technical Reports Server (NTRS)

    Friedmann, E. I.

    1988-01-01

    The recent discovery of microbial trace-fossil formation in the frigid Ross Desert of Antarctica suggests that early primitive life on Mars may have left behind similar signatures. These trace fossils are apparent as chemical or physical changes in rock (or sediment) structure (or chemistry) caused by the activity of organisms. Life on Mars, if it ever existed, almost certainly did not evolve above the level of microorganisms, and this should be considered in search for fossil life. For the reasons detailed here, microbial trace fossils seem to be a better and more realistic target for search than would be true microbial fossils (remnants of cellular structures).

  9. Toxicity test using medaka (Oryzias latipes) early fry and concentrated sample water as an index of aquatic habitat condition.

    PubMed

    Yamashita, H; Haribowo, R; Sekine, M; Oda, N; Kanno, A; Shimono, Y; Shitao, W; Higuchi, T; Imai, T; Yamamoto, K

    2011-08-01

    The aim of the present study was to show a relationship between toxicity of 100-fold concentrated water and aquatic habitat conditions. Environmental waters are 100-fold concentrated with solid-phase extraction. Medaka early fry was exposed in these waters for 48 h. The number of death and disorder was counted at 1, 2, 3, 6, 12, 24, and 48 h; toxicity was expressed using inverse median effect time and median lethal time (ET (50)(-1), LT (50)(-1)). Average score per taxon (ASPT) for benthic animals and Index of Biotic Integrity (IBI) for fish were applied as indices of aquatic habitat conditions. The results of toxicity test were compared using ASPT and IBI. The different levels of toxicity were detected in the seawater of Japan. At the Husino River area, toxicity cannot be detected. In rivers, high toxicity appeared at urban districts without sewerage. By Spearman coefficient, the relationship between toxicity and high biochemical oxygen demand (BOD) were obtained. BOD household wastewater contains hydrophobic toxic matters; otherwise, seawater in industrial area does not show clear relationship between toxicity and chemical oxygen demand. Gas chromatography to mass spectrometry simultaneous analysis database may give an answer for the source of toxicity, but further test is required. Ratio of clear stream benthic animal sharply decreased over 0.25 of LT (50)(-1) or 0.5 of ET (50)(-1). Tolerant fish becomes dominant over 0.3 of LT (50)(-1) or 0.5-1.0 of ET (50)(-1). By Pearson product-moment correlation coefficient, correlation coefficient between toxicity and ASPT was obtained at -0.773 (ET (50)(-1)) and -0.742 (LT (50)(-1)) at 1 % level of significance with a high negative correlation. Toxicity (LT (50)(-1) ) has strong correlation with the ratio of tolerant species. By Pearson product-moment correlation coefficient, correlation coefficient between toxicity and IBI obtained were -0.155 (ET (50)(-1)) and -0.190 (LT (50)(-1)) at 1 % level of significance and has a

  10. Habitability Conditions Constrained by Martian Meteorites: Implications for Microbial Colonization and Mars Sample Return

    NASA Astrophysics Data System (ADS)

    Shivak, J. N.; Banerjee, N.; Flemming, R. L.

    2013-12-01

    similarities to that of Nakhla. Fractures within Los Angeles and Zagami are fresh and unweathered, and no secondarily deposited phases were found. The environmental conditions suggested by the mineral phases in the Nakhla, Los Angeles, and Zagami meteorites can be used to assess their potential to act as microbial substrates for possible Martian life. Future Mars sample return missions have been proposed to involve the selection and caching of rock samples for return to the Earth. This will require intensive prioritization of samples on the surface and a need to vector towards areas with higher potential for astrobiologically interesting samples. The comparative methodologies developed here with Martian meteorites can be applied to unknown samples recovered from the surface of Mars to aid in mission operations and logistics. [1] J.C. Bridges et al., 2006. Journal of the Geological Society, London 163:229-251. [2] G. Southam et al., 2007. Treatise on Geophysics: Planets and Moons 10:421-438. [3] J.C. Bridges et al., 2001 Space Science Reviews 96: 365-392. [4] I.H. Thorseth et al., 1992. Geochimica et Cosmochimica Acta 56:845-850. [5] H.G. Changela et al., 2011 Meteoritics & Planetary Science 45(12):1847-1867.

  11. Detection of ostreid herpesvirus 1 microvariant DNA in aquatic invertebrate species, sediment and other samples collected from the Georges River estuary, New South Wales, Australia.

    PubMed

    Evans, Olivia; Paul-Pont, Ika; Whittington, Richard J

    2017-01-24

    Ostreid herpesvirus 1 microvariants (OsHV-1) present a serious threat to the Australian Crassostrea gigas industry. Of great concern is the propensity for mortality due to the virus recurring each season in farmed oysters. However, the source of the virus in recurrent outbreaks remains unclear. Reference strain ostreid herpesvirus 1 (OsHV-1 ref) and other related variants have been detected in several aquatic invertebrate species other than C. gigas in Europe, Asia and the USA. The aim of this study was to confirm the presence or absence of OsHV-1 in a range of opportunistically sampled aquatic invertebrate species inhabiting specific locations within the Georges River estuary in New South Wales, Australia. OsHV-1 DNA was detected in samples of wild C. gigas, Saccostrea glomerata, Anadara trapezia, mussels (Mytilus spp., Trichomya hirsuta), whelks (Batillaria australis or Pyrazus ebeninus) and barnacles Balanus spp. collected from several sites between October 2012 and April 2013. Viral loads in non-ostreid species were consistently low, as was the prevalence of OsHV-1 DNA detection. Viral concentrations were highest in wild C. gigas and S. glomerata; the prevalence of detectable OsHV-1 DNA in these oysters reached approximately 68 and 43%, respectively, at least once during the study. These species may be important to the transmission and/or persistence of OsHV-1 in endemically infected Australian estuaries.

  12. Microbial Indicator Profiling of Fresh Produce and Environmental Samples from Farms and Packing Facilities in Northern Mexico.

    PubMed

    Heredia, Norma; Caballero, Cindy; Cárdenas, Carmen; Molina, Karina; García, Rafael; Solís, Luisa; Burrowes, Vanessa; Bartz, Faith E; de Aceituno, Anna Fabiszewski; Jaykus, Lee-Ann; García, Santos; Leon, Juan

    2016-07-01

    To compare microbiological indicator and pathogen contamination among different types of fresh produce and environmental samples along the production chain, 636 samples of produce (rinsates from cantaloupe melons, jalapeño peppers, and tomatoes) and environmental samples (rinsates from hands of workers, soil, and water) were collected at four successive steps in the production process (from the field before harvest through the packing facility) on 11 farms in northern Mexico during 2011 and 2012. Samples were assayed for enteric pathogens (Escherichia coli O157:H7, other Shiga toxigenic E. coli, Salmonella, and Listeria monocytogenes) and microbial indicators (coliforms, other E. coli strains, and Enterococcus spp.). Salmonella was the only pathogen detected; it was found in one preharvest jalapeño sample (detection limits: 0.0033 CFU/ml in produce and hand samples, 0.0013 CFU/ml in water, and 0.04 CFU/g in soil). Microbial indicator profiles for produce, worker hands, and soil from jalapeño and tomato farms were similar, but cantaloupe farm samples had higher indicator levels (P < 0.05 for all comparisons) on fruit (6.5, 2.8, and 7.2 log CFU per fruit) and hands (6.6, 3.1, and 7.1 log CFU per hand) for coliforms, E. coli, and Enterococcus, respectively, and lower E. coli levels in soil (<1 CFU/g). In water from tomato farms, E. coli indicators were significantly more prevalent (70 to 89% of samples were positive; P = 0.01 to 0.02), and geometric mean levels were higher (0.3 to 0.6 log CFU/100 ml) than those in cantaloupe farm water (32 to 38% of samples were positive, geometric mean <1 CFU/100 ml). Microbial indicators were present during all production steps, but prevalence and levels were generally highest at the final on-farm production step (the packing facility) (P < 0.03 for significant comparisons). The finding that microbial contamination on produce farms is influenced by produce type and production step can inform the design of effective approaches to

  13. Microbial diversity in degraded and non-degraded petroleum samples and comparison across oil reservoirs at local and global scales.

    PubMed

    Sierra-Garcia, Isabel Natalia; Dellagnezze, Bruna M; Santos, Viviane P; Chaves B, Michel R; Capilla, Ramsés; Santos Neto, Eugenio V; Gray, Neil; Oliveira, Valeria M

    2017-01-01

    Microorganisms have shown their ability to colonize extreme environments including deep subsurface petroleum reservoirs. Physicochemical parameters may vary greatly among petroleum reservoirs worldwide and so do the microbial communities inhabiting these different environments. The present work aimed at the characterization of the microbiota in biodegraded and non-degraded petroleum samples from three Brazilian reservoirs and the comparison of microbial community diversity across oil reservoirs at local and global scales using 16S rRNA clone libraries. The analysis of 620 16S rRNA bacterial and archaeal sequences obtained from Brazilian oil samples revealed 42 bacterial OTUs and 21 archaeal OTUs. The bacterial community from the degraded oil was more diverse than the non-degraded samples. Non-degraded oil samples were overwhelmingly dominated by gammaproteobacterial sequences with a predominance of the genera Marinobacter and Marinobacterium. Comparisons of microbial diversity among oil reservoirs worldwide suggested an apparent correlation of prokaryotic communities with reservoir temperature and depth and no influence of geographic distance among reservoirs. The detailed analysis of the phylogenetic diversity across reservoirs allowed us to define a core microbiome encompassing three bacterial classes (Gammaproteobacteria, Clostridia, and Bacteroidia) and one archaeal class (Methanomicrobia) ubiquitous in petroleum reservoirs and presumably owning the abilities to sustain life in these environments.

  14. AQUATIC ECOSYSTEMS,

    EPA Science Inventory

    Aquatic ecosystems are a vital part of the urban water cycle (and of urban areas more broadly), and, if healthy, provide a range of goods and services valued by humans (Meyer 1997). For example, aquatic ecosystems (e.g., rivers, lakes, wetlands) provide potable water, food resou...

  15. IV. Aquatics

    Treesearch

    Michael K.. Young

    2011-01-01

    The problem of invasive aquatic species has long been recognized by scientists at the Rocky Mountain Research Station. Fausch and others (2006, 2009) recently overviewed this issue. A point that often distinguishes nonnative aquatic species from nonnatives in other environments is that the presence of some species is frequently prized by managers and the public. For...

  16. AQUATIC ECOSYSTEMS,

    EPA Science Inventory

    Aquatic ecosystems are a vital part of the urban water cycle (and of urban areas more broadly), and, if healthy, provide a range of goods and services valued by humans (Meyer 1997). For example, aquatic ecosystems (e.g., rivers, lakes, wetlands) provide potable water, food resou...

  17. Diversity of Microbial Carbohydrate-Active enZYmes (CAZYmes) Associated with Freshwater and Soil Samples from Caatinga Biome.

    PubMed

    Andrade, Ana Camila; Fróes, Adriana; Lopes, Fabyano Álvares Cardoso; Thompson, Fabiano L; Krüger, Ricardo Henrique; Dinsdale, Elizabeth; Bruce, Thiago

    2017-07-01

    Semi-arid and arid areas occupy about 33% of terrestrial ecosystems. However, little information is available about microbial diversity in the semi-arid Caatinga, which represents a unique biome that extends to about 11% of the Brazilian territory and is home to extraordinary diversity and high endemism level of species. In this study, we characterized the diversity of microbial genes associated with biomass conversion (carbohydrate-active enzymes, or so-called CAZYmes) in soil and freshwater of the Caatinga. Our results showed distinct CAZYme profiles in the soil and freshwater samples. Glycoside hydrolases and glycosyltransferases were the most abundant CAZYme families, with glycoside hydrolases more dominant in soil (∼44%) and glycosyltransferases more abundant in freshwater (∼50%). The abundances of individual glycoside hydrolase, glycosyltransferase, and carbohydrate-binding module subfamilies varied widely between soil and water samples. A predominance of glycoside hydrolases was observed in soil, and a higher contribution of enzymes involved in carbohydrate biosynthesis was observed in freshwater. The main taxa associated with the CAZYme sequences were Planctomycetia (relative abundance in soil, 29%) and Alphaproteobacteria (relative abundance in freshwater, 27%). Approximately 5-7% of CAZYme sequences showed low similarity with sequences deposited in non-redundant databases, suggesting putative homologues. Our findings represent a first attempt to describe specific microbial CAZYme profiles for environmental samples. Characterizing these enzyme groups associated with the conversion of carbohydrates in nature will improve our understanding of the significant roles of enzymes in the carbon cycle. We identified a CAZYme signature that can be used to discriminate between soil and freshwater samples, and this signature may be related to the microbial species adapted to the habitat. The data show the potential ecological roles of the CAZYme repertoire and

  18. Microbial and chemical analysis of illicit drugs samples confiscated from different areas of PakistanMicrobial and chemical analysis of illicit drugs samples confiscated from different areas of Pakistan.

    PubMed

    Hussain, Shahzad; Khattak, Zainab; Mahmood, Sidra; Malik, Farnaz; Riaz, Humayun; Raza, Syed Atif; Khan, Samiullah

    2016-09-01

    The microbial and chemical analysis of illicit drug samples from different areas of Pakistan i.e. Quetta, Karachi, Lahore and Islamabad was conducted in a cross-sectional study at National Institute of Health, Islamabad. The drug samples were confiscated by Anti Narcotics Force (ANF), Pakistan. Microbial analysis was done by estimating bioburden which revealed the presence of gram negative and positive bacteria's, fungus, Streptococcus, Staphylococcus species. Trypton soya agar was used for total aerobic count, MacConkey agar for gram-negative bacteria, Sabouraud dextrose agar for fungus and Vogel-Johnson agar for Streptococcus and Staphylococcus species. Colour tests were applied to identify the drug samples. Qualitative and quantitative analysis of suspected samples of Heroin, morphine, cocaine and acetic anhydride was made by employing different chromatographic techniques i.e. Thin-layer chromatography (TLC) and High-performance liquid chromatography (HPLC). The samples were found to be adulterated with paracetamol, diazepam and Dextromethorphen. Acetic anhydride was adulterated with hydrochloric acid (HCl). There is lack of information providing structured advice on responses to the consequences of illicit drug adulteration. Robust and rehearsed interventions and communication strategies would provide a basis for response for a wide variety of organisations. Research into the usefulness of media warnings about adulteration of illicit drugs is required.

  19. Aquatic dermatoses.

    PubMed

    Ulrich, Heidi; Landthaler, Michael; Vogt, Thomas

    2008-02-01

    Despite the fascination of the aquatic world, it can be associated with some medical risks. Natural bodies of water, swimming pools and aquariums contain many living organisms. Some of these aquatic species may cause traumatic or toxic skin reactions after contact as well as life-threatening systemic reactions. Most frequently, the injuries include poisonous bites and stings by coelenter-ates, worms,stinging fish,mollusks and echinoderms or skin infections. A basic knowledge regarding the flora and fauna that populate the aquatic world can help to avoid contact with such organisms,or if it happens,to promptly initiate appropriate therapy.

  20. Small-Scale DNA Sample Preparation Method for Field PCR Detection of Microbial Cells and Spores in Soil

    PubMed Central

    Kuske, Cheryl R.; Banton, Kaysie L.; Adorada, Dante L.; Stark, Peter C.; Hill, Karen K.; Jackson, Paul J.

    1998-01-01

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent–bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR. PMID:9647816

  1. Microbial diversity in bioaerosol samples causing ODTS compared to reference bioaerosol samples as measured using Illumina sequencing and MALDI-TOF.

    PubMed

    Madsen, Anne Mette; Zervas, Athanasios; Tendal, Kira; Nielsen, Jeppe Lund

    2015-07-01

    The importance of the microbial diversity of bioaerosols in relation to occupational exposure and work related health symptoms is not known. The aim of this paper is to gain knowledge on the bacterial and fungal communities in dust causing organic dust toxic syndrome (ODTS) and in reference dust not causing ODTS. Bacterial and fungal communities were described in personal exposure samples from grass seed workers developing ODTS, in dust generated from grass seeds causing ODTS and in dust generated from reference seeds not causing ODTS. Amplicon sequencing of the bacterial 16S rRNA gene and the fungal ITS region, as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) were used for identification of fungi and bacteria in personal exposure samples and in dust samples from grass seeds causing ODTS and in dust from reference grass seeds. Furthermore, activities of enzymes were measured in the same samples. The sequencing data revealed more than 150 bacterial and 25 fungal genera present in each sample. Streptomyces spp., Aspergillus fumigatus and Rhizopus microsporus were dominating in the dust causing ODTS but not in the reference dust. The dustiness in terms of Mucor sp. and R. microsporus were 100-1000 times higher for problematic seeds compared to reference seeds. The bacterial species in the dust causing ODTS included pathogenic species such as Klebsiella pneumonia and Streptomyces pneumonia, and it contained increased concentrations of total protein, serine protease, chitinase, and β-glucosidase. Twenty-three bacterial genera covered more than 50% of the total reads in the personal and problematic seed dust. These 23 genera accounted for less than 7% of the total reads in the reference seed dust. The microbial community of the dust from the problematic seeds showed great similarities to that from the personal air samples from the workers. In conclusion, we have shown for the first time a shift in the microbial community in aerosol

  2. THE EFFECTS OF DIFFERENT SAMPLE CONCENTRATIONS ON THE STRUCTURE OF MICROBIAL COMMUNITIES USING PHOSPHOLIPID FATTY ACID ANALYSIS

    EPA Science Inventory

    Phospholipid fatty acid (PLFA) analysis is a powerful tool for determination of microbial community structures in soils and sediments. However, accurate determination of total microbial biomass and structure of the microbial community may be dependent on the concentration of the...

  3. Development and Antarctic Testing of a Maneuverable Probe for Clean In-Situ Analysis and Sampling of Subsurface Ice and Subglacial Aquatic Ecosystems

    NASA Astrophysics Data System (ADS)

    Francke, G.; Dachwald, B.; Kowalski, J.; Digel, I.; Tulaczyk, S. M.; Mikucki, J.; Feldmann, M.; Espe, C.; Schöngarth, S.; Hiecker, S.; Blandfort, D.; Schüller, K.; Plescher, E.

    2016-12-01

    There is significant interest in sampling subglacial environments for geochemical and microbiological studies, but those environments are difficult to access. Such environments exist not only on Earth but are also expected beneath the icy crusts of some outer solar system bodies, like the Jovian moon Europa and the Saturnian moon Enceladus. Existing ice drilling technologies make it cumbersome to maintain microbiologically clean access for sample acquisition and environmental stewardship of potentially fragile subglacial aquatic ecosystems. The "IceMole" is a maneuverable subsurface ice probe for clean in-situ analysis and sampling of glacial ice and subglacial materials. The design is based on combining melting and mechanical propulsion, using an ice screw at the tip of the melting head to maintain firm contact between the melting head and the ice. It can change melting direction by differential heating of the melting head and optional side wall heaters. The first two prototypes were successfully tested between 2010 and 2012 on glaciers in Switzerland and Iceland, where they demonstrated downward, horizontal and upward melting, as well as curve driving and dirt layer penetration. Hence, the IceMole allows maneuvers which may be necessary for obstacle avoidance or target selection. Maneuverability, however, necessitates a sophisticated on-board navigation system capable of autonomous operations. Therefore, between 2012 and 2014, a more advanced probe was developed as part of the "Enceladus Explorer" (EnEx) project. The EnEx-IceMole offers systems for relative positioning based on in-ice attitude determination, acoustic positioning, ultrasonic obstacle and target detection, which is all integrated through a high-level sensor fusion. In December 2014, it was used for clean access into a unique subglacial aquatic environment at Blood Falls, Antarctica, where a subglacial brine sample was successfully obtained after about 17 meters of oblique melting. Particular

  4. Microbial community analysis and identification of alternative host-specific fecal indicators in fecal and river water samples using pyrosequencing.

    PubMed

    Jeong, Ju-Yong; Park, Hee-Deung; Lee, Kyong-Hee; Weon, Hang-Yeon; Ka, Jong-Ok

    2011-08-01

    It is important to know the comprehensive microbial communities of fecal pollution sources and receiving water bodies for microbial source tracking. Pyrosequencing targeting the V1-V3 hypervariable regions of the 16S rRNA gene was used to investigate the characteristics of bacterial and Bacteroidales communities in major fecal sources and river waters. Diversity analysis indicated that cow feces had the highest diversities in the bacterial and Bacteroidales group followed by the pig sample, with human feces having the lowest value. The Bacteroidales, one of the potential fecal indicators, totally dominated in the fecal samples accounting for 31%-52% of bacterial sequences, but much less (0.6%) in the river water. Clustering and Venn diagram analyses showed that the human sample had a greater similarity to the pig sample in the bacterial and Bacteroidales communities than to samples from other hosts. Traditional fecal indicators, i.e., Escherichia coli, were detected in the human and river water samples at very low rates and Clostridium perfringens and enterococci were not detected in any samples. Besides the Bacteroidales group, some microorganisms detected in the specific hosts, i.e., Parasutterella excrementihominis, Veillonella sp., Dialister invisus, Megamonas funiformis, and Ruminococcus lactaris for the human and Lactobacillus amylovorus and Atopostipes sp. for the pig, could be used as potential host-specific fecal indicators. These microorganisms could be used as multiple fecal indicators that are not dependent on the absence or presence of a single indicator. Monitoring for multiple indicators that are highly abundant and host-specific would greatly enhance the effectiveness of fecal pollution source tracking.

  5. The impact of different methods of DNA extraction on microbial community measures of BALF samples based on metagenomic data.

    PubMed

    Wen, Yan; Xiao, Fei; Wang, Chen; Wang, Zhen

    2016-01-01

    It is a challenge to find a better microorganisms DNA extraction method for samples taken from the lower airways for metagenomic sequencing, as the concentrations of bacteria in the alveoli and small airways are likely considerably less than that of the mouth or lower digestive tract. Background DNA from the host, and extraction biases can significantly interfere with microbiota assessment and increase the cost of sequencing. This study aimed to develop an optimized DNA extraction method, which would enable a higher concentration of microbial DNA to be extracted from the samples. We compared the microbiota profiles of the lower airway communities in twelve individuals with IIP. DNA was extracted using three different extraction methods: QIAamp UCP PurePathogen Blood Kit named kit3 in this study, QIAamp UCP Pathogen Mini Kit named kit2, and QIAamp DNA Microbiome Kit named kit1. DNA libraries were constructed according to the manufacturer's instructions (Illumina). The same workflows from Illumina were used to perform cluster generation, template hybridization, isothermal amplification, linearization, blocking, denaturing, and hybridization of the sequencing primers. Raw data was uploaded to MG-RAST v3 and analyzed. A great number of bacterium inhabits the lower airways of patients with IIP, though there is no airway infection. More bacterium was found in mouth or upper airway. DNA concentrations of DNA samples isolated with kit1 with Benzonase were significantly lower than those isolated with the other two kits for BALF and mouthwash samples. Moreover, the ratio of human genome in clean reads of samples isolated with kit1 with Benzonase was remarkably smaller than those isolated with kit2 and kit3. The relative abundance of total bacteria, the total number of taxa, and the relative abundance of taxa in BALF samples as opposed to mouthwash samples with kit1 were significantly higher than for those extracted the other kits. A microbial DNA extraction method with

  6. The impact of different methods of DNA extraction on microbial community measures of BALF samples based on metagenomic data

    PubMed Central

    Wen, Yan; Xiao, Fei; Wang, Chen; Wang, Zhen

    2016-01-01

    Purpose: It is a challenge to find a better microorganisms DNA extraction method for samples taken from the lower airways for metagenomic sequencing, as the concentrations of bacteria in the alveoli and small airways are likely considerably less than that of the mouth or lower digestive tract. Background DNA from the host, and extraction biases can significantly interfere with microbiota assessment and increase the cost of sequencing. This study aimed to develop an optimized DNA extraction method, which would enable a higher concentration of microbial DNA to be extracted from the samples. Methods: We compared the microbiota profiles of the lower airway communities in twelve individuals with IIP. DNA was extracted using three different extraction methods: QIAamp UCP PurePathogen Blood Kit named kit3 in this study, QIAamp UCP Pathogen Mini Kit named kit2, and QIAamp DNA Microbiome Kit named kit1. DNA libraries were constructed according to the manufacturer’s instructions (Illumina). The same workflows from Illumina were used to perform cluster generation, template hybridization, isothermal amplification, linearization, blocking, denaturing, and hybridization of the sequencing primers. Raw data was uploaded to MG-RAST v3 and analyzed. Results: A great number of bacterium inhabits the lower airways of patients with IIP, though there is no airway infection. More bacterium was found in mouth or upper airway. DNA concentrations of DNA samples isolated with kit1 with Benzonase were significantly lower than those isolated with the other two kits for BALF and mouthwash samples. Moreover, the ratio of human genome in clean reads of samples isolated with kit1 with Benzonase was remarkably smaller than those isolated with kit2 and kit3. The relative abundance of total bacteria, the total number of taxa, and the relative abundance of taxa in BALF samples as opposed to mouthwash samples with kit1 were significantly higher than for those extracted the other kits. Conclusion: A

  7. Application of Multi-Species Microbial Bioassay to Assess the Effects of Engineered Nanoparticles in the Aquatic Environment: Potential of a Luminous Microbial Array for Toxicity Risk Assessment (LumiMARA) on Testing for Surface-Coated Silver Nanoparticles

    PubMed Central

    Jung, YounJung; Park, Chang-Beom; Kim, Youngjun; Kim, Sanghun; Pflugmacher, Stephan; Baik, Seungyun

    2015-01-01

    Four different manufactured surface-coated silver nanoparticles (AgNPs) with coating of citrate, tannic acid, polyethylene glycol, and branched polyethylenimine were used in this study. The toxicity of surface-coated AgNPs was evaluated by a luminous microbial array for toxicity risk assessment (LumiMARA) using multi-species of luminescent bacteria. The salt stability of four different AgNPs was measured by UV absorbance at 400 nm wavelength, and different surface-charged AgNPs in combination with bacteria were observed using scanning electron microscopy (SEM). Both branched polyethylenimine (BPEI)-AgNPs and polyethylene glycol (PEG)-AgNPs were shown to be stable with 2% NaCl (non-aggregation), whereas both citrate (Cit)-AgNPs and tannic acid (Tan)-AgNPs rapidly aggregated in 2% NaCl solution. The values of the 50% effective concentration (EC50) for BPEI-AgNPs in marine bacteria strains (1.57 to 5.19 mg/L) were lower than those for the other surface-coated AgNPs (i.e., Cit-AgNPs, Tan-AgNPs, and PEG-AgNPs). It appears that the toxicity of AgNPs could be activated by the interaction of positively charged AgNPs with the negatively charged bacterial cell wall from the results of LumiMARA. LumiMARA for toxicity screening has advantageous compared to a single-species bioassay and is applicable for environmental samples as displaying ranges of assessment results. PMID:26184279

  8. Methyl tert-butyl ether biodegradation by microbial consortia obtained from soil samples of gasoline-polluted sites in Mexico.

    PubMed

    Morales, Marcia; Velázquez, Elia; Jan, Janet; Revah, Sergio; González, Uriel; Razo-Flores, Elías

    2004-02-01

    Microbial consortia obtained from soil samples of gasoline-polluted sites were individually enriched with pentane, hexane, isooctane and toluene. Cometabolism with methyl tert-butyl ether, (MTBE), gave maximum degradation rates of 49, 12, 32 and 0 mg g(-1)protein h(-1), respectively. MTBE was fully degraded even when pentane was completely depleted with a cometabolic coefficient of 1 mgMTBE mg(-1)pentane. The analysis of 16S rDNA from isolated microorganisms in the pentane-adapted consortia showed that microorganisms could be assigned to Pseudomonas. This is the first work reporting the cometabolic mineralization of MTBE by consortium of this genus.

  9. Influence of acid mine drainage on microbial communities in stream and groundwater samples at Guryong Mine, South Korea

    NASA Astrophysics Data System (ADS)

    Kim, Jaisoo; Koo, So-Yeon; Kim, Ji-Young; Lee, Eun-Hee; Lee, Sang-Don; Ko, Kyung-Seok; Ko, Dong-Chan; Cho, Kyung-Suk

    2009-10-01

    The effects of acid mine drainage (AMD) in a stream and groundwater near an abandoned copper mine were characterized by physicochemical properties, bacterial community structure using denaturing gel gradient electrophoresis (DGGE), and microbial activity/diversity using Ecoplate technique. Based on DGGE fingerprints, the eubacterial community structures grouped into the stream water (GRS1, GRS2 and GRS3) and groundwater samples (GW1 and GW2), apparently based on differences in water temperature and the concentrations of dissolved oxygen, nitrate and sulfate. The most highly AMD-contaminated sample (GRS1) had additional α-Proteobacteria whereas the groundwater samples included additional β-Proteobacteria, suggesting the development of populations resistant to AMD toxicity under aerobic and anaerobic conditions, respectively. Community level physiological activities on the 31 Ecoplate substrates suggested that the activities decreased with increasing concentrations of sulfate and heavy metals derived from AMD. The Shannon index showed that microbial diversity was greatest in GRS2, and lowest in GRS1, and was probably related to the level of AMD.

  10. Microbial profiling of cpn60 universal target sequences in artificial mixtures of vaginal bacteria sampled by nylon swabs or self-sampling devices under different storage conditions.

    PubMed

    Schellenberg, John J; Oh, Angela Yena; Hill, Janet E

    2017-05-01

    The vaginal microbiome is increasingly characterized by deep sequencing of universal genes. However, there are relatively few studies of how different specimen collection and sample storage and processing influence these molecular profiles. Here, we evaluate molecular microbial community profiles of samples collected using the HerSwab™ self-sampling device, compared to nylon swabs and under different storage conditions. In order to minimize technical variation, mixtures of 11 common vaginal bacteria in simulated vaginal fluid medium were sampled and DNA extracts prepared for massively parallel sequencing of the cpn60 universal target (UT). Three artificial mixtures imitating commonly observed vaginal microbiome profiles were easily distinguished and proportion of sequence reads correlated with the estimated proportion of the organism added to the artificial mixtures. Our results indicate that cpn60 UT amplicon sequencing quantifies the proportional abundance of member organisms in these artificial communities regardless of swab type or storage conditions, although some significant differences were observed between samples that were stored frozen and thawed prior to DNA extraction, compared to extractions from samples stored at room temperature for up to 7days. Our results indicate that an on-the-market device developed for infectious disease diagnostics may be appropriate for vaginal microbiome profiling, an approach that is increasingly facilitated by rapidly dropping deep sequencing costs. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. [Infrared Spectrum Studies of Hydrocarbon Generation and Structure Evolution of Peat Samples During Pyrolysis and Microbial Degradation].

    PubMed

    Bao, Yuan; Ju, Yi-wen; Wei, Chong-tao; Wang, Chao-yong; Li, Xiao-shi

    2015-03-01

    Hydrocarbon generation and structural evolution would be occurred in the process of from coal-forming material (i. e. peat sample) transforming to the coal. While Fourier Transform Infrared Spectroscopy (FTIR) have a special advantages in analyzing molecular structure of samples. For understanding the characteristics of hydrocarbon generation and structural evolution of coal-forming material during the process of pyrolysis and microbial degradation, based on the physical simulation experiments of closed pyrolysis and anaerobic microbial degradation, the generation potential of thermogenic gas and biogenic gas were studied in this paper, and characteristics of molecular structure evolution and its mechanism was analyzed by FTIR technology. Results show that cumulative gas yields of hydrocarbon gases (mainly for methane) increased with experiment temperature. The gas yield of non-hydrocarbon gas (mainly for CO2) exhibited two peaks at 250 and 375 degrees C. The degradation ability of anaerobe on coal samples weakened with the maturity increasing and there was no gas generation on the pyrolysis samples with maturity from 1.6% to 1.8%. After pyrolysis, the content of hydroxyl in peat sample decreased first and then increased with the pyrolysis temperature increasing. The content of aldehyde carbonyl, methylene and phosphate reduced. The content of aromatic esters decreased with nonlinear. The bone of S-O in stretching vibration appeared after 350 degrees C and its content increased with temperature. This shows that the sulfocompound restrains the activity of methanogenic bacteria. After degradation by anaerobe, the relative content of hydroxyl, aldehyde carbonyl, aromatic esters, methylene and phosphate in peat sample dropped significantly. It is shown that the intermolecular force between these groups weakened.

  12. Evidence of soluble microbial products accelerating chloramine decay in nitrifying bulk water samples.

    PubMed

    Bal Krishna, K C; Sathasivan, Arumugam; Chandra Sarker, Dipok

    2012-09-01

    The discovery of a microbially derived soluble product that accelerates chloramine decay is described. Nitrifying bacteria are believed to be wholly responsible for rapid chloramine loss in drinking water systems. However, a recent investigation showed that an unidentified soluble agent significantly accelerated chloramine decay. The agent was suspected to be either natural organic matter (NOM) or soluble microbial products (SMPs). A laboratory scale reactor was fed chloraminated reverse osmosis (RO) treated water to eliminate the interference from NOM. Once nitrification had set in, experiments were conducted on the reactor and feed waters to determine the identity of the component. The study showed the presence of SMPs released by microbes in severely nitrified waters. Further experiments proved that the SMPs significantly accelerated chloramine decay, probably through catalytic reaction. Moreover, application of common protein denaturing techniques stopped the reaction implying that the compound responsible was likely to be a protein. This significant finding will pave the way for better control of chloramine in the distribution systems. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Quality Sample Collection, Handling, and Preservation for an Effective Microbial Forensics Program.

    SciTech Connect

    Budowle, Bruce; Schutzer, Steven E.; Burans, James P.; Beecher, Douglas J.; Cebulla, Thomas; Chakraborty, Ranjit; Cobb, William T.; Fletcher, Jacqueline; Hale, Martha L.; Harris, Robert B.; Heitkamp, Michael; Keller, Frederick P.; Kuske, Cheryl; LeClerc, Joseph E.; Marrone, Babetta L.; McKenna, Thomas S.; Morse, Stephen A.; Rodriguez, Luis L.; Valentine, Nancy B.; Yadev, Jagjit

    2006-10-01

    Science can be part of an effective investigative response to a bioterrorism event or a biocrime by providing capabilities to analyze biological and associated signatures in collected evidence. Microbial forensics, a discipline comprised of several scientific fields, is dedicated to the analysis of evidence from such criminal acts to help determine the responsible party and to exonerate the innocent. A partnership has been formed amount a number of government agencies, academia, and the private sector to better respond and deter potential perpetrators of bioterrorism or biocrimes. This partnership leverages our national scientific and analytical capabilities to support activities of law enforcement agencies. The Department of Homeland Security (DHS), whose mission is, in part, to respond to and to prevent acts of terrorism against the United States, has established the national Bioforensics Analysis Center (NBFAC). The NBFAC, in partnership with the FBI, (1) provides a state-of-the-art central laboratory for the analysis of microbial forensic evidence; and (2) serves as a nexus for integrating the national resources to increase the effectiveness of law enforcement in obtaining the highest level of attribution possible in criminal cases where the weapon is a biological agent.

  14. Virioplankton: Viruses in Aquatic Ecosystems†

    PubMed Central

    Wommack, K. Eric; Colwell, Rita R.

    2000-01-01

    The discovery that viruses may be the most abundant organisms in natural waters, surpassing the number of bacteria by an order of magnitude, has inspired a resurgence of interest in viruses in the aquatic environment. Surprisingly little was known of the interaction of viruses and their hosts in nature. In the decade since the reports of extraordinarily large virus populations were published, enumeration of viruses in aquatic environments has demonstrated that the virioplankton are dynamic components of the plankton, changing dramatically in number with geographical location and season. The evidence to date suggests that virioplankton communities are composed principally of bacteriophages and, to a lesser extent, eukaryotic algal viruses. The influence of viral infection and lysis on bacterial and phytoplankton host communities was measurable after new methods were developed and prior knowledge of bacteriophage biology was incorporated into concepts of parasite and host community interactions. The new methods have yielded data showing that viral infection can have a significant impact on bacteria and unicellular algae populations and supporting the hypothesis that viruses play a significant role in microbial food webs. Besides predation limiting bacteria and phytoplankton populations, the specific nature of virus-host interaction raises the intriguing possibility that viral infection influences the structure and diversity of aquatic microbial communities. Novel applications of molecular genetic techniques have provided good evidence that viral infection can significantly influence the composition and diversity of aquatic microbial communities. PMID:10704475

  15. Status Report on the Microbial Characterization of Halite and Groundwater Samples from the WIPP

    SciTech Connect

    Swanson, Juliet S.; Reed, Donald T.; Ams, David A.; Norden, Diana; Simmons, Karen A.

    2012-07-10

    This report summarizes the progress made in the ongoing task of characterizing the microbial community structures within the WIPP repository and in surrounding groundwaters. Through cultivation and DNA-based identification, the potential activity of these organisms is being inferred, thus leading to a better understanding of their impact on WIPP performance. Members of the three biological domains - Bacteria, Archaea, and Eukarya (in this case, Fungi) - that are associated with WIPP halite have been identified. Thus far, their activity has been limited to aerobic respiration; anaerobic incubations are underway. WIPP halite constitutes the near-field microbial environment. We expect that microbial activity in this setting will proceed from aerobic respiration, through nitrate reduction to focus on sulfate reduction. This is also the current WIPP performance assessment (PA) position. Sulfate reduction can occur at extremely high ionic strengths, and sulfate is available in WIPP brines and in the anhydrite interbeds. The role of methanogenesis in the WIPP remains unclear, due to both energetic constraints imposed by a high-salt environment and substrate selectivity, and it is no longer considered in PA. Archaea identified in WIPP halite thus far fall exclusively within the family Halobacteriaceae. These include Halobacterium noricense, cultivated from both low- and high-salt media, and a Halorubrum-like species. The former has also been detected in other salt mines worldwide; the latter likely constitutes a new species. Little is known of its function, but it was prevalent in experiments investigating the biodegradation of organic complexing agents in WIPP brines. Bacterial signatures associated with WIPP halite include members of the phylum Proteobacteria - Halomonas, Pelomonas, Limnobacter, and Chromohalobacter - but only the latter has been isolated. Also detected and cultivated were Salinicoccus and Nesterenkonia spp. Fungi were also isolated from halite. Although

  16. Metal measurement in aquatic environments by passive sampling methods: Lessons learning from an in situ intercomparison exercise.

    PubMed

    Dabrin, A; Ghestem, J-P; Uher, E; Gonzalez, J-L; Allan, I J; Schintu, M; Montero, N; Balaam, J; Peinerud, E; Miège, C; Coquery, M

    2016-01-01

    Passive sampling devices (PS) are widely used for pollutant monitoring in water, but estimation of measurement uncertainties by PS has seldom been undertaken. The aim of this work was to identify key parameters governing PS measurements of metals and their dispersion. We report the results of an in situ intercomparison exercise on diffusive gradient in thin films (DGT) in surface waters. Interlaboratory uncertainties of time-weighted average (TWA) concentrations were satisfactory (from 28% to 112%) given the number of participating laboratories (10) and ultra-trace metal concentrations involved. Data dispersion of TWA concentrations was mainly explained by uncertainties generated during DGT handling and analytical procedure steps. We highlight that DGT handling is critical for metals such as Cd, Cr and Zn, implying that DGT assembly/dismantling should be performed in very clean conditions. Using a unique dataset, we demonstrated that DGT markedly lowered the LOQ in comparison to spot sampling and stressed the need for accurate data calculation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Retrospective bioindication of stratospheric ozone and ultraviolet radiation using hydroxycinnamic acid derivatives of herbarium samples of an aquatic liverwort.

    PubMed

    Otero, Saúl; Núñez-Olivera, Encarnación; Martínez-Abaigar, Javier; Tomás, Rafael; Huttunen, Satu

    2009-01-01

    We analyzed bulk UV absorbance of methanolic extracts and levels of five UV-absorbing compounds (hydroxycinnamic acid derivatives) in 135 herbarium samples of the liverwort Jungermannia exsertifolia subsp. cordifolia from northern Europe. Samples had been collected in 1850-2006 (96% in June-August). Both UV absorbance and compound levels were correlated positively with collection year. p-Coumaroylmalic acid (C1) was the only compound showing a significant (and negative) correlation with stratospheric ozone and UV irradiance in the period that real data of these variables existed. Stratospheric ozone reconstruction (1850-2006) based on C1 showed higher values in June than in July and August, which coincides with the normal monthly variation of ozone. Combining all the data, there was no long-term temporal trend from 1850 to 2006. Reconstructed UV showed higher values in June-July than in August, but again no temporal trend was detected in 1918-2006 using the joint data. This agrees with previous UV reconstructions.

  18. Effect of Nitrate Injection on the Microbial Community in an Oil Field as Monitored by Reverse Sample Genome Probing

    PubMed Central

    Telang, A. J.; Ebert, S.; Foght, J. M.; Westlake, D.; Jenneman, G. E.; Gevertz, D.; Voordouw, G.

    1997-01-01

    The reverse sample genome probe (RSGP) method, developed for monitoring the microbial community in oil fields with a moderate subsurface temperature, has been improved by (i) isolation of a variety of heterotrophic bacteria and inclusion of their genomes on the oil field master filter and (ii) use of phosphorimaging technology for the rapid quantitation of hybridization signals. The new master filter contains the genomes of 30 sulfate-reducing, 1 sulfide-oxidizing, and 16 heterotrophic bacteria. Most have been identified by partial 16S rRNA sequencing. Use of improved RSGP in monitoring the effect of nitrate injection in an oil field indicated that the sulfide-oxidizing, nitrate-reducing isolate CVO (a Campylobacter sp.) becomes the dominant community component immediately after injection. No significant enhancement of other community members, including the sulfate-reducing bacteria, was observed. The elevated level of CVO decayed at most sampling sites within 30 days after nitrate injection was terminated. Chemical analyses indicated a corresponding decrease and subsequent increase in sulfide concentrations. Thus, transient injection of a higher potential electron acceptor into an anaerobic subsurface system can have desirable effects (i.e., reduction of sulfide levels) without a permanent adverse influence on the resident microbial community. PMID:16535595

  19. Comparative characterization of reticular and duodenal digesta and possibilities of estimating microbial outflow from the rumen based on reticular sampling in dairy cows.

    PubMed

    Hristov, A N

    2007-10-01

    The objective of this experiment was to investigate the possibility of estimating the outflow of nutrients and microbial protein from the rumen based on sampling reticular contents as an alternative to duodenal sampling. Microbial protein flow estimates were also compared with a third method based on sampling of ruminal contents. Reticular and duodenal digesta and ruminal contents were recovered from 4 cows used in a 4 x 4 Latin square design experiment, in which the ruminal effects of 4 exogenous enzyme preparations were studied. Large and small particulate and fluid markers were used to estimate digesta flow in a triple-marker model; 15N was used as a microbial marker. Reticular and duodenal digesta were segregated into small and large particles (SP and LP, respectively) and a fluid phase, and ruminal digesta was segregated into particulate and fluid phases. Compared with digesta recovered at the duodenum, reticular digesta had lower OM and greater NDF contents. The proportion of microbial N was notably greater in the fluid phase of reticular digesta. Ruminal outflow of DM and OM was greater (by 17 and 28%) and that of NDF was lower (by 14%) when estimated from duodenal compared with reticular samples. There was no difference in the estimated flow of starch and nonammonia and microbial N between the reticular and duodenal techniques. Microbial N flow estimated based on ruminal sampling was similar to those based on duodenal and reticular sampling. The ruminal method, however, grossly overestimated flow of DM, OM, and NDF. This study supports the concept that microbial protein outflow from the rumen can be measured based on sampling of ruminal or reticular digesta. The reticular sampling technique can also provide reliable estimates for ruminal digestibility of OM, N, and fiber fractions. These findings need to be confirmed in experiments with basal diets varying in structure and forage-to-concentrate ratios.

  20. A passive sampling method for assessing the occurrence and risk of organophosphate flame retardants in aquatic environments.

    PubMed

    Yang, Cunman; Li, Ying; Zha, Daoping; Lu, Guanghua; Sun, Qin; Wu, Donghai

    2017-01-01

    A modified polar organic chemical integrative sampler (m-POCIS) was used to determine the occurrence of and risk posed by organophosphate flame retardants (OPFRs) in the Yangtze River in Nanjing. Laboratory calibrations were performed to determine sampling rates (Rs) in different situations. Rs values increased with the flow rate, but the effect of dissolved organic matter (DOM) on Rs was relatively small. The validation of Rs in the m-POCIS for 15 days at the Yangtze River in 2015 showed that the Rs values for most of the test compounds are considerably larger than those obtained in the laboratory. The aqueous concentrations of OPFRs were estimated by using Rs values obtained in the field. OPFRs were widely distributed in the Yangtze River, with total concentrations as determined by the m-POCIS ranging from 44.95 ng/L to 118.38 ng/L. These suggest their widespread use and persistence in the river. This study also provided a protocol for the assessment of risk posed by OPFRs. It showed that tripentyl phosphate (TPeP) could pose medium risk to daphnia in the Yangtze River.

  1. Composition of microbial communities in aerosol, snow and ice samples from remote glaciated areas (Antarctica, Alps, Andes)

    NASA Astrophysics Data System (ADS)

    Elster, J.; Delmas, R. J.; Petit, J.-R.; Řeháková, K.

    2007-06-01

    Taxonomical and ecological analyses were performed on micro-autotrophs (cyanobacteria and algae together with remnants of diatom valves), micro-fungi (hyphae and spores), bacteria (rod, cocci and red clusters), yeast, and plant pollen extracted from various samples: Alps snow (Mt. Blank area), Andean snow (Illimani, Bolivia), Antarctic aerosol filters (Dumont d'Urville, Terre Adélie), and Antarctic inland ice (Terre Adélie). Three methods for ice and snow sample's pre-concentration were tested (filtration, centrifugation and lyophilisation). Afterwards, cultivation methods for terrestrial, freshwater and marine microorganisms (micro-autotrophs and micro-fungi) were used in combination with liquid and solid media. The main goal of the study was to find out if micro-autotrophs are commonly transported by air masses, and later stored in snow and icecaps around the world. The most striking result of this study was the absence of culturable micro-autotrophs in all studied samples. However, an unusual culturable pigmented prokaryote was found in both alpine snow and aerosol samples. Analyses of many samples and proper statistical analyses (PCA, RDA- Monte Carlo permutation tests) showed that studied treatments highly significantly differ in both microbial community and biotic remnants composition F=9.33, p=0.001. In addition, GLM showed that studied treatments highly significantly differ in numbers of categories of microorganisms and remnants of biological material F=11.45, p=0.00005. The Antarctic aerosol samples were characterised by having red clusters of bacteria, the unusual prokaryote and yeasts. The high mountain snow from the Alps and Andes contained much more culturable heterotrophs. The unusual prokaryote was very abundant, as were coccoid bacteria, red clusters of bacteria, as well as yeasts. The Antarctic ice samples were quite different. These samples had higher numbers of rod bacteria and fungal hyphae. The microbial communities and biological remnants of

  2. Aquatic Sediments.

    ERIC Educational Resources Information Center

    Sanville, W. D.; And Others

    1978-01-01

    Presents a literature review of aquatic sediments and its effect upon water quality, covering publications of 1976-77. This review includes: (1) sediment water interchange; (2) chemical and physical characterization; and (3) heavy water in sediments. A list of 129 references is also presented. (HM)

  3. Aquatic conditions

    Treesearch

    Warren E. Heilman

    1999-01-01

    This publication provides citizens, private and public organizations, scientists, and others with information about the aquatic conditions in or near national forests in the Ozark-Ouachita Highlands: the Mark Twain in Missouri, the Ouachita in Arkansas and Oklahoma, and the Ozark-St. Francis National Forests in Arkansas. This report includes water quality analyses...

  4. Aquatic Sediments.

    ERIC Educational Resources Information Center

    Sanville, W. D.; And Others

    1978-01-01

    Presents a literature review of aquatic sediments and its effect upon water quality, covering publications of 1976-77. This review includes: (1) sediment water interchange; (2) chemical and physical characterization; and (3) heavy water in sediments. A list of 129 references is also presented. (HM)

  5. Microbial analysis of subgingival plaque samples compared to that of whole saliva in patients with periodontitis.

    PubMed

    Haririan, Hady; Andrukhov, Oleh; Bertl, Kristina; Lettner, Stefan; Kierstein, Sonja; Moritz, Andreas; Rausch-Fan, Xiaohui

    2014-06-01

    The detection of special bacterial species in patients with periodontitis is considered to be useful for clinical diagnosis and treatment. The collection of subgingival plaque samples is the common way for the determination of periodontopathic bacteria. However, recently, salivary analysis has been discussed as an advantageous future diagnostic method for periodontitis because it offers simple quantitative sampling and the possibility to assess various bacteria. The aim of this cross-sectional study is to investigate whether there is a correlation between the results of different bacterial species in saliva and subgingival plaque samples from individuals with aggressive periodontitis (AgP) and chronic periodontitis (CP). Whole saliva and subgingival plaque samples from the deepest pocket of each quadrant were collected from 43 patients with CP and 33 patients with AgP. Twenty different bacterial species from both samplings were determined by the 16S ribosomal RNA-based polymerase chain reaction with microarray technique. All bacterial species were detected in salivary and subgingival plaque samples. For Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, as well as Actinomyces viscosus, Campylobacter rectus/showae, Prevotella intermedia, Parvimonas micra, Eubacterium nodatum, and Campylobacter gracilis, a significant positive correlation between salivary and subgingival plaque samples was detected in patients with both types of periodontitis. There were no significant differences in bacteria in salivary and subgingival plaque samples between AgP and CP. Salivary analysis might be discussed as a potential alternative to subgingival plaque sampling for microbiologic analysis in both AgP and CP.

  6. Spatially resolved sampling reveals dynamic microbial communities in rising hydrothermal plumes across a back-arc basin.

    PubMed

    Sheik, Cody S; Anantharaman, Karthik; Breier, John A; Sylvan, Jason B; Edwards, Katrina J; Dick, Gregory J

    2015-06-01

    Within hydrothermal plumes, chemosynthetic processes and microbe-mineral interactions drive primary productivity in deep-ocean food webs and may influence transport of elements such as iron. However, the source of microorganisms in plumes and the factors governing how these communities assemble are poorly understood, in part due to lack of data from early stages of plume formation. In this study, we examined microbial community composition of rising hydrothermal plumes from five vent fields along the Eastern Lau Spreading Center. Seafloor and plume microbial communities were significantly dissimilar and shared few phylotypes. Plume communities were highly similar to each other with significant differences in community membership only between Kilo Moana and Mariner, two vents that are separated by extremes in depth, latitude and geochemistry. Systematic sampling of waters surrounding the vents revealed that species richness and phylogenetic diversity was typically highest near the vent orifice, implying mixing of microbial communities from the surrounding habitats. Above-plume background communities were primarily dominated by SAR11, SAR324 and MG-I Archaea, while SUP05, Sulfurovum, Sulfurimonas, SAR324 and Alteromonas were abundant in plume and near-bottom background communities. These results show that the ubiquitous water-column microorganisms populate plume communities, and that the composition of background seawater exerts primary influence on plume community composition, with secondary influence from geochemical and/or physical properties of vents. Many of these pervasive deep-ocean organisms are capable of lithotrophy, suggesting that they are poised to use inorganic electron donors encountered in hydrothermal plumes.

  7. Spatially resolved sampling reveals dynamic microbial communities in rising hydrothermal plumes across a back-arc basin

    PubMed Central

    Sheik, Cody S; Anantharaman, Karthik; Breier, John A; Sylvan, Jason B; Edwards, Katrina J; Dick, Gregory J

    2015-01-01

    Within hydrothermal plumes, chemosynthetic processes and microbe–mineral interactions drive primary productivity in deep-ocean food webs and may influence transport of elements such as iron. However, the source of microorganisms in plumes and the factors governing how these communities assemble are poorly understood, in part due to lack of data from early stages of plume formation. In this study, we examined microbial community composition of rising hydrothermal plumes from five vent fields along the Eastern Lau Spreading Center. Seafloor and plume microbial communities were significantly dissimilar and shared few phylotypes. Plume communities were highly similar to each other with significant differences in community membership only between Kilo Moana and Mariner, two vents that are separated by extremes in depth, latitude and geochemistry. Systematic sampling of waters surrounding the vents revealed that species richness and phylogenetic diversity was typically highest near the vent orifice, implying mixing of microbial communities from the surrounding habitats. Above-plume background communities were primarily dominated by SAR11, SAR324 and MG-I Archaea, while SUP05, Sulfurovum, Sulfurimonas, SAR324 and Alteromonas were abundant in plume and near-bottom background communities. These results show that the ubiquitous water-column microorganisms populate plume communities, and that the composition of background seawater exerts primary influence on plume community composition, with secondary influence from geochemical and/or physical properties of vents. Many of these pervasive deep-ocean organisms are capable of lithotrophy, suggesting that they are poised to use inorganic electron donors encountered in hydrothermal plumes. PMID:25489728

  8. Assessing the microbial activity of soil samples, its nutrient limitation and toxic effects of contaminants using a simple respiration test.

    PubMed

    Hollender, Juliane; Althoff, Katrin; Mundt, Matthias; Dott, Wolfgang

    2003-10-01

    Eight soil samples from five wells of a former gas plant site differing in the contamination with BTEX and PAHs as well as the nutrient content were investigated by soil respiration measurements. The basal, glucose as well as NH4+ and PO4(3-) induced cumulative oxygen consumption and carbon dioxide production in 72 and 120 h were determined and additionally the maximal turnover rates and the limitation quotients were calculated. Without additional carbon source only one of five investigated samples was clearly nutrient limited. After glucose supplementation four of seven investigated samples showed nutrient limitation that was in accordance with the available ammonium and phosphorous content. BTEX and PAHs did not exhibit an inhibiting effect on the respiration rate. In contrast, BTEX containing samples exhibited the highest oxygen consumption indicating biodegradation of the contaminants. The results show that oxygen consumption and carbon dioxide production as well as the kinetic of these processes are all informative parameters characterizing the whole microbial respiration potential and their nutrient limitation in soil samples. Therefore this fast respirometric method can be used for the decision if further detailed studies of the bioremediation are useful and if nutrient supplementation is recommended to enhance natural attenuation.

  9. Detection of ammonium-oxidizing bacteria of the beta-subclass of the class Proteobacteria in aquatic samples with the PCR.

    PubMed Central

    Voytek, M A; Ward, B B

    1995-01-01

    The PCR was used as the basis for the development of a sensitive and specific assay for the detection of ammonium-oxidizing bacteria belonging to the beta-subclass of the class Proteobacteria. PCR primers were selected on the basis of nucleic acid sequence data available for seven species of nitrifiers in this subclass. The specificity of the ammonium oxidizer primers was evaluated by testing known strains of nitrifiers, several serotyped environmental nitrifier isolates, and other members of the Proteobacteria, including four very closely related, nonnitrifying species (as determined by rRNA sequence analysis). DNA extracts from 19 bacterio-plankton samples collected from Lake Bonney, Antarctica, and the Southern California Bight were assayed for the presence of ammonium oxidizers. By using a two-stage amplification procedure, ammonium oxidizers were detected in samples collected from both sites. Chemical data collected simultaneously support the occurrence of nitrification and the presence of nitrifiers. This is the first report describing PCR primers specific for ammonium-oxidizing bacteria and the successful amplification of nitrifier genes coding for rRNA from DNA extracts from natural samples. This application of PCR is of particular importance for the detection and study of microbes, such as autotrophic nitrifiers, which are difficult or impossible to isolate from indigenous microbial communities. PMID:7538277

  10. Detection of ammonium-oxidizing bacteria of the beta-subclass of the class Proteobacteria in aquatic samples with the PCR.

    PubMed

    Voytek, M A; Ward, B B

    1995-04-01

    The PCR was used as the basis for the development of a sensitive and specific assay for the detection of ammonium-oxidizing bacteria belonging to the beta-subclass of the class Proteobacteria. PCR primers were selected on the basis of nucleic acid sequence data available for seven species of nitrifiers in this subclass. The specificity of the ammonium oxidizer primers was evaluated by testing known strains of nitrifiers, several serotyped environmental nitrifier isolates, and other members of the Proteobacteria, including four very closely related, nonnitrifying species (as determined by rRNA sequence analysis). DNA extracts from 19 bacterio-plankton samples collected from Lake Bonney, Antarctica, and the Southern California Bight were assayed for the presence of ammonium oxidizers. By using a two-stage amplification procedure, ammonium oxidizers were detected in samples collected from both sites. Chemical data collected simultaneously support the occurrence of nitrification and the presence of nitrifiers. This is the first report describing PCR primers specific for ammonium-oxidizing bacteria and the successful amplification of nitrifier genes coding for rRNA from DNA extracts from natural samples. This application of PCR is of particular importance for the detection and study of microbes, such as autotrophic nitrifiers, which are difficult or impossible to isolate from indigenous microbial communities.

  11. Sample prefractionation with liquid isoelectric focusing enables in depth microbial metaproteome analysis of mesophilic and thermophilic biogas plants.

    PubMed

    Kohrs, F; Heyer, R; Magnussen, A; Benndorf, D; Muth, T; Behne, A; Rapp, E; Kausmann, R; Heiermann, M; Klocke, M; Reichl, U

    2014-10-01

    Biogas production from energy crops and biodegradable waste is one of the major sources for renewable energies in Germany. Within a biogas plant (BGP) a complex microbial community converts biomass to biogas. Unfortunately, disturbances of the biogas process occur occasionally and cause economic losses of varying extent. Besides technical failures the microbial community itself is commonly assumed as a reason for process instability. To improve the performance and efficiency of BGP, a deeper knowledge of the composition and the metabolic state of the microbial community is required and biomarkers for monitoring of process deviations or even the prediction of process failures have to be identified. Previous work based on 2D-electrophoresis demonstrated that the analysis of the metaproteome is well suited to provide insights into the apparent metabolism of the microbial communities. Using SDS-PAGE with subsequent mass spectrometry, stable protein patterns were evaluated for a number of anaerobic digesters. Furthermore, it was shown that severe changes in process parameters such as acidification resulted in significant modifications of the metaproteome. Monitoring of changing protein patterns derived from anaerobic digesters, however, is still a challenge due to the high complexity of the metaproteome. In this study, different combinations of separation techniques to reduce the complexity of proteomic BGP samples were compared with respect to the subsequent identification of proteins by tandem mass spectrometry (MS/MS): (i) 1D: proteins were tryptically digested and the resulting peptides were separated by reversed phase chromatography prior to MS/MS. (ii) 2D: proteins were separated by GeLC-MS/MS according to proteins molecular weights before tryptic digestion, (iii) 3D: proteins were separated by gel-free fractionation using isoelectric focusing (IEF) conducted before GeLC-MS/MS. For this study, a comparison of two anaerobic digesters operated at mesophilic and at

  12. The Microbial Detection Array for Detection of Emerging Viruses in Clinical Samples - A Useful Panmicrobial Diagnostic Tool

    PubMed Central

    Rosenstierne, Maiken W.; McLoughlin, Kevin S.; Olesen, Majken Lindholm; Papa, Anna; Gardner, Shea N.; Engler, Olivier; Plumet, Sebastien; Mirazimi, Ali; Weidmann, Manfred; Niedrig, Matthias; Fomsgaard, Anders; Erlandsson, Lena

    2014-01-01

    Emerging viruses are usually endemic to tropical and sub-tropical regions of the world, but increased global travel, climate change and changes in lifestyle are believed to contribute to the spread of these viruses into new regions. Many of these viruses cause similar disease symptoms as other emerging viruses or common infections, making these unexpected pathogens difficult to diagnose. Broad-spectrum pathogen detection microarrays containing probes for all sequenced viruses and bacteria can provide rapid identification of viruses, guiding decisions about treatment and appropriate case management. We report a modified Whole Transcriptome Amplification (WTA) method that increases unbiased amplification, particular of RNA viruses. Using this modified WTA method, we tested the specificity and sensitivity of the Lawrence Livermore Microbial Detection Array (LLMDA) against a wide range of emerging viruses present in both non-clinical and clinical samples using two different microarray data analysis methods. PMID:24963710

  13. Fluorometric determination of DNA in aquatic microorganisms by use of hoechst 33258.

    PubMed

    Paul, J H; Myers, B

    1982-06-01

    A method for the determination of microbial DNA in aquatic environments by the use of Hoechst 33258 has been developed. With unsophisticated instrumentation and simple extraction procedures, it is possible to detect from 0.05 to 10 mug of DNA in bacterial cultures or natural water samples. The method is specific for DNA; DNase I treatment of extracts of natural microbial populations removed 95 to 100% of the observed fluorescence. DNA content ranged from 165 ng ml for relatively eutrophic Potomac River water to 27 ng ml for coastal Atlantic Ocean water and was correlated to an acridine orange direct count (r = 0.90).

  14. Mathematical Estimation of the Level of Microbial Contamination on Spacecraft Surfaces by Volumetric Air Sampling

    PubMed Central

    Oxborrow, G. S.; Roark, A. L.; Fields, N. D.; Puleo, J. R.

    1974-01-01

    Microbiological sampling methods presently used for enumeration of microorganisms on spacecraft surfaces require contact with easily damaged components. Estimation of viable particles on surfaces using air sampling methods in conjunction with a mathematical model would be desirable. Parameters necessary for the mathematical model are the effect of angled surfaces on viable particle collection and the number of viable cells per viable particle. Deposition of viable particles on angled surfaces closely followed a cosine function, and the number of viable cells per viable particle was consistent with a Poisson distribution. Other parameters considered by the mathematical model included deposition rate and fractional removal per unit time. A close nonlinear correlation between volumetric air sampling and airborne fallout on surfaces was established with all fallout data points falling within the 95% confidence limits as determined by the mathematical model. PMID:4151118

  15. Mathematical estimation of the level of microbial contamination on spacecraft surfaces by volumetric air sampling

    NASA Technical Reports Server (NTRS)

    Oxborrow, G. S.; Roark, A. L.; Fields, N. D.; Puleo, J. R.

    1974-01-01

    Microbiological sampling methods presently used for enumeration of microorganisms on spacecraft surfaces require contact with easily damaged components. Estimation of viable particles on surfaces using air sampling methods in conjunction with a mathematical model would be desirable. Parameters necessary for the mathematical model are the effect of angled surfaces on viable particle collection and the number of viable cells per viable particle. Deposition of viable particles on angled surfaces closely followed a cosine function, and the number of viable cells per viable particle was consistent with a Poisson distribution. Other parameters considered by the mathematical model included deposition rate and fractional removal per unit time. A close nonlinear correlation between volumetric air sampling and airborne fallout on surfaces was established with all fallout data points falling within the 95% confidence limits as determined by the mathematical model.

  16. Mathematical estimation of the level of microbial contamination on spacecraft surfaces by volumetric air sampling

    NASA Technical Reports Server (NTRS)

    Oxborrow, G. S.; Roark, A. L.; Fields, N. D.; Puleo, J. R.

    1974-01-01

    Microbiological sampling methods presently used for enumeration of microorganisms on spacecraft surfaces require contact with easily damaged components. Estimation of viable particles on surfaces using air sampling methods in conjunction with a mathematical model would be desirable. Parameters necessary for the mathematical model are the effect of angled surfaces on viable particle collection and the number of viable cells per viable particle. Deposition of viable particles on angled surfaces closely followed a cosine function, and the number of viable cells per viable particle was consistent with a Poisson distribution. Other parameters considered by the mathematical model included deposition rate and fractional removal per unit time. A close nonlinear correlation between volumetric air sampling and airborne fallout on surfaces was established with all fallout data points falling within the 95% confidence limits as determined by the mathematical model.

  17. The Impact of Selection, Gene Conversion, and Biased Sampling on the Assessment of Microbial Demography.

    PubMed

    Lapierre, Marguerite; Blin, Camille; Lambert, Amaury; Achaz, Guillaume; Rocha, Eduardo P C

    2016-07-01

    Recent studies have linked demographic changes and epidemiological patterns in bacterial populations using coalescent-based approaches. We identified 26 studies using skyline plots and found that 21 inferred overall population expansion. This surprising result led us to analyze the impact of natural selection, recombination (gene conversion), and sampling biases on demographic inference using skyline plots and site frequency spectra (SFS). Forward simulations based on biologically relevant parameters from Escherichia coli populations showed that theoretical arguments on the detrimental impact of recombination and especially natural selection on the reconstructed genealogies cannot be ignored in practice. In fact, both processes systematically lead to spurious interpretations of population expansion in skyline plots (and in SFS for selection). Weak purifying selection, and especially positive selection, had important effects on skyline plots, showing patterns akin to those of population expansions. State-of-the-art techniques to remove recombination further amplified these biases. We simulated three common sampling biases in microbiological research: uniform, clustered, and mixed sampling. Alone, or together with recombination and selection, they further mislead demographic inferences producing almost any possible skyline shape or SFS. Interestingly, sampling sub-populations also affected skyline plots and SFS, because the coalescent rates of populations and their sub-populations had different distributions. This study suggests that extreme caution is needed to infer demographic changes solely based on reconstructed genealogies. We suggest that the development of novel sampling strategies and the joint analyzes of diverse population genetic methods are strictly necessary to estimate demographic changes in populations where selection, recombination, and biased sampling are present. © The Author 2016. Published by Oxford University Press on behalf of the Society for

  18. The Impact of Selection, Gene Conversion, and Biased Sampling on the Assessment of Microbial Demography

    PubMed Central

    Lapierre, Marguerite; Blin, Camille; Lambert, Amaury; Achaz, Guillaume; Rocha, Eduardo P. C.

    2016-01-01

    Recent studies have linked demographic changes and epidemiological patterns in bacterial populations using coalescent-based approaches. We identified 26 studies using skyline plots and found that 21 inferred overall population expansion. This surprising result led us to analyze the impact of natural selection, recombination (gene conversion), and sampling biases on demographic inference using skyline plots and site frequency spectra (SFS). Forward simulations based on biologically relevant parameters from Escherichia coli populations showed that theoretical arguments on the detrimental impact of recombination and especially natural selection on the reconstructed genealogies cannot be ignored in practice. In fact, both processes systematically lead to spurious interpretations of population expansion in skyline plots (and in SFS for selection). Weak purifying selection, and especially positive selection, had important effects on skyline plots, showing patterns akin to those of population expansions. State-of-the-art techniques to remove recombination further amplified these biases. We simulated three common sampling biases in microbiological research: uniform, clustered, and mixed sampling. Alone, or together with recombination and selection, they further mislead demographic inferences producing almost any possible skyline shape or SFS. Interestingly, sampling sub-populations also affected skyline plots and SFS, because the coalescent rates of populations and their sub-populations had different distributions. This study suggests that extreme caution is needed to infer demographic changes solely based on reconstructed genealogies. We suggest that the development of novel sampling strategies and the joint analyzes of diverse population genetic methods are strictly necessary to estimate demographic changes in populations where selection, recombination, and biased sampling are present. PMID:26931140

  19. Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping

    PubMed Central

    Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A.; Debelius, Justine; Morton, James T.; Hyde, Embriette; Robbins-Pianka, Adam

    2017-01-01

    ABSTRACT The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria, grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., “bloomed”) at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called “Deblur” to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has

  20. Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping.

    PubMed

    Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A; Debelius, Justine; Morton, James T; Hyde, Embriette; Robbins-Pianka, Adam; Knight, Rob

    2017-01-01

    The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria, grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., "bloomed") at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called "Deblur" to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has been shown not to

  1. Draft Genome Sequences of 24 Microbial Strains Assembled from Direct Sequencing from 4 Stool Samples

    PubMed Central

    Hernández, Álvaro; White, Bryan A.; O’Brien, Daniel; Ahlquist, David; Boardman, Lisa

    2015-01-01

    The ability to assemble genomes from metagenomic sequencing avoids the need for culture and any associated culture biases. We assembled 24 essentially complete draft genomes from metagenomic pair-end and size-selected mate pair sequencing from 4 stool samples, 2 from subjects diagnosed with colorectal cancer and 2 from healthy controls. PMID:26021920

  2. Sampling and Selection Factors that Enhance the Diversity of Microbial Collections: Application to Biopesticide Development

    PubMed Central

    Park, Jun-Kyung; Lee, Seung-Hwan; Lee, Jang-Hoon; Han, Songhee; Kang, Hunseung; Kim, Jin-Cheol; Kim, Young Cheol; Gardener, Brian McSpadden

    2013-01-01

    Diverse bacteria are known to colonize plants. However, only a small fraction of that diversity has been evaluated for their biopesticide potential. To date, the criteria for sampling and selection in such bioprospecting endeavors have not been systematically evaluated in terms of the relative amount of diversity they provide for analysis. The present study aimed to enhance the success of bio-prospecting efforts by increasing the diversity while removing the genotypic redundancy often present in large collections of bacteria. We developed a multivariate sampling and marker-based selection strategy that significantly increase the diversity of bacteria recovered from plants. In doing so, we quantified the effects of varying sampling intensity, media composition, incubation conditions, plant species, and soil source on the diversity of recovered isolates. Subsequent sequencing and high-throughput phenotypic analyses of a small fraction of the collected isolates revealed that this approach led to the recovery of over a dozen rare and, to date, poorly characterized genera of plant-associated bacteria with significant biopesticide activities. Overall, the sampling and selection approach described led to an approximately 5-fold improvement in efficiency and the recovery of several novel strains of bacteria with significant biopesticide potential. PMID:25288941

  3. Microbial monitoring and methods of sample collection: a GITMO survey (Gruppo Trapianto di Midollo Osseo)

    PubMed Central

    Gori, Erica; Callea, Emanuela; Alberani, Francesca; Orlando, Laura

    2014-01-01

    The collection of microbiological samples represents an important aspect of care both for doctors as well as nurses. It is important to recognise and identify some key points, to avoid performing ‘unnecessary’ or ‘incorrect’ sampling, which may give useless or even misleading results, these are: the moment at which the sample is collected, the collection method and timing (if indicated). The comparison between the various nursing members of the Italian National bone marrow transplant group (GITMO), showed diversity of practice across all fields. A formal survey was therefore conducted within GITMO centres looking at the methods of microbiological sample collection. These results were compared with the literature, and in addition to the lack of homogeneity of practice within the centres, a lack of compliance with the recommendations was also observed. To evaluate the effectiveness of this survey in highlighting awareness of this issue and the presence of relevant guidelines, the questionnaire was repeated (with the same centres responding), which demonstrated no major changes in care practices. Conclusion: The survey has allowed us to highlight many critical issues regarding common procedures which are not commonly discussed. Considerable differences were noted between different transplant centres, which may be attributable to the lack of Italian guidelines that can be used as a starting point for clinical practice. The plenary discussion allowed for an exchange of findings with the medical staff, who are usually responsible for requesting microbiological samples. The ideal solution would be a unique field-based training programme, associated with the dissemination of a common procedural document for ensuring evidence-based practice. PMID:24834116

  4. Methodological tests of a heterotrophy index for aquatic ecosystems.

    PubMed

    Antonio, R M; Bianchini Júnior, I

    2003-08-01

    Experiments in glucose mineralization were carried out to investigate the effects caused by natural forcing functions on both the decomposition rates and heterotrophy capacity of aquatic ecosystems. In addition, the methodology used could show connections between mineralization rates measured in both laboratory and field work with those measured in aquatic ecosystems. Water samples from Infernão lagoon (21 degrees 35'S and 47 degrees 51'W) were collected, filtered, enriched with glucose, and incubated under aerobic and anaerobic conditions. The glucose concentration variation, dissolved oxygen (DO) consumption, pH, electric conductivity, and total CO2 amount in the water were determined for sixteen days. In the period with intense oxygen consumption there was also an evident glucose demand and the dissolved oxygen consumption rate was approximately the same as that for glucose mineralization. The process in the aerobic chambers was 2.2 times faster than that in the anaerobic chambers. An initial acidification of the water samples, probably due to microbial carbonic acid liberation, was noted. A rise in pH values was also observed at the end of the process. The electric conductivity was low for both aerobic and anaerobic chambers, indicating a probable ion uptake by microbial organisms due to the presence of carbon sources. The glucose content variations corresponded to both CO2 formation and dissolved oxygen consumption. It was estimated that 19.4% of the initial glucose content turned into CO2 and the remaining 80.6% into humic compounds and microbial biomass. This experiment showed that glucose can be used as a substrate indicating the heterotrophy of a given aquatic ecosystem.

  5. Comparison of Filters for Concentrating Microbial Indicators and Pathogens in Lake Water Samples

    PubMed Central

    Stelzer, Erin A.; Brady, Amie M. G.; Huitger, Carrie; Bushon, Rebecca N.; Ip, Hon S.; Ware, Michael W.; Villegas, Eric N.; Gallardo, Vicente; Lindquist, H. D. Alan

    2013-01-01

    Bacterial indicators are used to indicate increased health risk from pathogens and to make beach closure and advisory decisions; however, beaches are seldom monitored for the pathogens themselves. Studies of sources and types of pathogens at beaches are needed to improve estimates of swimming-associated health risks. It would be advantageous and cost-effective, especially for studies conducted on a regional scale, to use a method that can simultaneously filter and concentrate all classes of pathogens from the large volumes of water needed to detect pathogens. In seven recovery experiments, stock cultures of viruses and protozoa were seeded into 10-liter lake water samples, and concentrations of naturally occurring bacterial indicators were used to determine recoveries. For the five filtration methods tested, the highest median recoveries were as follows: glass wool for adenovirus (4.7%); NanoCeram for enterovirus (14.5%) and MS2 coliphage (84%); continuous-flow centrifugation (CFC) plus Virocap (CFC+ViroCap) for Escherichia coli (68.3%) and Cryptosporidium (54%); automatic ultrafiltration (UF) for norovirus GII (2.4%); and dead-end UF for Enterococcus faecalis (80.5%), avian influenza virus (0.02%), and Giardia (57%). In evaluating filter performance in terms of both recovery and variability, the automatic UF resulted in the highest recovery while maintaining low variability for all nine microorganisms. The automatic UF was used to demonstrate that filtration can be scaled up to field deployment and the collection of 200-liter lake water samples. PMID:23263948

  6. Comparison of filters for concentrating microbial indicators and pathogens in lake water samples.

    PubMed

    Francy, Donna S; Stelzer, Erin A; Brady, Amie M G; Huitger, Carrie; Bushon, Rebecca N; Ip, Hon S; Ware, Michael W; Villegas, Eric N; Gallardo, Vicente; Lindquist, H D Alan

    2013-02-01

    Bacterial indicators are used to indicate increased health risk from pathogens and to make beach closure and advisory decisions; however, beaches are seldom monitored for the pathogens themselves. Studies of sources and types of pathogens at beaches are needed to improve estimates of swimming-associated health risks. It would be advantageous and cost-effective, especially for studies conducted on a regional scale, to use a method that can simultaneously filter and concentrate all classes of pathogens from the large volumes of water needed to detect pathogens. In seven recovery experiments, stock cultures of viruses and protozoa were seeded into 10-liter lake water samples, and concentrations of naturally occurring bacterial indicators were used to determine recoveries. For the five filtration methods tested, the highest median recoveries were as follows: glass wool for adenovirus (4.7%); NanoCeram for enterovirus (14.5%) and MS2 coliphage (84%); continuous-flow centrifugation (CFC) plus Virocap (CFC+ViroCap) for Escherichia coli (68.3%) and Cryptosporidium (54%); automatic ultrafiltration (UF) for norovirus GII (2.4%); and dead-end UF for Enterococcus faecalis (80.5%), avian influenza virus (0.02%), and Giardia (57%). In evaluating filter performance in terms of both recovery and variability, the automatic UF resulted in the highest recovery while maintaining low variability for all nine microorganisms. The automatic UF was used to demonstrate that filtration can be scaled up to field deployment and the collection of 200-liter lake water samples.

  7. Comparison of filters for concentrating microbial indicators and pathogens in lake-water samples

    USGS Publications Warehouse

    Francy, Donna S.; Stelzer, Erin A.; Brady, Amie M.G.; Huitger, Carrie; Bushon, Rebecca N.; Ip, Hon S.; Ware, Michael W.; Villegas, Eric N.; Gallardo, Vincent; Lindquist, H.D. Alan

    2013-01-01

    Bacterial indicators are used to indicate increased health risk from pathogens and to make beach closure and advisory decisions; however, beaches are seldom monitored for the pathogens themselves. Studies of sources and types of pathogens at beaches are needed to improve estimates of swimming-associated health risks. It would be advantageous and cost-effective, especially for studies conducted on a regional scale, to use a method that can simultaneously filter and concentrate all classes of pathogens from the large volumes of water needed to detect pathogens. In seven recovery experiments, stock cultures of viruses and protozoa were seeded into 10-liter lake water samples, and concentrations of naturally occurring bacterial indicators were used to determine recoveries. For the five filtration methods tested, the highest median recoveries were as follows: glass wool for adenovirus (4.7%); NanoCeram for enterovirus (14.5%) and MS2 coliphage (84%); continuous-flow centrifugation (CFC) plus Virocap (CFC+ViroCap) for Escherichia coli (68.3%) and Cryptosporidium (54%); automatic ultrafiltration (UF) for norovirus GII (2.4%); and dead-end UF for Enterococcus faecalis (80.5%), avian influenza virus (0.02%), and Giardia (57%). In evaluating filter performance in terms of both recovery and variability, the automatic UF resulted in the highest recovery while maintaining low variability for all nine microorganisms. The automatic UF was used to demonstrate that filtration can be scaled up to field deployment and the collection of 200-liter lake water samples.

  8. Sample-based assessment of the microbial etiology of bovine necrotic vulvovaginitis.

    PubMed

    Blum, S; Mazuz, M; Brenner, J; Friedgut, O; Stram, Y; Koren, O; Goshen, T; Elad, D

    2007-07-15

    A semiquantitative evaluation of potential bacterial pathogens was correlated to the severity of lesions during an outbreak of bovine necrotic vulvovaginitis (BNVV) on an Israeli dairy herd. Bacteriologic examination of 287 vaginal swabs from 104 post-calving heifers showed a highly significant correlation between Porphyromonas levii colony forming unit numbers and the clinical scores of the lesions, when assessed by an ordinal regression statistical model. No such correlation was found for the other bacteria included in the study. Nineteen samples taken for virological examinations resulted negative for bovine herpes viruses 1, 2, 4 and 5. Thus the results of this study substantiate the essential role of P. levii in the etiology of BNVV and indicate that BHV4 is not required as a predisposing factor to the syndrome.

  9. Post-Flight Microbial Analysis of Samples from the International Space Station Water Recovery System and Oxygen Generation System

    NASA Technical Reports Server (NTRS)

    Birmele, Michele N.

    2011-01-01

    The Regenerative, Environmental Control and Life Support System (ECLSS) on the International Space Station (ISS) includes the the Water Recovery System (WRS) and the Oxygen Generation System (OGS). The WRS consists of a Urine Processor Assembly (UPA) and Water Processor Assembly (WPA). This report describes microbial characterization of wastewater and surface samples collected from the WRS and OGS subsystems, returned to KSC, JSC, and MSFC on consecutive shuttle flights (STS-129 and STS-130) in 2009-10. STS-129 returned two filters that contained fluid samples from the WPA Waste Tank Orbital Recovery Unit (ORU), one from the waste tank and the other from the ISS humidity condensate. Direct count by microscopic enumeration revealed 8.38 x 104 cells per mL in the humidity condensate sample, but none of those cells were recoverable on solid agar media. In contrast, 3.32 x lOs cells per mL were measured from a surface swab of the WRS waste tank, including viable bacteria and fungi recovered after S12 days of incubation on solid agar media. Based on rDNA sequencing and phenotypic characterization, a fungus recovered from the filter was determined to be Lecythophora mutabilis. The bacterial isolate was identified by rDNA sequence data to be Methylobacterium radiotolerans. Additional UPA subsystem samples were returned on STS-130 for analysis. Both liquid and solid samples were collected from the Russian urine container (EDV), Distillation Assembly (DA) and Recycle Filter Tank Assembly (RFTA) for post-flight analysis. The bacterium Pseudomonas aeruginosa and fungus Chaetomium brasiliense were isolated from the EDV samples. No viable bacteria or fungi were recovered from RFTA brine samples (N= 6), but multiple samples (N = 11) from the DA and RFTA were found to contain fungal and bacterial cells. Many recovered cells have been identified to genus by rDNA sequencing and carbon source utilization profiling (BiOLOG Gen III). The presence of viable bacteria and fungi from WRS

  10. An optical method for carbon dioxide isotopes and mole fractions in small gas samples: tracing microbial respiration from soil, litter, and lignin.

    Treesearch

    Steven J. Hall; Wenjuan Huang; Kenneth Hammel

    2017-01-01

    RATIONALE: Carbon dioxide isotope (Δ13C value) measurements enable quantification of the sources of soil microbial respiration, thus informing ecosystem C dynamics. Tunable diode lasers (TDLs) can precisely measure CO2 isotopes at low cost and high throughput, but are seldom used for small samples (≤5 mL). We developed a...

  11. Ecology, Microbial

    SciTech Connect

    Konopka, Allan

    2009-05-15

    Microbial ecology is a relatively young discipline within the field of microbiology. Its modern history spans just the past 60 years, and the field is defined by its emphasis on understanding the interactions of microbes with their environment, rather than their behavior under artificial laboratory conditions. Because microbes are ubiquitous, microbial ecologists study a broad diversity of habitats that range from aquatic to terrestrial to plant- or animal-associated. This has made it a challenge to identify unifying principles within the field. One approach is to recognize that although the activity of microbes in nature have effects at the macroscale, they interact with their physical, chemical and biological milieu at a scale of micrometers. At this scale, several different microbial ecosystems can be defined, based upon association with particles, the presence of environmental gradients and the continuous availability of water. Principles applicable to microbial ecology reflect not only their population ecology and physiological ecology, but also their broad versatility and quantitative importance in the biosphere as biogeochemical catalysts and capacity for rapid physiological and evolutionary responses.

  12. The development of point-of-use water filters as sampling devices in bioforensics: extent of microbial sorption and elution.

    PubMed

    Sedillo, Jennifer L; Quintana, Ayshea; Souza, Kathryn; Oshima, Kevin H; Smith, Geoffrey B

    2008-06-01

    The foundational idea for this project is that household faucet-mounted water filters may be used as bioforensic sampling devices to detect the extent of a potential bioagent release in domestic water supplies. An optimized eluent solution was determined experimentally by quantifying recoveries of microorganisms from point-of-use (POU) drinking water filters. The optimized extraction protocol was then used in mock bioagent release experiments to determine the feasibility of POU filters as bioforensic sampling devices. Bacillus atrophaeus spores, Escherichia coli and PP7 virus were exposed to filters and the number of attached organisms was determined by enumerating the unattached organisms on selective agar media. Subsequently, the filters were eluted and the percent of extracted organisms was determined based on the number of attached organisms. Two popular brands of carbon block filters retained 92%-99% of representative virus, spore and vegetative bacteria. In back-flush elutions of single filters, the most efficient eluent was identified as a combination of 1% peptone and 1% Tween-80, and extraction recovered 25.4% (+/-17.5%) of attached E. coli, 20.4% (+/-3.6%) of B. atrophaeus spores, and 9.4% (+/-5.2%) of PP7 virions (+/- standard deviations). In bioagent release studies in which filters were challenged with 100 agents mL(-1), greater than 99% of the spores were retained by the filters, and the percent of attached spores that were recovered ranged from 10.4% at day 0 to 4.3% five days after the release event (averaged from five separate experiments). In contrast, E. coli, Salmonella typhimurium and PP7 virus were rapidly inactivated in the chlorinated tap water, indicating their improbable survival in chlorinated water supplies. It is therefore concluded that household water filters can be used as microbial sampling devices for bioforensic applications in the event of a bioagent release in domestic drinking water supplies.

  13. Aquatic Acoustic Metrics Interface

    SciTech Connect

    2012-12-18

    Fishes and marine mammals may suffer a range of potential effects from exposure to intense underwater sound generated by anthropogenic activities such as pile driving, shipping, sonars, and underwater blasting. Several underwater sound recording (USR) devices have been built to acquire samples of the underwater sound generated by anthropogenic activities. Software becomes indispensable for processing and analyzing the audio files recorded by these USRs. The new Aquatic Acoustic Metrics Interface Utility Software (AAMI) is specifically designed for analysis of underwater sound recordings to provide data in metrics that facilitate evaluation of the potential impacts of the sound on aquatic animals. In addition to the basic functions, such as loading and editing audio files recorded by USRs and batch processing of sound files, the software utilizes recording system calibration data to compute important parameters in physical units. The software also facilitates comparison of the noise sound sample metrics with biological measures such as audiograms of the sensitivity of aquatic animals to the sound, integrating various components into a single analytical frame.

  14. Use of Spatial Sampling and Microbial Source-Tracking Tools for Understanding Fecal Contamination at Two Lake Erie Beaches

    USGS Publications Warehouse

    Francy, Donna S.; Bertke, Erin E.; Finnegan, Dennis P.; Kephart, Christopher M.; Sheets, Rodney A.; Rhoades, John; Stumpe, Lester

    2006-01-01

    Source-tracking tools were used to identify potential sources of fecal contamination at two Lake Erie bathing beaches: an urban beach (Edgewater in Cleveland, Ohio) and a beach in a small city (Lakeshore in Ashtabula, Ohio). These tools included identifying spatial patterns of Escherichia coli (E. coli) concentrations in each area, determining weather patterns that caused elevated E. coli, and applying microbial source tracking (MST) techniques to specific sites. Three MST methods were used during this study: multiple antibiotic resistance (MAR) indexing of E. coli isolates and the presence of human-specific genetic markers within two types of bacteria, the genus Bacteroides and the species Enterococcus faecium. At Edgewater, sampling for E. coli was done during 2003-05 at bathing-area sites, at nearshore lake sites, and in shallow ground water in foreshore and backshore areas. Spatial sampling at nearshore lake sites showed that fecal contamination was most likely of local origin; E. coli concentrations near the mouths of rivers and outfalls remote to the beach were elevated (greater than 235 colony-forming units per 100 milliliters (CFU/100 mL)) but decreased along transport pathways to the beach. In addition, E. coli concentrations were generally highest in bathing-area samples collected at 1- and 2-foot water depths, midrange at 3-foot depths, and lowest in nearshore lake samples typically collected 150 feet from the shoreline. Elevated E. coli concentrations at bathing-area sites were generally associated with increased wave heights and rainfall, but not always. E. coli concentrations were often elevated in shallow ground-water samples, especially in samples collected less than 10 feet from the edge of water (near foreshore area). The interaction of shallow ground water and waves may be a mechanism of E. coli storage and accumulation in foreshore sands. Infiltration of bird feces through sand with surface water from rainfall and high waves may be concentrating

  15. A Pilot Study on Integrating Videography and Environmental Microbial Sampling to Model Fecal Bacterial Exposures in Peri-Urban Tanzania.

    PubMed

    Julian, Timothy R; Pickering, Amy J

    2015-01-01

    Diarrheal diseases are a leading cause of under-five mortality and morbidity in sub-Saharan Africa. Quantitative exposure modeling provides opportunities to investigate the relative importance of fecal-oral transmission routes (e.g. hands, water, food) responsible for diarrheal disease. Modeling, however, requires accurate descriptions of individuals' interactions with the environment (i.e., activity data). Such activity data are largely lacking for people in low-income settings. In the present study, we collected activity data and microbiological sampling data to develop a quantitative microbial exposure model for two female caretakers in peri-urban Tanzania. Activity data were combined with microbiological data of contacted surfaces and fomites (e.g. broom handle, soil, clothing) to develop example exposure profiles describing second-by-second estimates of fecal indicator bacteria (E. coli and enterococci) concentrations on the caretaker's hands. The study demonstrates the application and utility of video activity data to quantify exposure factors for people in low-income countries and apply these factors to understand fecal contamination exposure pathways. This study provides both a methodological approach for the design and implementation of larger studies, and preliminary data suggesting contacts with dirt and sand may be important mechanisms of hand contamination. Increasing the scale of activity data collection and modeling to investigate individual-level exposure profiles within target populations for specific exposure scenarios would provide opportunities to identify the relative importance of fecal-oral disease transmission routes.

  16. Advanced instrument system for real-time and time-series microbial geochemical sampling of the deep (basaltic) crustal biosphere

    NASA Astrophysics Data System (ADS)

    Cowen, James P.; Copson, David A.; Jolly, James; Hsieh, Chih-Chiang; Lin, Huei-Ting; Glazer, Brian T.; Wheat, C. Geoffrey

    2012-03-01

    Integrated Ocean Drilling Program borehole CORK (Circulation Obviation Retrofit Kit) observatories provide long-term access to hydrothermal fluids circulating within the basaltic crust (basement), providing invaluable opportunities to study the deep biosphere. We describe the design and application parameters of the GeoMICROBE instrumented sled, an autonomous sensor and fluid sampling system. The GeoMICROBE system couples with CORK fluid delivery lines to draw large volumes of fluids from crustal aquifers to the seafloor. These fluids pass a series of in-line sensors and an in situ filtration and collection system. GeoMICROBE's major components include a primary valve manifold system, a positive displacement primary pump, sensors (e.g., fluid flow rate, temperature, dissolved O2, electrochemistry-voltammetry analyzer), a 48-port in situ filtration and fluid collection system, computerized controller, seven 24 V-40 A batteries and wet-mateable (ODI) communications with submersibles. This constantly evolving system has been successfully connected to IODP Hole 1301A on the eastern flank of the Juan de Fuca Ridge. Also described here is a mobile pumping system (MPS), which possesses many of the same components as the GeoMICROBE (e.g., pump, sensors, controller), but is directly powered and controlled in real time via submersible operations; the MPS has been employed repeatedly to collect pristine basement fluids for a variety of geochemical and microbial studies.

  17. GC and GC-MS characterization of crude oil transformation in sediments and microbial mat samples after the 1991 oil spill in the Saudi Arabian Gulf coast.

    PubMed

    Garcia de Oteyza, T; Grimalt, J O

    2006-02-01

    The massive oil discharge in the Saudi Arabian coast at the end of the 1991 Gulf War is used here as a natural experiment to study the ability of microbial mats to transform oil residues after major spills. The degree of oil transformation has been evaluated from the analysis of the aliphatic and aromatic hydrocarbons by gas chromatography (GC) and GC coupled to mass spectrometry (GC-MS). The oil-polluted microbial mat samples from coastal environments exhibited an intermediate degree of transformation between that observed in superficial and deep sediments. Evaporation, photo-oxidation and water-washing seemed to lead to more effective and rapid elimination of hydrocarbons than cyanobacteria and its associated microorganisms. Furthermore, comparison of some compounds (e.g. regular isoprenoid hydrocarbons or alkylnaphthalenes) in the oil collected in the area after the spill or in the mixtures retained by cyanobacterial growth gave rise to an apparent effect of hydrocarbon preservation in the microbial mat ecosystems.

  18. Microplastic in Aquatic Ecosystems.

    PubMed

    Ivleva, Natalia P; Wiesheu, Alexandra C; Niessner, Reinhard

    2017-02-06

    The contamination of marine and freshwater ecosystems with plastic, and especially with microplastic (MP), is a global ecological problem of increasing scientific concern. This has stimulated a great deal of research on the occurrence of MP, interaction of MP with chemical pollutants, the uptake of MP by aquatic organisms, and the resulting (negative) impact of MP. Herein, we review the major issues of MP in aquatic environments, with the principal aims 1) to characterize the methods applied for MP analysis (including sampling, processing, identification and quantification), indicate the most reliable techniques, and discuss the required further improvements; 2) to estimate the abundance of MP in marine/freshwater ecosystems and clarify the problems that hamper the comparability of such results; and 3) to summarize the existing literature on the uptake of MP by living organisms. Finally, we identify knowledge gaps, suggest possible strategies to assess environmental risks arising from MP, and discuss prospects to minimize MP abundance in aquatic ecosystems.

  19. Drastic changes in aquatic bacterial populations from the Cuatro Cienegas Basin (Mexico) in response to long-term environmental stress.

    PubMed

    Pajares, Silvia; Eguiarte, Luis E; Bonilla-Rosso, German; Souza, Valeria

    2013-12-01

    Understanding the changes of aquatic microbial community composition in response to changes in temperature and ultraviolet irradiation is relevant for predicting biogeochemical modifications in the functioning of natural microbial communities under global climate change scenarios. Herein we investigate shifts in the bacterioplankton composition in response to long-term changes in temperature and UV radiation. For this purpose, 15 mesocosms were seeded with composite aquatic microbial communities from natural pools within the Cuatro Cienegas Basin (Mexican Chihuahuan desert) and were subject to different temperatures and UV conditions. 16S rRNA gene clone libraries were obtained from water samples at the mid-point (4 months) and the end of the experiment (8 months). An increase in bacterial diversity over time was found in the treatment of constant temperature and UV protection, which suggests that stable environments promote the establishment of complex and diverse bacterial community. Drastic changes in the phylogenetic bacterioplankton composition and structure were observed in response to fluctuating temperature and increasing UV radiation and temperature. Fluctuating temperature induced the largest decrease of bacterial richness during the experiment, indicating that frequent temperature changes drive the reduction in abundance of several species, most notably autotrophs. The long-term impact of these environmental stresses reduced diversity and selected for generalist aquatic bacterial populations, such as Porphyrobacter. These changes at the community level occur at an ecological time scale, suggesting that under global warming scenarios cascade effects on the food web are possible if the microbial diversity is modified.

  20. Aquatic toxicity of triclosan.

    PubMed

    Orvos, David R; Versteeg, Donald J; Inauen, Josef; Capdevielle, Marie; Rothenstein, Arthur; Cunningham, Virginia

    2002-07-01

    The aquatic toxicity of triclosan (TCS), a chlorinated biphenyl ether used as an antimicrobial in consumer products, was studied with activated-sludge microorganisms, algae, invertebrates, and fish. Triclosan, a compound used for inhibiting microbial growth, was not toxic to wastewater microorganisms at concentrations less than aqueous solubility. The 48-h Daphnia magna median effective concentration (EC50) was 390 microg/L and the 96-h median lethal concentration values for Pimephales promelas and Lepomis macrochirus were 260 and 370 microg/L, respectively. A no-observed-effect concentration (NOEC) and lowest-observed-effect concentration of 34.1 microg/L and 71.3 microg/L, respectively, were determined with an early life-stage toxicity test with Oncorhynchus mykiss. During a 96-h Scenedesmus study, the 96-h biomass EC50 was 1.4 microg/L and the 96-h NOEC was 0.69 microg/L. Other algae and Lemna also were investigated. Bioconcentration was assessed with Danio rerio. The average TCS accumulation factor over the five-week test period was 4,157 at 3 microg/L and 2,532 at 30 microg/L. Algae were determined to be the most susceptible organisms. Toxicity of a TCS-containing wastewater secondary effluent to P. promelas and Ceriodaphnia was evaluated and no observed differences in toxicity between control and TCS-treated laboratory units were detected. The neutral form of TCS was determined to be associated with toxic effects. Ionization and sorption will mitigate those effects in the aquatic compartment.

  1. Mesocosms of aquatic bacterial communities from the Cuatro Cienegas Basin (Mexico): a tool to test bacterial community response to environmental stress.

    PubMed

    Pajares, Silvia; Bonilla-Rosso, German; Travisano, Michael; Eguiarte, Luis E; Souza, Valeria

    2012-08-01

    Microbial communities are responsible for important ecosystem processes, and their activities are regulated by environmental factors such as temperature and solar ultraviolet radiation. Here we investigate changes in aquatic microbial community structure, diversity, and evenness in response to changes in temperature and UV radiation. For this purpose, 15 mesocosms were seeded with both microbial mat communities and plankton from natural pools within the Cuatro Cienegas Basin (Mexico). Clone libraries (16S rRNA) were obtained from water samples at the beginning and at the end of the experiment (40 days). Phylogenetic analysis indicated substantial changes in aquatic community composition and structure in response to temperature and UV radiation. Extreme treatments with elevation in temperature or UV radiation reduced diversity in relation to the Control treatments, causing a reduction in richness and increase in dominance, with a proliferation of a few resistant operational taxonomic units. Each phylum was affected differentially by the new conditions, which translates in a differential modification of ecosystem functioning. This suggests that the impact of environmental stress, at least at short term, will reshape the aquatic bacterial communities of this unique ecosystem. This work also demonstrates the possibility of designing manageable synthetic microbial community ecosystems where controlled environmental variables can be manipulated. Therefore, microbial model systems offer a complementary approach to field and laboratory studies of global research problems associated with the environment.

  2. A Direct-Push Sample-Freezing Drive Shoe for Collecting Sediment Cores with Intact Pore Fluid, Microbial, and Sediment Distributions

    NASA Astrophysics Data System (ADS)

    Bekins, B. A.; Trost, J.; Christy, T. M.; Mason, B.

    2015-12-01

    Abiotic and biological reactions in shallow groundwater and bottom sediments are central to understanding groundwater contaminant attenuation and biogeochemical cycles. The laminar flow regime in unconsolidated surficial aquifers creates narrow reaction zones. Studying these reaction zones requires fine-scale sampling of water together with adjacent sediment in a manner that preserves in situ redox conditions. Collecting representative samples of these narrow zones with traditional subsurface sampling equipment is challenging. For example, use of a basket type core catcher for saturated, non-cohesive sediments results in loss of fluid and sediments during retrieval. A sample-freezing drive shoe designed for a wire line piston core sampler allowed collection of cores with intact sediment, microbial, and pore fluid distributions and has been the basis for studies documenting centimeter-scale variations in aquifer microbial populations (Murphy and Herkelrath, 1996). However, this freezing drive shoe design is not compatible with modern-day direct push sampling rigs. A re-designed sample-freezing drive shoe compatible with a direct-push dual-tube coring system was developed and field-tested. The freezing drive shoe retained sediment and fluid distributions in saturated sediment core samples by freezing a 10 centimeter plug below the core sample with liquid CO­2. Core samples collected across the smear zone at a crude oil spill site near Bemidji, Minnesota, were successfully extracted without loss of fluid or sediment. Multiple core sections from different depths in the aquifer were retrieved from a single hole. This new design makes a highly effective sampling technology available on modern-day direct push sampling equipment to inform myriad questions about subsurface biogeochemistry processes. The re-design of the freezing drive shoe was supported by the USGS Innovation Center for Earth Sciences. References: Murphy, Fred, and W. N. Herkelrath. "A sample

  3. Use of Homogeneously-Sized Carbon Steel Ball Bearings to Study Microbially-Influenced Corrosion in Oil Field Samples

    PubMed Central

    Voordouw, Gerrit; Menon, Priyesh; Pinnock, Tijan; Sharma, Mohita; Shen, Yin; Venturelli, Amanda; Voordouw, Johanna; Sexton, Aoife

    2016-01-01

    Microbially-influenced corrosion (MIC) contributes to the general corrosion rate (CR), which is typically measured with carbon steel coupons. Here we explore the use of carbon steel ball bearings, referred to as beads (55.0 ± 0.3 mg; Ø = 0.238 cm), for determining CRs. CRs for samples from an oil field in Oceania incubated with beads were determined by the weight loss method, using acid treatment to remove corrosion products. The release of ferrous and ferric iron was also measured and CRs based on weight loss and iron determination were in good agreement. Average CRs were 0.022 mm/yr for eight produced waters with high numbers (105/ml) of acid-producing bacteria (APB), but no sulfate-reducing bacteria (SRB). Average CRs were 0.009 mm/yr for five central processing facility (CPF) waters, which had no APB or SRB due to weekly biocide treatment and 0.036 mm/yr for 2 CPF tank bottom sludges, which had high numbers of APB (106/ml) and SRB (108/ml). Hence, corrosion monitoring with carbon steel beads indicated that biocide treatment of CPF waters decreased the CR, except where biocide did not penetrate. The CR for incubations with 20 ml of a produced water decreased from 0.061 to 0.007 mm/yr when increasing the number of beads from 1 to 40. CRs determined with beads were higher than those with coupons, possibly also due to a higher weight of iron per unit volume used in incubations with coupons. Use of 1 ml syringe columns, containing carbon steel beads, and injected with 10 ml/day of SRB-containing medium for 256 days gave a CR of 0.11 mm/yr under flow conditions. The standard deviation of the distribution of residual bead weights, a measure for the unevenness of the corrosion, increased with increasing CR. The most heavily corroded beads showed significant pitting. Hence the use of uniformly sized carbon steel beads offers new opportunities for screening and monitoring of corrosion including determination of the distribution of corrosion rates, which allows

  4. Influence of DNA Extraction Method, 16S rRNA Targeted Hypervariable Regions, and Sample Origin on Microbial Diversity Detected by 454 Pyrosequencing in Marine Chemosynthetic Ecosystems

    PubMed Central

    Cruaud, Perrine; Vigneron, Adrien; Lucchetti-Miganeh, Céline; Ciron, Pierre Emmanuel; Godfroy, Anne

    2014-01-01

    Next-generation sequencing (NGS) opens up exciting possibilities for improving our knowledge of environmental microbial diversity, allowing rapid and cost-effective identification of both cultivated and uncultivated microorganisms. However, library preparation, sequencing, and analysis of the results can provide inaccurate representations of the studied community compositions. Therefore, all these steps need to be taken into account carefully. Here we evaluated the effects of DNA extraction methods, targeted 16S rRNA hypervariable regions, and sample origins on the diverse microbes detected by 454 pyrosequencing in marine cold seep and hydrothermal vent sediments. To assign the reads with enough taxonomic precision, we built a database with about 2,500 sequences from Archaea and Bacteria from deep-sea marine sediments, affiliated according to reference publications in the field. Thanks to statistical and diversity analyses as well as inference of operational taxonomic unit (OTU) networks, we show that (i) while DNA extraction methods do not seem to affect the results for some samples, they can lead to dramatic changes for others; and (ii) the choice of amplification and sequencing primers also considerably affects the microbial community detected in the samples. Thereby, very different proportions of pyrosequencing reads were obtained for some microbial lineages, such as the archaeal ANME-1, ANME-2c, and MBG-D and deltaproteobacterial subgroups. This work clearly indicates that the results from sequencing-based analyses, such as pyrosequencing, should be interpreted very carefully. Therefore, the combination of NGS with complementary approaches, such as fluorescence in situ hybridization (FISH)/catalyzed reporter deposition (CARD)-FISH or quantitative PCR (Q-PCR), would be desirable to gain a more comprehensive picture of environmental microbial communities. PMID:24837380

  5. Multipurpose floating platform for hyperspectral imaging, sampling and sensing of surface water sources used in irrigation and recreation

    USDA-ARS?s Scientific Manuscript database

    The objective of this work was to design, construct, and test the self-propelled aquatic platform for imaging, multi-tier water sampling, water quality sensing, and depth profiling to document microbial content and environmental covariates in the interior of irrigation ponds and reservoirs. The plat...

  6. How mutualisms arise in phytoplankton communities: building eco-evolutionary principles for aquatic microbes.

    PubMed

    Kazamia, Elena; Helliwell, Katherine Emma; Purton, Saul; Smith, Alison Gail

    2016-07-01

    Extensive sampling and metagenomics analyses of plankton communities across all aquatic environments are beginning to provide insights into the ecology of microbial communities. In particular, the importance of metabolic exchanges that provide a foundation for ecological interactions between microorganisms has emerged as a key factor in forging such communities. Here we show how both studies of environmental samples and physiological experimentation in the laboratory with defined microbial co-cultures are being used to decipher the metabolic and molecular underpinnings of such exchanges. In addition, we explain how metabolic modelling may be used to conduct investigations in reverse, deducing novel molecular exchanges from analysis of large-scale data sets, which can identify persistently co-occurring species. Finally, we consider how knowledge of microbial community ecology can be built into evolutionary theories tailored to these species' unique lifestyles. We propose a novel model for the evolution of metabolic auxotrophy in microorganisms that arises as a result of symbiosis, termed the Foraging-to-Farming hypothesis. The model has testable predictions, fits several known examples of mutualism in the aquatic world, and sheds light on how interactions, which cement dependencies within communities of microorganisms, might be initiated.

  7. Aquatic Therapy for Children

    ERIC Educational Resources Information Center

    Kucher, Greta; Moore, Kelsey; Rodia, Rachel; Moser, Christy Szczech

    2015-01-01

    Aquatic therapy has long been highlighted in the literature as a potentially powerful therapeutic intervention. This review will highlight basic definitions of aquatic therapy, review salient research, and identify specific diagnoses that may benefit from aquatic therapy. Online resources, blogs, and books that occupational therapists may find…

  8. Aquatic Therapy for Children

    ERIC Educational Resources Information Center

    Kucher, Greta; Moore, Kelsey; Rodia, Rachel; Moser, Christy Szczech

    2015-01-01

    Aquatic therapy has long been highlighted in the literature as a potentially powerful therapeutic intervention. This review will highlight basic definitions of aquatic therapy, review salient research, and identify specific diagnoses that may benefit from aquatic therapy. Online resources, blogs, and books that occupational therapists may find…

  9. Lopez-Alegria with Passive Observatories for Experimental Microbial Systems in Micro-G (POEMS) sample container

    NASA Image and Video Library

    2006-10-03

    ISS014-E-05124 (3 Oct. 2006) --- Astronaut Michael E. Lopez-Alegria, Expedition 14 commander and NASA space station science officer, works with the Passive Observatories for Experimental Microbial Systems in Micro-G (POEMS) payload in the Minus Eighty Degree Laboratory Freezer for ISS (MELFI) in the Destiny laboratory of the International Space Station. MELFI is a low temperature freezer facility with nominal operating temperatures of -80, -26 and +4 degrees Celsius that will preserve experiment materials over long periods.

  10. A novel derivation of a within-batch sampling plan based on a Poisson-gamma model characterising low microbial counts in foods.

    PubMed

    Gonzales-Barron, Ursula; Zwietering, Marcel H; Butler, Francis

    2013-02-01

    This study proposes a novel step-wise methodology for the derivation of a sampling plan by variables for food production systems characterised by relatively low concentrations of the inspected microorganism. After representing the universe of contaminated batches by modelling the between-batch and within-batch variability in microbial counts, a tolerance criterion defining batch acceptability (i.e., up to a tolerance percentage of the food units having microbial concentrations lower or equal to a critical concentration) is established to delineate a limiting quality contour that separates satisfactory from unsatisfactory batches. The problem consists then of finding the optimum decision criterion - arithmetic mean of the analytical results (microbiological limit, m(L)) and the sample size (n) - that satisfies a pre-defined level of confidence measured on the samples' mean distributions from all possible true within-batch distributions. This is approached by obtaining decision landscape curves representing collectively the conditional and joint producer's and consumer's risks at different microbiological limits along with confidence intervals representing uncertainty due to the propagated between-batch variability. Whilst the method requires a number of risk management decisions to be made such as the objective of the sampling plan (GMP-based or risk-based), the modality of derivation, the tolerance criterion or level of safety, and the statistical level of confidence, the proposed method can be used when past monitoring data are available so as to produce statistically-sound dynamic sampling plans with optimised efficiency and discriminatory power. For the illustration of Enterobacteriaceae concentrations on Irish sheep carcasses, a sampling regime of n=10 and m(L)=17.5CFU/cm(2) is recommended to ensure that the producer has at least a 90% confidence of accepting a satisfactory batch whilst the consumer at least a 97.5% confidence that a batch will not be

  11. Comparison of Microbial Community Compositions of Injection and Production Well Samples in a Long-Term Water-Flooded Petroleum Reservoir

    PubMed Central

    Ren, Hong-Yan; Zhang, Xiao-Jun; Song, Zhi-yong; Rupert, Wieger; Gao, Guang-Jun; Guo, Sheng-xue; Zhao, Li-Ping

    2011-01-01

    Water flooding plays an important role in recovering oil from depleted petroleum reservoirs. Exactly how the microbial communities of production wells are affected by microorganisms introduced with injected water has previously not been adequately studied. Using denaturing gradient gel electrophoresis (DGGE) approach and 16S rRNA gene clone library analysis, the comparison of microbial communities is carried out between one injection water and two production waters collected from a working block of the water-flooded Gudao petroleum reservoir located in the Yellow River Delta. DGGE fingerprints showed that the similarities of the bacterial communities between the injection water and production waters were lower than between the two production waters. It was also observed that the archaeal composition among these three samples showed no significant difference. Analysis of the 16S rRNA gene clone libraries showed that the dominant groups within the injection water were Betaproteobacteria, Gammaproteobacteria and Methanomicrobia, while the dominant groups in the production waters were Gammaproteobacteria and Methanobacteria. Only 2 out of 54 bacterial operational taxonomic units (OTUs) and 5 out of 17 archaeal OTUs in the injection water were detected in the production waters, indicating that most of the microorganisms introduced by the injection water may not survive to be detected in the production waters. Additionally, there were 55.6% and 82.6% unique OTUs in the two production waters respectively, suggesting that each production well has its specific microbial composition, despite both wells being flooded with the same injection water. PMID:21858049

  12. Comparison of microbial community compositions of injection and production well samples in a long-term water-flooded petroleum reservoir.

    PubMed

    Ren, Hong-Yan; Zhang, Xiao-Jun; Song, Zhi-yong; Rupert, Wieger; Gao, Guang-Jun; Guo, Sheng-xue; Zhao, Li-Ping

    2011-01-01

    Water flooding plays an important role in recovering oil from depleted petroleum reservoirs. Exactly how the microbial communities of production wells are affected by microorganisms introduced with injected water has previously not been adequately studied. Using denaturing gradient gel electrophoresis (DGGE) approach and 16S rRNA gene clone library analysis, the comparison of microbial communities is carried out between one injection water and two production waters collected from a working block of the water-flooded Gudao petroleum reservoir located in the Yellow River Delta. DGGE fingerprints showed that the similarities of the bacterial communities between the injection water and production waters were lower than between the two production waters. It was also observed that the archaeal composition among these three samples showed no significant difference. Analysis of the 16S rRNA gene clone libraries showed that the dominant groups within the injection water were Betaproteobacteria, Gammaproteobacteria and Methanomicrobia, while the dominant groups in the production waters were Gammaproteobacteria and Methanobacteria. Only 2 out of 54 bacterial operational taxonomic units (OTUs) and 5 out of 17 archaeal OTUs in the injection water were detected in the production waters, indicating that most of the microorganisms introduced by the injection water may not survive to be detected in the production waters. Additionally, there were 55.6% and 82.6% unique OTUs in the two production waters respectively, suggesting that each production well has its specific microbial composition, despite both wells being flooded with the same injection water.

  13. Aquatic bacterial assemblage structure in Pozas Azules, Cuatro Cienegas Basin, Mexico: Deterministic vs. stochastic processes.

    PubMed

    Espinosa-Asuar, Laura; Escalante, Ana Elena; Gasca-Pineda, Jaime; Blaz, Jazmín; Peña, Lorena; Eguiarte, Luis E; Souza, Valeria

    2015-06-01

    The aim of this study was to determine the contributions of stochastic vs. deterministic processes in the distribution of microbial diversity in four ponds (Pozas Azules) within a temporally stable aquatic system in the Cuatro Cienegas Basin, State of Coahuila, Mexico. A sampling strategy for sites that were geographically delimited and had low environmental variation was applied to avoid obscuring distance effects. Aquatic bacterial diversity was characterized following a culture-independent approach (16S sequencing of clone libraries). The results showed a correlation between bacterial beta diversity (1-Sorensen) and geographic distance (distance decay of similarity), which indicated the influence of stochastic processes related to dispersion in the assembly of the ponds' bacterial communities. Our findings are the first to show the influence of dispersal limitation in the prokaryotic diversity distribution of Cuatro Cienegas Basin. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.

  14. Microbial reduction of uranium

    USGS Publications Warehouse

    Lovley, D.R.; Phillips, E.J.P.; Gorby, Y.A.; Landa, E.R.

    1991-01-01

    REDUCTION of the soluble, oxidized form of uranium, U(VI), to insoluble U(IV) is an important mechanism for the immobilization of uranium in aquatic sediments and for the formation of some uranium ores1-10. U(VI) reduction has generally been regarded as an abiological reaction in which sulphide, molecular hydrogen or organic compounds function as the reductant1,2,5,11. Microbial involvement in U(VI) reduction has been considered to be limited to indirect effects, such as microbial metabolism providing the reduced compounds for abiological U(VI) reduction and microbial cell walls providing a surface to stimulate abiological U(VI) reduction1,12,13. We report here, however, that dissimilatory Fe(III)-reducing microorganisms can obtain energy for growth by electron transport to U(VI). This novel form of microbial metabolism can be much faster than commonly cited abiological mechanisms for U(VI) reduction. Not only do these findings expand the known potential terminal electron acceptors for microbial energy transduction, they offer a likely explanation for the deposition of uranium in aquatic sediments and aquifers, and suggest a method for biological remediation of environments contaminated with uranium.

  15. Toward richer metadata for microbial sequences: replacing strain-level NCBI taxonomy taxids with BioProject, BioSample and Assembly records.

    PubMed

    Federhen, Scott; Clark, Karen; Barrett, Tanya; Parkinson, Helen; Ostell, James; Kodama, Yuichi; Mashima, Jun; Nakamura, Yasukazu; Cochrane, Guy; Karsch-Mizrachi, Ilene

    2014-06-15

    Microbial genome sequence submissions to the International Nucleotide Sequence Database Collaboration (INSDC) have been annotated with organism names that include the strain identifier. Each of these strain-level names has been assigned a unique 'taxid' in the NCBI Taxonomy Database. With the significant growth in genome sequencing, it is not possible to continue with the curation of strain-level taxids. In January 2014, NCBI will cease assigning strain-level taxids. Instead, submitters are encouraged provide strain information and rich metadata with their submission to the sequence database, BioProject and BioSample.

  16. Inflammatory potential in relation to the microbial content of settled dust samples collected from moisture-damaged and reference schools: results of HITEA study.

    PubMed

    Huttunen, K; Tirkkonen, J; Täubel, M; Krop, E; Mikkonen, S; Pekkanen, J; Heederik, D; Zock, J-P; Hyvärinen, A; Hirvonen, M-R

    2016-06-01

    Aiming to identify factors causing the adverse health effects associated with moisture-damaged indoor environments, we analyzed immunotoxicological potential of settled dust from moisture-damaged and reference schools in relation to their microbiological composition. Mouse RAW264.7 macrophages were exposed to settled dust samples (n = 25) collected from moisture-damaged and reference schools in Spain, the Netherlands, and Finland. After exposure, we analyzed production of inflammatory markers [nitric oxide (NO), tumor necrosis factor-α (TNF-)α, interleukin (IL)-6, and macrophage inflammatory protein (MIP)2] as well as mitochondrial activity, viability, apoptosis, and cell cycle arrest. Furthermore, particle counts, concentration of selected microbial groups as well as chemical markers such as ergosterol, 3-hydroxy fatty acids, muramic acid, endotoxins, and glucans were measured as markers of exposure. Dust from moisture-damaged schools in Spain and the Netherlands induced stronger immunotoxicological responses compared to samples from reference schools; the responses to Finnish samples were generally lower with no difference between the schools. In multivariate analysis, IL-6 and apoptosis responses were most strongly associated with moisture status of the school. The measured responses correlated with several microbial markers and numbers of particles, but the most important predictor of the immunotoxicological potential of settled dust was muramic acid concentration, a marker of Gram-positive bacteria.

  17. Mass Spectrometry and Multiplex Antigen Assays to Assess Microbial Quality and Toxin Production of Staphylococcus aureus Strains Isolated from Clinical and Food Samples

    PubMed Central

    Attien, Paul; Sina, Haziz; Moussaoui, Wardi; Zimmermann-Meisse, Gaëlle; Dadié, Thomas; Keller, Daniel; Riegel, Philippe; Edoh, Vincent; Kotchoni, Simeon O.; Djè, Marcellin; Prévost, Gilles

    2014-01-01

    The aim of our study was to investigate the microbial quality of meat products and on some clinical samples in Abidjan focused on Staphylococcus genus and the toxin production profile of Staphylococcus aureus (S. aureus) isolated. Bacteria were collected from 240 samples of three meat products sold in Abidjan and 180 samples issued from clinical infections. The strains were identified by both microbiological and MALDI-TOF-MS methods. The susceptibility to antibiotics was determined by the disc diffusion method. The production of Panton-Valentine Leukocidin, LukE/D, and epidermolysins was screened using radial gel immunodiffusion. The production of staphylococcal enterotoxins and TSST-1 was screened by a Bio-Plex Assay. We observed that 96/240 of meat samples and 32/180 of clinical samples were contaminated by Staphylococcus. Eleven species were isolated from meats and 4 from clinical samples. Forty-two S. aureus strains were isolated from ours samples. Variability of resistance was observed for most of the tested antibiotics but none of the strains displays a resistance to imipenem and quinolones. We observed that 89% of clinical S. aureus were resistant to methicillin against 58% for those issued from meat products. All S. aureus isolates issued from meat products produce epidermolysins whereas none of the clinical strains produced these toxins. The enterotoxins were variably produced by both clinical and meat product samples. PMID:24987686

  18. In situ chemistry and microbial community compositions in five deep-sea hydrothermal fluid samples from Irina II in the Logatchev field.

    PubMed

    Perner, Mirjam; Gonnella, Giorgio; Hourdez, Stephane; Böhnke, Stefanie; Kurtz, Stefan; Girguis, Peter

    2013-05-01

    We present data on the co-registered geochemistry (in situ mass spectrometry) and microbiology (pyrosequencing of 16S rRNA genes; V1, V2, V3 regions) in five fluid samples from Irina II in the Logatchev hydrothermal field. Two samples were collected over 24 min from the same spot and further three samples were from spatially distinct locations (20 cm, 3 m and the overlaying plume). Four low-temperature hydrothermal fluids from the Irina II are composed of the same core bacterial community, namely specific Gammaproteobacteria and Epsilonproteobacteria, which, however, differs in the relative abundance. The microbial composition of the fifth sample (plume) is considerably different. Although a significant correlation between sulfide enrichment and proportions of Sulfurovum (Epsilonproteobacteria) was found, no other significant linkages between abiotic factors, i.e. temperature, hydrogen, methane, sulfide and oxygen, and bacterial lineages were evident. Intriguingly, bacterial community compositions of some time series samples from the same spot were significantly more similar to a sample collected 20 cm away than to each other. Although this finding is based on three single samples only, it provides first hints that single hydrothermal fluid samples collected on a small spatial scale may also reflect unrecognized temporal variability. However, further studies are required to support this hypothesis.

  19. Counting viruses and bacteria in photosynthetic microbial mats.

    PubMed

    Carreira, Cátia; Staal, Marc; Middelboe, Mathias; Brussaard, Corina P D

    2015-03-01

    Viral abundances in benthic environments are the highest found in aquatic systems. Photosynthetic microbial mats represent benthic environments with high microbial activity and possibly high viral densities, yet viral abundances have not been examined in such systems. Existing extraction procedures typically used in benthic viral ecology were applied to the complex matrix of microbial mats but were found to inefficiently extract viruses. Here, we present a method for extraction and quantification of viruses from photosynthetic microbial mats using epifluorescence microscopy (EFM) and flow cytometry (FCM). A combination of EDTA addition, probe sonication, and enzyme treatment applied to a glutaraldehyde-fixed sample resulted in a substantially higher viral (5- to 33-fold) extraction efficiency and reduced background noise compared to previously published methods. Using this method, it was found that in general, intertidal photosynthetic microbial mats harbor very high viral abundances (2.8 × 10(10) ± 0.3 × 10(10) g(-1)) compared with benthic habitats (10(7) to 10(9) g(-1)). This procedure also showed 4.5- and 4-fold-increased efficacies of extraction of viruses and bacteria, respectively, from intertidal sediments, allowing a single method to be used for the microbial mat and underlying sediment. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Counting Viruses and Bacteria in Photosynthetic Microbial Mats

    PubMed Central

    Staal, Marc; Middelboe, Mathias; Brussaard, Corina P. D.

    2015-01-01

    Viral abundances in benthic environments are the highest found in aquatic systems. Photosynthetic microbial mats represent benthic environments with high microbial activity and possibly high viral densities, yet viral abundances have not been examined in such systems. Existing extraction procedures typically used in benthic viral ecology were applied to the complex matrix of microbial mats but were found to inefficiently extract viruses. Here, we present a method for extraction and quantification of viruses from photosynthetic microbial mats using epifluorescence microscopy (EFM) and flow cytometry (FCM). A combination of EDTA addition, probe sonication, and enzyme treatment applied to a glutaraldehyde-fixed sample resulted in a substantially higher viral (5- to 33-fold) extraction efficiency and reduced background noise compared to previously published methods. Using this method, it was found that in general, intertidal photosynthetic microbial mats harbor very high viral abundances (2.8 × 1010 ± 0.3 × 1010 g−1) compared with benthic habitats (107 to 109 g−1). This procedure also showed 4.5- and 4-fold-increased efficacies of extraction of viruses and bacteria, respectively, from intertidal sediments, allowing a single method to be used for the microbial mat and underlying sediment. PMID:25595761

  1. Metaproteomics to investigate the impact of sampling-site biogeochemistry on structure and functionality of leaf-litter degrading microbial communities

    NASA Astrophysics Data System (ADS)

    Schneider, Thomas; Keiblinger, Katharina; Gerrits, Bertran; Schmid, Emanuel; Eberl, Leo; Zechmeister-Boltenstern, Sophie; Riedel, Kathrin

    2010-05-01

    The composition of organic matter in natural ecosystems is strongly influenced by the microorganisms present. Conversely, bacteria and fungi are limited by the amount and type of organic matter available in a given environment, most of which is ultimately derived from plants. Changes in the stoichiometry and biochemical constituents of plant litter may therefore alter species composition and elicit changes in the activities of microbial communities and their component parts. The identification of the microbial proteins of a given habitat together with the analysis of their phylogenetic origin and their spatial and temporal distribution are expected to provide fundamentally new insights into the role of microbial diversity in biogeochemical processes. To relate structure and functionality of microbial communities involved in leaf-litter decomposition we determined biogeochemistry, community structure by phospholipid fatty acid (PLFA)-analyses, enzymatic activities, and analysed the protein complement of different litter types, which were collected in winter and spring at various Austrian sampling sites, in a semi-quantitative proteomics approach by one dimensional polyacrylamide gel electrophoresis (1-D-SDS-PAGE) combined with liquid chromatography/tandem mass-spectrometry (LC-MS/MS). Protein abundances were determined by counting the number of MS/MS spectra assigned to each protein. In samples with high manganese and phosphor content a significant increase of fungal proteins from February to May was observed, which was in good agreement with the PLFA-analyses showing similar trends towards an increase of the fungal community. In contrast, the PLFA analysis revealed no temporal changes in the community at Achenkirch and even a decrease in the fungal/bacterial ratio at Klausen-Leopoldsdorf, two sampling sites low in P and Mn; similar trends are reflected in our spectral counts. In conclusion, semi-quantitative proteome- and PLFA-analyses suggest that fungal and

  2. Soil Fertilization Leads to a Decline in Between-Samples Variability of Microbial Community δ13C Profiles in a Grassland Fertilization Experiment

    PubMed Central

    Veresoglou, Stavros D.; Thornton, Barry; Menexes, George; Mamolos, Andreas P.; Veresoglou, Demetrios S.

    2012-01-01

    Gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) was used to measure the 13C/12C ratios of PLFAs at natural abundance levels from a temperate grassland nitrogen (N) and phosphorus (P) factorial fertilization experiment in northern Greece. In each plot two rhizosphere samples were derived centred around individual Agrostis capillaris and Prunella vulgaris plants. It was hypothesized that the isotopic signal of microbes that preferentially feed on recalcitrant litter such as fungi would be modified by fertilization more strongly than that of opportunistic microbes using labile C. Microbial community δ13C was affected by both P and N fertilization regime and plant species identity. However, we have been unable to detect significant nutrient effects on individual groups of microbes when analyzed separately in contrast to our original hypothesis. Intra-treatment variability, as evaluated from Hartley’s Fmax tests in the five first PCA components axes as well as the size of the convex hulls in PCA scoreplots and Mahalanobis distances, was considerably higher in the non-fertilized controls. Moreover, a significant relationship was established between the change in PLFA abundances and their respective changes in δ13C for the aggregate of samples and those simultaneously fertilized with N and P. We conclude that use of compound specific isotope analysis in the absence of labelling represents a valuable and overlooked tool in obtaining an insight of microbial community functioning. PMID:22962602

  3. Phages across the biosphere: contrasts of viruses in soil and aquatic environments.

    PubMed

    Srinivasiah, Sharath; Bhavsar, Jaysheel; Thapar, Kanika; Liles, Mark; Schoenfeld, Tom; Wommack, K Eric

    2008-06-01

    Despite the predominance of aquatic environments on the planet Earth, microbial abundance and diversity within soil environments exceed that of the aquatic realm. Most of what we know of viral ecology within natural systems has come through investigations of aquatic environments. However, the 'aquatic-bias' in viral ecology is beginning to change as the cultivation-independent approaches, which revealed the extraordinary abundance and diversity of viruses within aquatic systems, are now being applied to soils. This review briefly summarizes recent investigations of viral abundance and diversity in soil environments.

  4. Which Microbial Communities Are Present? Using Fluorescence In Situ Hybridisation (FISH): Microscopic Techniques for Enumeration of Troublesome Microorganisms in Oil and Fuel Samples

    NASA Astrophysics Data System (ADS)

    Holmkvist, Lars; Østergaard, Jette Johanne; Skovhus, Torben Lund

    Enumeration of microbes involved in souring of oil fields and microbiologically influenced corrosion (MIC) with culture-based methods, usually yield inadequate and contradictory results. Any cultivation step will almost certainly alter the population structure of the sample and thus the results of cultivation analysis are not a good basis for mitigation decisions. The need for methods that are cultivation independent has over the past 10 years facilitated the development of several analytical methods for determination of bacterial identity, quantity, and to some extent function, applied directly to samples of the native population. In this chapter, we demonstrate the features and benefits of applying microscopic techniques to a situation often encountered in the oil and petroleum industry: Control of microbial growth in fuel storage tanks. The methods described in this chapter will focus on direct counts of specific groups of microorganisms with microscopy and these are based on the detection of genetic material and not on culturing.

  5. Ubiquitous Detection of Artificial Sweeteners and Iodinated X-ray Contrast Media in Aquatic Environmental and Wastewater Treatment Plant Samples from Vietnam, The Philippines, and Myanmar.

    PubMed

    Watanabe, Yuta; Bach, Leu Tho; Van Dinh, Pham; Prudente, Maricar; Aguja, Socorro; Phay, Nyunt; Nakata, Haruhiko

    2016-05-01

    Water samples from Vietnam, The Philippines, and Myanmar were analyzed for artificial sweeteners (ASs) and iodinated X-ray contrast media (ICMs). High concentrations (low micrograms per liter) of ASs, including aspartame, saccharin, and sucralose, were found in wastewater treatment plant (WWTP) influents from Vietnam. Three ICMs, iohexol, iopamidol, and iopromide were detected in Vietnamese WWTP influents and effluents, suggesting that these ICMs are frequently used in Vietnam. ASs and ICMs were found in river water from downtown Hanoi at concentrations comparable to or lower than the concentrations in WWTP influents. The ASs and ICMs concentrations in WWTP influents and adjacent surface water significantly correlated (r (2) = 0.99, p < 0.001), suggesting that household wastewater is discharged directly into rivers in Vietnam. Acesulfame was frequently detected in northern Vietnamese groundwater, but the concentrations varied spatially by one order of magnitude even though the sampling points were very close together. This implies that poorly performing domestic septic tanks sporadically leak household wastewater into groundwater. High acesulfame, cyclamate, saccharin, and sucralose concentrations were found in surface water from Manila, The Philippines. The sucralose concentrations were one order of magnitude higher in the Manila samples than in the Vietnamese samples, indicating that more sucralose is used in The Philippines than in Vietnam. Acesulfame and cyclamate were found in surface water from Pathein (rural) and Yangon (urban) in Myanmar, but no ICMs were found in the samples. The ASs concentrations were two-three orders of magnitude lower in the samples from Myanmar than in the samples from Vietnam and The Philippines, suggesting that different amounts of ASs are used in these countries. We believe this is the first report of persistent ASs and ICMs having ubiquitous distributions in economically emerging South Asian countries.

  6. Acute aquatic toxicity studies of Gulf of Mexico water samples collected following the Deepwater Horizon incident (May 12, 2010 to December 11, 2010).

    PubMed

    Echols, B S; Smith, A J; Gardinali, P R; Rand, G M

    2015-02-01

    The potential for the Deepwater Horizon MC-252 oil incident to affect ecosystems in the Gulf of Mexico (GOM) was evaluated using Americamysis bahia, Menidia beryllina and Vibrio fischeri (Microtox® assay). Organisms were exposed to GOM water samples collected in May-December 2010. Samples were collected where oil was visibly present on the water surface or the presence of hydrocarbons at depth was indicated by fluorescence data or reduced dissolved oxygen. Toxicity tests were conducted using water-accommodated fractions (WAFs), and oil-in-water dispersions (OWDs). Water samples collected from May to June 2010 were used for screening tests, with OWD samples slightly more acutely toxic than WAFs. Water samples collected in July through December 2010 were subjected to definitive acute testing with both species. In A. bahia tests, total PAH concentrations for OWD exposures ranged from non-detect to 23.0 μg L(-1), while WAF exposures ranged from non-detect to 1.88 μg L(-1). Mortality was >20% in five OWD exposures with A. bahia and three of the WAF definitive tests. Total PAH concentrations were lower for M. beryllina tests, ranging from non-detect to 0.64 μg L(-1) and non-detect to 0.17 μg L(-1) for OWD and WAF exposures, respectively. Only tests from two water samples in both the WAFs and OWDs exhibited >20% mortality to M. beryllina. Microtox® assays showed stimulatory and inhibitory responses with no relationship with PAH exposure concentrations. Most mortality in A. bahia and M. beryllina occurred in water samples collected before the well was capped in July 2010 with a clear decline in mortality associated with a decline in total PAH water concentrations.

  7. Resorcinol-formaldehyde xerogel as a micro-solid-phase extraction sorbent for the determination of herbicides in aquatic environmental samples.

    PubMed

    Es-haghi, Ali; Zare, Maryam; Piri-Moghadam, Hamed; Bagheri, Habib

    2015-07-01

    In this study, organic aerogels were synthesized by the sol-gel polycondensation of mixed cresol with formaldehyde in a slightly basic aqueous solution. Carbon aerogels and xerogels are generated by pyrolysis of organic aerogels. The novel sol-gel-based micro-solid-phase extraction sorbent, resorcinol-formaldehyde xerogel, was employed for preconcentration of some selected herbicides. Three herbicides of the aryloxyphenoxypropionate group, clodinafop-propargyl, haloxyfop-etotyl, and fenoxaprop-P-ethyl, were extracted from aqueous samples by micro-solid-phase extraction and subsequently determined by gas chromatography with mass spectrometry. The effect of different parameters influencing the extraction efficiency of these herbicides including sample flow rate, sample volume, and extraction time were investigated and optimized. Under optimum conditions, linear calibration curves in the range of 0.10-500 ng/L with R(2) > 0.99 were obtained. The relative standard deviation at 50 μg/L concentration level was lower than 10% (n = 5) and detection limits were between 0.05 and 0.20 μg/L. The proposed method was successfully applied to the sampling and extraction of herbicides from Zayanderood and paddy water samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Headspace solvent microextraction as a simple and highly sensitive sample pretreatment technique for ultra trace determination of geosmin in aquatic media.

    PubMed

    Bagheri, Habib; Salemi, Amir

    2006-01-01

    A headspace solvent microextraction method was developed for the trace determination of geosmin, an odorant compound, in water samples. After performing the extraction by a microdrop of an organic solvent, the microdrop was introduced directly into a GC-MS injection port. One-at-the-time optimization strategy was applied to investigate and optimize some important extraction parameters such as type of solvent, drop volume, temperature, stirring rate, ionic strength, sample volume, and extraction time. The analytical data exhibited an RSD of less than 5% (n = 5), a linear calibration range of 5-900 ng/L (r2 > 0.998), and a detection limit of 0.8 and 3.3 ng/L using two different sets of selected ions. The proposed method was successfully applied to the extraction and determination of geosmin in the spiked real water sample and reasonable recovery was achieved.

  9. Aquatic hyphomycete communities as potential bioindicators for assessing anthropogenic stress.

    PubMed

    Solé, M; Fetzer, I; Wennrich, R; Sridhar, K R; Harms, H; Krauss, G

    2008-01-25

    With a profound knowledge of how physico-chemical parameters affect these communities, microbial communities could be used as indicators for environmental changes and for risk assessment studies. We studied aquatic hyphomycete communities in rivers and aquifers from sites shaped by intense mining activities (namely the "Mansfeld region") and chemical industry (cities of Halle and Bitterfeld) in Central Germany. Environmental stress factors such as high concentrations of heavy metals, sulphate, and nitrate as well as low concentrations of oxygen significantly reduced the diversity and biomass of hyphomycetes in the investigated samples. Redundancy analysis (RDA) indicates that variations in water chemistry cause a significant proportion of the change in fungal community structure (86.2%). Fungi were negatively correlated with high metal and nutrient concentrations. RDA also showed a strong influence of organic matter on individual species, with Anguillospora longissima (Sacc. et Syd.), Clavatospora longibrachiata (Ingold), Clavariopsis aquatica (De Wild), Flagellospora curvula (Ingold), Heliscus lugdunensis (Sacc. et Thérry), Tumularia aquatica (Ingold) and Lemonniera aquatica (De Wild) being most sensitive. We propose that aquatic hyphomycete communities can be used as sensitive and integrative indicators for freshwater quality.

  10. Characterization of natural organic matter as major constituents in aquatic systems

    NASA Astrophysics Data System (ADS)

    Frimmel, F. H.

    1998-12-01

    Natural organic matter (NOM) is ubiquitous in global aquatic systems, the mass concentrations ranging from 0.5 to 100 mg/l of organic carbon. The polydispersity of molar masses and the chemical structures comprising NOM give it a multifunctional role in natural environment and in water treatment processes. Important functions include serving as an electron donor in metal complexation, sorption of xenobiotics and adsorption onto mineral phases and onto activated carbon. NOM is partially oxidized during microbial utilization and during water treatment in which it may also become substituted with chlorine leading to a suite of products with toxic relevance. Meaningful methods of NOM characterization would be useful for the development of a predictive capacity for NOM behaviour in different water sources. Among analytical characterization methods, those directly applicable to aqueous samples are most useful and in addition to classical spectroscopic methods, more advanced methods have become available within the last decade. High pressure liquid chromatography using gels have proved useful in combination with UV/vis, fluorescence, light scattering and sensitive dissolved organic carbon detection techniques, yielding information on molecular absorbance, size distribution, molar mass and reactivity. Information on biodegradability of NOM can be deduced from experimental measurement of bacterial growth under defined conditions. The nature and amount of biologically assimilable organic carbon (AOC) in combination with the bacterial cell number and growth rate constants can provide a meaningful characterization of microbial stability in aquatic systems. In addition, determination of directly available and acid or enzymatically hydrolysable amino acids and carbohydrates can add to the understanding of NOM biodegradability over different time scales. The paper gives the results obtained by the application of the different methods for the characterization of aquatic NOM and

  11. Interlaboratory comparison of three microbial source tracking quantitative polymerase chain reaction (qPCR) assays from fecal-source and environmental samples

    USGS Publications Warehouse

    Stelzer, Erin A.; Strickler, Kriston M.; Schill, William B.

    2012-01-01

    During summer and early fall 2010, 15 river samples and 6 fecal-source samples were collected in West Virginia. These samples were analyzed by three laboratories for three microbial source tracking (MST) markers: AllBac, a general fecal indicator; BacHum, a human-associated fecal indicator; and BoBac, a ruminant-associated fecal indicator. MST markers were analyzed by means of the quantitative polymerase chain reaction (qPCR) method. The aim was to assess interlaboratory precision when the three laboratories used the same MST marker and shared deoxyribonucleic acid (DNA) extracts of the samples, but different equipment, reagents, and analyst experience levels. The term assay refers to both the markers and the procedure differences listed above. Interlaboratory precision was best for all three MST assays when using the geometric mean absolute relative percent difference (ARPD) and Friedman's statistical test as a measure of interlaboratory precision. Adjustment factors (one for each MST assay) were calculated using results from fecal-source samples analyzed by all three laboratories and applied retrospectively to sample concentrations to account for differences in qPCR results among labs using different standards and procedures. Following the application of adjustment factors to qPCR results, ARPDs were lower; however, statistically significant differences between labs were still observed for the BacHum and BoBac assays. This was a small study and two of the MST assays had 52 percent of samples with concentrations at or below the limit of accurate quantification; hence, more testing could be done to determine if the adjustment factors would work better if the majority of sample concentrations were above the quantification limit.

  12. Modified RNA-seq method for microbial community and diversity analysis using rRNA in different types of environmental samples.

    PubMed

    Yan, Yong-Wei; Zou, Bin; Zhu, Ting; Hozzein, Wael N; Quan, Zhe-Xue

    2017-01-01

    RNA-seq-based SSU (small subunit) rRNA (ribosomal RNA) analysis has provided a better understanding of potentially active microbial community within environments. However, for RNA-seq library construction, high quantities of purified RNA are typically required. We propose a modified RNA-seq method for SSU rRNA-based microbial community analysis that depends on the direct ligation of a 5' adaptor to RNA before reverse-transcription. The method requires only a low-input quantity of RNA (10-100 ng) and does not require a DNA removal step. The method was initially tested on three mock communities synthesized with enriched SSU rRNA of archaeal, bacterial and fungal isolates at different ratios, and was subsequently used for environmental samples of high or low biomass. For high-biomass salt-marsh sediments, enriched SSU rRNA and total nucleic acid-derived RNA-seq datasets revealed highly consistent community compositions for all of the SSU rRNA sequences, and as much as 46.4%-59.5% of 16S rRNA sequences were suitable for OTU (operational taxonomic unit)-based community and diversity analyses with complete coverage of V1-V2 regions. OTU-based community structures for the two datasets were also highly consistent with those determined by all of the 16S rRNA reads. For low-biomass samples, total nucleic acid-derived RNA-seq datasets were analyzed, and highly active bacterial taxa were also identified by the OTU-based method, notably including members of the previously underestimated genus Nitrospira and phylum Acidobacteria in tap water, members of the phylum Actinobacteria on a shower curtain, and members of the phylum Cyanobacteria on leaf surfaces. More than half of the bacterial 16S rRNA sequences covered the complete region of primer 8F, and non-coverage rates as high as 38.7% were obtained for phylum-unclassified sequences, providing many opportunities to identify novel bacterial taxa. This modified RNA-seq method will provide a better snapshot of diverse microbial

  13. Aquatic species and habitats

    Treesearch

    Danny C. Lee; James R. Sedell; Bruce E. Rieman; Russell F. Thurow; Jack E. Williams

    1998-01-01

    Continuing human activities threaten the highly prized aquatic resources of the interior Columbia basin. Precipitous declines in native species, particularly Pacific salmon, and a large influx of introduced species have radically altered the composition and distribution of native fishes. Fortunately, areas of relatively high aquatic integrity remain, much of it on...

  14. Aquatic Activities for Youth.

    ERIC Educational Resources Information Center

    Greene, H. David; And Others

    Designed to meet the diverse educational needs of youth groups, this aquatic program consists of eight individual lesson units, each devoted to one aspect of the aquatic world. Unit topics include: fish aquariums; raising earthworms; simulation of coastal planning; entomology and water; rope; calculating stream flow; saltwater aquariums; and fish…

  15. Aquatic Activities for Youth.

    ERIC Educational Resources Information Center

    Greene, H. David; And Others

    Designed to meet the diverse educational needs of youth groups, this aquatic program consists of eight individual lesson units, each devoted to one aspect of the aquatic world. Unit topics include: fish aquariums; raising earthworms; simulation of coastal planning; entomology and water; rope; calculating stream flow; saltwater aquariums; and fish…

  16. Concentrations of cadmium, lead, and zinc in fish from mining-influenced waters of northeastern Oklahoma: Sampling of blood, carcass, and liver for aquatic biomonitoring

    USGS Publications Warehouse

    Brumbaugh, W.G.; Schmitt, C.J.; May, T.W.

    2005-01-01

    The Tri-States Mining District (TSMD) of Missouri (MO), Kansas (KS), and Oklahoma (OK), USA, was mined for lead (Pb) and zinc (Zn) for more than a century. Mining ceased more than 30 years ago, but wastes remain widely distributed in the region, and there is evidence of surface- and groundwater contamination in the Spring River-Neosho River (SR-NR) system of northeastern OK. In October 2001, we collected a total of 74 fish from six locations in the SR-NR system that included common carp (Cyprinus carpio), channel- and flathead catfish (Ictalurus punctatus and Pylodictis olivaris), largemouth- and spotted bass (Micropterus salmoides and Micropterus punctulatus), and white crappie (Pomoxis annularis). We obtained additional fish from locations in MO that included three reference sites and one site that served as a "positive control" (heavily contaminated by Pb). Blood, carcass (headed, eviscerated, and scaled) and liver (carp only) samples were analyzed for cadmium (Cd), Pb, and Zn. Our objectives were to assess the degree to which fish from the OK portion of the SR-NR system are contaminated by these elements and to evaluate fish blood sampling for biomonitoring. Concentrations of Cd and Pb in carp and catfish from OK sites were elevated and Pb concentrations of some approached those of the highly contaminated site in MO, but concentrations in bass and crappie were relatively low. For Zn, correlations were weak among concentrations in the three tissues and none of the samples appeared to reflect site contamination. Variability was high for Cd in all three tissues of carp; differences between sites were statistically significant (p < 0.05) only for blood even though mean liver concentrations were at least 100-fold greater than those in blood. Blood concentrations of Cd and Pb were positively correlated (r 2 = 0.49 to 0.84) with the concentration of the same element in carp and catfish carcasses or in carp livers, and the corresponding multiple regression models were

  17. Concentrations of cadmium, lead, and zinc in fish from mining-influenced waters of northeastern Oklahoma: sampling of blood, carcass, and liver for aquatic biomonitoring.

    PubMed

    Brumbaugh, William G; Schmitt, Christopher J; May, Thomas W

    2005-07-01

    The Tri-States Mining District (TSMD) of Missouri (MO), Kansas (KS), and Oklahoma (OK), USA, was mined for lead (Pb) and zinc (Zn) for more than a century. Mining ceased more than 30 years ago, but wastes remain widely distributed in the region, and there is evidence of surface- and groundwater contamination in the Spring River-Neosho River (SR-NR) system of northeastern OK. In October 2001, we collected a total of 74 fish from six locations in the SR-NR system that included common carp (Cyprinus carpio), channel- and flathead catfish (Ictalurus punctatus and Pylodictis olivaris), largemouth- and spotted bass (Micropterus salmoides and Micropterus punctulatus), and white crappie (Pomoxis annularis). We obtained additional fish from locations in MO that included three reference sites and one site that served as a "positive control" (heavily contaminated by Pb). Blood, carcass (headed, eviscerated, and scaled) and liver (carp only) samples were analyzed for cadmium (Cd), Pb, and Zn. Our objectives were to assess the degree to which fish from the OK portion of the SR-NR system are contaminated by these elements and to evaluate fish blood sampling for biomonitoring. Concentrations of Cd and Pb in carp and catfish from OK sites were elevated and Pb concentrations of some approached those of the highly contaminated site in MO, but concentrations in bass and crappie were relatively low. For Zn, correlations were weak among concentrations in the three tissues and none of the samples appeared to reflect site contamination. Variability was high for Cd in all three tissues of carp; differences between sites were statistically significant (p < 0.05) only for blood even though mean liver concentrations were at least 100-fold greater than those in blood. Blood concentrations of Cd and Pb were positively correlated (r2 = 0.49 to 0.84) with the concentration of the same element in carp and catfish carcasses or in carp livers, and the corresponding multiple regression models were

  18. Diversity of microbial eukaryotes in sediment at a deep-sea methane cold seep: surveys of ribosomal DNA libraries from raw sediment samples and two enrichment cultures.

    PubMed

    Takishita, Kiyotaka; Yubuki, Naoji; Kakizoe, Natsuki; Inagaki, Yuji; Maruyama, Tadashi

    2007-07-01

    Recent culture-independent surveys of eukaryotic small-subunit ribosomal DNA (SSU rDNA) from many environments have unveiled unexpectedly high diversity of microbial eukaryotes (microeukaryotes) at various taxonomic levels. However, such surveys were most probably biased by various technical difficulties, resulting in underestimation of microeukaryotic diversity. In the present study on oxygen-depleted sediment from a deep-sea methane cold seep of Sagami Bay, Japan, we surveyed the diversity of eukaryotic rDNA in raw sediment samples and in two enrichment cultures. More than half of all clones recovered from the raw sediment samples were of the basidiomycetous fungus Cryptococcus curvatus. Among other clones, phylotypes of eukaryotic parasites, such as Apicomplexa, Ichthyosporea, and Phytomyxea, were identified. On the other hand, we observed a marked difference in phylotype composition in the enrichment samples. Several phylotypes belonging to heterotrophic stramenopiles were frequently found in one enrichment culture, while a phylotype of Excavata previously detected at a deep-sea hydrothermal vent dominated the other. We successfully established a clonal culture of this excavate flagellate. Since these phylotypes were not identified in the raw sediment samples, the approach incorporating a cultivation step successfully found at least a fraction of the "hidden" microeukaryotic diversity in the environment examined.

  19. New nanostructure of polydimethylsiloxane coating as a solid-phase microextraction fiber: Application to analysis of BTEX in aquatic environmental samples.

    PubMed

    Zali, Sara; Jalali, Fahimeh; Es-Haghi, Ali; Shamsipur, Mojtaba

    2016-10-15

    Electrospinning technique was used to convert polydimethyl siloxane (PDMS) sol-gel solution to a new nanostructure on a stainless steel wire. The surface morphology of the fiber was observed by scanning electron microscopy (SEM). It showed a diameter range of 30-60nm for PDMS nanoparticles with a homogeneous and porous surface structure. The applicability of this coating was assessed for the headspace SPME (HS-SPME) of benzene, toluene, ethylbenzene and xylenes (BTEX) from water samples followed by gas chromatography-mass spectrometry. The important parameters affecting extraction efficiency such as extraction time and temperature, desorption conditions, agitation rate and ionic strength were investigated and optimized. Under the optimized conditions, LODs and LOQs of 0.3-5μgL(-1) and 1-10μgL(-1) were obtained, respectively. The method showed linearity in the broad range of 1-5000μgL(-1) with correlation coefficient of >0.99. Inter-day and intra-day precisions of the developed method ranged from 2.43% to 6.54% and from 5.24% to 13.73%, respectively. The thermal stability of the fiber was investigated on stainless steel wire. It was found to be durable at 260°C for more than 360min. Furthermore, the proposed method was successfully applied for quantification of BTEX in real water samples.

  20. Occurrence of microbial indicators and Clostridium perfringens in wastewater, water column samples, sediments, drinking water, and Weddell seal feces collected at McMurdo Station, Antarctica

    USGS Publications Warehouse

    Lisle, J.T.; Smith, J.J.; Edwards, D.D.; McFeters, G.A.

    2004-01-01

    McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered.

  1. Occurrence of microbial indicators and Clostridium perfringens in wastewater, water column samples, sediments, drinking water, and Weddell seal feces collected at McMurdo Station, Antarctica.

    PubMed

    Lisle, John T; Smith, James J; Edwards, Diane D; McFeters, Gordon A

    2004-12-01

    McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered.

  2. Occurrence of Microbial Indicators and Clostridium perfringens in Wastewater, Water Column Samples, Sediments, Drinking Water, and Weddell Seal Feces Collected at McMurdo Station, Antarctica

    PubMed Central

    Lisle, John T.; Smith, James J.; Edwards, Diane D.; McFeters, Gordon A.

    2004-01-01

    McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered. PMID:15574926

  3. Experimental Investigation of Microbially Induced Corrosion of Test Samples and Effect of Self-Assembled Hydrophobic Monolayers. Exposure of Test Samples to Continuous Microbial Cultures, Chemical Analysis, and Biochemical Studies

    SciTech Connect

    Laurinavichius, K.S.

    1998-09-30

    The study of biocorrosion of aluminum and beryllium samples were performed under conditions of continuous fermentation of thermophilic anaerobic microorganisms of different groups. This allowed us to examine the effect of various types of metabolic reactions of reduction-oxidation proceeding at different pH and temperatures under highly reduced conditions on aluminum and beryllium corrosion and effect of self-assembled hydrophobic monolayers.

  4. Which Microbial Communities are Present? Application of PCR-DGGE: Case Study on an Oilfield Core Sample

    NASA Astrophysics Data System (ADS)

    van der Kraan, Geert M.; Buijzen, Floris; Kuijvenhoven, Cor A. T.; Muyzer, Gerard

    In this case study a PCR-DGGE analysis has been performed on an oilfield core sample. In many scientific articles PCR-DGGE analysis is often referred to as one analysis technique, but in fact it is a combination of the PCR and DGGE techniques performed in succession (Muyzer et al., 1993).

  5. Water Quality Criteria for Human Health and Aquatic Life

    EPA Science Inventory

    Collaborative effort with the Office of Water to provide science in support of the development and implementation of new or revised ambient water quality criteria for microbial and chemical contaminants for human health and aquatic life. The research also addresses implementation...

  6. Water Quality Criteria for Human Health and Aquatic Life

    EPA Science Inventory

    Collaborative effort with the Office of Water to provide science in support of the development and implementation of new or revised ambient water quality criteria for microbial and chemical contaminants for human health and aquatic life. The research also addresses implementation...

  7. Selective preservation and origin of petroleum-forming aquatic kerogen

    USGS Publications Warehouse

    Hatcher, P.G.; Spiker, E. C.; Szeverenyi, N.M.; Maciel, G.E.

    1983-01-01

    Studies of a marine algal sapropel from Mangrove Lake, Bermuda, by 13C NMR and stable carbon isotopic methods show that precursors of aquatic kerogen (insoluble, macromolecular, paraffinic humic substances) are primary components of algae and possibly associated bacteria and that these substances survive microbial decomposition and are selectively preserved during early diagenesis. ?? 1983 Nature Publishing Group.

  8. Random sample consensus combined with partial least squares regression (RANSAC-PLS) for microbial metabolomics data mining and phenotype improvement.

    PubMed

    Teoh, Shao Thing; Kitamura, Miki; Nakayama, Yasumune; Putri, Sastia; Mukai, Yukio; Fukusaki, Eiichiro

    2016-08-01

    In recent years, the advent of high-throughput omics technology has made possible a new class of strain engineering approaches, based on identification of possible gene targets for phenotype improvement from omic-level comparison of different strains or growth conditions. Metabolomics, with its focus on the omic level closest to the phenotype, lends itself naturally to this semi-rational methodology. When a quantitative phenotype such as growth rate under stress is considered, regression modeling using multivariate techniques such as partial least squares (PLS) is often used to identify metabolites correlated with the target phenotype. However, linear modeling techniques such as PLS require a consistent metabolite-phenotype trend across the samples, which may not be the case when outliers or multiple conflicting trends are present in the data. To address this, we proposed a data-mining strategy that utilizes random sample consensus (RANSAC) to select subsets of samples with consistent trends for construction of better regression models. By applying a combination of RANSAC and PLS (RANSAC-PLS) to a dataset from a previous study (gas chromatography/mass spectrometry metabolomics data and 1-butanol tolerance of 19 yeast mutant strains), new metabolites were indicated to be correlated with tolerance within certain subsets of the samples. The relevance of these metabolites to 1-butanol tolerance were then validated from single-deletion strains of corresponding metabolic genes. The results showed that RANSAC-PLS is a promising strategy to identify unique metabolites that provide additional hints for phenotype improvement, which could not be detected by traditional PLS modeling using the entire dataset. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  9. Microbial community composition and endolith colonization at an Arctic thermal spring are driven by calcite precipitation

    USGS Publications Warehouse

    Starke, Verena; Kirshtein, Julie; Fogel, Marilyn L.; Steele, Andrew

    2013-01-01

    Environmental conditions shape community composition. Arctic thermal springs provide an opportunity to study how environmental gradients can impose strong selective pressures on microbial communities and provide a continuum of niche opportunities. We use microscopic and molecular methods to conduct a survey of microbial community composition at Troll Springs on Svalbard, Norway, in the high Arctic. Microorganisms there exist under a wide range of environmental conditions: in warm water as periphyton, in moist granular materials, and in cold, dry rock as endoliths. Troll Springs has two distinct ecosystems, aquatic and terrestrial, together in close proximity, with different underlying environmental factors shaping each microbial community. Periphyton are entrapped during precipitation of calcium carbonate from the spring's waters, providing microbial populations that serve as precursors for the development of endolithic communities. This process differs from most endolith colonization, in which the rock predates the communities that colonize it. Community composition is modulated as environmental conditions change within the springs. At Troll, the aquatic environments show a small number of dominant operational taxonomic units (OTUs) that are specific to each sample. The terrestrial environments show a more even distribution of OTUs common to multiple samples.

  10. Microbial community composition and endolith colonization at an Arctic thermal spring are driven by calcite precipitation.

    PubMed

    Starke, Verena; Kirshtein, Julie; Fogel, Marilyn L; Steele, Andrew

    2013-10-01

    Environmental conditions shape community composition. Arctic thermal springs provide an opportunity to study how environmental gradients can impose strong selective pressures on microbial communities and provide a continuum of niche opportunities. We use microscopic and molecular methods to conduct a survey of microbial community composition at Troll Springs on Svalbard, Norway, in the high Arctic. Microorganisms there exist under a wide range of environmental conditions: in warm water as periphyton, in moist granular materials, and in cold, dry rock as endoliths. Troll Springs has two distinct ecosystems, aquatic and terrestrial, together in close proximity, with different underlying environmental factors shaping each microbial community. Periphyton are entrapped during precipitation of calcium carbonate from the spring's waters, providing microbial populations that serve as precursors for the development of endolithic communities. This process differs from most endolith colonization, in which the rock predates the communities that colonize it. Community composition is modulated as environmental conditions change within the springs. At Troll, the aquatic environments show a small number of dominant operational taxonomic units (OTUs) that are specific to each sample. The terrestrial environments show a more even distribution of OTUs common to multiple samples.

  11. Biotechnological potential of aquatic plant-microbe interactions.

    PubMed

    Stout, L; Nüsslein, K

    2010-06-01

    The rhizosphere in terrestrial systems is the region of soil surrounding plant roots where there is increased microbial activity; in aquatic plants, this definition may be less clear because of diffusion of nutrients in water, but there is still a zone of influence by plant roots in this environment [1]. Within that zone chemical conditions differ from those of the surrounding environment as a consequence of a range of processes that were induced either directly by the activity of plant roots or by the activity of rhizosphere microflora. Recently, there are a number of new studies related to rhizospheres of aquatic plants and specifically their increased potential for remediation of contaminants, especially remediation of metals through aquatic plant-microbial interaction.

  12. Surface microbial contamination in hospitals: A pilot study on methods of sampling and the use of proposed microbiologic standards.

    PubMed

    Claro, Tânia; O'Reilly, Marese; Daniels, Stephen; Humphreys, Hilary

    2015-09-01

    Contamination of hospital surfaces by bacteria is increasingly recognized. We assessed commonly touched surfaces using contact plates and Petrifilms (3M, St. Paul, MN) and compared the results against proposed microbiology standards. Toilet door handles were the most heavily contaminated (7.97 ± 0.68 colony forming units [CFU]/cm(2)) and exceeded proposed standards on 74% of occasions. Petrifilms detected statistically higher CFU from bedside lockers. Further research is required on the use of standards and methods of sampling.

  13. Aquatic Life Criterion - Selenium

    EPA Pesticide Factsheets

    Documents pertaining to the 2016 Acute and Chronic Ambient Aquatic Life Water Quality Criteria for Selenium (Freshwater). These documents include what the safe levels of Selenium are in water for the majority of species.

  14. Aquatic Life Criteria - Atrazine

    EPA Pesticide Factsheets

    Documents pertaining to Acute and Chronic Ambient Water Quality Aquatic Life Criteria for Atrazine (Freshwater and Salt Water). This document contains the safe levels of Atrazine in water that should protect to the majority of species.

  15. Respiration in Aquatic Insects.

    ERIC Educational Resources Information Center

    MacFarland, John

    1985-01-01

    This article: (1) explains the respiratory patterns of several freshwater insects; (2) describes the differences and mechanisms of spiracular cutaneous, and gill respiration; and (3) discusses behavioral aspects of selected aquatic insects. (ML)

  16. Aquatic Life Criteria - Cadmium

    EPA Pesticide Factsheets

    Documents pertaining to 2016 Acute and Chronic Aquatic Life Ambient Water Quality Criteria for Cadmium (Freshwater, Estuarine/marine). These documents contain the safe levels of Cadmium in water that should protect to the majority of species.

  17. Aquatic Life Criteria - Carbaryl

    EPA Pesticide Factsheets

    Documents pertaining to 2012 Final National Recommended Aquatic Life Ambient Water Quality Criteria for Carbaryl (Freshwater, Estuarine/marine). These documents contain the safe levels of Carbaryl in water that should protect to the majority of species.

  18. Aquatic Life Criteria - Ammonia

    EPA Pesticide Factsheets

    Documents related to EPA's final 2013 Aquatic Life Ambient Water Quality Criteria for Ammonia (Freshwater). These documents pertain to the safe levels of Ammonia in water that should protect to the majority of species.

  19. Aquatic Life Criteria - Copper

    EPA Pesticide Factsheets

    Documents pertain to Aquatic Life Ambient Water Quality criteria for Copper (2007 Freshwater, 2016 Estuarine/marine). These documents contain the safe levels of Copper in water that should protect to the majority of species.

  20. Aquatic Life Criteria - Lead

    EPA Pesticide Factsheets

    References and documents pertaining to Acute and Chronic Ambient Aquatic Life Water Quality Criteria for Lead. These documents include the safe levels of Lead in water that should protect the majority of species.

  1. Respiration in Aquatic Insects.

    ERIC Educational Resources Information Center

    MacFarland, John

    1985-01-01

    This article: (1) explains the respiratory patterns of several freshwater insects; (2) describes the differences and mechanisms of spiracular cutaneous, and gill respiration; and (3) discusses behavioral aspects of selected aquatic insects. (ML)

  2. Developing a Collegiate Aquatics Program.

    ERIC Educational Resources Information Center

    Fawcett, Paul A.

    2001-01-01

    Presents suggestions for departments of health, physical education, and recreation that are planning to develop their own aquatics programs, focusing on: the prevalence of collegiate aquatics programs; course offerings in an aquatics minor; practicums and internships; graduate programs in aquatics; cross-disciplinary appeal; marketing the aquatics…

  3. Aquatic toxicology: fact or fiction

    SciTech Connect

    Macek, K.J.

    1980-02-01

    The science of aquatic toxicology is a relatively new science. The development of the field of aquatic toxicology since 1930 is traced. The state of the art of aquatic toxicology compared with that of classical toxicology is evaluated. The science of aquatic toxicology is expected to undergo a significant period of rapid growth and development, leading ultimately to the formation of a mature science.

  4. Developing a Collegiate Aquatics Program.

    ERIC Educational Resources Information Center

    Fawcett, Paul A.

    2001-01-01

    Presents suggestions for departments of health, physical education, and recreation that are planning to develop their own aquatics programs, focusing on: the prevalence of collegiate aquatics programs; course offerings in an aquatics minor; practicums and internships; graduate programs in aquatics; cross-disciplinary appeal; marketing the aquatics…

  5. Molecular microbial diversity of a soil sample and detection of ammonia oxidizers from Cape Evans, Mcmurdo Dry Valley, Antarctica.

    PubMed

    Shravage, Bhupendra V; Dayananda, Kannayakanahalli M; Patole, Milind S; Shouche, Yogesh S

    2007-01-01

    The aim of our study was to estimate the uncultured eubacterial diversity of a soil sample collected below a dead seal, Cape Evans, McMurdo, Antarctica by an SSU rDNA gene library approach. Our study by sequencing of clones from SSU rDNA gene library approach revealed high diversity in the soil sample from Antarctica. More than 50% of clones showed homology to Cytophaga-Flavobacterium-Bacteroides group; sequences also belonged to alpha, beta, gamma proteobacteria, Thermus-Deinococcus and high GC gram-positive group; Phylogenetic analysis of the SSU rDNA clones showed the presence of species belonging to Cytophaga spp., Vitellibacter vladivostokensis, Aequorivita lipolytica, Aequorivita crocea, Flavobacterium spp., Flexibacter sp., Subsaxibacter broadyi, Bacteroidetes, Roseobacter sp., Sphingomonas baekryungensis, Nitrosospira sp., Nitrosomonas cryotolerans, Psychrobacter spp., Chromohalobacter sp., Psychrobacter okhotskensis, Psychrobacter fozii, Psychrobacter urativorans, Rubrobacter radiotolerans, Marinobacter sp., Rubrobacteridae, Desulfotomaculum aeronauticum and Deinococcus sp. The presence of ammonia oxidizing bacteria in Antarctica soil was confirmed by the presence of the amoA gene. Phylogenetic analysis revealed grouping of clones with their respective groups.

  6. Biomarkers of sulfate reducing bacteria from a variety of different aged samples including a modern microbial mat

    NASA Astrophysics Data System (ADS)

    Pages, A.; Grice, K.; Lockhart, R.; Holman, A.; Melendez, I.; Van Kranendonk, M.; Jaraula, C.

    2011-12-01

    Most biomarkers present in sediments occur in only trace concentrations, trapped in kerogen or may be highly functionalised especially in recent sedimentary deposits making them difficult to chromatographically resolve, thus presenting considerable analytical challenges, especially for isotope studies. Innovative hydro (Hy) pyrolysis (Py) techniques are able to target or convert many of these compounds into free hydrocarbons more amenable to gas chromatography mass-spectrometry (GC-MS) and compound-specific isotope analysis (CSIA). HyPy has been applied to a modern layered smooth mat from Shark Bay, Western Australia. Saturate and aromatic fractions from different layers of the mat have been analysed by GC-MS and CSIA. After HyPy, an even-odd distribution of n-alkanes has been revealed as well as very long-chain n-alkanes up to n-C38. Stable carbon isotopic values of the n-alkanes indicated the presence of at least two bacterial communities. The short-chain n-alkanes were likely to be representative of a cyanobacteria community (δ13C, C15-C23, - 18 to -25 %VPDB) while the carbon isotopic values of the long-chain n-alkanes supported the presence of sulfate reducing bacteria (δ13C, C25-C33, - 30 to - 34 %VPDB). Long-chain fatty acids have been previously reported in sulfate reducing bacteria. It is hypothesised that this distribution and isotopic character representing sulfate reducing bacteria consortia may be preserved in the rock record. This hypothesis has been tested in Australian rocks: a Devonian carbonaceous concretion containing an exceptionally well preserved fossil invertebrate from the Canning Basin, Western Australia, a Paleoproterozoic sample (1.6 billion years old) from a lead-zinc ore deposit from the McArthur Basin, Northern Territories and a Paleoproterozoic chert (2.3 billion years old) from the Pilbara, Western Australia. Biomarkers of these samples showed a strong predominance of long-chain n-alkanes, up to n-C38 with an even-odd distribution

  7. Connecting Water Quality With Air Quality Through Microbial Aerosols

    NASA Astrophysics Data System (ADS)

    Dueker, M. Elias

    Aerosol production from surface waters results in the transfer of aquatic materials (including nutrients and bacteria) to air. These materials can then be transported by onshore winds to land, representing a biogeochemical connection between aquatic and terrestrial systems not normally considered. In urban waterfront environments, this transfer could result in emissions of pathogenic bacteria from contaminated waters. Despite the potential importance of this link, sources, near-shore deposition, identity and viability of microbial aerosols are largely uncharacterized. This dissertation focuses on the environmental and biological mechanisms that define this water-air connection, as a means to build our understanding of the biogeochemical, biogeographical, and public health implications of the transfer of surface water materials to the near-shore environment in both urban and non-urban environments. The effects of tidal height, wind speed and fog on coastal aerosols and microbial content were first quantified on a non-urban coast of Maine, USA. Culture-based, culture-independent, and molecular methods were used to simultaneously sample microbial aerosols while monitoring meteorological parameters. Aerosols at this site displayed clear marine influence and high concentrations of ecologically-relevant nutrients. Coarse aerosol concentrations significantly increased with tidal height, onshore wind speed, and fog presence. Tidal height and fog presence did not significantly influence total microbial aerosol concentrations, but did have a significant effect on culturable microbial aerosol fallout. Molecular analyses of the microbes settling out of near-shore aerosols provided further evidence of local ocean to terrestrial transport of microbes. Aerosol and surface ocean bacterial communities shared species and in general were dominated by organisms previously sampled in marine environments. Fog presence strengthened the microbial connection between water and land through

  8. Biogeochemical cycling at the aquatic-terrestrial interface is linked to parafluvial hyporheic zone inundation history

    NASA Astrophysics Data System (ADS)

    Goldman, Amy E.; Graham, Emily B.; Crump, Alex R.; Kennedy, David W.; Romero, Elvira B.; Anderson, Carolyn G.; Dana, Karl L.; Resch, Charles T.; Fredrickson, Jim K.; Stegen, James C.

    2017-09-01

    The parafluvial hyporheic zone combines the heightened biogeochemical and microbial interactions indicative of a hyporheic region with direct atmospheric/terrestrial inputs and the effects of wet-dry cycles. Therefore, understanding biogeochemical cycling and microbial interactions in this ecotone is fundamental to understanding biogeochemical cycling at the aquatic-terrestrial interface and to creating robust hydrobiogeochemical models of dynamic river corridors. We aimed to (i) characterize biogeochemical and microbial differences in the parafluvial hyporheic zone across a small spatial domain (6 lateral meters) that spans a breadth of inundation histories and (ii) examine how parafluvial hyporheic sediments respond to laboratory-simulated re-inundation. Surface sediment was collected at four elevations along transects perpendicular to flow of the Columbia River, eastern WA, USA. The sediments were inundated by the river 0, 13, 127, and 398 days prior to sampling. Spatial variation in environmental variables (organic matter, moisture, nitrate, glucose, % C, % N) and microbial communities (16S and internal transcribed spacer (ITS) rRNA gene sequencing, qPCR) were driven by differences in inundation history. Microbial respiration did not differ significantly across inundation histories prior to forced inundation in laboratory incubations. Forced inundation suppressed microbial respiration across all histories, but the degree of suppression was dramatically different between the sediments saturated and unsaturated at the time of sample collection, indicating a binary threshold response to re-inundation. We present a conceptual model in which irregular hydrologic fluctuations facilitate microbial communities adapted to local conditions and a relatively high flux of CO2. Upon rewetting, microbial communities are initially suppressed metabolically, which results in lower CO2 flux rates primarily due to suppression of fungal respiration. Following prolonged inundation

  9. Aquatic Plant Control Research Program. Allelopathic Aquatic Plants for Aquatic Plant Management: A Feasibility Study

    DTIC Science & Technology

    1989-10-01

    block number) FIELD GROUP SUB-GROUP Allelopathy "Bioassay . Growth inhibition. Aquatic macrophytes. Biocontrol Lena minor 19. ABSTRACT (Continue on...Bibliography of Aquatic Plant Allelopathy ........ Al 2 ALLELOPATHIC AQUATIC PLANTS FOR AQUATIC PLANT MANAGEMENT; A FEASIBILITY STUDY Introduction Background 1...nutrients, water, and other biotic effects could have overriding effects that appear as competition or allelopathy . These biotic factors must be

  10. Characterizing microbial community and geochemical dynamics at hydrothermal vents using osmotically driven continuous fluid samplers.

    PubMed

    Robidart, Julie; Callister, Stephen J; Song, Pengfei; Nicora, Carrie D; Wheat, Charles G; Girguis, Peter R

    2013-05-07

    Microbes play a key role in mediating aquatic biogeochemical cycles. However, our understanding of the relationships between microbial phylogenetic/physiological diversity and habitat physicochemical characteristics is restrained by our limited capacity to concurrently collect microbial and geochemical samples at appropriate spatial and temporal scales. Accordingly, we have developed a low-cost, continuous fluid sampling system (the Biological OsmoSampling System, or BOSS) to address this limitation. The BOSS does not use electricity, can be deployed in harsh/remote environments, and collects/preserves samples with daily resolution for >1 year. Here, we present data on the efficacy of DNA and protein preservation during a 1.5 year laboratory study as well as the results of two field deployments at deep-sea hydrothermal vents, wherein we examined changes in microbial diversity, protein expression, and geochemistry over time. Our data reveal marked changes in microbial composition co-occurring with changes in hydrothermal fluid composition as well as the temporal dynamics of an enigmatic sulfide-oxidizing symbiont in its free-living state. We also present the first data on in situ protein preservation and expression dynamics highlighting the BOSS's potential utility in meta-proteomic studies. These data illustrate the value of using BOSS to study relationships among microbial and geochemical phenomena and environmental conditions.

  11. Microbial Metalloproteomics

    PubMed Central

    Hagedoorn, Peter-Leon

    2015-01-01

    Metalloproteomics is a rapidly developing field of science that involves the comprehensive analysis of all metal-containing or metal-binding proteins in a biological sample. The purpose of this review is to offer a comprehensive overview of the research involving approaches that can be categorized as inductively coupled plasma (ICP)-MS based methods, X-ray absorption/fluorescence, radionuclide based methods and bioinformatics. Important discoveries in microbial proteomics will be reviewed, as well as the outlook to new emerging approaches and research areas. PMID:28248278

  12. [ANTIBIOTIC SENSITIVITY/RESISTANCY OF MICROBIAL STRAINS, ISOLATED FROM PUERPERAS, NEWBORNS AND SAMPLES OF MATERNITY WARD ENVIRONMENT].

    PubMed

    Kobeshavidze, D D; Chikviladze, D; Gachechiladze, Kh; Mikeladze, M

    2016-03-01

    In this article, there are given data of microorganisms sensitivity/resistancy investigation, to different groups of antibiotics. Microorganisms where isolated from puerperas, newborns and samples of maternity ward environment. Investigation was performed in L.T.D. "Imedis Clinica". Detection of microorganisms sensitivity/resistancy to antibiotics was performed by use of two methods: method of disc diffusion and serial dilution on solid breeding substratum. It was determined that gram positive, as well as gram negative microorganisms had sufficiently high level of resistancy to some penicillines, aminoglycosides, macrolides. Some species of gram negative microorganisms had resistancy to lincomicin in 100% of cases. High level of sensitivity was revealed to such antibiotics as amicalin, amoxiclav, cefepim, ciprofloxacin. Gram negative microorganisms had high level of sensitivity to imipenem-cilastatin and meropenem. Performed investigation confirms necessity of microbiological monitoring in different clinics, because it is one of the most significant components of infection control. It gives opportunity to perform exact antibiotic prophylaxis and if necessary - rational antibiotic therapy.

  13. Dissolved Organic Carbon Dynamics Along Terrestrial-aquatic Flowpaths in a Catchment Dominated by Sandy Soils

    NASA Astrophysics Data System (ADS)

    Wickland, K.; Walker, J. F.; Hood, K.; Butler, K. D.

    2015-12-01

    Aquatic systems receive significant amounts of terrestrially-derived dissolved organic carbon (DOC) from their watersheds. The amount and nature received depends on terrestrial carbon source strength, processing and losses of carbon during transport, and hydrologic connectivity between terrestrial and aquatic systems. While much research has been done on terrestrial DOC dynamics along terrestrial-aquatic flowpaths, there is still considerable uncertainty in many areas including the importance of different carbon sources, microbial metabolism and sorption of DOC, and processing of carbon in groundwater. Here we investigate DOC dynamics in soils, groundwater, and stream waters at the USGS Water, Energy, and Biogeochemical (WEBB) Program research site in northern Wisconsin. This site is well-suited for studying DOC dynamics as soils are sandy and homogenous with small DOC sorption potential, and previous work has characterized the hydrology of the region in detail. We collected water samples over two years from soil pit lysimeters along a series of hillslope transects, from shallow and deep groundwater wells, and from a first-order stream receiving these waters. We measured DOC concentration, DOC optical properties, and biodegradability of DOC. Combined with historical DOC and companion water chemistry data we characterize DOC generation and loss along the following flowpaths: 1) infiltration through the unsaturated zone to the groundwater table, 2) shallow groundwater flow, and 3) long groundwater flowpaths of different origin (lake-derived vs. terrestrial-derived water).

  14. Map showing bathymetry and aquatic plants of Lake Waramaug, Connecticut

    USGS Publications Warehouse

    Kulp, Kenneth P.

    1981-01-01

    The Bathymetry of Lake Waramaug is shown at 5-foot intervals, and the location and identification of significant concentrations of aquatic plants is indicated. The bathymetry shows the lake to be relatively steep-sided and flat-bottomed, with a maximum depth of between 40 and 45 feet. Aquatic plants were confined to the edges of the lake in depths of less than 15 feet. Nine species of aquatic plants occcur in significant numbers in the lake. Lake bathymetric data were collected using a recording fathometer and sounding rod; aquatic plant data were collected by visual survey; and the collection of samples using a weed rake. (USGS)

  15. Aquatic Plant Control Research Program. Field methods to measure aquatic plant treatment method efficacy. Final report

    SciTech Connect

    Killgore, K.J.; Payne, B.S.

    1984-04-01

    The Aquatic Plant Control Research Program (APCRP) of the U.S. Army Engineer Waterways Experiment Station (WES) is developing field techniques to measure treatment efficacy and to determine site characteristics that influence the treatment efficacy. Treatment efficacy is considered a quantitative determination of the extent and duration of changes in problem aquatic plant populations attributable to the use of a treatment method (i.e., chemical, mechanical, biological, environmental). Depending on the plant species, efficacy can be determined or indicated by changes in biomass, areal distribution, or height of an aquatic plant in response to treatment. Aquatic plant biomass is sampled with a WES aquatic biomass sampler; areal distribution of aquatic plants is determined by aerial photography or with an electronic positioning system; and submersed aquatic plant height is measured with a fathometer (depth recorder) used with an electronic positioning and repositioning system (AGNAV). The APCRP has also developed field techniques to determine site characteristics that influence efficacy using commercially available instrumentation. This instrumentation can be used to measure treatment efficacy and to determine site characteristics simultaneously.

  16. Linkages among geophysical facies, microbial composition, biogeochemical rates, and seasonal hydrology in the hyporheic zone

    NASA Astrophysics Data System (ADS)

    Stegen, J.

    2016-12-01

    The hyporheic zone is a critical ecosystem transition that links terrestrial, aquatic, and subsurface domains. To understand connections among physical, microbial, and biogeochemical components of the hyporheic zone, we obtained freeze cores along the Columbia River in the Hanford 300 Area and performed geologic, molecular, and microbial assays. Mud and sand content were found to be the primary drivers of microbial community attributes (in particular, of nitrite and carbon oxidizers). Microbial community analysis revealed an abundance of nitrifying Archaea (Thaumarchaea) and an absence of nitrifiying Bacteria. Network analysis revealed significant negative correlations between sand content and some statistical modules of microbial taxa, perhaps indicating the importance of pore water residence time on community composition. A similar set of microbial modules was positively correlated with total organic carbon. One such module that also positively correlated with aerobic metabolic rates was dominated by Thaumarchaea and Nitrospira, suggesting that ammonia oxidation was the dominant aerobic process. We also examined temporal changes in hyporheic microbial structure and activity through repeated sampling of attached and pore water microbes across a spatial gradient. We found that microbial communities remained distinct in river, hyporheic, and inland zones across seasonal variation in hydrologic mixing conditions. One reason was temperature-driven increases in microbial species richness in the hyporheic zone. We show that the relative importance of ecological selection and dispersal varied across environments and across geographic zones. Our results also indicated that while selection imposed short-term constraints on microbial community structure, hyporheic sediment communities did not respond to short-term hydrologic variation. Importantly, we demonstrated that the influence of selective pressures varied with phylogenetic affiliation, which may have been responsible

  17. Possibility of using a lithotrophic iron-oxidizing microbial fuel cell as a biosensor for detecting iron and manganese in water samples.

    PubMed

    Tran, Phuong Hoang Nguyen; Luong, Tha Thanh Thi; Nguyen, Thuy Thu Thi; Nguyen, Huy Quang; Duong, Hop Van; Kim, Byung Hong; Pham, Hai The

    2015-10-01

    Iron-oxidizing bacterial consortia can be enriched in microbial fuel cells (MFCs) operated with ferrous iron as the sole electron donor. In this study, we investigated the possibility of using such lithotrophic iron-oxidizing MFC (LIO-MFC) systems as biosensors to monitor iron and manganese in water samples. When operated with anolytes containing only ferrous iron as the sole electron donor, the experimented LIO-MFCs generated electrical currents in response to the presence of Fe(2+) in the anolytes. For the concentrations of Fe(2+) in the range of 3-20 mM, a linear correlation between the current and the concentration of Fe(2+) could be achieved (r(2) = 0.98). The LIO-MFCs also responded to the presence of Mn(2+) in the anolytes but only when the Mn(2+) concentration was less than 3 mM. The presence of other metal ions such as Ni(2+) or Pb(2+) in the anolytes reduced the Fe(2+)-associated electricity generation of the LIO-MFCs at various levels. Organic compounds, when present at a non-excessive level together with Fe(2+) in the anolytes, did not affect the generation of electricity, although the compounds might serve as alternative electron donors for the anode bacteria. The performance of the LIO-MFCs was also affected to different degrees by operational parameters, including surrounding temperature, pH of the sample, buffer strength and external resistance. The results proved the potential of LIO-MFCs as biosensors sensing Fe(2+) in water samples with a significant specificity. However, the operation of the system should be in compliance with an optimal procedure to ensure reliable performance.

  18. Sequential extractions on mine tailings samples after and before bioassays: implications on the speciation of metals during microbial re-colonization

    NASA Astrophysics Data System (ADS)

    García-Meza, J. V.; Carrillo-Chávez, A.; Morton-Bermea, O.

    2006-01-01

    Mine tailings may be remediated using metal tolerant microorganisms as they may solve the limiting conditions for healthy development of plants (i.e., low organic mater content and poor physical conditions). The aim of this study was to investigate the consequences of microbial colonization on the chemical speciation of trace metals. Surface samples from the Valenciana mine tailings (Guanajuato, Mexico) were used for long-term bioassays (BA), which consisted in the promotion of microorganisms, development on tailings material under stable laboratory conditions (humidity, temperature, and light exposure). A five-step sequential extraction method (exchangeable, carbonate/specifically adsorbed, Fe-Mn oxides, organic matter (OM)/sulfide, and residual fractions) was performed before and after BA. Extraction solutions and leachates were analyzed by inductively coupled plasma-mass spectrometry. OM content, cationic exchange capacity, and pH values were also assessed before and after BA. The results indicate that trace elements are generally present in nonresidual fractions, mainly in the Fe-Mn oxides fraction. The concentration of total Zn, As, Se, Pb, and exchangeable Cu and Pb is above the recommendable limits for soils. Despite the high bioavailability of the former elements, biofilms successfully colonized the tailing samples during the BA. Cyanobacteria and green algae, heterotrophic fungi, aerobic bacteria, and anaerobic bacteria composed the developed biofilms. Chemical controls of trace elements could be attributed to absorption onto inorganic complexes (carbonates, metal oxides), while biofilm occurrence seems to enhance complexation and immobilization of Cr, Ni, Cu, Zn, As, and Pb. The biofilm developed does not increase the bioavailable forms and the leaching of the trace elements, but significantly improves the OM contents (natural fertilization). The results suggest that biofilms are useful during the first steps of the mine tailings remediation.

  19. Subglacial Lake Whillans microbial biogeochemistry: a synthesis of current knowledge.

    PubMed

    Mikucki, J A; Lee, P A; Ghosh, D; Purcell, A M; Mitchell, A C; Mankoff, K D; Fisher, A T; Tulaczyk, S; Carter, S; Siegfried, M R; Fricker, H A; Hodson, T; Coenen, J; Powell, R; Scherer, R; Vick-Majors, T; Achberger, A A; Christner, B C; Tranter, M

    2016-01-28

    Liquid water occurs below glaciers and ice sheets globally, enabling the existence of an array of aquatic microbial ecosystems. In Antarctica, large subglacial lakes are present beneath hundreds to thousands of metres of ice, and scientific interest in exploring these environments has escalated over the past decade. After years of planning, the first team of scientists and engineers cleanly accessed and retrieved pristine samples from a West Antarctic subglacial lake ecosystem in January 2013. This paper reviews the findings to date on Subglacial Lake Whillans and presents new supporting data on the carbon and energy metabolism of resident microbes. The analysis of water and sediments from the lake revealed a diverse microbial community composed of bacteria and archaea that are close relatives of species known to use reduced N, S or Fe and CH4 as energy sources. The water chemistry of Subglacial Lake Whillans was dominated by weathering products from silicate minerals with a minor influence from seawater. Contributions to water chemistry from microbial sulfide oxidation and carbonation reactions were supported by genomic data. Collectively, these results provide unequivocal evidence that subglacial environments in this region of West Antarctica host active microbial ecosystems that participate in subglacial biogeochemical cycling. © 2015 The Author(s).

  20. Capabilities of Seven Species of Aquatic Macrophytes for Phytoremediation of Pentachlorophenol Contaminated Sediment

    NASA Astrophysics Data System (ADS)

    Zhao, Liangyuan; Guo, Weijie; Li, Qingyun; Li, Huan; Zhao, Weihua; Cao, Xiaohuan

    2017-01-01

    Sediments are regarded as the ultimate sink of pentachlorophenol(PCP) in aquatic environment, and capabilities of seven species of aquatic macrophytes for remediating PCP contaminated sediment were investigated. Seven species of aquatic macrophytes could significantly accelerate the degradation of PCP in sediments. Among all, canna indica L., Acorus calamus L. and Iris tectorum Maxim. can be used as efficient alternative plants for remediation of PCP contaminated sediment, which attained 98%, 92% and 88% of PCP removal in sediments, respectively. PCP was detected only in root tissues and the uptake was closely related to the root lipid contents of seven plants. The presence of seven aquatic macrophytes significantly increased microbial populations and the activities of dehydrogenase compared with control sediments, indicating that rhizosphere microorganism played important role in the remediation process. In conclusion, seven species of aquatic macrophytes may act as promising tools for the PCP phytoremediation in aquatic environment, especially Canna indica L., Acorus calamus L. and Iris tectorum Maxim.

  1. Microbial effects on sorption and transport of actinides in tuff samples from the Nevada Test Site and soils from McGuire AFB, NJ

    NASA Astrophysics Data System (ADS)

    Fisher, J. C.; Gostic, R.; Gostic, J.; Czerwinski, K.; Moser, D. P.

    2009-12-01

    The sorption and behavior of various actinides were examined for two sets of environmental samples. The Nevada Test Site (NTS) harbors a variety of radionuclides resulting from atomic weapons testing from the 1950s-1990s. Modeling the transport of radionuclides at the NTS is difficult because each detonation cavity is a unique environment with distinct hydrologic characteristics, chemical composition, and microbial community structure. McGuire AFB was the site of an explosion that resulted in the burning of a BOMARC nuclear missile and deposition of particles containing high-fired oxides of Am, Pu, and U in soils on the base. Analysis of the NTS samples focused on sorption/desorption of 233-U and 241-Am in the presence/absence of bacteria, and work on the BOMARC cores addressed the potential role of microorganisms in mediating particle degradation and movement. Batch experiments with various NTS tuff samples and strains of bacteria showed that sorption of actinides may be enhanced by >25% under certain conditions by bacteria. Sorption of 233-U was highly dependent on carbonate concentrations in the liquid matrix, while 241-Am was unaffected. Different bacterial species also affected sorption differently. Sorption kinetics for both actinides were rapid, with maximum sorption usually occurring within 4 hours. Actinides bound tightly to tuff and little desorption occurred in carbonate-free batch experiments. Column experiments showed that bacterial cultures in minimal salts buffer desorbed significantly more 233-U from tuff than low carbonate NTS water, but less than 30 mM bicarbonate buffer. Hot particles in the BOMARC cores were located using CT mapping and were extracted from the soil prior to analysis of core sections by gamma spectroscopy. Subcores for DNA extraction and culturing were collected from soil in direct contact with hot particles. The extracted particles consisted of a mixture of weapons-grade Pu, 241-Am and 235-U and ranged in activity from 5-66 k

  2. Assessing the soil microbial carbon budget: Probing with salt stress

    NASA Astrophysics Data System (ADS)

    Rath, Kristin; Rousk, Johannes

    2014-05-01

    The amount of carbon stored as soil organic matter (SOM) constitutes a pool more than double the size of the atmospheric carbon pool. Soil respiration represents one of the largest fluxes of carbon between terrestrial ecosystems and the atmosphere. A large fraction of the CO2 released by soils is produced by the microbial decomposition of SOM. The microbial carbon budget is characterized by their carbon use efficiency, i.e. the partitioning of substrate into growth and respiration. This will shape the role of the soil as a net source or sink for carbon. One of the canonical factors known to influence microbial processes in soil is pH. In aquatic systems salinity has been found to have a comparably strong influence as pH. However salinity remains understudied in soil, despite its growing relevance due to land use change and agricultural practices. The aim of this project is to understand how microbial carbon dynamics respond to disturbance by changing environmental conditions, using salinity as a reversible stressor. First, we compiled a comparative analysis of the sensitivity of different microbial processes to increasing salt concentrations. Second, we compared different salts to determine whether salt toxicity depended on the identity of the salt. Third, we used samples from a natural salinity gradient to assess if a legacy of salt exposure can influence the microbial response to changing salt concentrations. If salt had an ecologically significant effect in shaping these communities, we would assume that microbial processes would be less sensitive to an increase in salt concentrations. The sensitivity of microbial processes to salt was investigated by establishing inhibition curves in order to estimate EC50 values (the concentration resulting in 50% inhibition). These EC50 values were used to compare bacterial and fungal growth responses, as well as catabolic processes such as respiration and nitrogen mineralisation. Initial results suggest that growth related

  3. Aquatic Life Benchmarks

    EPA Pesticide Factsheets

    The Aquatic Life Benchmarks is an EPA-developed set of criteria for freshwater species. These benchmarks are based on toxicity values reviewed by EPA and used in the Agency's risk assessments developed as part of the decision-making process for pesticide registration.

  4. Investigating Aquatic Dead Zones

    ERIC Educational Resources Information Center

    Testa, Jeremy; Gurbisz, Cassie; Murray, Laura; Gray, William; Bosch, Jennifer; Burrell, Chris; Kemp, Michael

    2010-01-01

    This article features two engaging high school activities that include current scientific information, data, and authentic case studies. The activities address the physical, biological, and chemical processes that are associated with oxygen-depleted areas, or "dead zones," in aquatic systems. Students can explore these dead zones through both…

  5. Aquatic Microbiology Laboratory Manual.

    ERIC Educational Resources Information Center

    Cooper, Robert C.; And Others

    This laboratory manual presents information and techniques dealing with aquatic microbiology as it relates to environmental health science, sanitary engineering, and environmental microbiology. The contents are divided into three categories: (1) ecological and physiological considerations; (2) public health aspects; and (3)microbiology of water…

  6. Investigating Aquatic Dead Zones

    ERIC Educational Resources Information Center

    Testa, Jeremy; Gurbisz, Cassie; Murray, Laura; Gray, William; Bosch, Jennifer; Burrell, Chris; Kemp, Michael

    2010-01-01

    This article features two engaging high school activities that include current scientific information, data, and authentic case studies. The activities address the physical, biological, and chemical processes that are associated with oxygen-depleted areas, or "dead zones," in aquatic systems. Students can explore these dead zones through both…

  7. Aquatics [Chapter 6

    Treesearch

    F. A. Vertucci; M. A. Conrad

    1994-01-01

    Within the GLEES boundary there are three alpine lakes and several streams and ponds. The selection of GLEES as a research site for investigating of the effects of chemical and physical climate change was in part based on the accessibility of these low alkalinity "sensitive" aquatic ecosystems. This chapter provides a brief description of the physical,...

  8. Adapted Aquatics and Inclusion.

    ERIC Educational Resources Information Center

    Block, Martin E.; Conatser, Phillip

    2002-01-01

    Presents strategies and techniques to help instructors and directors promote successful inclusive aquatics programs for students with disabilities, discussing the importance of considering issues related to: teaching style, collaborative planning, goal determination, appropriate inclusive placement, personnel preparation, curriculum adaptation,…

  9. Aquatic Equipment Information.

    ERIC Educational Resources Information Center

    Sova, Ruth

    Equipment usually used in water exercise programs is designed for variety, intensity, and program necessity. This guide discusses aquatic equipment under the following headings: (1) equipment design; (2) equipment principles; (3) precautions and contraindications; (4) population contraindications; and (5) choosing equipment. Equipment is used…

  10. Aquatic Microbiology Laboratory Manual.

    ERIC Educational Resources Information Center

    Cooper, Robert C.; And Others

    This laboratory manual presents information and techniques dealing with aquatic microbiology as it relates to environmental health science, sanitary engineering, and environmental microbiology. The contents are divided into three categories: (1) ecological and physiological considerations; (2) public health aspects; and (3)microbiology of water…

  11. Overview of microbial biofilms.

    PubMed

    Costerton, J W

    1995-09-01

    As the success of this two-issue special section of the Journal of Industrial Microbiology attests, the study of microbial biofilms is truly burgeoning as the uniqueness and the importance of this mode of growth is increasingly recognized. Because of its universality the biofilm concept impacts virtually all of the subdivisions of Microbiology (including Medical, Dental, Agricultural, Industrial and Environmental) and these two issues incorporate contributions from authors in all of these disciplines. Some time ago we reasoned that bacteria cannot possibly be aware (sic) of their precise location, in terms of this spectrum of anthrocentric subspecialties, and that their behavior must be dictated by a standard set of phenotypic responses to environmental conditions in what must seem to them (sic) to be a continuum of very similar aquatic ecosystems. In this overview I will, therefore, stress the common features of microbial biofilms that we should bear in mind as we use this simple universal concept to seek to understand bacterial behavior in literally hundreds of aquatic ecosystems traditionally studied by dozens of subspecies of microbiologists reared in sharply different scientific and academic conventions.

  12. Composition of free and adherent ruminal bacteria: inaccuracy of the microbial nutrient supply estimates obtained using free bacteria as reference samples and (15)N as the marker.

    PubMed

    González, J; Arroyo, J M; Ouarti, M; Guevara-González, J; Rodríguez, C A; Alvir, M R; Moya, V J; Piquer, O

    2012-03-01

    .5% and 59.6%, respectively, as an average of the four experiments). Large differences in AA profile were observed between SAB and LAB. The prediction equation obtained using (15)N as the marker may be used to correct the errors associated with the traditional use of LAB as the reference sample, and therefore to obtain more accurate estimates of the microbial nutrient supply to the ruminants.

  13. Aquatic Plants and their Control.

    ERIC Educational Resources Information Center

    Michigan State Dept. of Natural Resources, Lansing.

    Aquatic plants can be divided into two types: algae and macrophytes. The goal of aquatic plant management is to maintain a proper balance of plants within a lake and still retain the lake's recreational and economic importance. Aquatic plant management programs have two phases: long-term management (nutrient control), and short-term management…

  14. Methods for Aquatic Resource Assessment

    EPA Science Inventory

    The Methods for Aquatic Resource Assessment (MARA) project consists of three main activities in support of assessing the conditions of the nation’s aquatic resources: 1) scientific support for EPA Office of Water’s national aquatic resource surveys; 2) spatial predications of riv...

  15. Aquatic Pest Control. Manual 99.

    ERIC Educational Resources Information Center

    Missouri Univ., Columbia. Agricultural Experiment Station.

    This training manual provides information needed to meet the minimum EPA standards for certification as a commercial applicator of pesticides in the aquatic pest control category. The text discusses various water use situations; aquatic weed identification; herbicide use and effects; and aquatic insects and their control. (CS)

  16. Aquatic Plants and their Control.

    ERIC Educational Resources Information Center

    Michigan State Dept. of Natural Resources, Lansing.

    Aquatic plants can be divided into two types: algae and macrophytes. The goal of aquatic plant management is to maintain a proper balance of plants within a lake and still retain the lake's recreational and economic importance. Aquatic plant management programs have two phases: long-term management (nutrient control), and short-term management…

  17. Introduced aquatic plants and algae

    USDA-ARS?s Scientific Manuscript database

    Non-native aquatic plants such as waterhyacinth and hydrilla severely impair the uses of aquatic resources including recreational faculties (lakes, reservoirs, rivers) as well as timely delivery of irrigation water for agriculture. Costs associated with impacts and management of all types of aquatic...

  18. Aquatic Pest Control. Manual 99.

    ERIC Educational Resources Information Center

    Missouri Univ., Columbia. Agricultural Experiment Station.

    This training manual provides information needed to meet the minimum EPA standards for certification as a commercial applicator of pesticides in the aquatic pest control category. The text discusses various water use situations; aquatic weed identification; herbicide use and effects; and aquatic insects and their control. (CS)

  19. Using fine-scale high-resolution sampling to link Fe oxide-dominated hydrothermal vent-generated microbial mat morphology with community structure composition at Loihi Seamount, Hawaii

    NASA Astrophysics Data System (ADS)

    Hager, K. W.; Hilton, T. S.; Kimber, J.; Jesser, K. J.; Fullerton, H.; McAllister, S.; Chan, C. S.; Emerson, D.; Moyer, C. L.

    2013-12-01

    The scale at which sampling can be carried out in the deep ocean has largely been determined by the limits of robotic instrument manipulation at depth. Bulk sampling, via push cores, suction, or scoop samplers, collects mat material from heterogeneous microbial communities living in environments variable not only laterally with respect to an active vent (direct versus diffuse vent flow), but also with respect to depth across the steep redox gradient separating reduced hydrothermal fluid from oxygenated seawater. While initially unavoidable, these kinds of sampling strategies can only go so far in describing the intricate microbial ecology interactions occurring at hydrothermal vents. For this reason, a syringe sampler was developed for this study to sample targeted observable mat morphologies of surface mat material at small spatial scales. Multiple 'BioMat' samples (~ 20) were collected in 2013 from active, Fe oxide-dominated mats from several vent sites at Loihi Seamount, Hawaii, all experience temperatures ranging from 20 to 46°C. Quantitative-PCR (Q-PCR) primers were designed to detect the relative abundance of the marine iron-oxidizing Zetaproteobacteria. At Pohaku Vents, which is perched on the upper lip of the caldera, the average relative abundance of Zetaproteobacteria was significantly higher at 79.2% of the total microbial mat community. Two other sites within the caldera (Hiolo North and Hiolo South) had 26.8% and 36.9% Zetaproteobacteria, respectively. In addition, Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis of these samples showed that the Pohaku Vent microbial mat community fingerprints all grouped together into a single cluster much more closely and with much less variability than with those from either Hiolo North or South. Finally, it was also observed that there were site-specific gross morphological characteristics associated with these microbial mats including veils, curds, and amorphous particles. It is presently

  20. Antibiotic resistant Pseudomonas spp. in the aquatic environment: A prevalence study under tropical and temperate climate conditions.

    PubMed

    Devarajan, Naresh; Köhler, Thilo; Sivalingam, Periyasamy; van Delden, Christian; Mulaji, Crispin K; Mpiana, Pius T; Ibelings, Bastiaan W; Poté, John

    2017-05-15

    Microbial populations which are resistant to antibiotics are an emerging environmental concern with potentially serious implications for public health. Thus, there is a growing concern in exploring the occurrence of antibiotic resistance in the environment with no limitations to the factors that contribute to their emergence. The aquatic environment is considered to be a hot-spot for the acquisition and spread of antibiotic resistance due to pollution with emerging contaminants derived from anthropogenic activities. In this study, we report on the isolation and characterization of 141 Pseudomonas spp. from aquatic sediments receiving partially (un)treated hospital and communal effluents from three distinct geographical locations: Democratic Republic of the Congo (DRC), India (IN), and Switzerland (CH). P. putida (42%) and P. aeruginosa (39%) were the dominant Pseudomonas species. The highest frequency of antibiotic resistance against eight anti-pseudomonas agents was found among IN isolates (35-60%), followed by DRC (18-50%) and CH (12-54%). CTX-M was the most frequent β-lactamase found in CH (47% of isolates), while VIM-1 was dominant in isolates from DRC (61%) and IN (29%). NDM-1 was found in 29% of the total IN isolates and surprisingly also in 6% of CH isolates. Chromosomally-encoded efflux mechanisms were overexpressed in P. aeruginosa isolates from all three geographic locations. In vitro conjugative transfers of antibiotic resistance plasmids occurred more frequently under tropical temperatures (30 and 37 °C) than under temperate conditions (10 °C). The presence of Extended Spectrum β-lactamases (ESBLs) and Metallo β-lactamases (MBLs) in the isolates from environmental samples has important implications for humans who depend on public water supply and sanitation facilities. To our knowledge, this is the first study to demonstrate a comparison between treated/untreated effluents from urban and hospital settings as a source of microbial resistance

  1. Manuals Used in the National Aquatic Resource Surveys

    EPA Pesticide Factsheets

    Various manuals are used to communicate the methods and guidelines for the National Aquatic Resource Surveys. The Field Operations Manual: outlines the field protocols that crews will utilize to sample sites.

  2. The fate of eroded soil organic carbon along a European transect - controls after deposition in terrestrial and aquatic systems

    NASA Astrophysics Data System (ADS)

    Kirkels, Frédérique; Cammeraat, Erik; Kalbitz, Karsten; Van Oost, Kristof; Ellerbrock, Ruth; Folain, Stéphane; Gerke, Horst; Heckrath, Goswin; Kögel-Knabner, Ingrid; Kuhn, Nikolaus; Quinton, John; Salvador-Blanes, Sébastien; Sommer, Michael; Steffens, Markus

    2014-05-01

    The potential fate of eroded soil organic carbon (SOC) after deposition is key to understand carbon cycling in eroding landscapes. Globally, large quantities of sediments and SOC are redistributed by soil erosion on agricultural land, particularly after heavy precipitation events. Deposition of eroded SOC takes place on downslope soils within the catchment and in adjacent inland waters, i.e. substantial amounts of SOC are transferred from terrestrial to aquatic ecosystems. However, the net effect on C exchange between soils, atmosphere and inland waters is unknown. We hypothesize that the turnover of deposited C is significantly affected by soil and organic matter properties, and whether deposition occurs in terrestrial or aquatic environments. We sampled topsoils from 10 agricultural sites along a European transect, spanning a wide range of SOC and soil characteristics (e.g. texture, aggregation, C content, etc.). Turnover of SOC was determined for terrestrial and aquatic depositional conditions in a 10-week incubation study. Moreover, we studied the impact of labile carbon inputs ('priming') on SOC stability using 13C labelled cellulose. We evaluated potentially important controls on the fate of SOC such as amounts and composition of soil organic matter (SOM), distribution of SOC in density fractions and aggregates as well as soil physical and chemical properties. NMR analysis provided an in-depth characterization of SOM quality, showing large similarities in chemical composition among the sites. The role of the microbial biomass was specifically assessed in relation to SOC turnover. The results of our study broadly enhanced our knowledge about controls on SOC decomposition/stabilization after its deposition in terrestrial and aquatic environments. We envisage that our quantitative relationships will contribute to obtain better estimates of the impact of soil erosion on carbon budgets and reduce uncertainties in the linkage between terrestrial and aquatic carbon

  3. An inexpensive, accurate, and precise wet-mount method for enumerating aquatic viruses.

    PubMed

    Cunningham, Brady R; Brum, Jennifer R; Schwenck, Sarah M; Sullivan, Matthew B; John, Seth G

    2015-05-01

    Viruses affect biogeochemical cycling, microbial mortality, gene flow, and metabolic functions in diverse environments through infection and lysis of microorganisms. Fundamental to quantitatively investigating these roles is the determination of viral abundance in both field and laboratory samples. One current, widely used method to accomplish this with aquatic samples is the "filter mount" method, in which samples are filtered onto costly 0.02-μm-pore-size ceramic filters for enumeration of viruses by epifluorescence microscopy. Here we describe a cost-effective (ca. 500-fold-lower materials cost) alternative virus enumeration method in which fluorescently stained samples are wet mounted directly onto slides, after optional chemical flocculation of viruses in samples with viral concentrations of <5×10(7) viruses ml(-1). The concentration of viruses in the sample is then determined from the ratio of viruses to a known concentration of added microsphere beads via epifluorescence microscopy. Virus concentrations obtained by using this wet-mount method, with and without chemical flocculation, were significantly correlated with, and had precision equivalent to, those obtained by the filter mount method across concentrations ranging from 2.17×10(6) to 1.37×10(8) viruses ml(-1) when tested by using cultivated viral isolates and natural samples from marine and freshwater environments. In summary, the wet-mount method is significantly less expensive than the filter mount method and is appropriate for rapid, precise, and accurate enumeration of aquatic viruses over a wide range of viral concentrations (≥1×10(6) viruses ml(-1)) encountered in field and laboratory samples.

  4. An Inexpensive, Accurate, and Precise Wet-Mount Method for Enumerating Aquatic Viruses

    PubMed Central

    Cunningham, Brady R.; Brum, Jennifer R.; Schwenck, Sarah M.; Sullivan, Matthew B.

    2015-01-01

    Viruses affect biogeochemical cycling, microbial mortality, gene flow, and metabolic functions in diverse environments through infection and lysis of microorganisms. Fundamental to quantitatively investigating these roles is the determination of viral abundance in both field and laboratory samples. One current, widely used method to accomplish this with aquatic samples is the “filter mount” method, in which samples are filtered onto costly 0.02-μm-pore-size ceramic filters for enumeration of viruses by epifluorescence microscopy. Here we describe a cost-effective (ca. 500-fold-lower materials cost) alternative virus enumeration method in which fluorescently stained samples are wet mounted directly onto slides, after optional chemical flocculation of viruses in samples with viral concentrations of <5 × 107 viruses ml−1. The concentration of viruses in the sample is then determined from the ratio of viruses to a known concentration of added microsphere beads via epifluorescence microscopy. Virus concentrations obtained by using this wet-mount method, with and without chemical flocculation, were significantly correlated with, and had precision equivalent to, those obtained by the filter mount method across concentrations ranging from 2.17 × 106 to 1.37 × 108 viruses ml−1 when tested by using cultivated viral isolates and natural samples from marine and freshwater environments. In summary, the wet-mount method is significantly less expensive than the filter mount method and is appropriate for rapid, precise, and accurate enumeration of aquatic viruses over a wide range of viral concentrations (≥1 × 106 viruses ml−1) encountered in field and laboratory samples. PMID:25710369

  5. A long-term, multitrophic level study to assess pulp and paper mill effluent effects on aquatic communities in four US receiving waters: characteristics of the study streams, sample sites, mills, and mill effluents.

    PubMed

    Hall, Timothy J; Ragsdale, Renee L; Arthurs, William J; Ikoma, Joan; Borton, Dennis L; Cook, Diana L

    2009-04-01

    Watershed characteristics, study streams, sample sites, mills, and mill effluents are provided for 4 streams included in a long-term study to assess potential effects of pulp and paper mill effluents on US receiving waters. The study streams are Codorus Creek (Pennsylvania, USA), Leaf River (Mississippi, USA) and McKenzie and Willamette rivers (Oregon, USA) and were chosen to represent a blend of mill process types, effluent concentrations, and coldwater/warmwater stream systems. The described effluent quality, water quality, and habitat data sets encompass the initial 7 to 8 y of a study anticipated to continue >10 y and provide a backdrop to a series of articles describing periphyton, macroinvertebrate, and fish community properties in these same streams. The mean in-stream waste concentration (IWC) for these 4 effluent discharges was 32.4%, 2.0%, 0.5%, and 0.2% v/v for Codorus Creek and Leaf, McKenzie, and Willamette rivers, respectively, as compared with a median of 0.4% for US mills. Effluent quality measurements included Selenastrum capricornutum, Ceriodaphnia dubia, and Pimephales promelas chronic bioassays as sanctioned by the US Environmental Protection Agency for estimating effluent effects on receiving-water aquatic communities. Based on mean bioassay inhibition concentration for a 25% effect and on mean IWC, a margin of safety against adverse biological effects of 2, 25, 137, and 150 times was indicated for Codorus Creek and Leaf, McKenzie, and Willamette rivers, respectively. Habitat and water quality assessment was carried out over a gradient of sample sites above and below the effluent discharge to determine nonmill-related conditions that might interfere with interpretation of effluent effects. Noneffluent related localized differences in conditions for some parameters, including current velocity (McKenzie River), and surface incident photosynthetically active radiation (Codorus Creek and Willamette River) occurred at the sample stations immediately

  6. Risk screening of pharmaceutical compounds in Romanian aquatic environment.

    PubMed

    Gheorghe, Stefania; Petre, Jana; Lucaciu, Irina; Stoica, Catalina; Nita-Lazar, Mihai

    2016-06-01

    The aquatic environment is under increased pressure by pharmaceutically active compounds (PhACs) due to anthropogenic activities. In spite of being found at very low concentrations (ng/L to μg/L) in the environment, PhACs represent a real danger to aquatic ecosystems due to their bioaccumulation and long-term effects. In this study, the presence in the aquatic environment of six non-steroidal anti-inflammatory drugs (ibuprofen, diclofenac, acetaminophen, naproxen, indomethacin, and ketoprofen), caffeine, and carbamazepine were monitored. Moreover, their aquatic risk and ecotoxicity by three biological models were evaluated. The monitoring studies performed in Romania showed that all studied PhACs were naturally present at concentrations >0.01 μg/L, pointing out the necessity to perform further toxicity tests for environmental risk assessment. The toxicity studies were carried out on aquatic organisms or bacteria and they indicated, for most of the tested PhACs, an insignificant or low toxicity effects: lethal concentrations (LC50) on fish Cyprinus carpio ranged from 42.60 mg/L to more than 100 mg/L; effective concentrations (EC50) on planktonic crustacean Daphnia magna ranged from 11.02 mg/L to more than 100 mg/L; inhibitory concentrations (IC50)/microbial toxic concentrations (MTC) on Vibrio fischeri and other bacterial strains ranged from 7.02 mg/L to more than 100 mg/L. The PhAC aquatic risk was assessed by using the ratio between measured environmental concentration (MEC) and predicted no effect concentration (PNEC) calculated for each type of organism. The average of quotient risks (RQs) revealed that the presence of these compounds in Romania's aquatic environment induced a lower or moderate aquatic risk.

  7. Metabolism of polycyclic aromatic hydrocarbons in the aquatic environment

    SciTech Connect

    Varanasi, U.

    1989-01-01

    During the past decade, knowledge of polycyclic aromatic hydrocarbons (PAH) in the aquatic environment has advanced substantially to encompass studies of bioavailability, metabolism, subsequent toxic effects, and their ecological consequences. In this book, recent advances in the areas of PAH biogeochemistry and bioaccumulation, microbial degradation, enzymes of activation and detoxication, metabolism of PAH, and laboratory and field studies on carcinogenic/toxic effects, are presented. Additionally, important similarities and differences in metabolism of PAH by aquatic and terrestrial organisms are discussed. These chapters also illustrate that although considerable progress has been made in certain areas of PAH metabolism in the aquatic environment, the field is relatively unexplored and many exciting possibilities exist for future investigations. Separate abstracts are included for 9 chapters in this book for inclusion in the appropriate data bases.

  8. Scaling macroscopic aquatic locomotion

    NASA Astrophysics Data System (ADS)

    Gazzola, Mattia; Argentina, Mederic; Mahadevan, Lakshminarayanan

    2014-11-01

    Inertial aquatic swimmers that use undulatory gaits range in length L from a few millimeters to 30 meters, across a wide array of biological taxa. Using elementary hydrodynamic arguments, we uncover a unifying mechanistic principle characterizing their locomotion by deriving a scaling relation that links swimming speed U to body kinematics (tail beat amplitude A and frequency ω) and fluid properties (kinematic viscosity ν). This principle can be simply couched as the power law Re ~ Swα , where Re = UL / ν >> 1 and Sw = ωAL / ν , with α = 4 / 3 for laminar flows, and α = 1 for turbulent flows. Existing data from over 1000 measurements on fish, amphibians, larvae, reptiles, mammals and birds, as well as direct numerical simulations are consistent with our scaling. We interpret our results as the consequence of the convergence of aquatic gaits to the performance limits imposed by hydrodynamics.

  9. Scaling macroscopic aquatic locomotion

    NASA Astrophysics Data System (ADS)

    Gazzola, Mattia; Argentina, Médéric; Mahadevan, L.

    2014-10-01

    Inertial aquatic swimmers that use undulatory gaits range in length L from a few millimetres to 30 metres, across a wide array of biological taxa. Using elementary hydrodynamic arguments, we uncover a unifying mechanistic principle characterizing their locomotion by deriving a scaling relation that links swimming speed U to body kinematics (tail beat amplitude A and frequency ω) and fluid properties (kinematic viscosity ν). This principle can be simply couched as the power law Re ~ Swα, where Re = UL/ν >> 1 and Sw = ωAL/ν, with α = 4/3 for laminar flows, and α = 1 for turbulent flows. Existing data from over 1,000 measurements on fish, amphibians, larvae, reptiles, mammals and birds, as well as direct numerical simulations are consistent with our scaling. We interpret our results as the consequence of the convergence of aquatic gaits to the performance limits imposed by hydrodynamics.

  10. Aquatic Nuisance Species Locator

    EPA Pesticide Factsheets

    Data in this map has been collected by the United States Geological Survey's Nonindigenous Aquatic Species program located in Gainesville, Florida (http://nas.er.usgs.gov/default.aspx). This dataset may have some inaccuracies and is only current to June 15, 2012. The species identified in this dataset are not inclusive of all aquatic nuisance species, but rather a subset identified to be at risk for transport by recreational activities such as boating and angling. Additionally, the locations where organisims have been identified are also not inclusive and should be treated as a guide. Organisms are limited to the following: American bullfrog, Asian clam, Asian shore crab, Asian tunicate, Australian spotted jellyfish, Chinese mitten crab, New Zealand mudsnail, Colonial sea squirt, Alewife, Bighead carp, Black carp, Flathead catfish, Grass carp, Green crab, Lionfish, Northern snakehead, Quagga mussel, Round Goby, Ruffe, Rusty crayfish, Sea lamprey, Silver carp, Spiny water flea, Veined rapa whelk, Zebra mussel

  11. Cytochromes of Aquatic Fungi

    PubMed Central

    Gleason, Frank H.; Unestam, Torgny

    1968-01-01

    The cytochrome systems of two classes of aquatic fungi, the Oomycetes and Chytridiomycetes, were studied by means of reduced-minus-oxidized difference spectra at room and at low temperature. At room temperature, all of these fungi have a c-type cytochrome with an absorption maximum at 551 mμ and a b-type cytochrome at 564 mμ. The Oomycetes have a-type cytochromes at 605 mμ, and the Chytridiomycetes have a-type cytochromes at 606 mμ (Blastocladiales) or at 609 mμ (Monoblepharidales). Additional b-type cytochromes are found at 557 mμ in the Oomycetes and at approximately 560 mμ in the Chytridiomycetes. The data obtained from spectra at low temperature are consistent with these conclusions. Thus, the difference spectra reveal variation between the cytochrome systems of these two classes of aquatic fungi. PMID:5650068

  12. Characterizing Microbial Community and Geochemical Dynamics at Hydrothermal Vents Using Osmotically Driven Continuous Fluid Samplers

    SciTech Connect

    Robidart, Julie C.; Callister, Stephen J.; Song, Peng F.; Nicora, Carrie D.; Wheat, Charles G.; Girguis, Peter R.

    2013-05-07

    Microbes play a key role in mediating all aquatic biogeochemical cycles, and ongoing efforts are aimed at better understanding the relationships between microbial phylogenetic and physiological diversity, and habitat physical and chemical characteristics. Establishing such relationships is facilitated by sampling and studying microbiology and geochemistry at the appropriate spatial and temporal scales, to access information on the past and current environmental state that contributes to observed microbial abundances and activities. A modest number of sampling systems exist to date, few of which can be used in remote, harsh environments such as hydrothermal vents, where the ephemeral nature of venting underscores the necessity for higher resolution sampling. We have developed a robust, continuous fluid sampling system for co-registered microbial and biogeochemical analyses. The osmosis-powered bio-osmosampling system (BOSS) use no electricity, collects fluids with daily resolution or better, can be deployed in harsh, inaccessible environments and can sample fluids continuously for up to five years. Here we present a series of tests to examine DNA, RNA and protein stability over time, as well as material compatability, via lab experiments. We also conducted two field deployments at deep-sea hydrothermal vents to assess changes in microbial diversity and protein expression as a function of the physico-chemical environment. Our data reveal significant changes in microbial community composition co-occurring with relatively modest changes in the geochemistry. These data additionally provide new insights into the distribution of an enigmatic sulfur oxidizing symbiont in its free-living state. Data from the second deployment reveal differences in the representation of peptides over time, underscoring the utility of the BOSS in meta-proteomic studies. In concert, these data demonstrate the efficacy of this approach, and illustrate the value of using this method to study

  13. Aquatic sports massage therapy.

    PubMed

    Bell, G W

    1999-04-01

    Athletic trainers are continually bombarded with requests to assist aquatic athletes with the management of musculoskeletal concerns involved with training and overtraining. The trainer has options for initial training management through the administration of massage, cryotherapy, thermotherapy, and injury-preventative strengthening exercises. This article describes and illustrates athletic training techniques such as massage, cryotherapy or cold applications, thermotherapy or heat applications, and proprioceptive neuromuscular facilitation or strengthening exercise.

  14. Adaption of the microbial community to continuous exposures of multiple residual antibiotics in sediments from a salt-water aquacultural farm.

    PubMed

    Xi, Xiuping; Wang, Min; Chen, Yongshan; Yu, Shen; Hong, Youwei; Ma, Jun; Wu, Qian; Lin, Qiaoyin; Xu, Xiangrong

    2015-06-15

    Residual antibiotics from aquacultural farming may alter microbial community structure in aquatic environments in ways that may adversely or positively impact microbially-mediated ecological functions. This study investigated 26 ponds (26 composited samples) used to produce fish, razor clam and shrimp (farming and drying) and 2 channels (10 samples) in a saltwater aquacultural farm in southern China to characterize microbial community structure (represented by phospholipid fatty acids) in surface sediments (0-10 cm) with long-term exposure to residual antibiotics. 11 out of 14 widely-used antibiotics were quantifiable at μg kg(-1) levels in sediments but their concentrations did not statistically differ among ponds and channels, except norfloxacin in drying shrimp ponds and thiamphenicol in razor clam ponds. Concentrations of protozoan PLFAs were significantly increased in sediments from razor clam ponds while other microbial groups were similar among ponds and channels. Both canonical-correlation and stepwise-multiple-regression analyses on microbial community and residual antibiotics suggested that roxithromycin residuals were significantly related to shifts in microbial community structure in sediments. This study provided field evidence that multiple residual antibiotics at low environmental levels from aquacultural farming do not produce fundamental shifts in microbial community structure. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Methane emissions to the atmosphere through aquatic plants

    NASA Technical Reports Server (NTRS)

    Sebacher, D. I.; Harriss, R. C.; Bartlett, K. B.

    1985-01-01

    The movement of methane (CH4) from anaerobic sediments through the leaves, stems, and flowers of aquatic plants and into the atmosphere was found to provide a significant pathway for the emission of CH4 from the aquatic substrates of flooded wetlands. Methane concentrations well above the surrounding ambient air levels were found in the mesophyll of 16 varies of aquatic plants and are attributed to transpiration, diffusion, and pressure-induced flow of gaseous CH4 from the roots when they are embedded in CH4-saturated anaerobic sediments. Methane emissions from the emergent parts of aquatic plants were measured using floating chamber techniques and by enclosing the plants in polyethylene bags of known volume. Concentration changes were monitored in the trapped air using syringes and gas chromatographic techniques. Vertical profiles of dissolved CH4 in sediment pore water surrounding the aquatic plants' rhizomes were obtained using an interstitial sampling technique. Methane emissions from the aquatic plants studied varied from 14.8 mg CH4/d to levels too low to be detectable. Rooted and unrooted freshwater aquatic plants were studied as well as saltwater and brackish water plants. Included in the experiment is detailed set of measurements on CH4 emissions from the common cattail (Typha latifolia). This paper illustrates that aquatic plants play an important gas exchange role in the C cycle between wetlands and the atmosphere.

  16. Methane emissions to the atmosphere through aquatic plants

    NASA Technical Reports Server (NTRS)

    Sebacher, D. I.; Harriss, R. C.; Bartlett, K. B.

    1985-01-01

    The movement of methane (CH4) from anaerobic sediments through the leaves, stems, and flowers of aquatic plants and into the atmosphere was found to provide a significant pathway for the emission of CH4 from the aquatic substrates of flooded wetlands. Methane concentrations well above the surrounding ambient air levels were found in the mesophyll of 16 varies of aquatic plants and are attributed to transpiration, diffusion, and pressure-induced flow of gaseous CH4 from the roots when they are embedded in CH4-saturated anaerobic sediments. Methane emissions from the emergent parts of aquatic plants were measured using floating chamber techniques and by enclosing the plants in polyethylene bags of known volume. Concentration changes were monitored in the trapped air using syringes and gas chromatographic techniques. Vertical profiles of dissolved CH4 in sediment pore water surrounding the aquatic plants' rhizomes were obtained using an interstitial sampling technique. Methane emissions from the aquatic plants studied varied from 14.8 mg CH4/d to levels too low to be detectable. Rooted and unrooted freshwater aquatic plants were studied as well as saltwater and brackish water plants. Included in the experiment is detailed set of measurements on CH4 emissions from the common cattail (Typha latifolia). This paper illustrates that aquatic plants play an important gas exchange role in the C cycle between wetlands and the atmosphere.

  17. Occurrence of 4-tert-butylphenol (4-t-BP) biodegradation in an aquatic sample caused by the presence of Spirodela polyrrhiza and isolation of a 4-t-BP-utilizing bacterium.

    PubMed

    Ogata, Yuka; Toyama, Tadashi; Yu, Ning; Wang, Xuan; Sei, Kazunari; Ike, Michihiko

    2013-04-01

    Although 4-tert-butylphenol (4-t-BP) is a serious aquatic pollutant, its biodegradation in aquatic environments has not been well documented. In this study, 4-t-BP was obviously and repeatedly removed from water from four different environments in the presence of Spirodela polyrrhiza, giant duckweed, but 4-t-BP persisted in the environmental waters in the absence of S. polyrrhiza. Also, 4-t-BP was not removed from autoclaved pond water with sterilized S. polyrrhiza. These results suggest that the 4-t-BP removal from the environmental waters was caused by biodegradation stimulated by the presence of S. polyrrhiza rather than by uptake by the plant. Moreover, Sphingobium fuliginis OMI capable of utilizing 4-t-BP as a sole carbon and energy source was isolated from the S. polyrrhiza rhizosphere. Strain OMI degraded 4-t-BP via a meta-cleavage pathway, and also degraded a broad range of alkylphenols with linear or branched alkyl side chains containing two to nine carbon atoms. Root exudates of S. polyrrhiza stimulated 4-t-BP degradation and cell growth of strain OMI. Thus, the stimulating effects of S. polyrrhiza root exudates on 4-t-BP-degrading bacteria might have contributed to 4-t-BP removal in the environmental waters with S. polyrrhiza. These results demonstrate that the S. polyrrhiza-bacteria association may be applicable to the removal of highly persistent 4-t-BP from wastewaters or polluted aquatic environments.

  18. Using HSDP Samples to Evaluate the Staudigel et al. Model of Euendolithic Microbial Alteration of Marine Basalt Glass: A Peek at the Unseen Majority

    NASA Astrophysics Data System (ADS)

    Walton, A. W.

    2008-12-01

    Staudigel et al. (2008, Earth Science Reviews 89 156-176.) recognize separate alteration processes of basalt glass in oceanic crust and elsewhere: Formation of palagonite and pore-filling cements is abiotic, while microbes form granular and tubular textures. Granular alteration begins shortly after glass cools to a habitable state and is a product of organisms that etch the surface of basalt glass by adjusting pH. These organisms ultimately dissolve the glass surface to form an agglomeration of micrometer-scale sub-spherical cavities. Smectite, zeolites, Fe-oxide, and Ti-rich structures occupy the cavities. Together with microbes, they form a bio- and mineral film that ultimately shuts down the system. Tubular structures reflect the activity of hypha-bearing organisms that mine the glass to get nutrients. Hyphae allow the organisms to remain in contact with circulating water while penetrating any biofilm and mining nutrients from unaltered glass. Tubules are about 1 micrometer in diameter and 10s to 100s of micrometers long. Both suites of organisms use reduced elements from the glass as sources of electrons for their metabolism. Activity ceases when secondary minerals reduce permeability to circulating seawater and availability of some nutrients. HSDP results, from an ocean island, show some areas of agreement. Petrographic studies indicate that initiation of tubules post-dates crushing of grains during early burial and formation of isopachous smectite grain coatings. Growth of tubules may be very slow; they grow during a time window of about 30000 to 60000 years, although duration of growth of individual tubules may be much shorter. Tubules and other textures of alteration are commonly coated with ferric hydroxide, perhaps a product of microbial oxidation. However, some features of biotic alteration in HSDP hyaloclastites are inconsistent with the proposed model. Smectitic grain replacement, which resembles granular alteration, forms in HSDP samples after and in

  19. Organic carbon composition and thermodynamics indicate preferential carbon sequestration at a terrestrial-aquatic interface

    NASA Astrophysics Data System (ADS)

    Graham, E.; Crump, A.; Kennedy, D.; Tfaily, M. M.

    2016-12-01

    Terrestrial carbon (C) inputs into aquatic systems have increased by up to 1 PgC yr-1 in the anthropogenic era, yet considerable uncertainty remains in the flux of organic matter across terrestrial-aquatic linkages. Previous research has demonstrated impacts of organic matter composition on C burial efficiency and stabilization within sediments. Here, we investigated preferential sediment C sequestration along two elevation transects in the hyporheic zone of the Columbia River in eastern Washington State. We sampled depth profiles at 10 cm intervals from 0-60 cm using liquid nitrogen freeze cores to maintain spatial integrity. At each depth, we used Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) to yield the elemental composition and Gibbs free energy of C oxidation (dGCox) in C profiles. We expected dGCox to increase with depth, reflecting declines in labile C and in redox conditions. Instead, we observed the opposite trend in freeze cores with high rates of aerobic activity, while those sampled locations with low aerobic activity did not show thermodynamic trends. Preliminary analysis of C composition profiles supports this relationship, with enhanced labile C pools at lower depths in high-activity freeze cores. We also found spatial variation in C processing within depth profiles across <10 m horizontal distances. We highlight the importance of sediment microbiomes in determining C concentration and composition, as microorganisms can both consume and produce labile C. In turn, these reactions define the energetic environment under which subsequent biogeochemical cycling operates. Our results generate new insights into preferential C burial rates, microbially-mediated transformations of organic matter, and thermodynamic constraints on C sequestration at terrestrial-aquatic interfaces.

  20. Development of aquatic animal experiment facility, Aquatic Habitat (AQH)

    NASA Astrophysics Data System (ADS)

    Uchida, S.; Kono, Y.; Sakimura, T.; Nishikawa, W.; Fujimoto, N.; Murakami, K.; Nakamura, T.

    We have been performing technical studies to develop aquatic animal experiment facility, Aquatic Habitat (AQH), for both of short-term experiments in the Space Shuttle middeck and long-term experiments in the Space Station including the Centrifuge Accommodation Module (CAM). The AQH will have the capabilities to accommodate three-generations of small freshwater fish (medaka and zebrafish) and egg through metamorphosis of amphibian (African clawed frog). For these purposes, the AQH will have the following brand-new capabilities that the previous facilities have never had; 90days experiment duration, automatic feeding according to specimen types and their developmental stages, separation of generations for fish, specimen sample collection in various developmental stages, air/water interface control for amphibian, continuous monitoring of specimen behavior even in dark condition, and so on. We have already performed preliminary breeding tests for medaka and zebrafish with a breeding system prototype. Their mating behavior was performed successfully in the small closed chamber and the hatched larvae grew and started spawning on the 45-47th day after hatching. These results demonstrated that three generational breeding of medaka and zebrafish within 90days would be possible based on this breeding system prototype. Also, we have developed almost of the above new mechanisms, that is, an automatic feeding system, an egg separation mechanism for fish, an air stabilizer to control air/water interface, and a continuous specimen monitoring system through light/dark cycle. Based on these results, we have manufactured a BBM of AQH water circulation system and performed biological compatibility tests as a next step. For African clawed frog breeding, some problems have been revealed through the preliminary tests with the breeding system prototype. Currently, we are performing the investigations to resolve the problems and preparing to proceed to the next step.

  1. Production of extracellular nucleic acids by genetically altered bacteria in aquatic-environment microcosms

    SciTech Connect

    Paul, J.H.; David, A.W.

    1989-01-01

    Factors which affect the production of extracellular DNA by genetically altered strains of Escherichia coli, Pseudomonas aeruginosa, Pseudomonas cepacia, and Bradyrhizobium japonicum in aquatic environments were investigated. The presence or absence of the ambient microbial community had little effect on the production of extracellular DNA. Results indicate the extracellular-DNA production by genetically altered bacteria released into aquatic environments is more strongly influenced by physiochemical factors than biotic factors; extracellular-DNA production rates are usually greater for organisms released in freshwater than marine environments; and ambient microbial populations can readily utilize materials released by these organisms.

  2. Conceptual Framework for Aquatic Interfaces

    NASA Astrophysics Data System (ADS)

    Lewandowski, J.; Krause, S.

    2015-12-01

    Aquatic interfaces are generally characterized by steep gradients of physical, chemical and biological properties due to the contrast between the two adjacent environments. Innovative measurement techniques are required to study the spatially heterogeneous and temporally variable processes. Especially the different spatial and temporal scales are a large challenge. Due to the steep biogeochemical gradients and the intensive structural and compositional heterogeneity, enhanced biogeochemical processing rates are inherent to aquatic interfaces. Nevertheless, the effective turnover depends strongly on the residence time distribution along the flow paths and in sections with particular biogeochemical milieus and reaction kinetics. Thus, identification and characterization of the highly complex flow patterns in and across aquatic interfaces are crucial to understand biogeochemical processing along exchange flow paths and to quantify transport across aquatic interfaces. Hydrodynamic and biogeochemical processes are closely coupled at aquatic interfaces. However, interface processing rates are not only enhanced compared to the adjacent compartments that they connect; also completely different reactions might occur if certain thresholds are exceeded or the biogeochemical milieu differs significantly from the adjacent environments. Single events, temporal variability and spatial heterogeneity might increase overall processing rates of aquatic interfaces and thus, should not be neglected when studying aquatic interfaces. Aquatic interfaces are key zones relevant for the ecological state of the entire ecosystem and thus, understanding interface functioning and controls is paramount for ecosystem management. The overall aim of this contribution is a general conceptual framework for aquatic interfaces that is applicable to a wide range of systems, scales and processes.

  3. Automated sample area definition for high-throughput microscopy.

    PubMed

    Zeder, M; Ellrott, A; Amann, R

    2011-04-01

    High-throughput screening platforms based on epifluorescence microscopy are powerful tools in a variety of scientific fields. Although some applications are based on imaging geometrically defined samples such as microtiter plates, multiwell slides, or spotted gene arrays, others need to cope with inhomogeneously located samples on glass slides. The analysis of microbial communities in aquatic systems by sample filtration on membrane filters followed by multiple fluorescent staining, or the investigation of tissue sections are examples. Therefore, we developed a strategy for flexible and fast definition of sample locations by the acquisition of whole slide overview images and automated sample recognition by image analysis. Our approach was tested on different microscopes and the computer programs are freely available (http://www.technobiology.ch). Copyright © 2011 International Society for Advancement of Cytometry.

  4. Automated Sampling Procedures Supported by High Persistence of Bacterial Fecal Indicators and Bacteroidetes Genetic Microbial Source Tracking Markers in Municipal Wastewater during Short-Term Storage at 5°C

    PubMed Central

    Mayer, R. E.; Vierheilig, J.; Egle, L.; Reischer, G. H.; Saracevic, E.; Mach, R. L.; Kirschner, A. K. T.; Zessner, M.; Farnleitner, A. H.

    2015-01-01

    Because of high diurnal water quality fluctuations in raw municipal wastewater, the use of proportional autosampling over a period of 24 h at municipal wastewater treatment plants (WWTPs) to evaluate carbon, nitrogen, and phosphorus removal has become a standard in many countries. Microbial removal or load estimation at municipal WWTPs, however, is still based on manually recovered grab samples. The goal of this study was to establish basic knowledge regarding the persistence of standard bacterial fecal indicators and Bacteroidetes genetic microbial source tracking markers in municipal wastewater in order to evaluate their suitability for automated sampling, as the potential lack of persistence is the main argument against such procedures. Raw and secondary treated wastewater of municipal origin from representative and well-characterized biological WWTPs without disinfection (organic carbon and nutrient removal) was investigated in microcosm experiments at 5 and 21°C with a total storage time of 32 h (including a 24-h autosampling component and an 8-h postsampling phase). Vegetative Escherichia coli and enterococci, as well as Clostridium perfringens spores, were selected as indicators for cultivation-based standard enumeration. Molecular analysis focused on total (AllBac) and human-associated genetic Bacteroidetes (BacHum-UCD, HF183 TaqMan) markers by using quantitative PCR, as well as 16S rRNA gene-based next-generation sequencing. The microbial parameters showed high persistence in both raw and treated wastewater at 5°C under the storage conditions used. Surprisingly, and in contrast to results obtained with treated wastewater, persistence of the microbial markers in raw wastewater was also high at 21°C. On the basis of our results, 24-h autosampling procedures with 5°C storage conditions can be recommended for the investigation of fecal indicators or Bacteroidetes genetic markers at municipal WWTPs. Such autosampling procedures will contribute to better

  5. Microbial growth under humic-free conditions in a supraglacial stream system on the Cotton Glacier, Antarctica

    NASA Astrophysics Data System (ADS)

    Foreman, Christine M.; Cory, Rose M.; Morris, Cindy E.; SanClements, Michael D.; Smith, Heidi J.; Lisle, John T.; Miller, Penney L.; Chin, Yu-Ping; McKnight, Diane M.

    2013-09-01

    During the austral summers of 2004 and 2009, we sampled a supraglacial stream on the Cotton Glacier, Antarctica. The stream dissolved organic matter (DOM) was low (44-48 μM C) and lacked detectable humic fluorescence signatures. Analysis of the excitation emissions matrices (EEMs) indicated that amino-acid fluorophores dominated, consistent with DOM of microbial origin, with little humic-like fluorescence. In most aquatic ecosystems, humic DOM attenuates harmful UV radiation and its absence may represent an additional stressor influencing the microbial community. Nonetheless, the stream contained an active microbial assemblage with bacterial cell abundances from 2.94 × 104 to 4.97 × 105 cells ml-1, and bacterial production ranging from 58.8 to 293.2 ng C l-1 d-1. Chlorophyll-a concentrations ranged from 0.3 to 0.53 μg l-1 indicating that algal phototrophs were the probable source of the DOM. Microbial isolates produced a rainbow of pigment colors, suggesting adaptation to stress, and were similar to those from other cryogenic systems (Proteobacteria and Bacteroidetes lineages). Supraglacial streams provide an example of contemporary microbial processes on the glacier surface and a natural laboratory for studying microbial adaptation to the absence of humics.

  6. Microbial Load Monitor

    NASA Technical Reports Server (NTRS)

    Gibson, S. F.; Royer, E. R.

    1979-01-01

    The Microbial Load Monitor (MLM) is an automated and computerized system for detection and identification of microorganisms. Additionally, the system is designed to enumerate and provide antimicrobic susceptibility profiles for medically significant bacteria. The system is designed to accomplish these tasks in a time of 13 hours or less versus the traditional time of 24 hours for negatives and 72 hours or more for positives usually required for standard microbiological analysis. The MLM concept differs from other methods of microbial detection in that the system is designed to accept raw untreated clinical samples and to selectively identify each group or species that may be present in a polymicrobic sample.

  7. Aquatic species program

    SciTech Connect

    Bollmeier, W.S.; Sprague, S.

    1989-09-01

    Researchers have learned that many species of aquatic microalgae produce lipids, or oils, when stimulated by environmental stress. These oils can then be processed into diesel fuel or gasoline. Scientists in the Department of Energy (DOE)/Solar Energy Research Institute (SERI) Aquatics Species Program have collected and screened more than 3,000 strains of microalgae from desert and saline environments. The most promising of these strains are maintained in a culture collection at SERI, and research is now focusing on applying genetic techniques to enhance lipid production of microalgae. Researchers are also studying ways to optimize microalgae lipid production by growing the microalgae in intensive cultures of large outdoor ponds. Because microalgae require large amounts of carbon dioxide as a nutrient, these microalgae facilities could be coupled with a power plant or other source of carbon dioxide. Thus, this technology offers not only the potential of producing renewable liquid fuels, but a possible way to improve the environment at the same time. 135 refs., 25 figs., 29 tabs.

  8. Nitrogen in aquatic ecosystems.

    PubMed

    Rabalais, Nancy N

    2002-03-01

    Aquatic ecosystems respond variably to nutrient enrichment and altered nutrient ratios, along a continuum from fresh water through estuarine, coastal, and marine systems. Although phosphorus is considered the limiting nutrient for phytoplankton production in freshwater systems, the effects of atmospheric nitrogen and its contribution to acidification of fresh waters can be detrimental. Within the estuarine to coastal continuum, multiple nutrient limitations occur among nitrogen, phosphorus, and silicon along the salinity gradient and by season, but nitrogen is generally considered the primary limiting nutrient for phytoplankton biomass accumulation. There are well-established, but nonlinear, positive relationships among nitrogen and phosphorus flux, phytoplankton primary production, and fisheries yield. There are thresholds, however, where the load of nutrients to estuarine, coastal and marine systems exceeds the capacity for assimilation of nutrient-enhanced production, and water-quality degradation occurs. Impacts can include noxious and toxic algal blooms, increased turbidity with a subsequent loss of submerged aquatic vegetation, oxygen deficiency, disruption of ecosystem functioning, loss of habitat, loss of biodiversity, shifts in food webs, and loss of harvestable fisheries.

  9. The microbial nitrogen cycle.

    PubMed

    Jetten, Mike S M

    2008-11-01

    This special issue highlights several recent discoveries in the microbial nitrogen cycle including the diversity of nitrogen-fixing bacteria in special habitats, distribution and contribution of aerobic ammonium oxidation by bacteria and crenarchaea in various aquatic and terrestrial ecosystems, regulation of metabolism in nitrifying bacteria, the molecular diversity of denitrifying microorganisms and their enzymes, the functional diversity of freshwater and marine anammox bacteria, the physiology of nitrite-dependent anaerobic methane oxidation and the degradation of recalcitrant organic nitrogen compounds. Simultaneously the articles in this issue show that many questions still need to be addressed, and that the microbes involved in catalyzing the nitrogen conversions still harbour many secrets that need to be disclosed to fully understand the biogeochemical nitrogen cycle, and make future predictions and global modelling possible.

  10. Microbial pesticides

    Treesearch

    Michael L. McManus

    1991-01-01

    Interest in the use of microbial pesticides has intensified because of public concern about the safety of chemical pesticides and their impact in the environment. Characteristics of the five groups of entomopathogens that have potential as microbial pesticides are briefly discussed and an update is provided on research and development activities underway to enhance the...

  11. Antibiotics promote aggregation within aquatic bacterial communities

    PubMed Central

    Corno, Gianluca; Coci, Manuela; Giardina, Marco; Plechuk, Sonia; Campanile, Floriana; Stefani, Stefania

    2014-01-01

    The release of antibiotics (AB) into the environment poses several threats for human health due to potential development of AB-resistant natural bacteria. Even though the use of low-dose antibiotics has been promoted in health care and farming, significant amounts of AB are observed in aquatic environments. Knowledge on the impact of AB on natural bacterial communities is missing both in terms of spread and evolution of resistance mechanisms, and of modifications of community composition and productivity. New approaches are required to study the response of microbial communities rather than individual resistance genes. In this study a chemostat-based experiment with 4 coexisting bacterial strains has been performed to mimicking the response of a freshwater bacterial community to the presence of antibiotics in low and high doses. Bacterial abundance rapidly decreased by 75% in the presence of AB, independently of their concentration, and remained constant until the end of the experiment. The bacterial community was mainly dominated by Aeromonas hydrophila and Brevundimonas intermedia while the other two strains, Micrococcus luteus and Rhodococcus sp. never exceed 10%. Interestingly, the bacterial strains, which were isolated at the end of the experiment, were not AB-resistant, while reassembled communities composed of the 4 strains, isolated from treatments under AB stress, significantly raised their performance (growth rate, abundance) in the presence of AB compared to the communities reassembled with strains isolated from the treatment without AB. By investigating the phenotypic adaptations of the communities subjected to the different treatments, we found that the presence of AB significantly increased co-aggregation by 5–6 fold. These results represent the first observation of co-aggregation as a successful strategy of AB resistance based on phenotype in aquatic bacterial communities, and can represent a fundamental step in the understanding of the effects of

  12. Investigating microbial transformations of soil organic matter: synthesizing knowledge from disparate fields to guide new experimentation

    NASA Astrophysics Data System (ADS)

    Billings, S. A.; Tiemann, L. K.; Ballantyne, F., IV; Lehmeier, C. A.; Min, K.

    2015-04-01

    Discerning why some soil organic matter (SOM) leaves soil profiles relatively quickly while other compounds, especially at depth, can be retained for decades to millennia is challenging for a multitude of reasons. Simultaneous with soil-specific advances, multiple other disciplines have enhanced their knowledge bases in ways potentially useful for future investigations of SOM decay. In this article, we highlight observations highly relevant for those investigating SOM decay and retention but often emanating from disparate fields and residing in literature seldom cited in SOM research. We focus on recent work in two key areas. First, we turn to experimental approaches using natural and artificial aquatic environments to investigate patterns of microbially mediated OM transformations as environmental conditions change, and highlight how aquatic microbial responses to environmental change can reveal processes likely important to OM decay and retention in soils. Second, we emphasize the importance of establishing intrinsic patterns of decay kinetics for purified substrates commonly found in soils to develop baseline rates. These decay kinetics - which represent the upper limit of the reaction rates - can then be compared to substrate decay kinetics observed in natural samples, which integrate intrinsic decay reaction rates and edaphic factors essential to the site under study but absent in purified systems. That comparison permits the site-specific factors to be parsed from the fundamental decay kinetics, an important advance in our understanding of SOM decay (and thus persistence) in natural systems. We then suggest ways in which empirical observations from aquatic systems and purified substrate-enzyme reaction kinetics can be used to advance recent theoretical efforts in SOM-focused research. Finally, we suggest how the observations in aquatic and purified substrate-enzyme systems could be used to help unravel the puzzles presented by oft-observed patterns of SOM

  13. Does aquatic foraging impact head shape evolution in snakes?

    PubMed

    Segall, Marion; Cornette, Raphaël; Fabre, Anne-Claire; Godoy-Diana, Ramiro; Herrel, Anthony

    2016-08-31

    Evolutionary trajectories are often biased by developmental and historical factors. However, environmental factors can also impose constraints on the evolutionary trajectories of organisms leading to convergence of morphology in similar ecological contexts. The physical properties of water impose strong constraints on aquatic feeding animals by generating pressure waves that can alert prey and potentially push them away from the mouth. These hydrodynamic constraints have resulted in the independent evolution of suction feeding in most groups of secondarily aquatic tetrapods. Despite the fact that snakes cannot use suction, they have invaded the aquatic milieu many times independently. Here, we test whether the aquatic environment has constrained head shape evolution in snakes and whether shape converges on that predicted by biomechanical models. To do so, we used three-dimensional geometric morphometrics and comparative, phylogenetically informed analyses on a large sample of aquatic snake species. Our results show that aquatic snakes partially conform to our predictions and have a narrower anterior part of the head and dorsally positioned eyes and nostrils. This morphology is observed, irrespective of the phylogenetic relationships among species, suggesting that the aquatic environment does indeed drive the evolution of head shape in snakes, thus biasing the evolutionary trajectory of this group of animals.

  14. PLFA analyses of microbial communities associated with PAH-contaminated riverbank sediment.

    PubMed

    Pratt, Brenda; Riesen, Roland; Johnston, Carl G

    2012-10-01

    Sediment contaminated with polycyclic aromatic hydrocarbons (PAHs) is widely distributed in aquatic ecosystems. The microbial community structure of riverbank PAH-contaminated sediments was investigated using phospholipid-derived fatty acid (PLFA) analysis. Surface and subsurface riverbank sediment was collected from a highly contaminated site and from an uncontaminated site along the Mahoning River, OH. PAH concentrations, physical sediment characteristics, and other microbial community parameters (biomass as phospholipid phosphate (PLP) and activity) were also measured. PAHs were detected in all samples but were only quantifiable in the contaminated (250 μg/g g(-1)) subsurface sediment. Subsurface samples from both locations showed very similar PLP values and distribution of PLFAs, with 27-37 % of the microbial community structure being composed of sulfate reducing and other anaerobic bacteria. Principal components analysis indicated no correlation between PAH contamination and PLFA diversity. Although PLP and phospholipid fatty acid measurements of bacterial communities did not reflect the environmental differences among sites, the highly PAH-contaminated sediment showed the highest measured microbial activity (reduction of 1,200 nmol INT g(-1) h(-1)), likely from a population adapted to environmental pollutants, rates that are much higher than measured in many uncontaminated soil and sediment systems. These data warrant further investigation into community structure at the genetic level and indicate potential for bioremediation by indigenous microbes.

  15. Assessing impacts of unconventional natural gas extraction on microbial communities in headwater stream ecosystems in Northwestern Pennsylvania

    PubMed Central

    Trexler, Ryan; Solomon, Caroline; Brislawn, Colin J.; Wright, Justin R.; Rosenberger, Abigail; McClure, Erin E.; Grube, Alyssa M.; Peterson, Mark P.; Keddache, Mehdi; Mason, Olivia U.; Hazen, Terry C.; Grant, Christopher J.; Lamendella, Regina

    2014-01-01

    Hydraulic fracturing and horizontal drilling have increased dramatically in Pennsylvania Marcellus shale formations, however the potential for major environmental impacts are still incompletely understood. High-throughput sequencing of the 16S rRNA gene was performed to characterize the microbial community structure of water, sediment, bryophyte, and biofilm samples from 26 headwater stream sites in northwestern Pennsylvania with different histories of fracking activity within Marcellus shale formations. Further, we describe the relationship between microbial community structure and environmental parameters measured. Approximately 3.2 million 16S rRNA gene sequences were retrieved from a total of 58 samples. Microbial community analyses showed significant reductions in species richness as well as evenness in sites with Marcellus shale activity. Beta diversity analyses revealed distinct microbial community structure between sites with and without Marcellus shale activity. For example, operational taxonomic units (OTUs) within the Acetobacteracea, Methylocystaceae, Acidobacteriaceae, and Phenylobacterium were greater than three log-fold more abundant in MSA+ sites as compared to MSA− sites. Further, several of these OTUs were strongly negatively correlated with pH and positively correlated with the number of wellpads in a watershed. It should be noted that many of the OTUs enriched in MSA+ sites are putative acidophilic and/or methanotrophic populations. This study revealed apparent shifts in the autochthonous microbial communities and highlighted potential members that could be responding to changing stream conditions as a result of nascent industrial activity in these aquatic ecosystems. PMID:25408683

  16. Assessing impacts of unconventional natural gas extraction on microbial communities in headwater stream ecosystems in Northwestern Pennsylvania.

    PubMed

    Trexler, Ryan; Solomon, Caroline; Brislawn, Colin J; Wright, Justin R; Rosenberger, Abigail; McClure, Erin E; Grube, Alyssa M; Peterson, Mark P; Keddache, Mehdi; Mason, Olivia U; Hazen, Terry C; Grant, Christopher J; Lamendella, Regina

    2014-01-01

    Hydraulic fracturing and horizontal drilling have increased dramatically in Pennsylvania Marcellus shale formations, however the potential for major environmental impacts are still incompletely understood. High-throughput sequencing of the 16S rRNA gene was performed to characterize the microbial community structure of water, sediment, bryophyte, and biofilm samples from 26 headwater stream sites in northwestern Pennsylvania with different histories of fracking activity within Marcellus shale formations. Further, we describe the relationship between microbial community structure and environmental parameters measured. Approximately 3.2 million 16S rRNA gene sequences were retrieved from a total of 58 samples. Microbial community analyses showed significant reductions in species richness as well as evenness in sites with Marcellus shale activity. Beta diversity analyses revealed distinct microbial community structure between sites with and without Marcellus shale activity. For example, operational taxonomic units (OTUs) within the Acetobacteracea, Methylocystaceae, Acidobacteriaceae, and Phenylobacterium were greater than three log-fold more abundant in MSA+ sites as compared to MSA- sites. Further, several of these OTUs were strongly negatively correlated with pH and positively correlated with the number of wellpads in a watershed. It should be noted that many of the OTUs enriched in MSA+ sites are putative acidophilic and/or methanotrophic populations. This study revealed apparent shifts in the autochthonous microbial communities and highlighted potential members that could be responding to changing stream conditions as a result of nascent industrial activity in these aquatic ecosystems.

  17. Aquatic facility design-designing for Atlantis?

    PubMed

    Bartlett, Doreen H

    2005-01-01

    The requirements for aquatic facility design differ greatly from those of a rodent facility. The author discusses factors to consider when planning new construction of an aquatic facility or renovating space to house aquatic species.