Environmental Microbiology and Microbial Ecology Commons^™

Methanogen Diversity Evidenced By Molecular Characterization Of Methyl Coenzyme M Reductase A (Mcra) Genes In Hydrothermal Sediments Of The Guaymas Basin, Ashita Dhillon, Mark Lever, Karen Lloyd, Daniel B. Albert, Mitchell L. Sogin, Andreas Teske

Karen Lloyd

The methanogenic community in hydrothermally active sediments of Guaymas Basin (Gulf of California, Mexico) was analyzed by PCR amplification, cloning, and sequencing of methyl coenzyme M reductase (mcrA) and 16S rRNA genes. Members of the Methanomicrobiales and Methanosarcinales dominated the mcrA and 16S rRNA clone libraries from the upper 15 cm of the sediments. Within the H2/CO2- and formate-utilizing family Methanomicrobiales, two mcrA and 16S rRNA lineages were closely affiliated with cultured species of the genera Methanoculleus and Methanocorpusculum. The most frequently recovered mcrA PCR amplicons within the Methanomicrobiales did not branch with any cultured genera. Within the nutritionally versatile …

Characterization Of Microbial Activity, Mark P. Buttner, Patricia Cruz, Klaus J. Stetzenbach, Abe Van Luik, Thomas Williams, Amy J. Smiecinski

Publications (YM)

The overall goal of this study is to investigate the phenomena that affect the fate and transport of radionuclides in the environment. The objective of this task, “Characterization of Microbial Activity”, is to develop a molecular biological method for the characterization of the microbial population indigenous to the Yucca Mountain Project site, with emphasis in detection and measurement of species or groups of microorganisms that could be involved in actinide and/or metal reduction, and subsurface transport. Subtasks consist of QA planning and preparation, and literature review. This task is part of a cooperative agreement between the UNLV Research Foundation and …

The Uptake Of Amino Acids By Microbes And Trees In Three Cold-Temperate Forests, Adrien C. Finzi, Sean T. Berthrong

Scholarship and Professional Work - LAS

Amino acids are emerging as a critical component of the terrestrial N cycle, yet there is little understanding of amino acid cycling in temperate forests. This research studied the uptake and turnover of amino acid N by soil microbes and the capacity of forest trees to take up the amino acid glycine in comparison to NH₄⁺ and NO₃⁻. This research was conducted in three temperate forests located in northwest Connecticut, USA. The three forests differed in soil parent material and canopy tree species composition. At all three sites, amino acids were released from soil organic …

Effects Of Dissolved Sulfide, Ph, And Temperature On Growth And Survival Of Marine Hyperthermophilic Archaea, Karen Lloyd, Virginia P. Edgcomb, Stephen J. Molyneaux, Simone Böer, Carl O. Wirsen, Michael S. Atkins, Andreas Teske

Microbiology Publications and Other Works

The ability of metabolically diverse hyperthermophilic archaea to withstand high temperatures, low pHs, high sulfide concentrations, and the absence of carbon and energy sources was investigated. Close relatives of our study organisms, Methanocaldococcus jannaschii, Archaeoglobus profundus, Thermococcus fumicolans, and Pyrococcus sp. strain GB-D, are commonly found in hydrothermal vent chimney walls and hot sediments and possibly deeper in the subsurface, where highly dynamic hydrothermal flow patterns and steep chemical and temperature gradients provide an ever-changing mosaic of microhabitats. These organisms (with the possible exception of Pyrococcus strain GB-D) tolerated greater extremes of low pH, high sulfide concentration, and high temperature …

Periodic Selection And Ecological Diversity In Bacteria, Frederick Cohan

Frederick M. Cohan

No abstract provided.

Effects Of Dissolved Sulfide, Ph, And Temperature On Growth And Survival Of Marine Hyperthermophilic Archaea, Karen Lloyd, Virginia P. Edgcomb, Stephen J. Molyneaux, Simone Böer, Carl O. Wirsen, Michael S. Atkins, Andreas Teske

Karen Lloyd

The ability of metabolically diverse hyperthermophilic archaea to withstand high temperatures, low pHs, high sulfide concentrations, and the absence of carbon and energy sources was investigated. Close relatives of our study organisms, Methanocaldococcus jannaschii, Archaeoglobus profundus, Thermococcus fumicolans, and Pyrococcus sp. strain GB-D, are commonly found in hydrothermal vent chimney walls and hot sediments and possibly deeper in the subsurface, where highly dynamic hydrothermal flow patterns and steep chemical and temperature gradients provide an ever-changing mosaic of microhabitats. These organisms (with the possible exception of Pyrococcus strain GB-D) tolerated greater extremes of low pH, high sulfide concentration, and high temperature …

The Effects Of Dams In The Big Sandy Watershed Using A Novel Bacteria-Based Bioindicator Of Water Quality, Kathleen Riha Loughman

Theses, Dissertations and Capstones

During 2003, water samples from the Big Sandy watershed were collected in conjunction with the United States Army Corps of Engineers (USACE). Objectives were to determine the effects of dams on bacterial populations and to test a bioindicator of water quality based on antibiotic-resistant and fecal indicator bacteria. Thirty-five samples were taken each season within the Big Sandy Watershed, which includes six USACE dams. Total cultivable, ciprofloxacin-resistant, erythromycin-resistant, tetracycline- resistant, total coliform, and fecal coliform bacteria were enumerated. Data on water chemistry and physical parameters were collected by the USACE in the spring and summer seasons. Antibiotic-resistant bacteria and fecal …

Copepods As Microbial Hotspots In The Ocean: Effects Of Host Feeding Activities On Attached Bacteria, Kw Tang

VIMS Articles

Through ingestion a copepod introduces rich organic substrates into its guts and fecal pellets, where dense bacteria may exploit them and show fast growth. Thus, a copepod and its fecal pellets may be regarded as microbial hotspots in the ocean. This study investigated the effects of copepods' feeding activities on the associated bacteria, using the Most Probable Number (MPN) method. Starved Acartia tonsa (calanoid copepod) carried a background bacteria population of 103 to 10(4) copepod(-1). When fed axenic cultures of algae Rhodomonas salina or Dunaliella tertiolecta, the bacterial abundance increased curvilinearly with the copepods' ingestion rates. When fed axenic cultures …

High-Temperature Fluorescent In Situ Hybridization For Detecting Escherichia Coli In Seawater Samples, Using Rrna-Targeted Oligonucleotide Probes And Flow Cytometry, Ying Zhong Tang, Karina Yew Hoong Gin, Tok Hoon Lim

OES Faculty Publications

Fluorescence in situ hybridization (FISH) is a widely used method to detect environmental microorganisms. The standard protocol is typically conducted at a temperature of 46°C and a hybridization time of 2 or 3 h, using the fluorescence signal intensity as the sole parameter to evaluate the performance of FISH. This paper reports our results for optimizing the conditions of FISH using rRNA-targeted oligonucleotide probes and flow cytometry and the application of these protocols to the detection of Escherichia coli in seawater spiked with E. coli culture. We obtained two types of optimized protocols for FISH, which showed rapid results with …

Microbial Diversity In Hot Spring Cyanobacterial Mats: Pattern And Prediction, Frederick M. Cohan, D. M. Ward

Frederick M. Cohan

No abstract provided.

Which Species Concept For Bacteria?—An E-Debate, S. Godreuil, Frederick M. Cohan, H. Shah, M. Tibayrenc

Frederick M. Cohan

No abstract provided.

Real-Time Pcr For Detection And Quantification Of The Protistan Parasite Perkinsus Marinus In Environmental Waters, Corinne Audemard, Kimberly S. Reece, Em Burreson

VIMS Articles

The protistan parasite Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica along the east coast of the United States. Very few data have been collected, however, on the abundance of the parasite in environmental waters, limiting our understanding of P. marinus transmission dynamics. Real-time PCR assays with SybrGreen I as a label for detection were developed in this study for quantification of P. marinus in environmental waters with P. marinus species-specific primers and of Perkinsus spp. with Perkinsus genus-specific primers. Detection of DNA concentrations as low as the equivalent of 3.3 x 10(-2) cell per 10-mul reaction …

Transport Of The Harmful Bloom Alga Aureococcus Anophagefferens By Oceangoing Ships And Coastal Boats, Martina A. Doblin, Linda C. Popels, Kathryn J. Coyne, David A. Hutchins, S. Craig Cary, Fred C. Dobbs

OES Faculty Publications

It is well established that cyst-forming phytoplankton species are transported in ships' ballast tanks. However, there is increasing evidence that other phytoplankton species which do not encyst are also capable of surviving ballast transit. These species have alternative modes of nutrition (hetero- or mixotrophy) and/or are able to survive long-term darkness. In our studies of no-ballast-on-board vessels arriving in the Great Lakes, we tested for the presence of the harmful algal bloom species Aureococcus anophagefferens (brown tide) in residual (i.e., unpumpable) ballast water using methods based on the PCR. During 2001, the brown tide organism was detected in 7 of …

Dinitrogen Fixation And Release Of Ammonium And Dissolved Organic Nitrogen By Trichodesmium Ims101, Mr Mulholland, Da Bronk, Dg Capone

VIMS Articles

Two methods used to measure dinitrogen (N-2) fixation (acetylene reduction and N-15(2) uptake) often result in different N-2 fixation rates. Part of the discrepancy may arise from the observation that Trichodesmium can release a fraction of their recently fixed N-2 as dissolved organic nitrogen (DON) and/or ammonium (NH4+). To resolve outstanding issues regarding N-2 fixation and the production of dissolved combined nitrogen (N) by Trichodesmium, we conducted a comprehensive analysis of N-2 fixation and the production of DON and NH4+ in cultures of Trichodesmium IMS101. We performed N-15(2) uptake experiments in parallel with acetylene (C2H2) reduction assays, and measured production …

Light Dependence Of [H-3]Leucine Incorporation In The Oligotrophic North Pacific Ocean, Mj Church, Hw Ducklow, Da Karl

VIMS Articles

The influence of irradiance on bacterial incorporation of [H-3] leucine was evaluated at Station ALOHA in the oligotrophic North Pacific subtropical gyre. Six experiments were conducted on three cruises to Station ALOHA to examine how [H-3]leucine incorporation varied as a function of irradiance. Two experiments were also conducted to assess the photoautotrophic response to irradiance (based on photosynthetic uptake of [C-14] bicarbonate) in both the upper and lower photic zones. Rates of [H-3]leucine incorporation responded to irradiance in a photosynthesis-like manner, increasing sharply at low light and then saturating and sometimes declining with increasing light intensity. The influence of irradiance …

Methanogenic Archaea And Human Periodontal Disease, Paul W. Lepp, Mary M. Brinig, Cleber C. Ouverney, Katherine Palm, Gary C. Armitage, David A. Relman

Faculty Publications, Biological Sciences

Archaea have been isolated from the human colon, vagina, and oral cavity, but have not been established as causes of human disease. In this study, we reveal a relationship between the severity of periodontal disease and the relative abundance of archaeal small subunit ribosomal RNA genes (SSU rDNA) in the subgingival crevice by using quantitative PCR. Furthermore, the relative abundance of archaeal small subunit rDNA decreased at treated sites in association with clinical improvement. Archaea were harbored by 36% of periodontitis patients and were restricted to subgingival sites with periodontal disease. The presence of archaeal cells at these sites was …

Sulfide Ameliorates Metal Toxicity For Deep-Sea Hydrothermal Vent Archaea, Virginia P. Edgcomb, Stephen J. Molyneaux, Mak A. Saito, Karen Lloyd, Simone Böer, Carl O. Wirsen, Michael S. Atkins, Andreas Teske

Microbiology Publications and Other Works

The chemical stress factors for microbial life at deep-sea hydrothermal vents include high concentrations of heavy metals and sulfide. Three hyperthermophilic vent archaea, the sulfur-reducing heterotrophs Thermococcus fumicolans and Pyrococcus strain GB-D and the chemolithoautotrophic methanogen Methanocaldococcus jannaschii, were tested for survival tolerance to heavy metals (Zn, Co, and Cu) and sulfide. The sulfide addition consistently ameliorated the high toxicity of free metal cations by the formation of dissolved metal-sulfide complexes as well as solid precipitates. Thus, chemical speciation of heavy metals with sulfide allows hydrothermal vent archaea to tolerate otherwise toxic metal concentrations in their natural environment.

DOI: 10.1128/AEM.70.4.2551–2555.2004

Physiology Of The Thermophilic Acetogen Moorella Thermoacetica, Harold L. Drake, Steven L. Daniel

Steven L. Daniel

Moorella thermoacetica (originally isolated as Clostridium thermoaceticum) has served as the primary acetogenic bacterium for the resolution of the acetyl coenzyme A (acetyl-CoA) orWood–Ljungdahl pathway, a metabolic pathway that (i) autotrophically assimilates CO2 and (ii) is centrally important to the turnover of carbon in many habitats. The purpose of this article is to highlight the diverse physiological features of this model acetogen and to examine some of the consequences of its metabolic capabilities.

Sulfide Ameliorates Metal Toxicity For Deep-Sea Hydrothermal Vent Archaea, Virginia P. Edgcomb, Stephen J. Molyneaux, Mak A. Saito, Karen Lloyd, Simone Böer, Carl O. Wirsen, Michael S. Atkins, Andreas Teske

Karen Lloyd

The chemical stress factors for microbial life at deep-sea hydrothermal vents include high concentrations of heavy metals and sulfide. Three hyperthermophilic vent archaea, the sulfur-reducing heterotrophs Thermococcus fumicolans and Pyrococcus strain GB-D and the chemolithoautotrophic methanogen Methanocaldococcus jannaschii, were tested for survival tolerance to heavy metals (Zn, Co, and Cu) and sulfide. The sulfide addition consistently ameliorated the high toxicity of free metal cations by the formation of dissolved metal-sulfide complexes as well as solid precipitates. Thus, chemical speciation of heavy metals with sulfide allows hydrothermal vent archaea to tolerate otherwise toxic metal concentrations in their natural environment. DOI: 10.1128/AEM.70.4.2551–2555.2004

Physiology Of The Thermophilic Acetogen Moorella Thermoacetica, Harold Drake, Steven Daniel

Faculty Research & Creative Activity

Moorella thermoacetica (originally isolated as Clostridium thermoaceticum) has served as the primary acetogenic bacterium for the resolution of the acetyl coenzyme A (acetyl-CoA) orWood–Ljungdahl pathway, a metabolic pathway that (i) autotrophically assimilates CO2 and (ii) is centrally important to the turnover of carbon in many habitats. The purpose of this article is to highlight the diverse physiological features of this model acetogen and to examine some of the consequences of its metabolic capabilities.

Physiology Of The Thermophilic Acetogen Moorella Thermoacetica, Harold L. Drake, Steven L. Daniel

Faculty Research & Creative Activity

Moorella thermoacetica (originally isolated as Clostridium thermoaceticum) has served as the primary acetogenic bacterium for the resolution of the acetyl coenzyme A (acetyl-CoA) orWood–Ljungdahl pathway, a metabolic pathway that (i) autotrophically assimilates CO2 and (ii) is centrally important to the turnover of carbon in many habitats. The purpose of this article is to highlight the diverse physiological features of this model acetogen and to examine some of the consequences of its metabolic capabilities.

Particle-Associated Flagellates: Swimming Patterns, Colonization Rates, And Grazing On Attached Bacteria, T Kiorboe, Hp Grossart, H Ploug, K Tang, B Auer

VIMS Articles

Some pelagic flagellates colonize particles, such as marine snow, where they graze on bacteria and thus impact the dynamics of the attached microbial communities. Particle colonization is governed by motility. Swimming patterns of 2 particle-associated flagellates, Bodo designis and Spumella sp., are very different, the former swimming slowly in an erratic, random pattern, and the latter faster and along smooth helixes of variable amplitude and frequency. At spatial scales exceeding ca. 50 mum, the motility of B. designis can be described as a random walk and modeled as diffusion. Spumella sp. shows directional persistence of the helical axes up to …

Mycobacterium-Inducible Nramp In Striped Bass (Morone Saxatilis), Erin J. Burge, David T. Gauthier, Christopher A. Ottinger, Peter A. Van Veld

VIMS Articles

In mammals, the natural resistance-associated macrophage protein 1 gene, Nramp1, plays a major role in resistance to mycobacterial infections. Chesapeake Bay striped bass (Morone saxatilis) is currently experiencing an epizootic of mycobacteriosis that threatens the health of this ecologically and economically important species. In the present study, we characterized an Nramp gene in this species and obtained evidence that there is induction following Mycobacterium exposure. The striped bass Nramp gene (MsNramp) and a 554-amino-acid sequence contain all the signal features of the Nramp family, including a topology of 12 transmembrane domains (TM), the transport protein-specific binding-protein-dependent transport system inner membrane …

Oxalate Metabolism By The Acetogenic Bacterium Moorella Thermoacetica, Steven L. Daniel, Christine Pilsl, Harold L. Drake

Steven L. Daniel

Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction …

Oxalate Metabolism By The Acetogenic Bacterium Moorella Thermoacetica, Steven Daniel, Christine Pilsl, Harold Drake

Faculty Research & Creative Activity

Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction …

Oxalate Metabolism By The Acetogenic Bacterium Moorella Thermoacetica, Steven L. Daniel, Christine Pilsl, Harold L. Drake

Faculty Research & Creative Activity

Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction …

High Phosphate (Up To 600 Mm) Induces Pseudohyphal Development In Five Wild Type Candida Albicans, Jacob M. Hornby, Raluca Dumitru, Kenneth Nickerson

Kenneth Nickerson Papers

A method is described for the formation of nearly 100% pseudohyphae populations of wild-type Candida albicans A72. The method employs fungal growth at 37 °C (ca. 5×10⁶ cells/ml) in a glucose–proline–N-acetyl-glucosamine medium supplemented with up to 600 mM phosphate (KH₂PO₄/K₂HPO₄ 1:1) at pH 6.5. Four other strains of C. albicans (MEN, 10261, SG5314 and CAI-4) also formed pseudohyphae under these conditions, although the phosphate response profiles differed in the concentration required for each strain to form pseudohyphae.

A Survey Of Antibiotic Resistance Among Coliform Bacteria Isolated From The Missouri River, Sara E. Mcdonnell, Amy M. Treonis

Biology Faculty Publications

The prevalence of antibiotic resistant microorganisms in the environment is not well known but could represent a challenge to maintaining public health in the future. Fecal waste from livestock facilities, where antibiotics are routinely used, is recognized as a significant source of pollution to surface waters in the United States. We collected water from the Missouri River across a winter to spring seasonal change in order to survey the density of coliform bacteria. We tested bacterial isolates for resistance to penicillin, tetracycline, ampicillin, erythromycin, and ciprofloxacin. Coliform density in Missouri River water generally was high throughout the study (0-405 cfu …

Microbial Endemism And Biogeography, Brian P. Hedlund, James T. Staley

Life Sciences Faculty Research

The topic of microbial biogeography is almost 100 years old, however, when confronted with questions about the existence and extent of endemism in the microbial world, many microbiologists respond with opinions and theoretical arguments rather than examples of well-conducted studies. We begin this chapter with an overview of this debate as it applies to free-living prokayotes in part because there are relatively few good microbial biogeography studies. Furthermore, the arguments help to frame microbial biogeography in the larger context of biodiversity in that if endemism is common, then many more species exist.

Endocrine-Disrupting Effects Of Cattle Feedlot Effluent On An Aquatic Sentinel Species, The Fathead Minnow, Edward Orlando, Alan Kolok, Gerry Binzcick, Jennifer Gates, Megan Horton, Christy Lambright, L. Earl Gray Jr., Ana M. Soto, Louis J. Guilette

Biology Faculty Publications

Over the last decade, research has examined the endocrine-disrupting action of various environmental pollutants, including hormones, pharmaceuticals, and surfactants, in sewage treatment plant effluent. Responding to the growth of concentrated animal feeding operations (CAFOs) and the pollutants present in their wastewater (e.g., nutrients, pharmaceuticals, and hormones), the U.S. Environmental Protection Agency developed a new rule that tightens the regulation of CAFOs. In this study, we collected wild fathead minnows (Pimephales promelas) exposed to feedlot effluent (FLE) and observed significant alterations in their reproductive biology. Male fish were demasculinized (having lower testicular testosterone synthesis, altered head morphometrics, and smaller …