Wednesday, December 11, 2013

Chapter Outline: Phylogenomic Networks of Microbial Genome Evolution


Chapter Outline:  Phylogenomic Networks of Microbial Genome Evolution

 
Author(s): Tal Dagan (PhD),* Ovidiu Popa (Dipl), Thorsten Klösges (Dipl), Giddy Landan

(PhD)


Chapter no.: _________


*Genomic Microbiology Group, Institute of Microbiology

Christian-Albrechts-University Kiel

ZMB, Am Botanischen Garten 11

24118 Kiel, Germany


Tel: +49 431 880 5712

Fax: +49 431 880 5747




Proposed topics


1. Microbial Evolution by Lateral Gene Transfer

1.1 Transformation

1.2 Conjugation

1.3 Cytoplasmic bridges and nanotubes

1.4 Transduction

1.5 Gene transfer agents (GTA)

1.6 Outer membrane vesicles


2 Microbial Phylogenomics

2.1 Networks

2.2 Phylogenetic networks

2.3 Phylogenomic networks of shared genes

2.4 Phylogenomic LGT networks from shared genes

2.5 Phylogenomic LGT networks from trees

2.6 Structural properties of phylogenomic networks



Chapter Highlights


The following concepts will be conveyed in this chapter:


1. Mechanisms for lateral gene transfer during microbial evolution

2. Microbial phylogeny

3. Phylogenetic networks

4. Constructing and analyzing phylogenomic networks

Monday, November 18, 2013

Chapter Outline: The Microbiology of Extremely Acidic Environments (Johnson and Aguilera)


The Microbiology of Extremely Acidic Environments

D. Barrie Johnson1 and Angeles Aguilera2
 
1College of Natural Sciences, Bangor University, Deiniol Road, Bangor, LL57 2UW, UK

e-mail: d.b.johnson@bangor.ac.uk
Tel: +44 1248 382358

2Centro de Astrobiología (INTA-CSIC), Carretera de Ajalvir Km4, 28850 Madrid,Spain.
e-mail: aguileraba@cab.inta-csic.es
Tel: +34 520 6461

Contents
Abstract & Highlights

1.     Nature and diversity of extremely acidic environments

2.     Diversity of acidophiles, and adaptation to living in low pH environments

      3.   Diversity of eukaryotic acidophiles

      4.   Physiological and phylogenetic diversity of prokaryotic acidophiles

      5.   Ecological and biotechnological aspects    
 

Abstract

 

Extremely acidic environments, defined having a pH of <3, are found in locations as diverse as the Arctic and the Tropics. While these can be natural phenomena, human activity, most notoriously mining of metals and coals, is often responsible for the severe acidification of localized environments. The indigenous microflora in extremely acidic environments includes species of prokaryotes and eukaryotes, many of which are obligately acidophilic. Acidophiles are widely distributed throughout the “tree of life” and include species of Bacteria, Archaea, and Eukarya that are often only very distantly related to each other. Various mechanisms are used by acidophiles to adapt to the challenges they face, which include contending with elevated concentrations of transition metals and metalloids, and severely limited bioavailability of macronutrients such as phosphate. Inorganic energy sources (reduced iron and sulfur) are highly abundant in many extremely acidic environments. Chemolithotrophic acidophiles are the basis of food webs in subterranean and also contribute to net primary production in deep submarine geothermal vents. However, where solar energy is available phototrophic acidophiles, predominantly species of acidophilic eukaryotic microalgae, proliferate and assume the dominant role of primary producers. Acidophilic microorganisms interact with each other in various ways, including via redox transformations of iron and sulfur, generating electron donors and acceptors for prokaryotic metabolisms, and via provision of organic compounds (supporting heterotrophic species) or inorganic carbon (supporting autotrophs). Acidophiles have long been used to extract metals from ores (biomining) and biotechnologies are emerging that harness their abilities to remediate polluted waters and recover metals.

 

 

Highlights (major points covered)

·        The characteristics and origins of extremely acidic environments

·        How acidophiles adapt to acid stress and elevated concentrations of metals

·        An overview of the physiological and phylogenetic diversities of prokaryotic and eukaryotic microorganisms

·        The microbial ecology of extremely acidic environments

·        Application of acidophiles in established and emerging biotechnologies

 

Wednesday, September 11, 2013

Chapter Outline: Fecal Indicator Bacteria Monitoring in Environmental Waters: Overview of Existing Modeling Efforts (Nevers et al.)


Fecal Indicator Bacteria Monitoring in Environmental Waters: Overview of Existing Modeling Efforts                                                               Chapter: 5  

Authors:
      Meredith Nevers,1 Murulee Byappanahalli,1 Phanikumar Mantha,2 and Richard Whitman1
            1US Geological Survey, 1100 N. Mineral Springs Road, Porter, IN 46304
      2Michigan State University, Department of Civil and Environmental Engineering, East Lansing,  MI 48824

Chapter Outline
      1.      Introduction
a.      Description of models
b.      Different uses for models
2.      Sources of indicator bacteria to recreational surface waters
a.      Environmental sources (sand, algae and vegetation, detritus, soil/sediments, planktonic materials, and macroinvertebrates)
b.      Distinguishing sewage sources using microbial source tracking
c.      Characterizing human health risk based on source origin
3.      Mechanistic/dynamic modeling of microbial movement and survival and application to source identification
a.      Salient physical and biological processes
b.      Plume dynamics and extent of influence on coastal areas
c.      Influence of beach morphometry/situation on sources and transport
4.      Predicting pathogens/pathogen indicators’ concentrations to protect public health; using knowledge of microbial movement, survival, and source to predict beach water quality in real time
a.      Model types: simple, regression, advanced statistical methods and improvements in accuracy
b.      Development of individual and multi-beach models
c.      Integration of mechanistic and empirical models; database models
d.      Importance of validation and multi-year studies
5.      Modeling indicator bacteria/pathogens for surface water monitoring: applied use
a.      Examples of current applications
b.      Assessment of reduction in health risk
c.      Potential for signaling contamination events and opportunities for remediation
6.      Comparison with other potential microbiological monitoring techniques
a.      Table showing pros and cons of each monitoring approach, including accuracy, health protection, and practical use
7.      Summary and Conclusions

Chapter Outline: Validating MST Methods (Stoeckel et al.)


Validating MST Methods (Stoeckel et al.)                                         Chapter 2       

Authors: Don Stoeckel, John Griffith and Asja Korajkic
Background

  • What do we mean by method validation?
    • Validation needs depend on study objectives
      • Resolution of sources (categories; human/nonhuman)
      • To quantify or to categorize?
    • Validation includes three levels – source identifier, measurement protocol, and interpretation paradigm
  • Summary of source identifiers
    • Technique used to develop the method (e.g. metagenomics, subtractive hybridization, etc)
    • Target characteristic (e.g. 16S or other genes)
    • Target host (General fecal/ruminant/bird or species-level association)
  • Summary of validation literature
    • Lessons learned from FC/FS ratio
    • Mention observed changes in application from library-dependent to library-independent methods following USGS and SCCWRP validation attempts
    • Existing compiled sensitivity and specificity information for markers
      • Stoeckel and Harwood
      • Series by Shanks
      • SCCWRP Study – Boehm et al., Layton et al.
MST method evaluations

    • Validation of the Source Identifier
      • Theoretical tests
        • If genetic, check sequences in GenBank  for specificity and evenness within host population
      • Controlled tests for sensitivity and specificity
        • For sensitivity, serial dilutions of the “real-life” intended target (i.e. sewage for human markers) (use of aged or partially-treated material for enhanced relevance)
        • For specificity, test method against non-targets (use of pooled material for efficiency)
    • Protocol Performance
      • Analytical limit of detection and quantifiable range of the target
      • Inter-lab performance comparisons to validate transferability of methodology
    • Interpretation Paradigm
      • Test method against known levels of target seeded into ambient water
      • Field test at sites known to be contaminated versus “pristine”
Considerations for Effective Interpretation

(Each topic carries specific examples of how to obtain the needed information to “validate” the use of a source identifier, protocol, and interpretation paradigm.)

    • Coverage across “intended” hosts – has variability been fully captured?
      • Understanding of population of source required in order to detect contamination (e.g., septic systems vs WWTP losses)
      • Understanding of units needed to measure “fecal contamination” (i.e., targets per gram fecal material, per unit of pathogen carried, or per unit of fecal indicator carried depending on the objective of the study)
    • Coverage across “alternate” hosts – has potential for false-positive results been characterized?
    • Limits of detection and quantifiable ranges – does the protocol deliver data that meet the goals of the study
    • Geographic/temporal stability – is the source identifier and protocol used to measure the source identifier verified in the study area/study lab?
    • Effect of aging on marker stability and relationship to regulated FIB
    • Quantitation – have all sources of loss been mathematically corrected?
      • Recovery efficiency
      • Extraction efficiency
      • Volumetric losses during processing
      • Matrix inhibition/detection efficiency
      • Degradation during storage
      • Standard curve
    • Reporting – are the results reported in relevant units?
      • Targets per liter of water
      • Targets per nanogram DNA extracted
      • Targets per E. coli or enterococci cells (to allocate fecal contamination and meet regulatory criteria)
      • Effect of aging on marker stability and relationship to regulated FIB

Chapter Outline: MST: Field Study Planning and Implementation


Proposed title: MST: Field Study Planning and Implementation
(Kinzelman and Ahmed)        

Chapter no.: 6
 
Authors:
Julie Kinzelman (Ph.D.)
                  Racine Health Department Laboratory
                  730 Washington Avenue, Room 8
                  Racine, WI 53403 USA
                  Phone: (+001) 262-636-9501
                  Fax: (+001) 262-636-9576
                  E-mail: julie.kinzelman@cityofracine.org
 
                  Warish Ahmed (Ph.D.)
                  CSIRO Land and Water
                  Water for a Healthy Country Flagship
                  Queensland Biosciences Precinct
                  306 Carmody Rd. St. Lucia Qld 4067 AUSTRALIA
                  Phone: (+ 617) 3833 5582
                  Fax: (+617) 3833 5503
                  E- mail: Warish.Ahmed@csiro.au

Notes

  • Define the anticipated outcome in order to determine study design
  • Multiple lines of evidence may be necessary, i.e., a “tool-box” approach. Refer to methods presented in other chapters (analytical and field based)
  • Weight of evidence approach towards implementation—correlation, not causation
  • A multi-barrier approach should be stressed in communication of results/implementation, i.e., public education in addition to naturalized or engineered solutions
      Proposed Topics

1)     Define (or refer back to) aquatic environments, routes of exposure, human exposure interventions.
      2)     Regulatory approaches: TMDLs, bathing water, permitting

3)     Determining study outcome/objectives.

4)     Field study design: sampling duration/frequency, accounting for temporal/spatial variation, leveraging historic data.

5)     Considerations for method choice. Stress the use of sanitary surveys/inspections and spatial distribution studies to focus more costly, targeted MST methods on the probable source. Use of both analytical methods and models. Determine level of detail required, i.e. human vs. nonhuman or species specific. Multiple methods may be necessary.

6)     What do you do with all the data? Data analysis and interpretation.

7)     Stakeholder engagement. Translation of study results into actionable items (implementation).

8)     Case studies.

9)     Barriers and confounding factors, i.e., cost, access to analytical facilities, assay inhibition, lack of specificity/sensitivity, changes in land use, climate change, transferability of technologies to local water authorities, etc.

     Chapter Highlights

The following concepts will be conveyed in this chapter:

  1. Significant advances have occurred in both the development of MST tools and the application of these to tools in assessing fecal pollution in aquatic environments.
  2. Host specificity—can Baye’s theorem be applied based on the published data on the host specificity of a particular marker such as HF183?
  3. A single MST tool may provide adequate information, multiple lines of evidence may be necessary in order to strengthen the association between measures environmental parameters and pollution sources.
  4. What magnitude of fecal pollution can pose a risk? Is the presence of a marker enough? Importance of quantitative data over presence/absence. 
  5. A weight of evidence approach, in the presence or absence of a definite source, has been successfully applied to the mitigation of pollution sources.
  6. Results of MST studies must be taken in context. Confounding factors exist and must be addressed.

 

 

Chapter Outline: Methods of Targeting Animal Sources of Fecal Pollution in Water (Blanch et al.)


Proposed title: Methods of Targeting Animal Sources of Fecal Pollution in Water
Chapter number: 4
Authors
Anicet R Blanch. Department of Microbiology, University of Barcelona. Diagonal 643. 08028 Barcelona (Spain), ablanch@ub.edu Phone: +34 934029012 (AB)
Elisenda Ballesté. Department of Microbiology, University of Barcelona. Diagonal 643. 08028 Barcelona (Spain), elisballeste@gmail.com Phone: +34 934039044 (EB)
Jennifer Weidhaas. Civil and Environmental Engineering, 647 Engineering Sciences Building, PO Box 6103, Morgantown, WV 26506. jennifer.weidhaas@mail.wvu.edu Phone: +1 304-293-9952 (JW)
Jorge Santodomingo. U.S. EPA, NRMRL/WSWRD/MCCB, 26 W. Martin Luther King Dr. MS 387 Cincinnati, OH  45268, Santodomingo.Jorge@epamail.epa.gov Phone: +1 5135697085 (JS)
Hodon Ryu. U.S. EPA, NRMRL/WSWRD/MCCB, 26 W. Martin Luther King Dr. MS 387 Cincinnati, OH  45268 ryu.hodon@epamail.epa.gov Phone: XXXXXXX (HR)
Notes
Chemical indicators are briefly overviewed by avoiding overlap with chapter 5.
Chapter Highlights
The determination of fecal pollution sources in waters is an essential subject in the management of catchments. Municipal sewage, slaughterhouse wastewaters, manure and different biowaste disposal, wildlife and undetermined runoff are some of the different fecal pollution sources. Although traditional microbiological water analyses using indicator microorganisms have showed to be highly useful for water-health management for more than a century, it is known that they are not providing information about the origin of fecal pollution. The distinction between anthropogenic and non-anthropogenic (animal) fecal pollution would greatly support assessment of health risks associated with the host-specificity of many pathogens. Human sewage could constitute a higher health risk to humans than wastewater of animal origin. However, there are some exceptions because of some pathogens (named zoonosis) can infect and cause clinical disease in both humans and animals. Therefore, the fecal pollution source assessment could support and determine different water management strategies, treatment measures and policies to prevent or decrease fecal inputs in water based on the principles of precaution, prevention and remediation of environmental contaminant at the source.
At the beginning, most of the proposed indicators on Microbial Source Tracking were defined and developed to target human fecal pollution sources. However, the distinction of different animal species sources was enforced to QMRA studies, to resolve complex mixtures from several distinct animal species or to identify diffuse pollution sources.
In this chapter, proposed chemical and biological MST indicators for the determination of animal fecal sources are firstly described and analyzed. The biological indicators are grouped based on the phylogenetic adscription of the proposed target (eukarya, bacteria, and virus). A comprehensive description for each proposed target is provided and the developed methodologies employed in their respective analyses are presented, referred and analyzed. Special emphasis on validation and applicability for each proposed method and animal-MST indicator is quoted.  Moreover, each proposed target is critically reviewed concerning environmental factors such prevalence, resistance to different water treatments, and environmental persistence. New molecular approaches for animal-NST targets based on metagenomics are also presented in a specific section by analyzing limitations and strengths at the present stage of the methodology. Finally, MST assay implementation on practical cases, their contribution to the assessment of maximum fecal load of water bodies and their relationship to traditional microbial indicators and waterborne pathogens is examined.
 Proposed topics
1.      Introduction (JS & AB). Short description of initial studies and factors requesting the animal fecal source markers on MST studies, and state of the art.
2.      MST targets for animal sources:
2.1.   Chemical targets (JW). Comprehensive presentation of proposed chemical markers for animal fecal sources, avoiding overlap with chapter 5.
2.2.   Biological targets (each proposed marker/target will be comprehensively described; guidelines for selecting the best procedure will be indicated explaining applications or circumstances supported by reference. Validation studies and environmental factors such prevalence, resistance to different water treatments, and environmental persistence should be also considered):
2.2.1. Eukaryotic targets:
2.2.1.1.            DNA Mitochondrial Targets (JW).
2.2.1.2.            Parasites: Cryptosporidium and Giardia (HR & JS)
2.2.2. Bacterial targets:
2.2.2.1.            Bacteroides and related genera (EB & AB). Also reported as Order Bacteroidales (Bacteroides, Prevotella, Porphyromonas and Parabacteroides).
2.2.2.2.            Bifidobacterium and related genera (EB & AB). Including the recently described genus Neoscardovia associated with porcine.
2.2.2.3.            Brevibacterium (JW)
2.2.2.4.            Rodococcus coprophilus (JW)
2.2.2.5.            Enterococcus, Streptococcus, Catellicoccus, and Helicobacter (JS & HR)
2.2.2.6.            Phenotypic library-depending methods based on microbial antibiotic resistance or carbon source utilization (JS & HR). Avoiding overlapping to chapter 3 being focused on animal target.
2.2.2.7.            Genotypic library-depending methods (rep-PCR, RAPD, AFLP, PFGE, rybotypin) (JS & HR). Avoiding overlapping to chapter 3 being focused on animal targets.
2.2.3. Viral targets:
2.2.3.1.            Bacteriophages (EB & AB). Including mainly bacteriophages of F-RNA and Bacteroides.
2.2.3.2.            Animal viruses (EB & AB).
3.      New molecular approaches and future perspectives: metagenomics (HR & JS).
4.      Implementation in routine MST analyses. Relationship to traditional FIB and pathogens (JS & JW).
5.      Discussion (AB, EB, JW, JS, HR). All co-authors should provide their respective contributions to this section. A table of methods and targets with pro/cons could be also included in this section.