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01 Microbial World - Coggle Diagram

- - - - Suspect Must be Present in All Cases of Disease & Absent from Healthy Animals
      - Suspect Must be Grown in Pure Culture
      - Pure Culture Must Cause Disease in Healthy Animal
      - Suspect Must Be Reisolated & Same as Original
      - Limitations
        
        Difficult to Culture
        
        Host or Condition Preference
        
        Asymptomatic Carriers
  - - - Enrichment Culture Technique
      - Isolation by Manipulating Nutrient & Incubation Conditions
  - - - Frederick Griffith
        
        Transforming Principle
        
        Stable Material Can Alter Another Cell's Inherited Traits
      - Avery-Macleod-McCarty
        
        Destruction of DNA, Not Protein or RNA, Eliminated Transformation
      - Watson, Crick, Franklin
        
        Double Helical Structure of DNA
      - Zuckerkandl & Pauling
        
        Molecular Sequences & Evolutionary Relationships in DNA & RNA
  - - - Methanogens
      - Discovered Archaea
      - Ribosomal RNA
        
        Non-Coding
        
        Same Function Across Life
      - Phylogenetic Tree
        
        Culture
        
        Experiments, Phenotyping, Antimicrobial Testing, Recovery
        
        Sequencing
        
        Detects Uncultured Organisms & Describes Community Composition
- - - - Genetically Engineered Microbes Make High-Value Products
    - - Microbial Growth on Submerged Surfaces
    - - Massive Growth of Naturally Occurring Microbes to Make Low-Cost Products
    - - Microbial Use in Cleaning Pollutants, Wastewater, Prescriptions
    - - Cellulose & Cornstarch Converted to Glucose & Ethanol
- - - - Makes Object Larger
    - - Distinguishes Two Adjacent Objects
      - Limit 0.2um at 1000X
    - - Differences in Contrast (Density) Between Specimen & Surroundings
      - Pigmented Microbes
    - - Improves Contrast
      - Organic Compounds Bind to Specific Materials
      - Basic Dyes
        
        Positively Charged Dyes Bind to Negatively Charged Components
      - Fixed or Dried Cells
      - Procedure
        
        Spread Thin Film Over Slide
        
        Dry in Air
        
        Pass Through Flame
        
        Flood with Stain
        
        Oil & View
      - Differential Stains
        
        Gram Stain
        
        Hans Christian Gram 1884
        
        Gram-Positive Bacteria
        
        Appear Purple
        
        Thick Peptidoglycan Cell Wall
        
        Gram-Negative Bacteria
        
        Appear Pink/Red
        
        Thin Peptidoglycan Cell Wall
        
        Crystal Violet, Iodine, Alcohol, Safranin
    - - Unstained, Live Cells
      - Phase Ring
      - Dark Cells on Light Background
      - Observes Motility, Division, Morphology
    - - Light from Sides
      - Light Cells on Dark Background
      - Thin or Delicate Organisms
      - Unstained, Live Cells
    - - Fluorescent Specimens
        
        Emit Light After Illumination With Different Wavelengths
      - Separates Excitation & Emission
      - Autoflouorescence
      - DAPI, GFP, RFP
      - Enumerates, Detects Taxa, Locate Internal Molecules
    - - Computerized Microscope
      - 3D Image Generation
    - - Greater Resolution
      - Fixed, Dehydrated, Stained, Coated, Imaged in Vacuum
      - Electron Micrograph
      - Transmission Electron Microscope
        
        2D Internal Structures, Viruses, Macromolecular Complexes
        
        Must be Very Thin (20-60nm)
        
        High Atomic Weight Stains Scatter Electrons & Improve Contrast
        
        Negative Staining Allows Observation of Intact Cells
      - Scanning Electron Microscopy
        
        3D Surface Topology, Cell Arrangements, Larger Specimens
        
        Coated with Thin Film of Heavy Metal
        
        No Internal Structures Unless Fractured