MT: Chapter 3 - Culturing Microbes [PART I-III]

• The Five “I’s

  Inoculation, Isolation, Incubation, Inspection, Identification
• General growth media

  Nutrient Agar, Tryptic Soy Agar
• Differential agar


  MacConkey, Eosin Methylene Blue, Salmonella Shigella Agar 
• rich with nutrients, can see a difference, thus differential; much more later

  Blood agar
• Allow organisms to grow under the optimal conditions

  Incubation
• is an instrument used in the production of an anaerobic environment.

  Anaerobic jar
• Correlating data from all observations to ID organism to species

  Identification
• Microscopy: Visible Light is the energy source


  Light microscope
• Bounces off (Mirror)

  Reflected
• Passes through (GLASS)

  Transmitted
• Soaked (black colored paper)

  Absorbed
• Scattered as it passes through

  Diffracted
• Bent as it passes (objects seen under water) Glass lenses

  Refracted
• degree of bending, based on lens material and shape of lens

  Refractive index
• Lens system with two magnifying lenses, magnification is calculated by multiplying the power of the two lenses (10 X 10 = 100 power)

  Compound Light Microscope
• Stains are colored dyes (____) that increase contrast.

  chromophores
• The ability of a lens system to allow an observer to see fine detail. 

  Resolution
• The best magnification on our scopes is achieved with the?

  Oil immersion objective
• most common, objects are dark against a bright background

  Brightfield
• special condenser, objects are light against a dark background – used to see live microbes unstained (spirochetes in fluid)

  Darkfield
• expensive condenser and internal lens components, change “phase of light”, so live specimens appear with more internal contrast

  Phase contrast
• fluorescent dyes and UV light

  Fluorescence
• energy source for magnification is a beam of electrons (negative charged subatomic particles

  Electron Microscope
• very high magnification (100,000 X)

  Transmission electron microscope
• tremendous surface detail

  Scanning electron microscope
• Dying: complex procedure, see difference between cells

  Differential stain
• is a fundamental unit of biological classification. It refers to a group of organisms that are capable of interbreeding and producing fertile offspring

  Species
• refers to a genetic variant or subtype within a species. It's a term commonly used in microbiology to describe organisms that are closely related but exhibit minor genetic differences.

  Strain
• Variant prokaryotic strains characterized by biochemical or physiological differences.

  Biovar
• Differ morphologically

  Morphovar
• Have distinctive antigenic properties

  Serovar
• Usually one of first strains of a species studied, often most fully characterized than other strains.

  Type Strain
• Deals with the biological nomenclature for plants

  International Code of Botanical Nomenclature
• Deals with the biological nomenclature of animals.

  International Code of Zoological Nomenclature
• a tentative identification

  Clinical signs of the disease
• Definitive identification

  Laboratory examination
• Laboratory examination consists of:

  Microscopy, Culturing, Phage Typing, Biochemical tests, Serological tests, Molecular Technique
• Microscopy consist of:

  Wet mount microscopy, Gram staining, acid-fast staining, Negative staining, spore staining, Motility staning
• Culturing consist of

  Differential media, Selective media, Enriched media, Enrichment media
• Biochemical consist of:

  Catalase test, Coagulase test, Oxidase test, Sugar fermenting test, Indole test, Citrate test, Urease test